Advancing butterfly systematics through genomic analysis Author Zhang, Jing Author Cong, Qian Author Shen, Jinhui Author Song, Leina grishin@chop.swmed.edu Author Grishin, Nick V. text The Taxonomic Report of the International Lepidoptera Survey 2025 2025-05-27 12 5 1 201 journal article 311708 10.5281/zenodo.16642576 934f018d-7eae-4ad4-bf4e-5f27e94cf045 2643-4806 16642576 Entheus venezuelius Grishin , new species http://zoobank.org/ 3DE8615E-5BAB-43AB-8EDD-DEA0B421FCF3 ( Figs. 36p–z , 44 part, 47, 50 part, 51o–p) Definition and diagnosis. Genomic analysis reveals that specimens from Venezuela that are traditionally identified as Entheus aequatorius Mabille & Boullet, 1919 , stat. rest. ( type locality given in the original description as “ Bolivia ” likely by mistake, and may be in Ecuador : Bolivar), are indeed its sister, but are genetically differentiated from it at the species level ( Figs. 44 , 50 ); e.g., their COI barcodes differ by 1.2% (8 bp), thus representing a new species. This new species keys to “ Entheus matho aequatorius ”(B.10.5(c)) in Evans (1952) who included it into this taxon, but noted that Venezuelan specimens were “smaller, ♀ upf with spot mid costa,” and differs from its relatives by a combination of the following characters: smaller in size; males with more extensive rusty overscaling on both sides of the wings, the color of the forewing bands and spots is yellower, less orange; the tibial tuft is tawny, paler than in E. aequatorius ; the pale area of the anal fold is smaller and yellower; females with a white area on the hindwing more restricted towards the anal margin and a pale spot by mid-costa on the forewing aligned with the discal cell white spot, not offset basad. Due to the cryptic nature of this species and unexplored individual variation, most reliable identification is achieved by DNA, and a combination of the following base pairs is diagnostic in the nuclear genome: aly2653.1.9:C117G, aly2130.3.5:T153C, aly997.1.7:T42A, aly671.50.4: C147 T , aly 2874.5.3 :G177A; and COI barcode: T49 C, T115 C, T226 C, A373G, G506G, T646 C. Fig. 47. Entheus venezuelius sp. n. in dorsal (left) and ventral (right) views, data in text: a) holotype ♂ NVG-15026F10, inset shows hindtibial tuft enlarged two times compared to specimens (scale not given) and b) paratype ♀ NVG-15026F11. Barcode sequence of the holotype . Sample NVG-15026F10, GenBank PV550001, 658 base pairs: AACTTTATATTTTATTTTCGGAATCTGAGCAGGAATAGTAGGTACTTCCCTAAGATTATTAATTCGAACTGAATTAGGAACTCCTGGATCATTAATTGGAGATGATCAAATTTACAATACT ATTGTTACTGCTCATGCTTTTATTATAATTTTTTTTATAGTTATACCAATTATAATTGGAGGTTTTGGAAATTGATTAGTACCTTTAATATTAGGAGCCCCTGACATAGCTTTTCCTCGAA TAAATAATATAAGTTTTTGACTTCTACCCCCATCATTAACATTATTAATTTCTAGAAGAATTGTTGAAAATGGAGCTGGAACAGGATGAACAGTTTACCCCCCTTTATCTGCTAACATTGC CCACCAAGGGTCTTCAGTAGATTTAGCTATTTTTTCCCTTCATTTAGCTGGTATTTCATCAATTTTAGGAGCTATTAATTTTATTACAACAATTATTAATATACGAATTAGAAATTTATCA TTTGATCAAATACCTTTATTTGTTTGAGCAGTAGGTATTACTGCACTACTTTTATTATTATCTTTACCTGTATTAGCAGGTGCTATTACTATACTTTTAACAGATCGAAATTTAAATACAT CATTTTTTGATCCTGCGGGAGGAGGAGATCCAATTCTTTACCAACATTTATTT Type material. Holotype : ♂ deposited in the McGuire Center for Lepidoptera and Biodiversity Collection, Gainesville, FL, USA ( MGCL ), illustrated in Figs. 47a and 51o (genitalia Fig. 36p–w ), bears the following five printed (text in italics handwritten) rectangular labels, four white: [ Rancho Grande . 1200 m . | Parque Nac. Henri Pittier | Edo. Aragua , Venezuela | T.E. Pliske VI-29-75 ], [Genit. Prep. | SRS- 1278 ], [Allyn Museum | Acc. 19 80-29 ], [DNA sample ID: | NVG-15026F10 | c/o Nick V. Grishin], and one red [HOLOTYPE ♂ | Entheus | venezuelius Grishin]. Paratypes : 2♂♂ and 2♀♀ from Venezuela , Argua : 1♀ NVG-15026F11 the same data as the holotype, genitalia SRS-1279 ( Figs. 47b , 51p ) and 1♀ NVG-14062C01 Eancho Grande , 1100 m , 19-Jun-1985 , S. S. Nicolay leg. [ USNM ]; 1♂ NVG-14113A05 Yuracay , Hacienda Tropicale ca. 10 km S of San Felipe , 100–400 m , GPS 10.2917 , −68.6667 , 26-Jan-23-Feb-1993 , Kareofelas & Witham leg. [ LACM ]; and 1♂ NVG-15032C11 (leg DNA extraction, sequenced), NVG-24028H11 (abdomen DNA extraction and dissection) Carabobo , Puerto Cabello , no date [around 1900], Hahnel leg., genitalia vial NVG241114 -17 ( Fig. 36x–z ) [ MFNB ] . Type locality. Venezuela : Aragua , Henri Pittier National Park , Rancho Grande , elevation 1200 m . Etymology. The name is given for the country with the type locality and to rhyme with its related species, E. aequatorius . The name is treated as an adjective. Distribution. Venezuela .