More than Olceclostera bifenestrata : New species and morphology of immature stages of Olceclostera Butler, 1879 (Lepidoptera: Bombycoidea, Apatelodidae)

Apatelodidae is a family of Neotropical bombycoids that still needs to be studied in several aspects, as many groups of species have not yet undergone a careful systematic revision. On the other hand, recent studies showed that some species known to be widely distributed in fact form species complexes. Until now, Olceclostera bifenestrata Schaus, 1912, described from Costa Rica, supposedly has a wide distribution, reaching the south of Brazil. We reviewed specimens from South America identified as O. bifenestrata . Using morphological data and DNA barcodes, we discovered that South American specimens belong to four new species: Olceclostera jairana sp. nov. , Olceclostera quilombola sp. nov. , Olceclostera xeta sp. nov. and Olceclostera wayana sp. nov. The four species can be distinguished mainly by characters of genitalia, mostly in aedeagus structures. However, wing characters (number of hyaline spots) and abdomen characters (number of patches of larger scales) may be useful for classifying species groups in Olceclostera . Additionally, we illustrate and describe for the first time all the life stages of an Olceclostera species, including its chaetotaxy and life history, and provide the first SEM images of the egg of an Apatelodidae species. We present a discussion about the morphological characters of adults and immatures and their relevance to the systematics of Apatelodidae.


Introduction
Apatelodidae is a family of Neotropical bombycoids (Lepidoptera: Bombycoidea) that lacks conspicuous colors or economic or medical importance.As a consequence, the group has been largely neglected in biodiversity studies, especially if compared to other large-sized Bombycoidea (e. g.Saturniidae and Sphingidae) (Kitching et al. 2018;Ballesteros Mejia et al. 2020).More recently, some taxonomic efforts carried out mainly by Herbin (Herbin 2015(Herbin , 2017(Herbin , 2018(Herbin , 2021a(Herbin , 2021b;;Herbin and Monzon Sierra 2015;Herbin and Beccacece 2018;Herbin and Mielke 2018;Herbin et al. 2021) have shown that the family is more diverse than previously thought.The study of the male and female genitalia structures and the use of molecular tools has shown that many species comprise cryptic species complexes (Herbin 2017;Herbin et al. 2021;Orlandin and Carneiro 2021).Currently, the family comprises 200 species and ten subspecies, distributed in 13 genera (Kitching et al. 2018;Hamilton et al. 2019;Herbin 2021aHerbin , 2021b;;Herbin et al. 2021;Orlandin and Carneiro 2021) but studies testing the monophyly of the genera are lacking.
The immature stages of Olceclostera species are also poorly known.Only the larvae of two North American species have been described (Lintner 1874;Packard 1895;Franclemont 1973;Stehr 1987;Peigler 1994;Wagner 2005), and additional images of some species from Central America can be found in online databases (Janzen and Hallwachs 2009).Also, host plant records are only available for a few species (Franclemont 1973;Biezanko et al. 1974;Wagner 2005;Janzen and Hallwachs 2009;Robinson et al. 2010;Diniz et al. 2013).However, none of the studies provides a detailed description of all immature stages, including the larval chaetotaxy, an important set of characters that contribute to the systematics of many groups of Lepidoptera (Peterson 1962;Scoble 1992) and which have shown substantial differences when comparing the immatures of some genera of Apatelodidae (Orlandin et al. 2021(Orlandin et al. , 2022a(Orlandin et al. , 2022b)).
Aiming to test if the specimens from South America recognized as O. bifenestrata belong to this species, the morphology of males and females, including genitalia, was studied.To confirm if this morphological similarity reflects a close relationship, we perform a molecular analysis based on COX1 (barcode region, 658pb).Additionally, we describe in detail all immature stages from one of these new species adding a discussion of its relevance to the systematics of Apatelodidae.In addition, we accessed all original descriptions of Olceclostera species (Walker 1855(Walker , 1865;;Grote 1864;Packard 1864;Butler 1879;Edwards 1886;Druce 1890;Schaus 1892Schaus , 1894Schaus , 1900Schaus , 1905Schaus , 1910Schaus , 1912Schaus , 1920Schaus , 1927;;Dyar 1906;Jones 1908;Draudt 1929;Herbin 2018Herbin , 2021;;Herbin and Mielke 2018).In the section "Material examined" where the information on the types is provided, the labels are separated from one another by trans verse bars.

Morphology
Immature.The first instar was illustrated with and without the long setae that ornamented the body.These setae were carefully removed using tweezers to allow the visualization of its insertions and other structures present along the body.Images of the eggs were taken using scanning electron microscopy (SEM-JEOL JSM 6360-LV).Images of the egg 1st, and 2nd instars, and pupa were taken using an optical stereomicroscope adapted with focus stacking (Leica Application Suite Version 4.12.0 [Build 86]), and subsequent instars were recorded with a Sony DSC-HX100V digital camera.We used ImageJ Software (Schneider et al. 2012) to measure the eggs, cephalic capsules, larvae, and pupae.The terminology follows Peterson (1962) and Stehr et al. (1987).
Adults.For the description of adults, the head, palpi, legs, and abdomen were detached, soaked in hot 10% potassium hydroxide solution (KOH) for approximately 3 min, washed in water, examined, illustrated, and then stored in glass vials containing glycerin.Images were taken using an optical stereomicroscope adapted with focus stacking (Leica Application Suite Version 4.12.0 [Build 86]) and a Sony DSCHX100V digital camera.The terminology follows Kristensen (2003), but the term "fultura inferior" (= juxta) is applied sensu Petersen (1904); the term aedeagus is applied sensu Snodgrass (1935); and followed Zwick (2009) for the terminology of the semioval sclerotized plates located ventrally of the subscaphium [gnathos] (Fig. 1).were offered to the larvae.These hostplant species were chosen based on previous records of Apatelodidae host plants (Biezanko et al. 1974;Janzen and Hallwachs 2009;Robinson et al. 2010;Orlandin et al. 2021Orlandin et al. , 2022aOrlandin et al. , 2022b)).The larvae were reared with the chosen hostplant species until pupae.Whenever necessary, the branches were replaced to maintain fresh, abundant, and high-quality leaves for the immatures.Immediately after each moult, the dry head capsules were retained and stored in glass vials.From the fifth instar on, the larvae were transferred to similar containers with the hostplant branches and a layer of dark soil beneath.A few specimens from each immature stage were killed through immersion in boiling Figure 1.Terminology adopted for Olceclostera genitalia: A male, ventral view; B male, lateral view; C male, phallus, lateral view; D female, lateral view.

Immatures rearing
water, fixed in Kahle-Dietrich 10% (Borror and deLong 1971) for 72 h, subsequently preserved in ethanol 70%, and then deposited in the Immature Collection of Lepidoptera from the Entomological Collection "Pe.Jesus Santiago Moure, Universidade Federal do Paraná" under voucher number DZUPIL 0170.

Genetic divergence and phylogenetic inference
A series of 30 Apatelodidae specimens were selected for DNA sequencing.S1).
Alignment was then performed with MAFFT (Katoh and Standley 2013).The most optimal model of nucleotide evolution was selected by ModelFinder (Kalyaanamoorthy et al. 2017) using the AICc criterion, and the best fit model according to AICc was GTR+F+I+G4.The tree was rooted with Drepatelodes friburgensis (Schaus, 1924) following Hamilton et al. (2019) that recovered Drepatelodes as the sister group of other Apatelodidae genera.Maximum likelihood phylogenies were inferred using IQ-TREE v. 1.6.8 (Nguyen et al. 2015), with branch supports reported as 5,000 standard bootstraps.Alignment, model selection, and phylogenetic analyses were performed in the multifunctional platform PhyloSuite (Zhang et al. 2020).
Genetic distances of the "O.bifenestrata group" were calculated by the Tamura-Nei distance model (Tamura and Nei 1993) in MEGA11 (Tamura et al. 2021).

"Olceclostera bifenestrata group"
Comments.The "Olceclostera bifenestrata group" comprises eight species, namely: Olceclostera angelica (Grote, 1864), Olceclostera bifenestrata, Olceclostera indistincta (Edwards, 1886) Diagnosis.This group of species is easily distinguished from other species of Olceclostera by the following com-bination of characters: forewings grayish, with sinuous brownish lines, two subapical hyaline spots, and one pair of large patches of broader scales laterally on the abdomen.The South American species are reliably separated from the North American species O. angelica, O. indistincta and O. seraphica by their larger size, less grayish coloring (the North American species are light grey) (see Franclemont 1973: plate 3, figs 14-16), molecular distance (Table 1), and geographic distribution (Schaus 1912;Franclemont 1973).They can be distinguished from O. bifenestrata (Fig. 2A, B) by the sclerotized subscaphium region with a narrow median ventral hook (Fig. 3).al portion of the vesica is covered by small cornuti (Figs 10,19,21).2C, D, 4A, B, 5, 6A, 7-11).Head.MALE.Vertex and frons pinkish beige; compound eyes naked; antenna approximately 1/3 the length of the costal margin of the anterior wing, bipectinate from the base to apex, ventrally naked, dorsally covered by pearl scales; labial palpus dorsally covered by brown scales, ventrally covered by a compact block of pinkish beige scales (Fig. 5A); first and second segment thick and slightly curved, the second about twice the length of the first, third segment reduced and oval (Fig. 5B); proboscis absent.FEMALE.Similar to the male, slightly smaller than the male's head; antenna, pecten shorter than the male's pecten; palpus thinner and shorter than the male's palpus (Fig. 4A, B). -Thorax.MALE.Dorsally and ventrally gray; dorsally with a longitudinal medial brownish line.-Legs.All legs smooth, femur and tibia gray; tibias with a band of long scales with light brown apex projecting towards the posterior region (Fig. 6A); tarsomeres brownish.Foreleg: coxa thick, developed, approximately 3/4 the length of the femur; femur about the same length as the tibia; tibia anteriorly flattened, epiphysis approximately half-length of the tibia (Fig. 7A).Midleg: femur slightly smaller than the tibia; distally tibia bears a pair of spurs, the inner one longer than the outer one (Fig. 7B).Hindleg: femur smaller than the tibia; distal half of tibia bears two pair of spurs, the inner longer than the outer ones (Fig. 7C).All tarsomeres have a similar length, posteriorly with scattered small spines; tarsal claws simple.FEMALE.Similar to the male, epiphysis approximately 1/3 the length of the tibia, narrower than the male; ventral surface distitarsus of fore, mid, and hindlegs with a patch of small spines and an inner row with four conspicuous spines (Fig. 7D).-Wings shape and venation.Venation (Fig. 8), same as that described for Olceclostera sp. by Costa-Lima (1950).MALE.Forewing length: 14-19.5 mm (n = 10); elongated triangular shape.Upperside: Ground color grayish with scattered black scales; basal region limited by a fine irregular black line; discal region marked between two sinuous brownish lines; on dcm, a patch of black scales surrounded by a patch of light gray scales; post-discal region with a thin irregular black line; two rounded subapical hyaline spots, between M 1 and M 3 ; posterior to the hyaline spots a sinuous Table 1.Pairwise genetic distances (%) for COX1 gene (barcode region, 658pb) sequences among Olceclostera bifenestrata group and the new species of Olceclostera.band of light gray scales; apical region light gray; between M 2 and CuA 2 marginal region with black scales and outer margin slightly crenulated and marked with black scales.Underside: Ground color similar to the upperside but more brownish; post-discal region with a sinuous brownish line followed by a thin, sinuous black line.Hindwing.Frenulum long and simple (Fig. 8).Upperside: Ground color grayish brown; on dcm, a faint black dot; postdiscal region with two faint sinuous brownish lines; outer margin, between M 3 and CuA 2 , slightly crenulated and marked with black scales.Underside: Similar to the upperside, but with scattered black scales; dots and lines more marked; inner margin lighter.FEMALE.Forewing length: 19-22 mm (n = 06); general color similar to the male, the apex is more acute than in males; on dcm, the patch of black scales is smaller than in the male and absent on the underside; hyaline spot between M 2 -M 3 reduced.Hindwing.Frenulum grayish brown, as multiple bristles, similar to other female Apatelodidae species (Orlandin et al. 2021(Orlandin et al. , 2022a)).Similar to the male, but on dcm the dot of black scales is smaller than in the male.-Abdomen.MALE.Dorsally coppery and gray, ventrally gray with scattered black scales; pleural region of segments A1 to A3 with patches of broader scales, creamy at the base and apically coppery (Fig. 9A); into this patch, on A2 and A3, a row of white scales (Fig. 9B).FEMALE.Dorsally grayer than the male; at the posterior region of each segment, a band of light gray scales; ventral and pleural region similar to the male.-Genitalia.MALE (Fig. 10).Tegumen shield-like, sclerotized in all its extension; anterior projection of saccus tapered, dorsally curved; ventrally, the posterior margin of saccus has a semicircular projection projected posteriorly; uncus dorsoventrally flattened, ventrally projected, 3/4 bifid V-shaped, with a pair of laterally flattened socii at the base; subscaphium sclerotized, with a median ventral hook, proximal region, ventrally with two semi-oval sclerotized plates [gnathos] with numerous tiny spines; valva less sclerotized than uncus and tegumen, rectangular, tongue-like, laterally covering the uncus apex; costa (= processus superior) forming a slightly sclerotized process bearing some setae; sacculus (= processus inferior) slightly more sclerotized than valva; process of sacculus finger-like, projected at 90º from the valva; fultura inferior sclerotized and rectangular; aedeagus cylindrical with truncated caecum, ventrally projected forming a protuberance; in lateral view, apex with a projection bearing one to three spines (more frequently two); these structures are best seen when the vesica is everted.FEMALE (Fig. 11).Eighth tergum formed by a trapezoidal plate bearing wide lateral arms ventrally reaching the lamella antevaginalis; anterior apophysis thin and cylindrical, slightly spatulate at the tip; lamella antevaginalis flat, almost as long as the lamella postvaginalis; lamella postvaginalis sclerotized, rectangular; ductus bursae about two times the length of corpus bursae, approximately the same diameter for the entire length; corpus bursae ovoid, membranous with a signum heart-shaped, bearing inward-pointing spines and with the central portion less sclerotized with fewer spines; papilla analis dorsoventrally elongated, sclerotized and covered with setae.12-16).Eggs (Fig. 12).Average duration (from oviposition to hatching): approximately 15 days (n = 110); pie-shaped 1.5-   1.7 mm in diameter and 0.4 mm in height (n = 10); chorion transparent without visible ornamentation; yolk greenish during the first days of development, becoming pale greenish-yellow before eclosion (Fig. 12A-C).The micropylar area bears seven petal-shaped rosette cells of distinct sizes; around the micropylar rosette, there are ten secondary cells (Fig. 12D); these micropylar cells surround the six openings (Fig. 12E).Aeropyles rounded, organized forming geometric shapes, in this region the egg is quite rough (Fig. 12F, G).First instar (Fig. 13A, B).Average duration: approximately five days (n = 94); body length 4.8-5.1 mm (n = 3); head capsule width 0.54-0.57mm (n = 4), hypognathous, slightly flattened in front, yellowish with brownish spots and stripes, stemma 3 larger than the other stemmata, ventral margin of labrum moderately notched; head bears only primary setae (see chaetotaxy below).Body yellow, cylindrical, dorsally with faint black stripes, laterally, with black stripes on subdorsal, lateral, and subventral regions; long setae cover the whole body, except ventrally; dorsally and subdorsally covered with black and yellow setae, laterally and subventrally only with yellow setae, most of them grouped over verrucae; spiracles elliptical, T1 and A8 ones larger than the others; abdominal prolegs fully developed on A3-A6 and A10, with two sclerotized plates, one anterior and one posterior; crochets in biordinal mesoseries.-Chaetotaxy.Head.(Fig. 14) One pair of frontal setae (F1), between them one pair of frontal pores (Fa); two pairs of adfrontal setae (AF1, AF2), between them one pair of adfrontal pores (AFa); three anterior pairs of setae (A1, A2, A3) and one pair of pores (Aa) above A2; two pairs of posterior setae (P1, P2) and two pairs of pores (Pa, Pb); three pairs of microdorsal setae (MD1, MD2, MD3), and a pair of pores (MDa) between MD2 and MD3; on clypeus, two pairs of setae (C1, C2); on labrum three central (Lrm1, Lrm2, Lrm3) and three lateral pairs of setae (Lrl1, Lrl2, Lrl3).Laterally, one pair of lateral setae (L1) and one pair of pores (La); three stemmatal pairs of setae (S1, S2, S3), and two pairs of stemmatal pore (Sa, Sb); three substemmatal pairs of setae (SS1, SS2, and SS3), and a pair of microgenal pores (MGa).-Body (Fig. 15).Thorax.T1. pronotal plate brownish, weakly sclerotized; XD setae located in the anterior area of the pronotal plate close to four secondary setae; D1 and D2 setae solitary, located in the posterior region of the pronotal plate; SD1 and SD2 setae solitary, SD1 seta at the margin of the pronotal plate; L setae grouped with numerous secondary setae on a conspicuous spherical grayish yellow verruca; SV setae grouped with numerous secondary setae on a large yellowish verruca; V setae grouped with some secondary setae on a sclerotized grayish plate.T2.D1 seta solitary on a reduced grayish yellow plate, D2 seta grouped with a secondary seta on a grayish yellow plate; SD setae grouped with numerous secondary setae on a grayish yellow verruca; L setae grouped with numerous secondary setae on a grayish verruca; SV setae grouped with numerous secondary setae on a grayish verruca; V setae grouped with some secondary setae on a sclerotized grayish plate.

Description of immature stages (Figs
T3. D1 seta grouped with a secondary seta on a grayish yellow plate, D2 as D1; SD setae grouped with numerous secondary setae on a grayish verruca; L, SV, and V setae  as in T2.-Abdomen.A1, A2 and A7.D1 seta grouped with two secondary setae on a dark gray plate, D2 seta grouped with six secondary setae on a dark gray verruca; SD setae grouped with eight secondary setae on a dark gray verruca; L setae grouped with seven secondary setae on a dark gray verruca; SV setae grouped with seven secondary setae on dark gray verruca; V seta grouped with a secondary seta on a grayish plate.A3-A6.D1, D2, and L setae as in segments A1, A2, and A7; SD setae grouped with five secondary setae on a dark gray verruca; SV and V setae on the prolegs on two dark grey plates with some secondary setae.A8.D1 seta grouped with a secondary seta on a grayish yellow plate, D2 seta solitary; SD setae grouped with numerous secondary setae on a grayish yellow verruca; L setae as in the other abdominal segments; SV setae not visible; V seta isolated.A9.D1 and D2 setae grouped with a secondary seta on a grayish yellow plate; SD setae grouped with numerous secondary setae on a  grayish yellow verruca; SV and V setae not visible.A10.D1, D2, and SD setae arranged on a large grayish-yellow anal plate; L and SV setae on the proleg on two grayish plates with some secondary setae. Second instar (Fig. 13C, D).Average duration: 6-8 days (n = 68); head capsule width 1.1-1.2mm (n = 5); body length 14.2-14.7 mm (n = 2); head brownish with yellowish spots and stripes; with numerous secondary setae.Body gray with yellow stripes; setae grayish shorter than the first instar; T1-T3 and A8-A10 dorsally darker; between T1 and T2, dorsally with a sparse tuft of reddish setae; A1-A7, D1, and D2 verrucae reddish; legs and abdominal prolegs reddish.
Life history.After being collected, the female was placed in a plastic container where it oviposited 110 eggs on the container wall.After about 15 days, the larvae hatched from the egg by cutting a small semicircular opening on the dorsal surface of the chorion.The larvae preferred to feed on Handroanthus sp.(Bignoniaceae).During the first and second instars, larvae fed by scraping both the adaxial and abaxial surfaces of the leaf; from the third on, larvae ate the entire leaf.
During the first and second instars, the larvae were very active all day and night, and when threatened, they released themselves from the host plant to the ground by a silk strand.From the third instar on, the larvae were less active during the day, spending this period resting on the branches of the host plant.In the last instar, when threatened, the larvae bristled the dorsal setae of the thoracic segments, making visible the arrangement of the red setae located in this region.
When larvae reached the prepupa, they left the host plant and walked on the soil of the container, burying in it after a few hours to pupate.The underground chamber was not found.After pupating, the earlier imagos emerged in about 60 days, while the latter in about 120 days.There was no difference between the emergence time of females and males.When both emerged on the same night, they immediately mated.An unmated female oviposited about 130 eggs, while a female that had copulated oviposited about 270 eggs.
Distribution.Based on museum records, the species is present in the Argentina state of Entre Ríos and the Brazilian states of Espírito Santo, Minas Gerais, Paraná, Rio de Janeiro, Santa Catarina, and São Paulo (Fig. 22).
Etymology.This species is named in honor of the parents of the first author, who always encouraged the first and second authors to persist in their studies; the specific epithet is a combination of their first names, Jair and Ana.2C, D, 4A, B).Olceclostera quilombola sp.nov.has all tibias with a band of long scales with dark brown apex, while O. jairana sp.nov.presents these scales with light brown apex (Fig. 6A, B).In the genitalia, O. quilombola sp.nov.presents, in lateral view, a more flat tegumen, wider socii, and a less elongated uncus, and, in ventral view, the anterior projection of the saccus is wider than O. jairana sp.nov.(Figs 17,19,21).2E, F, 4C, D, 6B, 17, 18).Head.MALE.As in O. jairana sp.nov.Female.Similar to the male, slightly smaller than the male's head; antenna, pecten shorter than the male's pecten; palpus, thinner and shorter than the male's palpus.-Thorax.MALE and FEMALE.As in O. jairana sp.nov.-Legs.As in O. jairana sp.nov., but all tibias with a band of long scales with dark brown apex projecting towards the posterior region (Fig. 6B).-Wings shape and venation.As in O. jairana sp.nov.(Figs 2E,F,4C,D).MALE.Forewing length: 18-20.5 mm (n = 10).Upperside: As in O. jairana sp.nov., but ground color gray; subapical region with two rectangular hyaline spots between M 1 and M 3 ; outer margin crenulated.Underside: Similar to O. jairana sp.nov.Hindwing.Upperside and underside: Similar to O. jairana sp.nov., but the outer margin crenulated between M 3 and CuA 2 .FEMALE.Forewing length: 25.8 mm (n = 01); similar to O. jairana sp.nov.; general color similar to the male, the apex is more acute than in males; on dcm, the patch of black scales and the hyaline spots are broader than in the male.Hindwing.Similar to the male, but on dcm, the dot of black scales is broader than in the male.-Abdomen.MALE and FEMALE.As in O. jairana sp.nov.-Genitalia.MALE (Fig. 17).Similar to O. jairana sp.nov., but the aedeagus is thinner and does not have spines at the apex.FEMALE (Fig. 18).As in O. jairana sp.nov., but the eighth tergum and the ostium bursae are narrower.

Description (Figs
Distribution.Based on museum records, the species is present in the Brazilian states of Brasília, Espírito Santo, Minas Gerais, Paraná, Rio de Janeiro, and São Paulo (Fig. 22).
Etymology.This species is named in honor of the quilombola communities that inhabit the region of the "Parque Estadual das Lauráceas" the type locality of this species.Diagnosis.Externally, O. xeta sp.nov.can be distinguished from O. jairana sp.nov.by all tibias with a band of long scales with grayish brown apex, while O. jairana sp.nov.presents these scales with light brown apex (Fig. 6A, C).Dorsally, the abdomen of O. xeta sp.nov.has a grayish color, while O. jairana sp.nov.has a copper color (Fig. 2C, D, G, H).The genitalia of O. xeta sp.nov.presents, in lateral view, a wider socci than O. jairana sp.nov., and the process of sacculus is half the length of that of O. jairana sp.nov.The aedeagus of O. xeta sp.nov., does not have spines, and the basal portion of the vesica is covered by small cornuti (Figs 10,19).
Olceclostera xeta sp.nov.differs from O. quilombola sp.nov.by external and genitalia characters: on the forewing of O. xeta sp.nov. the two subapical hyaline spots, between M 1 and M 3 , are rounded and smaller.Otherwise, these spots are rectangular in O. quilombola sp.nov.Dorsally, the abdomen of O. xeta sp.nov.has a grayish color, while O. quilombola sp.nov.has a copper color (Fig. 2E-H).Olceclostera xeta sp.nov.has all tibias with a band of long scales with grayish brown apex, while O. quilombola sp.nov.presents these scales with dark brown apex (Fig. 6B, C).The genitalia of O. xeta sp.nov.presents the process of sacculus with half of the length of O. quilombola sp.nov., and the basal portion of the vesica is covered by small cornuti, while the aedeagus of O. quilombola sp.nov.has vesica without cornuti (Figs 17,19).
Olceclostera xeta sp.nov. is similar to O. wayana sp.nov., but slightly darker gray and smaller in size (17 mm forewing length).The subapical hyaline spots are slightly larger.In the genitalia, O. xeta sp.nov.differs from O. wayana sp.nov.by the presence of a protrusion on the apex of the aedeagus.2G, H, 4E, F   Etymology.This species is named after the Xetá indigenous people.The Xetá were original inhabitants of the northwestern Paraná and the last ethnic group in the state to come into contact with white colonizers (in 1954) who invaded their territory and decimated the Xetá population.Etymology.This species is named after the Wayana indigenous people, inhabiting along the Maroni River, at the border between French Guiana and Suriname.

Maximum Likelihood Tree
Based on sequences of the mitochondrial COX1 gene (barcode region, 658pb  23).This relationship, however, shows low support.The topology obtained between the Olceclostera new species supports their distinctiveness, whose genetic distances ranged from 3.4% to 10% (Table 1).forewings and patches of broader scales on the abdomen are also present in the other Olceclostera species but with different configurations.These characters, together with characters of the genitalia (uncus bifid V-shaped, gnathos converted to sclerotized plates, and subscaphium sclerotized), are proposed as diagnostic of the genus.

Discussion
The configuration of the hyaline spots and the number of patches of broader scales laterally on the abdomen can be used to distinguish groups of species of Olceclostera.For example, some species have only a yellowish hyaline spot on their forewings and one pair of large patches of broader scales laterally on each abdominal segment (e. g.Olceclostera guanduna Drautd, 1929, Olceclostera dentata Schaus, 1910, O. irrorata, Olceclostera leticiensis Herbin, 2021, Olceclostera microps (Walker, 1855) and Olceclostera reperta (Walker, 1865)), while others have only a whitish hyaline spot on their forewings and two pairs of large patches of broader scales laterally on the proximal abdominal segments (e.  these spots are not present in all the species of the genus.Therefore the phylogenetic significance of this character requires further studies. Knowledge about immature traits is essential to understanding the Lepidoptera evolution (Balcázar-Lara and Wolfe 1997; Freitas and Brown 2004).In this study, we describe for the first time the morphology of all immature stages of one Olceclostera species and illustrate for the first time images of the egg of an Apatelodidae species using SEM.In dorsal view, the egg of O. jairana sp.nov. is similar to that described for O. seraphica, Apatelodes, and Thelosia species (Packard 1895;Peigler 1994;Thomazini 2010;Herbin 2020;Orlandin et al. 2022aOrlandin et al. , 2022b) ) appearing to be flattened as cited for these species.However, the lateral view of the eggs of O. jairana sp.nov.shows that the micropylar pole is concave (pie-shaped) and not flattened (coin-shaped).This finding is important because it demonstrates that there are a greater variety of egg shapes in Apatelodidae than previously thought since we found dome-shaped eggs in Drepatelodes (Orlandin et al. 2021) and coin-shaped eggs in Apatelodes (Packard 1895;Orlandin et al. 2022aOrlandin et al. , 2022b)).To infer the evolutionary significance of the egg morphology in Apatelodidae, eggs of more species of the different genera should be obtained, and these eggs must be studied in detail (e. g. using SEM), including records in lateral view.
The chaetotaxy of O. jairana sp.nov. is very similar to the chaetotaxy of other Apatelodidae species, which were described in detail (Orlandin et al. 2021(Orlandin et al. , 2022a(Orlandin et al. , 2022b)).The head shows only primary setae, as the other species already studied.On the body, the known species have secondary setae, mostly arranged on large verrucae.However, the most striking difference in the chaetotaxy of O. jairana sp.nov. is the lack of L2 on A1-A8 isolated, while in Apatelodes je Orlandin, Piovesan & Carneiro, 2022, Apatelodes kotzschi Draudt, 1929, and D. friburgensis, L2 is isolated (Orlandin et al. 2021, 2022a, 2022b).Additionally, first instar larvae of Apatelodidae also seem to present a great variation in the number of secondary setae grouped with D1 and D2 on the thorax and the abdomen.Olceclostera jairana sp.nov.has on T2-T3 segments, D1 seta solitary and D2 seta arranged with only one secondary seta, while A. je also has D1 seta solitary but D2 seta on a verruca with about six secondary setae (Orlandin et al. 2022a).On the other hand, in A. kotzschi, D1 seta is on a verruca with three secondary setae, and D2 seta is on a verruca with about six secondary setae (Orlandin et al. 2022b), while D. friburgensis has D1 seta on a verruca with two or three secondary setae, and D2 seta next to a secondary seta (Orlandin et al. 2021).Between A1-A7, O. jairana sp.nov.presents D1 seta on a verruca with two more secondary setae and D2 seta on a verruca with about six secondary setae.Unlike this configuration, A. je and A. kotzschi have both D1 and D2 setae on each verruca with five to seven secondary setae, and D. friburgensis has D1 seta on a verruca with about five secondary setae and D2 seta solitary (Orlandin et al. 2021(Orlandin et al. , 2022a(Orlandin et al. , 2022b)).In A8, the configuration of dorsal setae is very similar, and all species have the D1 seta with one to three secondary setae and solitary D2 seta.Here we emphasize that the chaetotaxy of Apatelodiade still requires a broader study to evaluate its diagnostics and phylogenetic relevance to delimit monophyletic groups in the family.
The available information on the immature stages of Olceclostera allows us to make some comparisons between the species of the genus.In the last larval instars, O. jairana sp.nov. is very similar to O. bifenestrata and O. seraphica, differing from them by the more brownish color, by the dorsal region of the body with smaller yellow spots (larger in O. bifenestrata) and by the absence of orange spots on the dorsal region of the body (present in O. seraphica) (Peigler 1994;Janzen and Hallwachs 2009).The color pattern, the slightly flattened dorsoventrally body, and the pattern in the setae distribution make these species very similar to larvae of some species of Euglyphis (Lasiocampidae) (Janzen and Hallwachs 2009).Euglyphis caterpillars are known for having urticating setae (Specht et al. 2008), absent in the Olceclostera genus, which may constitute a Batesian-type mimicry.In Apatelodidae, this type of mimicry seems to be common, with caterpillars mimicking larvae of Megalopygidae, Arctiinae, and Lasiocampidae (Orlandin et al. 2021(Orlandin et al. , 2022a)).
Olceclostera jairana sp.nov.pupates in the soil like other known Apatelodidae species.This trait appears to be shared by most species in the family, as, so far, only Prothysana is known to make cocoons (Becker and Orlandin in press).The pupa of O. jairana sp.nov. is similar to that described for O. seraphica and to the other species of Apatelodidae (Peigler 1994;Janzen and Hallwachs 2009;Thomazini 2010;Orlandin et al. 2021Orlandin et al. , 2022aOrlandin et al. , 2022b)).However, the absence of a rudimentary cremaster and the almost smooth body (few punctuations) makes it much more similar to the pupae of D. friburgensis (Orlandin et al. 2021).
Apatelodidae, like most moth groups, has been neglected in worldwide collections, although they belong to one of the most collectible moth groups in the world, the Bombycoidea (e. g.Saturniidae, Sphingidae, Brahmaeidae).As for these groups, detailed investigation on male genitalia showed that some old recognized wide-spread species comprise a set of cryptic species whose geographic distribution is much more restricted than previously thought.This is the case of South American specimens formerly identified as O. bifenestrata, that currently are the type series of O. jairana sp.nov., O. quilombola sp.nov., O. xeta sp.nov., and O. wayana sp.nov., whose differences in genitalia morphology are corroborated by great genetic distances in COX1 (barcode region, 658pb) sequences.The accumulated knowledge, especially in recent years, shows that there is still much to be studied to understand the evolutionary relationships between Apatelodidae lineages.

Competing interests
The authors have declared that no competing interests exist.

Figure 6 .
Figure 6.Lateral view of the scales that cover the tibia of Olceclostera.A Olceclostera jairana sp.nov.; B Olceclostera quilombola sp.nov.; C Olceclostera xeta sp.nov.The arrow points to the apex of the scales, which shows a different brown hue in each species.

Figure 9 .
Figure 9. Olceclostera jairana sp.nov.Large patch of broader scales on the abdomen.A lateral view; B detail.

Figure 10 .
Figure 10.Male genitalia of Olceclostera jairana sp.nov.A ventral view; B lateral view; C aedeagus, lateral view, apex with two spines; D aedeagus, apex with one spine; E aedeagus, apex with three spines.

Figure 11 .
Figure 11.Female genitalia of Olceclostera jairana sp.nov.A ventral view; B dorsal view; C lateral view; D signum detail.

Figure 12 .
Figure 12.Egg of Olceclostera jairana sp.nov.A three days after oviposition, dorsal view; B close to eclosion, lateral view; under scanning electronic microscopy (SEM) C dorsal view; D rosette cells; E micropylar openings; F aeropyles; G magnification of the aeropyles.

Figure 13 .
Figure 13.Immature stages of Olceclostera jairana sp.nov.A first instar, dorsal view; B first instar, lateral view; C second instar, dorsal view; D second instar, lateral view; E third instar, dorsal view; F third instar, lateral view; G fourth instar, dorsal view; H fourth instar, lateral view; I fifth instar, dorsal view; J fifth instar, lateral view.

Figure 14 .
Figure 14.Head chaetotaxy of first instar of Olceclostera jairana sp.nov.A frontal view; B lateral view.

Figure 15 .
Figure 15.Body chaetotaxy of first instar of Olceclostera jairana sp.nov.A dorsal view; B lateral view.

Figure 16 .
Figure 16.Pupa of Olceclostera jairana sp.nov.A dorsal view; B lateral view; C ventral view; D details of male genital scars; E details of female genital scars.

Figures
Figures 2E, F, 4C, D, 17, 18 Type locality.Parque Estadual das Lauráceas, Adria nópolis Municipality, Paraná State, Brazil.Diagnosis.Olceclostera quilombola sp.nov.differs from O. jairana sp.nov.by the following characters: on the forewing of O. quilombola sp.nov. the two subapical hyaline spots between M 1 and M 3 are rectangular and always visible, while these spots are rounded, with the one between M 2 -M 3 almost unnoticeable in some cases in O. jairana sp.nov.(Figs2C, D, 4A, B).Olceclostera quilombola sp.nov.has all tibias with a band of long scales with dark brown apex, while O. jairana sp.nov.presents these scales with light brown apex (Fig.6A, B).In the genitalia, O. quilombola sp.nov.presents, in lateral view, a more flat tegumen, wider socii, and a less elongated uncus, and, in ventral view, the anterior projection of the saccus is wider than O. jairana sp.nov.The aedeagus of O. quilombola sp.nov. is thinner than that of O. jairana sp.nov.and does not have spines at the apex (Figs 10, 17).Olceclostera quilombola sp.nov.differs from O. xeta sp.nov.and O. wayana sp.nov.by external and genitalia characters: on the forewing of O. quilombola sp.nov., the two subapical hyaline spots, between M 1 and M 3 , are rectangular and always visible, while in O. xeta sp.nov.and O. wayana sp.nov., these spots are rounded and

Figure 18 .
Figure 18.Female genitalia of Olceclostera quilombola sp.nov.A ventral view; B dorsal view; C lateral view; D signum detail.
, 6C, 19, 20).Head.MALE.As in O. jairana sp.nov.FEMALE.Antenna, pecten shorter than the male's pecten.-Thorax.MALE and FEMALE.As in O. jairana sp.nov.-Legs.All tibias with a band of long scales with grayish-brown apex projecting towards the posterior region (Fig. 6C).-Wings shape and venation.MALE and FEMALE.As in O. jairana sp.nov.(Figs 2G, H, 4E, F).MALE.Forewing length: 16.5-17 mm (n = 03).Upperside and underside: Similar to O. jairana sp.nov.Hindwing.Upperside and underside: Similar to O. jairana sp.nov.FEMALE.Forewing length: 17-21 mm (n = 03); general color similar to the male, the apex is more acute than in males; lighter grey than the female of O. jairana sp.nov.and O. quilombola sp.nov.The hyaline spot between M 1 -M 2 is small than in males, and the hyaline spot between M 2 -M 3 is reduced or absent.Hindwing.As in male.Abdomen.MALE and FEMALE.Similar to O. jairana sp.nov., but dorsally gray.-Genitalia.MALE.(Fig. 19).Similar to O. jairana sp.nov., but aedeagus without spines; vesica with the basal portion covered by small cornuti.FEMALE.(Fig. 20).Similar to O. jairana sp.nov., but the ostium bursae are broader and the signum leaf-shaped.Distribution.Based on museum records, the species is present in the Brazilian states of Paraná and the Para-

Figure 21 .
Figure 21.Male genitalia of Olceclostera wayana sp.nov.A ventral view; B lateral view; C aedeagus lateral view; D detail of vesica with a plate with minute teeth.

Figure 22 .
Figure 22.Geographical distribution of new species of Olceclostera.

Figure 23 .
Figure 23.Maximum likelihood phylogenetic tree, inferred with IQ-TREE, based on cytochrome c oxidase subunit I (COX1, barcode region, 658pb) showing the relationship among new species of Olceclostera, twenty other species of Olceclostera, and representatives of ten genera of Apatelodidae as outgroups.Numbers indicate ML ≥ 70, and * indicates maximum support of 100.Low support values were not shown.The scale bar represents the expected number of nucleotide substitutions per site.

xeta sp. nov. and O. wayana sp. nov., and
the process of sacculus is about twice as big as in O.

xeta sp. nov. and O. wayana sp. nov. The aedeagus of O. jairana sp. nov. has
an apex with spines and vesica without cornuti, while in O.

xeta sp. nov. and O. wayana sp. nov. the
aedeagus does not have spines and the bas- The aedeagus of O. quilombola sp.nov. is thinner than that of O. jairana sp.nov.and does not have spines at the apex (Figs 10, 17).Olceclostera quilombola sp.nov.has all tibias with a band of long scales with dark brown apex, while O. xeta sp.nov.and O. wayana sp.nov.present these scales with grayish brown apex (Fig. 6B, C).Dorsally, the abdomen of O. quilombola sp.nov.has a copper color, while O. xeta sp.nov.and O. wayana sp.nov.have a grayish color (Fig. 2E-J).The genitalia of O. quilombola sp.nov.presents, in lateral view, the socci apex narrower and triangular, while O. xeta sp.nov.and O. wayana sp.nov.have the socci apex rounded.The process of sacculus of O. quilombola sp.nov. is about twice as big as that of O. xeta sp.nov.and O. wayana sp.nov.The aedeagus of O. quilombola sp.nov.has vesica without cornuti, while in O. xeta sp.nov.and O. wayana sp.nov. the basal portion of the vesica is covered by small cornuti Olceclostera quilombola sp.nov.differs from O. xeta sp.nov.and O. wayana sp.nov.by external and genitalia characters: on the forewing of O. quilombola sp.nov., the two subapical hyaline spots, between M 1 and M 3 , are rectangular and always visible, while in O. xeta sp.nov.and O. wayana sp.nov., these spots are rounded and shorter.

3.1.4. Olceclostera wayana Herbin, Orlandin and Carneiro sp. nov.
The two subapical hyaline spots between M 1 and M 3 are tiny.In the genitalia, O. wayana sp.nov.differs from O. xeta sp.nov.by the absence of a protrusion on the apex of the aedeagus and vesica with a plate with minute teeth.