Description of three new species of Apatelodidae from the southern neotropical region (Lepidoptera, Bombycoidea)

Abstract Three new species of Apatelodidae are described from Argentina, Bolivia, Brazil, and Paraguay: Apatelodesnavarroisp. n., Apatelodeschalupaesp. n., and Apatelodesulfisp. n., and are figured with their genitalia. Detailed examination of primary types leads to the establishment of a new synonymy: A.florisa Schaus, 1929 = A.schreiteri Schaus, 1924, syn. n., and a revised status for another Apatelodidae species previously misplaced in the Bombycidae: Apatelodesbrunnea (Dognin, 1916), comb. n.


Introduction
The adults of the American silkworm moths (Lepidoptera, Bombycoidea, Apatelodidae) are small to medium size with earthen tones. The larvae are exposed feeders on trees and shrubs and several species are polyphagous (Gillott 2005). This family is mainly Neotropical with a few representatives in North America (Kitching et al. 2018).
An extensive revision of Apatelodidae was published by Draudt (1929). More recently, a checklist was issued by Becker in Heppner (1996), and a Bombycoidea checklist by Kitching et al. (2018). The status of Apatelodidae as a family belonging to the Bombycoidea has been confirmed by morphological and molecular studies (Lemaire andMinet 1998, Zwick et al. 2011).
Within the Apatelodidae, the most diverse genus is Apatelodes Packard, 1864, which contains 115 species of the total of 214 species in the family, as per the most recent report by Kitching et al. (2018). The caterpillars of Apatelodes are generally hairy, with long hairs, and pupate under ground. A large variety of food plants seem to be used by species in the genus, as found by extensive rearing conducted in Costa Rica, Guanacaste, and reported on a dedicated Internet site by Janzen and Hallwachs (2009). Hosts of Apatelodes include plants from the families: Malvaceae, Chrysobalanaceae, Myrtaceae, Fabaceae, Salicaceae, Meliaceae, Annonaceae, Asteraceae, Piperaceae, Amaranthaceae, Convolvulaceae, Verbenaceae, and Bignoniaceae. An Apatelodes species from Colombia has recently been reared in the laboratory in Europe, and accepted a Salix Linneaus sp. (Salicaceae) as a replacement food plant (Herbin unpublished), and some other Apatelodes species are reported as pests of banana trees (Musa Linneaus, Musaceae) in Venezuela (Dominguez et al. 2002).
In this study, three new species of Apatelodidae from Argentina and neighboring countries (Brazil, Bolivia and Paraguay) are described in Apatelodes based on their habitus, genitalia, and DNA barcoding.

Materials and methods
Materials and methods are as per Herbin (2017) and Herbin and Mielke (2018).
Figures were manipulated with Adobe Photoshop CS4. Green labels in figures relate to a voucher number in CDH (see below for collection abbreviations). White labels with the format "BC-Her####" relate to barcode reference numbers from specimens in CDH. All other labels shown belong to the holotype.
All species treated here were subjected to DNA analysis using the DNA barcode region of the mitochondrial COI gene in BOLD (Barcode of Life Data System: http:// www.boldsystems.org, see also Ratnasingham and Hebert (2007)). Sequences were aligned using the tools provided in BOLD (BOLD Aligner: Amino Acid Based HMM), exported in fasta format, and imported in MEGA6 (Tamura et al. 2013). The evolutionary history of the taxa was inferred using the Neighbor-Joining method (Saitou and Nei 1987). The phenograms are drawn to scale, with branch lengths (next to the branches) in the same units as the evolutionary distances used to infer the Neighbor-Joining tree. The evolutionary distances were computed using the Kimura 2-distances (Kimura 1980) and measure the number of base substitutions per site. All codon positions were included and all positions containing gaps and missing data were excluded.
Description of colors in the descriptions refers to the RAL color standard, see https://www.ral-farben.de/en/home/.  (Fig. 3). In CDH.

Abbreviations
Etymology. Apatelodes navarroi sp. n. is named after the late Dr. Fernando Navarro (IFML), who directed the expedition that enabled the discovery of this new species and the following one.
Diagnosis. Apatelodes navarroi sp. n. belongs to a small group of Apatelodidae showing very developed socii. This group includes A. hierax Dognin, 1924, A. schreiteri Schaus, 1924, A. florisa Schaus, 1939, A. zikani Draudt, 1929, and A. combi Herbin, 2015. Apatelodes navarroi sp. n. is similar to A. florisa and A. schreiteri, both described from Argentina, Tucumán (see taxonomical notes hereafter). Apatelodes navarroi sp. n. is easily distinguished by its very dark brown color, its greater size, the elongated shape of the forewings, and the slightly concave termen of the forewing below apex. The male genitalia exhibit large socii with well-developed teeth.  Description. Male. Antennae: antennae bipectinate to the tip. Scape, pedicel, and antennomeres beige, rami brown. Head: brown, labial palpi thick, brown, slightly ascending and reaching beyond the front. Eyes naked, dark brown. Thorax: coloration terra brown (RAL8028) with a median longitudinal black line. Legs: coloration as for thorax, with tibia appearing very thick as covered by long light brown hair like  scales. Femora densely pilose beige with dark brown scales; Abdomen: sepia brown (RAL8014). Forewing dorsum: Forewing length: 17 mm, wingspan: 34 mm. Triangular, apex acute, outer margin slightly concave below apex. Coloration terra brown (RAL8028), antemedial line black, a basal rectangular black mark in antemedial area, postmedial line wavy, black. Two small hyaline spots near apex and costa: one tiny spot near costa, bordered proximally with small black triangular mark, second spot posterior to first slightly larger, triangular, and bordered proximally with black scales. Between costa and first tiny hyaline spot, a black comma marking present. Forewing ventrum: Similar to dorsum, but lighter in color: pale brown (RAL8025), with postmedial line lighter in color, darker terra brown patch at apex. Hindwing dorsum: homogenous terra brown coloration, slightly lighter than forewing. Medially, a curved transverse line lighter brown. Hindwing ventrum: Dark terra brown with a pale brown transverse line. Genitalia ( Figure 2).
Uncus strongly sclerotized, with wide base and single bent hook-like apex. Base of uncus with a pair of large socii made of a two folded sclerotized sheet bordered with numerous strong teeth. Valves elongated, not reaching the uncus, rather narrow, with apex rounded and a strong sclerotized spine at the dorsal base of the valve. Aedeagus short and cylindrical, with no teeth or cornuti. Caecum penis present. Vesica with a ventral diverticulum. No cornuti on vesica.
Papillae anales hemispherical, slightly bulbous and covered with setae. Apophyses thin and cylindrical, slightly spatulate at tip, anteriores about the same length as posteriores. Ductus bursae as a very long (about 5 mm, twice the length of corpus bursae), narrow, sinuous ribbon, of equal diameter for entire length. Ductus bursae sclerotized at entry near ostium bursae. Bursa copulatrix smooth and ovoid with a horse-shoe shaped, semi circular, single signum at extremity of bursa, signum equipped with minute teeth, inward pointing. Distribution. A. navarroi sp. n. is presently only known from Argentina, Jujuy, at medium altitude. Etymology. Apatelodes chalupae is named after Dr. Adriana Chalup (IFML), Geometridae and Noctuidae lepidopterist specialist.
Diagnosis. Apatelodes chalupae sp. n. is similar to A. navarroi sp. n., but is much lighter in color (see comparison in Figs 10-11), bears two small hyaline spots near apex bordered proximally by black scales. The male genitalia also show structures similar to  those of A. navarroi sp. n., but reduced in size, and with less developed socii, including less developed teeth bordering the socii. A distance in COI barcodes of 4.3% is found between these two species, as shown in the tree in Figure 16.
Description. Male. Antennae: Bipectinate to the tip. Scape, pedicel and antennomeres beige, rami beige brown. Head: beige brown, labial palpi thick, beige brown, slightly ascending and reaching beyond the front. Eyes naked, dark brown. Thorax: ochre brown with longitudinal black central line. Legs: beige brown, tibia appearing thick due to long beige brown scales. Forewing dorsum: Forewing length: 16 mm, wingspan: 32 mm. Triangular, apex acute, outer margin slightly concave below apex.
Ground color light ochre brown (RAL8001), maculation similar to previous species with two small hyaline spots near apex, bordered proximally by tiny black marking. A black comma-shaped mark exists between costa and the smaller hyaline spot. A dark brown diffuse marking present basally in antemedian area. Antemedial and postmedial lines faint. Forewing ventrum: Ground color beige (RAL1001). Outer margin copper brown (RAL8004). Postmedial line light brown. Hindwing dorsum: coloration uniform light ochre brown, a faint lighter beige longitudinal line present. Hindwing ventrum: coloration rather uniform copper brown with contrasting beige distal longitudinal line inwardly bent at CuA2 and diffuse brown proximal longitudinal line. Abdomen: a black collar at interface of thorax/abdomen. First two abdominal segments dorsally reddish brown, remaining segments ochre brown. Black spot present dorsally on each abdominal segment. Genitalia (Figure 6).
Uncus heavily sclerotized, wide, bent mesally, with a simple hooked apex. Base of uncus with a pair of medium size socii made of a two folded sclerotized sheet bordered with few small teeth. Valves elongated, rather narrow, not reaching the uncus, with apex rounded and a strongly sclerotized spine at the dorsal base of the valve. Aedeagus short and cylindrical, with no teeth or cornuti. Caecum penis present. Vesica with a ventral finger like diverticulum and a smaller lateral diverticulum. No cornuti on vesica.

Distribution. A. chalupae sp. n. is only known from the type locality in Argentina.
Remarks. Initially, we thought that the type specimen of A. chalupae sp. n. was a lighter colored representative of A. navarroi sp. n., however the COI barcodes showed a significant distance between the two species (4.3%), further confirmed by the differences found in the male genitalia. Figure 7 presents a side-by-side comparison between the genitalia of both species, from a photo taken under a microscope with both male genitalia taken in the same picture, therefore at the same scale. Smaller genitalia size for A. chalupae sp. n., the most striking difference is in the relative size of the socii, and the much stronger teeth present on the edge in A. navarroi sp. n.
In Figure 16, we present the tree built with MEGA6, with the new taxa A. navarroi sp. n., A. chalupae sp. n., and the most similar previously described species: A. schreiteri.    Etymology. Apatelodes ulfi sp. n. is named after Ulf Drechsel in Paraguay, who collected the majority of the known specimens.

Apatelodes ulfi
Diagnosis. Apatelodes ulfi sp. n. is a rather small species, with the basal half of the forewing dorsum dark reddish brown very contrasting with the light colored (grey beige) postmedial and marginal area. A single hyaline preapical spot. The ventrum with inverted contrasting area compared to dorsum: darker on the marginal area, and lighter in median and basal area. The particular feature of the male genitalia lies in the socii, showing two ventral projections, one very short and truncated, another slightly longer.
Description. Male. Antennae: Bipectinate to the tip. Scape, pedicel, antennomeres and rami beige. Head: brown red (RAL3011) with some beige-tipped scales, labial palpi thick, brown, projected forward, eyes dark brown. Thorax: prothoracic collar brown red, thorax vinaceous red (RAL3005). Legs: Tibia thick with long hair like scales, brown and whitish for prothoracic legs, brown red for mesothoracic and metathoracic legs. Abdomen: brown red. Forewing dorsum: Forewing length (n = 13): 13-17 mm, wingspan: 28-35 mm (holotype: length 16 mm and wingspan 34 mm). Ground coloration grey beige (RAL1019), with basal half oxide red (RAL3009) con- trasting with two grey beige undulating antemedial lines. Single elliptic hyaline spot, bordered proximally with an oxide red small triangle, distally by a small oxide red spot present near apex. Termen bordered with narrow oxide red. Postmedial line crenulated, oxide red. Forewing ventrum: Ground color beige (RAL1001) with some oxide red scales near costa. Marginal area oxide red with beige triangle above hyaline spot. Hindwing dorsum: Ground color red brown (RAL8012) with beige median line. Termen bordered by nut brown (RAL8011). Hindwing ventrum: Marginal area oxide red with beige postmedial line, antemedial and medial areas of lighter color due to beige scales interlaced with oxide red scales. Genitalia (Figure 9): Uncus downcurved, strongly sclerotized, wide, with a single small spine at apex. At base of uncus, socii exhibit two finger-like extensions, one short and truncated and one longer (see Figure 9c). Valves elongated, rather narrow, with apex rounded. Process at base of valve strongly sclerotized with two small apical teeth. Aedeagus short and cylindrical, caecum penis present, with no teeth or cornuti. Vesica with a small ventral finger like diverticulum, then a very small diverticulum laterally, slightly sclerotized (appearing orange in Figure 9d).
Female. Unknown. Distribution. Specimens of A. ulfi sp. n. have been collected in various localities in northern Argentina, southern Bolivia, central western Brazil and north to south Paraguay. Possible extension of the range to Peru remains to be investigated.
Remarks. DNA barcoding of specimens from various localities reveal that these populations are all perfectly aligned (i.e. 0% distance between specimens of northern Argentina, southern Bolivia and Paraguay. Specimens from Brazil in MHNG not barcoded) despite some variation in wingspan.
A similar specimen in CDH, from northern Bolivia (Nor Yungas, Coroico area), shows a larger size and some differences in habitus, and has therefore not been included in paratype series. Similar specimens are likely to be found in southern or central Peru, this will then enable to verify identity or not with A.ulfi sp. n.

Additional taxonomic notes
Note 1: A comparison of species similar to A. schreiteri. As illustrated in Figures 10-12, where a specimen of A. schreiteri is figured together with A. navarroi sp. n. and A. chalupae sp. n., differences in size of the specimens is obvious. The figured A. schreiteri specimen (Figure 12) was also collected in Argentina, Jujuy, Parc Calilegua, 1028 m (specimen CDH3.334, barcode BC-Her4940, barcoded and belonging to the A. schreiteri clade shown in Figure 16). Apart from the smaller size of this species compared to A. navarroi sp. n. and A. chalupae sp. n., the shape of the forewing in A. schreiteri is such that the termen is rather rounded, without the truncated apex found in the two newly described species. A significant difference also lies in the black marking proximal to the larger hyaline spot: in A. schreiteri this marking is more elongated than in the other two species.  Re-examination of the types in the USNM reveals exactly the same configuration of moth size, termen, and black markings in the syntype male of A. schreiteri, and in the female holotype of A. florisa Schaus, 1939. Both type specimens originate from Argentina, Tucumán. Figures 13-14 present the holotypes of both A. schreiteri and A. florisa: the female is slightly larger than the male, as observed in all other Apatelodidae species. We can observe the exact identical configuration of the forewing dark markings and hyaline spot in the male and female, this being generally a key in all species of Apatelodes to correctly allocate a female to the corresponding male.
We therefore consider that A. florisa Schaus is a subjective junior synonym of A. schreiteri Schaus and propose: Apatelodes florisa Schaus, 1929 = Apatelodes schreiteri Schaus, 1924, syn. n.  Note: the genitalia of these holotypes have not been examined, but being the case of a male compared to a female this would not have actually helped in demonstration.

Note 2:
The examination of various primary types enables the recognition of an anomaly in the present classification of Bombycoidea: Carthara brunnea Dognin, 1916 (holotype male examined in USNM) was previously placed in the genus Quentalia Schaus, 1929(Becker 1996, and more recently with the same combination in the global Bombycoidea checklist (Kitching et al. 2018). The genus Quentalia being now included in Bombycidae (Zwick 2011, Kitching et al. 2018) and separated from Apatelodidae, this species should instead be classified in the apatelodid genus Apatelodes. Figure 15 illustrates the type specimen of Carthara brunnea (photo courtesy USNM), supporting the obvious proposed reclassification. The habitus, venation, antennae, and patterning, all suggest that this species belongs in Apatelodidae. Draudt (1928: 681) established the new genus Quentalia and indicated: "This genus will contain the many species described under Carthara Wkr which was first used in Cat. Lep. Het. B.M. 33, p.914....", and this is likely the root of the transfer of brunnea to Quentalia, as Dognin originally placed this species in Carthara.
The taxon brunnea is not addressed in the text nor in the color plates by Draudt (1928), but can be found in the same work under: Alphabetical List of the American Bombycidae on page 710 under Q. brunnea. It is likely that since Draudt (1928), no one has re-examined the holotype, and the mistaken classification has been propagated. Here, the following taxonomical rearrangement is proposed: Carthara brunnea Dognin, 1916= Quentalia brunnea (Dognin, 1916 = Apatelodes brunnea (Dognin, 1916), comb. n.
We notice a similarity of A. brunnea to the species newly described above, but no hyaline spot exists on the forewings in A. brunnea.