Three new species of Byrsopteryx Flint microcaddisflies from Peru (Insecta: Trichoptera) including DNA-based larval associations

Specimen collecting and morphological study

Adults and immatures of Byrsopteryx studied here were collected with aspirators, with finger moistened in alcohol (adults) or with aid of entomological forceps (immatures). In addition, adults were also collected with a Malaise trap set at one of the collecting sites. Specimens were collected under permit RD-0297-2012-AG-DGFFS-DGEFFS, issued by Dirección General Forestal y de Fauna Silvestre, Peru. Larvae and most adults were fixed and preserved in 96% ethanol, however some adults were killed in jars with ethyl acetate and pinned dry, to better preserve coloration. Byrsopteryx specimens were found in four sites, in Cusco and Puno provinces, Peru. Collecting sites exhibited the typical environment for Byrsopteryx species: rocky streams and small waterfalls (Fig. 1).

Male and female genital structures were analyzed after the clearing procedure with a heated solution of 10% KOH, as described in Ross (1944). Abdomens were mounted in temporary slides and observed under a compound microscope equipped with camera lucida. Then, pencil sketches were made and used as templates in Adobe Illustrator (Adobe Systems Inc.) to create vector graphics, as detailed in Holzenthal (2008). Adult male holotypes and larvae were photographed with a digital camera attached to a Leica stereomicroscope. Photographs at different focal planes were obtained and then stacked with Leica Application Suite or Helicon focus Lite software and then edited in Adobe Photoshop (Adobe Systems Inc.). After observation, abdomens were stored in microvials with ethanol, for specimens in alcohol, or glycerin for pinned specimens. Terminology used here follows that of Harris & Holzenthal (1994), except for the inferior appendages, for which we used the interpretation of Santos, Nessimian & Takiya (2016). Holotypes of the newly described species are deposited at Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima (MUSM). Paratypes are in the same institution and also in Coleção Prof. José Alfredo Pinheiro Dutra, Departamento de Zoologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro (DZRJ) and Coleção de Invertebrados, Instituto Nacional de Pesquisas da Amazônia, Manaus (INPA). Extracted DNA solutions are deposited at DZRJ.

DNA extraction, amplification, and sequencing

DNA was extracted from the entire body of specimens using the Wizard SV Genomic DNA Purification System (Promega Corporation, Madison, WI, USA), without tissue maceration. After extraction, specimens were returned to ethanol and deposited in the original collection as a DNA voucher. COI fragments were amplified using different pair of primers (Folmer et al., 1994; Simon et al., 1994): HCO-2198 (5′-TAAACTTCAGGGTGACCAAAAAATCA) in combination with LCO- 1490 (5′-GGTCAACAAATCATAAAGATATTGG); HCO-2198 with C1-J-1718 (5′-GGAGGATTTGGAAATTGATTAGTTCC); or Ron (5′-GGATCACCTGATA- TAGCATTCCC) and Nancy (5′-CCCGGTAAAATTAAAATATAAACTTC).

Polymerase chain reaction (PCR) conditions were as follows: when using HCO-2198 and LCO-1490, initial denaturation at 95 °C for 3 min; 5 cycles of denaturation at 95 °C for 1 min, annealing at 45 °C for 1,5 min, and extension at 72 °C for 1 min, then 35 cycles of 95 °C for 40 s, 51 °C for 1 min, and 72 °C for 1 min; and final extension at 72 °C for 7 min. When using other primer combinations, initial denaturation at 94 °C for 3 min; then 35 cycles of 94 °C for 1 min, 50 °C for 1 min, and 72 °C for 2 min; and final extension at 72 °C for 7 min. PCR products were sent to Macrogen Inc., Seoul, for purification and sequencing reactions.

Phylogenetic analysis

Forward and reverse electropherograms of each sample were assembled and manually edited in Sequencher 4.1 (Gene Codes, Ann Arbor, Michigan, USA). Consensus sequences were verified with the Blast tool in GenBank to check for contamination. Additional sequences of Byrsopteryx and of Celaenotrichia edwardsi Mosely, 1934 as outgroup, were obtained from Bold Systems (Table 1). COI sequences were aligned using ClustalW algorithm implemented in MEGA X (Kumar et al., 2018) and translated into amino-acid sequences to check for stop codons. Final alignment resulted in a matrix with 653 bp and 20 sequences (File S1). All COI sequences are available in GenBank (accession numbers are provided in Table 1). MEGA X was also used to calculate Kimura 2-Parameter (K2P) divergences (Kimura, 1980), with pairwise deletion of missing information. A maximum likelihood analysis was conducted in RAxML 8.2.11 (Stamatakis, 2014) with 500 search replicates and model GTR+G+I, as selected in jModeltest 2 (Darriba et al., 2012) using Akaike Information Criterion (Akaike, 1974). Branch support tree was assessed with 1,000 pseudoreplicates of non-parametric bootstrap (BS, Felsenstein, 1985).

New species names

The electronic version of this article in Portable Document Format (PDF) will represent a published work according to the International Commission on Zoological Nomenclature (ICZN), and hence the new names contained in the electronic version are effectively published under that Code from the electronic edition alone. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix http://zoobank.org/. The LSID for this publication is: urn:lsid:zoobank.org:pub:71531CB9-F919-4DB9-8885-B8207F0A82F4. The online version of this work is archived and available from the following digital repositories: PeerJ, PubMed Central and CLOCKSS.

Phylogenetic analysis and species divergences

The maximum likelihood tree (-lnL= 3322.784548, Fig. 2) recovered two of the new species described herein, B. inti sp. nov. and B. mamaocllo sp. nov., as sister species (BS = 79) and related to a clade (BS = 52) including B. gomezi, B. tapanti, and B. esparta. The other new species described, B. mancocapac sp. nov., was found as sister to B. abrelata, but with no significant bootstrap support.

Concerning species interspecific COI K2P divergences, Byrsopteryx species varied from 12.3–26.1% (see Table S1), minimum divergence (K2P distances) was between B. tapanti and B. gomezi, both from Costa Rica. Among the Peruvian species, minimum interspecific divergence was 14.9%, between B. inti sp. nov. and B. mamaocllo sp. nov.. Furthermore, maximum K2P intraspecific distances observed were 1.4% for B. mancocapac sp. nov. (n = 8), 0.7% for B. mamaocllo sp. nov. (n = 3), and 0% for B. gomezi (n = 2). Based on these divergences and phylogenetic placement (Fig. 2), we were able to associate males and larval specimens of B. mamaocllo sp. nov. and B. mancocapac sp. nov.

Taxonomy

Byrsopteryx inti new species

urn:lsid:zoobank.org:act:9B5FAC2D-A98F-4B88-97BC-CE7CA5105535

- Byrsopteryx sp. PE1 Santos, Nessimian & Takiya, 2016:461. Phylogenetic placement.

Figs. 3–5

Description. Adult male. Coloration. General color dark brown; head dorsum and antennal base densely covered with white setae; mesoscutum mostly covered with white setae; forewing with four distinct maculae of white setae: an oval band at base of medial area, a trapezoidal macula near midcostal margin, a small subapical spot near posterior margin of wing, and an apical spot (Fig. 3). Length. Total length 2.4–2.6 mm (n = 3). Head. Unmodified. Antennae 19-articulated. Ocelli 3. Maxillary palpi 5-articulated; articles I and II very short and globular, article III very long, as long as articles IV and V combined; articles IV and V with similar lengths, each one about twice as long as wide. Thorax. Forewing venation strongly reduced; with small, oblique basal lobe; line of weakness distinct, bordered by row of very long, thin setae; with semi-dome formed by a thickening of wing membrane, to which apices of very long setae converge; retinaculum distinct (Fig. 4A, Fig. 4B). Hind wing venation reduced; frenulum distinct, with row of four short, hooked setae (Fig. 4C). Tibial spur formula 0, 3, 4. Abdomen. Segment VII without ventromesal process. Male genitalia. Segment VIII shorter dorsally than ventrally. Sternum with median digitate process posteriorly, slightly capitate (Fig. 5A); with pair of posteroventral processes, diverging apically in ventral view (Fig. 5A), slightly upturned in lateral view, each one with a thin rounded membrane at apex (Figs. 5B, 5D); and with pair of dorsolateral processes, crossing each other apically in dorsal view (Fig. 5C), upturned in lateral view (Fig. 5D). Very long stout setae covering apical portions of both sternum and tergum. Segment IX recessed within segment VIII, projecting anteriorly through posterior portion of segment VII; open ventrally; with deep mesal incision anteriorly in dorsal and ventral views (Figs. 5A, 5C); narrowing anteriorly in lateral view (Fig. 5D). Subgenital plate slender, with rounded apex in ventral view (Fig. 5A), downturned and with apical semicircular incision (Fig. 5D). Inferior appendages positioned dorsolaterally; elongate, digitate, converging apically in ventral and dorsal view (Figs. 5A, 5C); downturned in lateral view (Fig. 5D); covered with very short setae. Tergum X membranous; covered with minute setae; with sclerotized lateral bars. Phallus tubular, with slight constriction between basal and apical portions; basal portion short, about half length of apical portion and slightly wider; sclerotized region of apex with deep median incision in dorsal view, with pair of short triangular projections at basal margin of the incision (Fig. 5E), with a V-shaped incision in lateral view (Fig. 5F); membranous at apex.

Adult female. Unknown.

Larva. Unknown.

Type material. Holotype male. PERU: Cusco, 19 rd km W Quincemil, Río Araza tributary. 13°20′10″S 70°50′57″W 874 m. 23-28.viii.2012. Malaise trap. RR Cavichioli, JA Rafael, APM Santos, DM Takiya leg. Alcohol (MUSM). Paratypes. Same data as holotype, except 1 male, pinned (DZRJ); same data as holotype, manual collecting, 2 males, in alcohol (MUSM), 1 male, in alcohol (DZRJ).

Etymology. The species name (used as a noun in apposition) is an allusion to the popular Inca god Apu Inti, the sun god. The Inca people appeared in the Andes region and established the Inca Empire in pre-Colombian America. Cusco, where the type material comes from, was the center of the Inca Empire.

Distribution. Peru (Cusco).

Remarks. Among the three new species described here from Peru, Byrsopteryx inti sp. nov. was the most rarely collected. Only five male specimens were collected and, although Byrsopteryx larvae and adults were abundantly seen and collected in explored localities, both female and immature specimens of this new species were not found and remain unknown.

Males of B. inti sp. nov. share with those of B. mamaocllo sp. nov. a feature not observed in any other species in the genus: presence of a sclerotized semi-dome on forewings (Figs. 3A, 4A). Also, the general aspect of the male genitalia of these two new species is also most similar (Figs. 5B, 5D), in that they share both a pair of dorsolateral and a pair of posteroventral processes on sternum VIII. Other Byrsopteryx species that also have dorsolateral processes on segment VIII are B. chaconi Harris & Holzenthal, 1994, B. cuchilla Harris & Holzenthal, 1994, B. esparta Harris & Holzenthal, 1994, B. solisi Harris & Holzenthal, 1994, B. tapanti Harris & Holzenthal, 1994, and B. tica Harris & Holzenthal, 1994. However, B. inti sp. nov. can be further distinguished from all others based on (1) the sternum VIII with a median digitate process, slightly capitate posteriorly (Fig. 5A); and (2) longer dorsolateral processes from segment VIII, which cross each other apically in dorsal view (Fig. 5C).

Byrsopteryx mamaocllo new species

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- Byrsopteryx sp. PE2 Santos, Nessimian & Takiya, 2016:461. Phylogenetic placement.

Figs. 6–10

Description. Adult male. Coloration. General color dark brown; head dorsum and antennal base densely covered with white setae; mesoscutum mostly covered with white setae; forewing with three maculae of white setae, a longitudinal band along basal third of costal margin, a subapical spot near posterior margin of wing, and an apical spot (Fig. 6). Length. Total length 2.6–3.0 mm (n = 15). Head. Unmodified. Antennae 20-articulated. Ocelli 3. Maxillary palpi 5-articulated; articles I and II very short and globular, article III very long, as long as articles IV and V combined; articles IV and V with similar lengths, each one about twice as long as wide. Thorax. Forewing venation strongly reduced; with small, oblique basal lobe; line of weakness distinct, bordered by row of very long, thin setae; with semi-dome formed by thickening of wing membrane (absent in females), to which apices of very long setae converge; retinaculum distinct (Fig. 7A, Fig. 7B). Hind wing venation reduced; frenulum distinct, with row of five short, hooked setae (Fig. 7C). Tibial spur formula 0, 3, 4. Abdomen. Segment VII without ventromesal process. Male genitalia. Segment VIII shorter dorsally than ventrally. Sternum with a pair of short spinelike posteroventral processes, curved inwards in ventral and caudal views (Figs. 8A, 8B), upturned in lateral view (Fig. 8D), each one with a thin rounded membrane; and with pair of short dorsolateral digitate processes, slightly upturned in lateral view (Fig. 8D). Very long stout setae covering apical portions of both sternum and tergum. Segment IX recessed within segment VIII, projecting anteriorly through posterior portion of segment VII; with deep mesal incision anteriorly in ventral and dorsal views (Figs. 8A, 8C); narrowing anteriorly in lateral view (Fig. 8D). Subgenital plate subtriangular in ventral and dorsal views, apex with a small darkened digitate projection (Figs. 8A, 8C); apex downturned in lateral view (Fig. 8D). Inferior appendages positioned dorsolaterally; elongate, almost straight, parallel in dorsal and ventral views (Figs. 8A, 8C); apex downturned in lateral view (Fig. 8D); covered with very short setae. Tergum X membranous; covered with minute setae or sensilla; with sclerotized lateral bars. Phallus tubular; with slight constriction between basal and apical portions; basal portion short, about half length of apical portion; sclerotized region of apex rounded in dorsal view, with a short bifid projection subapically in dorsal view (Fig. 8E), trifid in lateral view (Fig. 8F); membranous at apex.

Adult female. Coloration and general morphology of head and thorax as in male, except forewing simple, without semi-dome structure or any other modification. Length. Total length 2.8–3.8 mm (n = 13). Abdomen. Segment VI without ventromesal process. Segment VII elongate, ventrally with short plumose setae (Fig. 9A). Segment VIII approximately as long as wide, with row of long setae on posterior margin, internally with pair of elongate lateral apodemes extending to middle of segment VII (Fig. 9A). Segment IX short, slightly sclerotized, internally with pair of elongate lateral apodemes extending to anterior margin of segment VIII (Fig. 9A). Segment X very short, narrowed apically, with a pair of digitate papillae (Fig. 9A). Vaginal apparatus slightly sclerotized, inconspicuous in cleared specimens; elongate, with anterior region rounded (Fig. 9B).

Larva (final instar). Length 1.8–2.5 mm (n = 29). Head brown to dark brown, unpigmented around eyes; quadrangular; frontoclypeal and coronal sutures indistinct; dorsum with reticulate pattern formed by microscopic setae; setae 9 very long; antennae 1-articulated, apical seta indistinct; mandibles without distinct teeth. Thoracic nota strongly sclerotized, brown, with lateral and posterior margins dark brown, with short stout setae (Fig. 10); pro- and mesonota with transverse mesal sclerotized ridge, less developed on metanotum; pronotum with middorsal ecdysial line, pair of anterolateral depressed areas (Fig. 10C); meso- and metanota slightly broader than long, without middorsal ecdysial lines (Fig. 10C); meso- and metathoracic pleural sclerite large, sclerotized; ventrally, prothorax with two pairs of thin, elongate sclerites, which converge medially; with a ventral thin intersegmental sclerite between meso- and metathorax; thoracic legs short, stout, similar in size, shape, and setation (Fig. 10B). Abdomen slightly compressed (Figs. 10B, 10C); all abdominal segments with sclerotized tergites: segments I and II with large tergite, covering most of dorsal area of the segment (Fig. 10C); segments III–VII with transverse tergite, lightly sclerotized, bearing many setae; segments VIII and IX with platelike tergite, heavily sclerotized, setose, forming a dorsal, almost circular operculum (Fig. 10D), which has a typical color pattern, with tergites dark brown with a pair of oval lighter areas (Fig. 10D); operculum closing posterior opening of the case. Pleurites of segments I and II small, with three and two setae respectively (Fig. 10B, in detail); those of segments III–VIII absent, but region with two small setae each one. Anal proleg short, setose with pair of long posterior setae; anal claw stout, curved to approximately 90° , with basal peglike setae.

Laval case. Length 1.6–2.8 mm (n = 29). Made entirely of silk with bits of mineral material incorporated, translucid; slightly compressed laterally; with poorly closed dorsal seam (Fig. 10A); dorsal margin irregular; anterior and posterior openings circular.

Type material. Holotype male. PERU: Cusco, 20 rd km W Quincemil, Pte. Saucipata, Río Araza tributary, 13°20′13″S 70°51′15″W 893 m. 22.viii.2012. APM Santos, DM Takiya leg., pinned (MUSM). Paratypes. Same data as holotype, except 5 males, 7 females, pinned (DZRJ); Same data as holotype, except 5 males, in alcohol (MUSM); PERU: Cusco, 19 rd km W Quincemil, Rio Araza tributary, 13°20′10″S 70°50′57″W 874 m. 23-28.viii.2012. APM Santos, DM Takiya leg., 16 males, 8 females, in alcohol (MUSM); same data, except 5 males, in alcohol (DZRJ); same data, except malaise trap, 4 males, in alcohol (INPA); PERU: Cusco, 3 rd km E Quincemil, 13°13′03″S 70°43′40″W 633 m. 20.viii.2012. APM Santos, DM Takiya leg., 1 female, pinned (DZRJ); PERU: Puno, 6 rd km W Mazuko, Pte. La Cigarra 13°08′27″S 70°23′14″W 353 m. 01.ix.2012. APM Santos, DM Takiya leg., 5 females, pinned (DZRJ); same data, except 1 female, in alcohol (MUSM).

Additional material examined. Same data as holotype, 3 larvae, in alcohol (MUSM); PERU: Cusco, 19 rd km W Quincemil, Rio Araza tributary, 13°20′10″S 70°50′57″W 874 m. 23–28.viii.2012. APM Santos, DM Takiya leg., 11 larvae, in alcohol (INPA); same data, except 18 larvae, 2 male pharate adult, in alcohol (DZRJ); same data, except 10 larvae, in alcohol (MUSM).

Etymology. The species name is an allusion to the Inca goddess Mama Ocllo (used as a noun in apposition), the “mother fertility”. According to Inca mythology, Mama Ocllo was daughter of Apu Inti and Mama Quilla, and she married her brother Manco Capac. Together, Mama Ocllo and Manco Capac founded Cusco and guided the people, enabling the beginning of the Inca civilization.

Distribution. Peru (Cusco, Puno).

Remarks. In the field, B. mamaocllo sp. nov. can be recognized from other two Peruvian species of Byrsopteryx by the color pattern with a long longitudinal white band on each forewing (Fig. 6). Byrsopteryx species show no sexual dimorphism in color pattern (Harris & Holzenthal, 1994), which have allowed male–female associations presented here. Byrsopteryx mamaocllo sp. nov. can be easily distinguished from other Byrsopteryx species also by its male genitalia; with segment VIII bearing a pair of short, posterior, spinelike processes on sternum, which are curved inwards and bordered by rounded, membranous structure (Figs. 8A, 8B); and a pair of short, heavily sclerotized, dorsolateral processes (Figs. 8A, 8D). See further comments on above Remarks of B. inti sp. nov. Females of B. mamaocllo sp. nov. also differ from those of other Byrsopteryx species by genitalia with the vaginal apparatus only slightly sclerotized, with anterior region rounded, and a posterior region mostly membranous (Fig. 9B).

Larvae of B. mamaocllo sp. nov. and of B. mancocapac sp. nov. are very similar to those previously described for other Byrsopteryx species, including general aspect of the case, the setation pattern of the head, and the sclerites. General aspects of Byrsopteryx larval morphology have been described in detail by Holzenthal & Harris (1991) and Santos & Nessimian (2010). Considering the two Peruvian larvae described here, B. mamaocllo sp. nov. differ from B. mancocapac sp. nov. by the: (1) almost circular operculum with two pairs of oval lighter areas, a pair on tergite VIII and another on IX (Fig. 10D), in B. mancocapac sp. nov. larvae the operculum is more rectangular; (2) case generally smaller than larva (Fig. 10A), in B. mancocapac sp. nov. its case is as long as larva; and (3) abdominal segment II with small pleurite bearing two setae (Fig. 10B), whereas in B. mancocapac sp. nov., it has three setae.

Byrsopteryx mancocapac new species

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- Byrsopteryx sp. PE3 Santos, Nessimian & Takiya, 2016:461. Phylogenetic placement.

Figs. 11–14

Description. Adult male. Coloration. General color dark brown; head dorsum and mesoscutum densely covered with black setae, with no white setae; forewing with four distinct maculae of white setae: a longitudinal oval band at base of medial area, a trapezoidal macula near midcostal margin, a subapical spot near posterior margin of wing, and an apical spot, spreading towards costal margin (Fig. 11). Length. Total length 2.6–3.6 mm (n = 23). Head. Unmodified. Antennae 18-articulated. Ocelli 3. Maxillary palpi 5-articulated; articles I and II very short and globular, article III very long, as long as articles IV and V combined; articles IV and V with similar lengths, each one about twice as long as wide. Thorax. Forewing venation strongly reduced; with distinct line of weakness; basal lobe apparently absent; semi-dome or other wing modification absent; retinaculum distinct (Fig. 12A). Hind wing venation strongly reduced; frenulum distinct, with two rows of three to five short, hooked setae (Fig. 12B). Tibial spur formula 0, 3, 4. Abdomen. Segment VII without ventromesal process. Male genitalia. Segment VIII shorter dorsally than ventrally. Sternum with deep mesal incision posteriorly, forming pair of platelike lobes in ventral view, each one with rounded corners and bearing longer setae on the posterolateral quadrant and smaller setae surrounding the internal margin (Fig. 13A). Tergum with transverse row of long and stout setae near posterior margin (Fig. 13B). Segment IX recessed within segment VIII, projecting anteriorly through posterior portion of segment VII; with deep mesal incision anteriorly in dorsal and ventral views (Figs. 13A, 13B); narrowing anteriorly in lateral view (Fig. 13C); open dorsally; with pair of small, rounded posterior projections in ventral view, each one with acute beak on internal margin (Fig. 13A). Subgenital plate with lateral arms converging posteriorly to a single darkened apex, strongly downturned in lateral view (Fig. 13C); with strong horn-like process emerging from base of each arm (Fig. 13C). Inferior appendages positioned dorsolaterally; elongate, slender, almost parallel in ventral and dorsal views (Figs. 13A, 13B); in lateral view, each one with dorsal platelike projection at basal portion, narrowing to apex and downturned (Fig. 13C). Tergum X with a basal trapezoid sclerite, with shallow incision in anterior margin in dorsal view (Fig. 13B); posterior portion conical and membranous. Phallus tubular, with slight constriction between basal and apical portions; basal portion short, less than half length of apical portion; with an acute sclerotized apical projection (Figs. 13C, 13D); membranous at apex (Fig. 13C).

Adult female. Unknown.

Larva (final instar). Length 2.0–3.8 mm (n = 28). Head brown to dark brown, unpigmented around eyes; quadrangular; frontoclypeal and coronal sutures indistinct; dorsum with reticulate pattern formed by microscopic setae; setae 9 very long; antennae 1-articulated, apical seta indistinct; mandibles without distinct teeth. Thoracic nota strongly sclerotized, brown, with lateral and posterior margins dark brown, with short stout setae; pro- and mesonota with transverse mesal sclerotized ridge, less developed on metanotum; pronotum with middorsal ecdysial line, pair of anterolateral depressed areas (Fig. 14C); meso- and metanota slightly broader than long, without middorsal ecdysial lines (Fig. 14C); meso- and metathoracic pleural sclerite large, sclerotized (Fig. 14B); ventrally, prothorax with two pairs of thin, elongate sclerites, which converge medially; with ventral thin intersegmental sclerite between meso- and metathorax; thoracic legs short, stout, similar in size, shape, and setation (Fig. 14B). Abdomen slightly compressed (Fig. 14C); all abdominal segments with sclerotized tergites: segments I and II with large tergite, covering most of dorsal area of the segment (Fig. 14C); segments III-VII with transverse tergite, lightly sclerotized, bearing many setae; segments VIII and IX with platelike tergite, heavily sclerotized, setose, forming a dorsal, subrectangular operculum (Fig. 14D), which has a typical color pattern, with tergites brown, with mesal, longitudinal darker area on tergite VIII and darker line near anterolateral margins, and darker border on anterior and lateral margins of tergite IX (Fig. 14D); operculum closing posterior opening of case. Pleurites of segments I and II small, each one with three setae (Fig. 14B, in detail); those of segments III-VIII absent, but region with two small setae each one. Anal proleg short, setose with pair of long posterior setae; anal claw stout, curved to approximately 90° , with basal peglike setae.

Laval case. Length 1.8–4.0 mm (n = 28). Made entirely of silk with bits of mineral material incorporated, slightly compressed laterally, with poorly closed dorsal seam (Fig. 14A); dorsal margin irregular; anterior and posterior openings circular.

Type material. Holotype male. PERU: Cusco, 20 rd km W Quincemil, Pte. Saucipata, Río Araza tributary, 13°20′13″S 70°51′15″W 893 m. 22.viii.2012. APM Santos, DM Takiya leg., pinned (MUSM). Paratypes. Same data as holotype, except 9 males, pinned (DZRJ); same data as holotype, except 7 males, in alcohol (MUSM); same data as holotype, except 1 male, in alcohol (DZRJ). PERU: Cusco, 3 rd km E Quincemil, 13°13′03″S 70°43′40″W 633 m. 20.viii.2012. APM Santos, DM Takiya leg., 3 males, in alcohol (DZRJ). PERU: Puno, 6 rd km W Mazuko, Pte. La Cigarra, 13°08′27″S 70°23′14″W 353 m. 01.ix.2012. APM Santos, DM Takiya leg., 4 males, pinned (DZRJ), 4 males, pinned (MUSM); same data, except, 3 males, in alcohol (INPA), 6 males, in alcohol (DZRJ).

Additional material examined. Same data as holotype, except 4 larvae, in alcohol (MUSM), 3 larvae, in alcohol (INPA). PERU: Cusco, 19 rd km W Quincemil, Rio Araza tributary, 13°20′10″S 70°50′57″W 874 m. 26.viii.2012. APM Santos, DM Takiya leg., 8 larvae, in alcohol (DZRJ), 7 larvae, in alcohol (MUSM), 2 larvae, in alcohol (INPA). PERU: Puno, 6 rd km W Mazuko, Pte. La Cigarra, 13°08′27″S 70°23′14″W 353 m. 01.ix.2012. APM Santos, DM Takiya leg., 4 larvae, in alcohol (DZRJ); same data, except 15 larvae, in alcohol (DZRJ); 15 larvae, in alcohol (INPA); 18 larvae, in alcohol (MUSM).

Etymology. The species name is an allusion to Manco Capac (used as a noun in apposition), son of Inti and Mama Quilla, according to some legends. Manco Capac and Mama Ocllo, his sister and wife, were sent to Earth to find the best place to begin a civilization. They travelled to the region which later became Cusco, where they established the center of the Inca Empire.

Remarks. Byrsopteryx mancocapac sp. nov. is very distinctive from the previous and other species described in the genus. In the field, this new species can be easily recognized from others occurring at same site by its coloration, whereas adults from both B. inti sp. nov. and B. mamaocllo sp. nov. have head and mesoscutum densely covered by white setae (Figs. 3, 6), and adults of B. mancocapac sp. nov. show only black setae on head and mesoscutum (Fig. 11). Furthermore, males of B. mancocapac sp. nov. do not show the wing modification which is seen in males of both B. inti sp. nov. and B. mamaocllo sp. nov. Male genitalia are also unique among Byrsopteryx species, particularly due to the conspicuous spine-like, curved processes projecting from dorsal region of subgenital plate (Figs. 13B, 13C). These processes superficially resemble those of B. loja Harris & Holzenthal, 1994, but in B. loja the paired processes arise from segment IX, instead of subgenital plate, as in the new species described here. In addition to the spine-like processes, this new species is also easily distinguished from B. loja and other Byrsopteryx species by sternum VIII with posterior margin divided into two plate-like lobes (Fig. 13A). Although a relatively high number of males and larvae of B. mancocapac sp. nov. were collected, females were not found.

Larvae of this new species show typical features of Byrsopteryx, for example, lateral depressed areas on pronotum, tergites VIII and IX large, forming an operculum, and case made of silk, poorly sealed dorsally (Fig. 14). As mentioned before, general aspects of larval morphology of this new species are similar to those previously described for other Byrsopteryx species (Holzenthal & Harris, 1991; Santos & Nessimian, 2010). Comparing with larvae of B. mamaocllo sp. nov., the only other Byrsopteryx larva known from Peru, the larvae of B. mancocapac sp. nov. can be recognized by the following features, as mentioned before: (1) operculum subrectangular, with rounded corners (Fig. 14D); (2) case usually as long as larva and, when compared with that of B. mamaocllo sp. nov., opaquer (Fig. 14A); and (3) abdominal segment II with a small pleurite bearing three setae, instead of two in B. mamaocllo sp. nov. (Fig. 14B).