Research Article |
Corresponding author: Emmanuel D. Delocado ( edelocado@ateneo.edu ) Academic editor: Christopher Majka
© 2021 Emmanuel D. Delocado, Hendrik Freitag.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Delocado ED, Freitag H (2021) Two new species of Byrrhinus Motschulsky, 1858 (Coleoptera, Limnichidae, Limnichinae) from Negros, Philippines. ZooKeys 1070: 51-72. https://doi.org/10.3897/zookeys.1070.70531
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Two new species of Limnichidae beetles, Byrrhinus negrosensis sp. nov. and Byrrhinus villarini sp. nov., are described from the Island of Negros in the Philippines. The adult specimens of the new species can be differentiated by patterns of body punctation, colour and orientation of elytral pubescence, posterolateral angle of pronotum, tarsomere length ratio and aedeagal form. Two clades, representing the two new species, were retrieved in the Maximum Likelihood gene tree using the 3’-end of the COI gene. Maximum genetic divergence within B. negrosensis sp. nov. and B. villarini sp. nov. were recorded to be 2.3% and 1.3%, respectively, while the mean interspecific divergence between the two new species was 19.7%. Morphological descriptions, digital photographs and COI sequences were provided for the two species. The state of knowledge of Byrrhinus is reviewed and an updated Philippine checklist is provided. By coupling morphological and molecular data, this paper provides the first additional new species of Philippine Byrrhinus in the last 28 years.
Biodiversity assessment, COI, integrative taxonomy, minute marsh-loving beetle, Negros Island, new species
Byrrhinus Motschulsky, 1858 is the most speciose limnichid genus with currently at least 87 species (Yoshitomi, unpublished data). Approximately 20% of known limnichid species belong to the genus Byrrhinus. The distribution of the genus is pantropical, but it is lacking or not yet recorded in some regions (
The distinguishing features of Byrrhinus include its elongate oval habitus and deeply bisinuate pronotum and elytral base (
The genus was originally erected in 1858 to contain Byrrhinus latus Motschulsky, 1858 from India. Later on,
Currently, five Byrrhinus species have been recorded in the Philippines (
Species identification in limnichid beetles is challenging due to their cryptic nature and subtle interspecific phenetic differences. Congeneric species are typically differentiated through details in genitalia, pubescence and punctation. Thus, to accelerate species discovery of the almost cryptic Byrrhinus fauna in a megadiverse locality, this study uses DNA sequences to complement morphological descriptions. This approach, referred to as integrative taxonomy (
This contribution presents two new species, Byrrhinus negrosensis sp. nov. and Byrrhinus villarini sp. nov., as supported by morphological and genetic data. Both species were collected from the Island of Negros in the central Philippine island group of Visayas. Recent extensive and collaborative sampling by the Ateneo Biodiversity Research Laboratory on this Island has led to the discovery of new aquatic insect species (
The organismic material used in this study was primarily retrieved in the scope of the School of Science and Engineering Industry 4.0 Research Fund (SI4-013) project on the freshwater macroinvertebrate diversity inventory by the Ateneo Biodiversity Research Laboratory of the Ateneo de Manila University, Quezon City, Philippines (AdMU). A light trap was set on riverbanks between 6:00 pm to 8:00 pm. Insects attracted to the black light were manually collected and stored in vials with 95% ethanol. Specimens were stored in a freezer (–20 °C) prior to their examination. Collections of the Ateneo Biodiversity Research Laboratory from previous field works of the second author were also examined and used for the molecular analysis of this study.
Isolation of DNA was performed using the Qiagen DNeasy kit (Qiagen, Hilden, Germany) following the instructions of the manufacturer for animal tissues (
The sequences of both complementary strands were traced manually using CHROMAS (
GenBank and BOLD accession numbers of COI-3’ mtDNA sequences of specimens used in this analysis.
Species | Specimen code | Locality | Sex | GenBank | BOLD | GenSeq nomenclature |
---|---|---|---|---|---|---|
Byrrhinus negrosensis sp. nov. | EDD116 | Negros | female | OK316812 | COLPH052-21 | genseq-2 COI |
EDD119 | Negros | male | OK316811 | COLPH053-21 | genseq-2 COI | |
EDD122 | Negros | male | OK316808 | COLPH054-21 | genseq-1 COI | |
EDD123 | Negros | male | OK316803 | COLPH055-21 | genseq-2 COI | |
EDD127 | Negros | male | OK316809 | COLPH056-21 | genseq-2 COI | |
EDD270 | Negros | male | OK316807 | COLPH057-21 | genseq-2 COI | |
Byrrhinus villarini sp. nov. | EDD113 | Negros | female | OK316817 | COLPH058-21 | genseq-2 COI |
EDD114 | Negros | male | OK316815 | COLPH059-21 | genseq-2 COI | |
EDD115 | Negros | male | OK316804 | COLPH060-21 | genseq-2 COI | |
EDD121 | Negros | male | OK316813 | COLPH061-21 | genseq-1 COI | |
EDD124 | Negros | male | OK316818 | COLPH062-21 | genseq-2 COI | |
EDD126 | Negros | male | OK316805 | COLPH063-21 | genseq-2 COI | |
Byrrhinus ferax Wooldridge, 1993 | EDD067 | Mindoro | male | OK316810 | COLPH064-21 | genseq-4 COI |
Byrrhinus sp. A | EDD057 | Palawan | male | OK316816 | COLPH065-21 | genseq-4 COI |
Byrrhinus sp. B | EDD105 | Luzon | male | OK316806 | COLPH066-21 | genseq-4 COI |
Byrrhinus sp. C | EDD112 | Mindanao | female | OK316814 | COLPH067-21 | genseq-4 COI |
Byrrhinus sp. | UPOL RK0663 | Cameroon | KX092882 | GBCL40978-19 | ||
Byrrhinus sp. | UPOL RK0664 | Indonesia | KX092889 | GBCL40979-19 | ||
Byrrhinus sp. | UPOL RK0727 | Malaysia | KX092888 | GBCL40980-19 | ||
Limnichus sp. (outgroup) | UPOL RK0666 | Indonesia | KX092883 | GBCL40986-19 | ||
Limnichus sp. (outgroup) | UPOL RK0725 | Malaysia | KX092886 | GBCL40987-19 |
Generation of a Maximum Likelihood (ML) tree with bootstrap analysis for 1000 replicates, as well as genetic distance computation using Kimura-2-paramter (K2P) model (
External morphology was examined by using an OLYMPUS SZ61 stereomicroscope (Olympus, Tokyo, Japan). The terminal abdominal portion of the specimens was dissected and treated overnight with lactic acid. The male aedeagi were then observed under an OLYMPUS CX21 compound microscope (Olympus, Tokyo, Japan). The specimen and its genitalia were glued on to entomologic paper for vouchering purposes. Photographs of vouchers were taken using a Canon EOS 6D with a macro lens and were stacked in Helicon Focus Pro v.7.6.1 (Helicon Soft, Kharkiv, Ukraine) and further processed using Adobe Photoshop CS6 (Adobe, San Jose, CA, USA). The aedeagi were digitally drawn over microscopic photograph underlays using Inkscape (GNU GPL, Boston, MA, USA).
Congeneric vouchers and type material at the Natural History Museum, Vienna, Austria (
Terminologies of the species’ descriptions follow the Limnichidae chapter of the Handbook of Zoology/Coloeptera (
The following abbreviations were used:
a.s.l. above sea level;
BS bootstrap value;
EL elytra length;
EW elytra width;
PL pronotum length;
PW pronotum width;
TL total length.
The alignment of COI-3’ sequences is composed of 723 bases with 221 parsimony-informative sites and 281 variable sites. No sequence in the matrix contains gaps, insertion, deletions or ambiguous sites. Sequences of Byrrhinus have low G-C concentration (16.5–20.2% C, 15.6–16.2% G).
Two clades were retrieved from the Byrrhinus specimens from Negros (Fig.
Mean genetic distance of Byrrhinus specimens, based on partial COI-3’ sequences (in %).
# | Identity | n | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | Byrrhinus negrosensis sp. nov. | 6 | 1.3 | ||||||||
2 | Byrrhinus villarini sp. nov. | 6 | 19.7 | 0.4 | |||||||
3 | Byrrhinus ferax Wooldridge, 1993 | 1 | 13.2 | 13.8 | – | ||||||
4 | Byrrhinus sp. A | 1 | 19.8 | 16.2 | 19.5 | – | |||||
5 | Byrrhinus sp. B | 1 | 16.6 | 15.6 | 19.4 | 17.7 | – | ||||
6 | Byrrhinus sp. C | 1 | 18.2 | 17.5 | 21.4 | 10.6 | 19.6 | – | |||
7 | Byrrhinus sp. (Malaysia) | 1 | 20.0 | 5.8 | 17.0 | 16.4 | 17.0 | 13.7 | – | ||
8 | Byrrhinus sp. (Indonesia) | 1 | 21.6 | 20.0 | 21.7 | 22.2 | 22.4 | 17.9 | 20.2 | – | |
9 | Byrrhinus sp. (Cameroon) | 1 | 25.16 | 28.2 | 28.6 | 24.7 | 25.0 | 26.5 | 28.9 | 23.4 | – |
Subfamily Limnichinae Erichson, 1847
Philippines • Negros Island, Negros Oriental, Valencia, Casaroro River, in secondary vegetation; ca. 09°18'N, 123°14'E; ca.150 m a.s.l.
Holotype: Philippines • ♂ (
Body: (Fig.
Head: obscurely rugulose; broadly laminate over eyes; margins of frons grooved over eyes; sides of frons with deep and well–marked pit-like depressions. Punctation minute, sparse, slightly coarser near epistomal suture. Pubescence dense and erect in anterior regions, sparse and recumbent posteriad. Eyes slightly convex, visible from above; upper margin of eyes strongly bordered, margin anteriorly almost reaching insertion of antennae, extending posterior of eyes although weaker. Surface of head posterior to eyes flat, without depressions or fossae; surface with fine and sparse punctation, denser and coarser on clypeus; surface between punctures smooth and shiny. Antennae moniliform, strongly pubescent; pedicel oblong, brown, slightly darker than adjacent antennomeres; antennomeres longer than wide, brown, darker distally, terminal antennomeres asymmetrical and darker than pedicel; pubescence brown, of two series: first series composed of one to two pairs of long and erect pubescence per antennomere, about as long as antennomere and second series composed of denser, shorter, paler, recumbent pubescence.
Pronotum: transverse, slightly paler on sides than on disc, distinctly wider at base; anterior margin of pronotum slightly concave, but almost straight between eyes, without crenulations, bordered; lateral margins only slightly arched, posterolateral angle 75–80°, with prominent borders; posterior margin with a distinct double sinuation; PL/PW = 0.42 (0.40–0.43); PW/PL = 2.40 (2.30–2.51). Punctation dense, minute and shallowly impressed; punctures larger than that of the head, denser at median line and posterior margins, sparse near suture and anteriad; surface between punctures rugose. Pubescence similar to the erect series of the head, denser on the anterior one-third of the lateral margin, evenly spaced on the rest of the margin, very sparse to almost obsolete and slightly decumbent proximally. Hypomeron flat, without depressions or fossae.
Elytra: EL/EW = 1.36 (1.30–1.43); EL/PL = 3.63 (3.63–3.85); EW/PW = 1.23 (1.16–1.27); TL/EW = 1.78 (1.66–1.78), slightly less than 4.0 times longer than the pronotum, widest at anterior 0.25; anterior margin of elytra bordered, bi-sinuately articulated with pronotum; lateral margins slightly explanate in anterior half; apices jointly rounded; humeral callus weak. Elytra with two series of punctation; first series with nine or more perceptible and irregular rows of large, deeply impressed punctures; punctures of medial five rows denser and more strongly impressed in anterior half; increasingly scattered, finer and more shallowly impressed laterally and posteriorly; intervals and interstices distinctly broader than punctures; second series of punctures much smaller, densely and evenly distributed over entire elytra. Pubescence long, yellowish-brown to brown, of two distinct types (erect and recumbent): erect series on sides, slightly recumbent series on disc; erect series longer and denser; recumbent series shorter, very sparse. Scutellum subtriangular, with irregular and densely punctured surface; with two series of punctation, few large and deeply impressed punctures, numerous finer and shallowly impressed ones; pubescence erect, sparse. Metathoracic wings well developed. Epipleura almost flat.
Ventral surface: punctation dense and almost uniform; pubescence brown, minute, long, finer than on dorsal side, recumbent, dense and evenly distributed. Prosternum slightly impressed at the process; process narrow, punctation more distinct at tip. Mesosternal ridge along posterior margins of mesosternum distinct. Metasternum perforate at sides, with raised triangular, rugulose area behind mesocoxal cavities; raised area comprising nearly half of surface; metasternal ridge along posterior margin of metasternum faint laterally, well-developed medially. Abdominal punctation finer at mid-line than at sides; surface between punctation with polygonal network, with median pore. Intermetacoxal plate on ventrite I triangular, strongly acuminate. Abdominal ventrite I with depressions for reception of metafemora and metatibiae; ventrites I–III connate, fused; ventrites IV–V without polygonal network; ventrite V distinctly emarginate.
Legs: less than half of body length. Tibia brown, lateral margins darker, distal margin with comb of long spines; protibia very short, a little longer than half of either mesotibia and metatibia, lateral margin slightly concave, setae denser than on mesotibia and metatibia; mesotibia with lateral margin curved more prominently in interior margins, setae evenly distributed; metatibia almost twice as long as protibia, slightly longer than mesotibia, lateral margins almost parallel, setae sparse and almost recumbent; apex of mesotibia and metatibia smoothly and broadly curved. Tarsi 5-5-5, brown, paler towards the apex, almost half as long as mesotibia; tarsomere length ratio ca. 1.0:1.0:1.0:1.0:4.0 (0.9–1.1: 0.9–1.1: 0.9–1.1: 0.9–1.1: 3.5–4.7); tarsomere 1 widest towards the apex, distal margin almost double the width of proximal margin, with dense comb of setae; tarsomeres 2–4 similar to tarsomere 1, but outer edge with long yellow spiny setae on both sides, remaining portions with sparse minute setae; tarsomere 5 widest towards the apex, almost triangular, with long robust spiny setae. Tarsal claws long, narrow, symmetrical.
Male genitalia: (Figs
Female genitalia: ovipositor relatively short (0.58–0.62 mm long), straight.
In the elongate oval shape, the new species resembles several species, including B. ferax and B. tarawakanus. Amongst the Philippine species, the range of size overlaps with B. subtestaceus and B. ferax. The protibia of B. negrosensis sp. nov. is notably smaller than the mesotibia and metatibia. The male genitalia of B. negrosensis sp. nov. resembles that of B. ferax due to the medially parallel paramere apices, which is quite uncommon in the Oriental members of the genus. Despite numerous similarities, B. negrosensis sp. nov. differs from B. ferax in the dorsally V-shaped basal fusion point of the parameres (Fig.
This species is only recorded from the Island of Negros in the Philippines.
No external sexual dimorphism is observed. Teneral specimens are significantly paler brown.
The species is named after the Island of Negros from where the specimens were collected.
Philippines • Negros Island, Occidental Mindoro, Murcia, Pandanon River in secondary vegetation; ca. 10°34'54"N, 123°10'30"E; ca. 440 m a.s.l.
Holotype: Philippines • ♂ (
Body: (Fig.
Head: obscurely rugulose; broadly laminate over eyes; margins of frons grooved over eyes; sides of frons with deep and well–marked pit-like depressions. Punctation minute, slightly coarser near epistomal suture. Pubescence brown, fine, quite long, erect, more numerous and denser on the anterior region. Eyes slightly convex, visible from above; upper margin of eyes bordered; anterior margin almost reaching insertion of antennae, extending posterior of eyes although weaker. Surface of head posterior to eyes flat, without depressions or fossae; surface with fine and sparse punctation, denser and coarser on clypeus; surface between punctures smooth and shiny. Antennae moniliform, strongly pubescent; pedicel globular, brown, darker than adjacent antennomeres; antennomeres longer than wide, yellow-brown; pubescence brown, uniform, erect, mostly as long as antennomere.
Pronotum: transverse, black, with dark brown colouration on the sides, distinctly narrower at base; anterior margin of pronotum straight, without crenulations, bordered; lateral margins strongly arched, posterolateral angle ca. 50°, with prominent borders; posterior margin with distinct double sinuation; PL/PW = 0.42 (0.40–0.44); PW/PL = 2.40 (2.28–2.50). Punctation strong and deeply impressed, but sparse; punctures stronger than that of the head, larger at posterior margins, sparse near suture and anteriad, surface very depressed at projections along posterior margin. Pubescence similar to that on the head, slightly decumbent near the median line, denser at sides. Hypomeron flat, without depressions or fossae.
Elytra: EL/EW = 1.41 (1.30–1.41); EL/PL = 4.14 (4.11–4.14); EW/PW = 1.20 (1.20–1.21); TL/EW = 1.75 (1.68–1.78); elytra slightly more than 4.0 times longer than the pronotum; widest at anterior 0.2; anterior margin of elytra bordered, strongly bi-sinuately articulated with the pronotum; lateral margins pronounced, finer towards apex; apices jointly rounded; humeral callus weak. Elytra punctation of two series; first series with nine or ten distinct and almost regular rows of large deeply impressed punctures; increasingly scattered, finer, not as strongly impressed as in rows laterally and posteriorly; intervals and interstices distinctly broader than punctures; second series of small punctures moderately dense only and less conspicuous than in previous species. Pubescence long, brown, with yellowish shade depending on illumination, of two distinct types: erect series on sides, slightly recumbent series on disc; erect series longer and denser; disc series shorter, sparse. Scutellum subtriangular, with few punctures and pubescence similar to surrounding area of elytra. Metathoracic wings well developed. Epipleura almost flat.
Ventral surface: punctation dense and uniform; pubescence brown, minute, long, finer than on dorsal side, recumbent, dense and evenly distributed. Prosternum slightly impressed at the process; process narrow, punctation more distinct at tip. Mesosternal ridge along posterior margins distinct. Metasternum minutely perforate at sides, with raised triangular, rugulose area behind cavities; raised area comprising nearly half of surface; metasternal ridge along posterior margin of metasternum faint laterally, well-developed medially. Abdominal punctation finer at mid-line than at sides; surface between punctation with polygonal network, with median pore. Intermetacoxal plate on ventrite I triangular, strongly acuminate. Abdominal ventrite I with depressions for reception of metafemora and metatibiae; ventrites I–III connate, fused; ventrites IV–V without polygonal network; ventrite V distinctly emarginate apically.
Legs: length less than half of body length. Tibia brown, lateral margins darker, curved, with pre-apical comb of spines; metatibia slightly longer than protibia and mesotibia; apex of mesotibia and metatibia slightly curved. Tarsi 5-5-5, dark yellow to light brown, paler towards the apex, about two-thirds of length of tibia; tarsomere length ratio ca. 1.2:1.0:1.0:1.0:2.9 (1.0–1.5: 0.8–1.0: 0.8–1.0: 0.8–1.0: 2.7–3.3); tarsomere 1 brown, with parallel margins, widest towards the apex, with dense comb of setae; tarsomeres 2–4 almost globular, outer edge with long yellow spiny setae on both sides, remaining portions with sparse minute setae; tarsomere 5 more yellow than brown, widest towards the apex, with at least three pairs of long robust spiny setae. Tarsal claws long, narrow, symmetrical.
Male genitalia: (Figs
Female genitalia: ovipositor relatively short (0.50–0.56 mm long), straight.
In the ovoid shape, the new species resembles B. vestitus (Sharp, 1902), B. maculatus Wooldridge, 1987 and B. magnus Wooldridge, 1987. Compared to other Philippine species, the range of size overlaps with B. punctatus and B. tarawakanus. The new species is remarkably different from these two in the smaller posterolateral angle (ca. 50°) of the pronotum in B. villarini sp. nov. compared to B. punctatus and B. tarawakanus, as well as B. negrosensis sp. nov. (75–80°). In addition to the posterolateral angle measure of pronotum, B. villarini sp. nov. is notably different from B. negrosensis sp. nov. in the length of tarsomere 5. Tarsomere 5 of B. villarini sp. nov. is as long as tarsomeres 2–4, while tarsomere 5 of B. negrosensis sp. nov. is almost as long as tarsomeres 1–4. Additionally, erect series of elytral pubescence is present on the posterior end of both species, but covers the distal one-third only of elytra in B. villarini sp. nov., while covering the distal one-half in B. negrosensis sp. nov.
Males of B. villarini sp. nov. are easily recognisable because the parameres are dorsally fused forming a rather shallow “U” (Fig.
B. villarini sp. nov. varies by 5.8% mean genetic distance (723 bp COI-3’ mtDNA barcode) from an unidentified, but presumably closely-related Malaysian species and by at least 13.4% from any other Philippine congeners with available barcodes (Suppl. material
This species is only recorded from the Island of Negros in the Philippines.
No external sexual dimorphism is observed.
The new species is named and dedicated to the immediate past president of the Ateneo de Manila University, Fr Jose Ramon T. Villarin, SJ, PhD, who finished his term last year. During his reign for the past decade, Fr Villarin showed ardent support for research activities on the environment and sustainability. He is also a member of the Intergovernmental Panel on Climate Change which was conferred the 2007 Nobel Peace Prize for their study and recommendations on counteracting the global climate crisis.
Byrrhinus ferax Wooldridge, 1993: 359–360 (orig. descr.).
Philippines • ♂ (AdMU), Occ. Mindoro, Sablayan, small limestone river; rootpacks, dist. primary forest; ca. 12°47'49"N, 120°54'33"E; ca. 100 m a.s.l.; 01 Jan. 1995, leg. Mendoza “(365)M”; GenBank: OK316810; BOLD: COLPH064-21.
Assignment of the specimen to this taxon was based primarily on genital characters. No remarkable difference was noted compared to the original description. The material used for the molecular analysis in this study was collected 90 km south of one of the localities of the paratypes, but on the same island. Known distribution of B. ferax includes the Philippine Islands of Mindoro and Mindanao.
The three specimens listed below have an interspecific divergence of 10.6 to 19.6% (Table
Byrrhinus sp. A: Philippines • 1 ♂, Palawan, P. Princesa, Irawan River, 6 km NW of PPC, 0.5 km upstream of water plant ca. 9°49'50"N, 118°39'46"E; 105 m a.s.l.; 06 Aug. 2019, leg. H. Freitag “(60b)M”; GenBank: OK316816; BOLD: COLPH065-21.
Byrrhinus sp. B: Philippines • 1 ♂, Camarines Sur, Lupi, Brgy. Sooc, Sooc River, Bicol National Park; ca. 13°52'28"N, 122°56'38"E; 90 m a.s.l.; 09 Aug. 1996; leg. Mendoza “(M585)L”; GenBank: OK316806; BOLD: COLPH066-21.
Byrrhinus sp. C: Philippines • 1 ♂, Mindanao, Agusan N, R.T.R, Panaytayon, paddy field, ca.10 m a.s.l. 9°02'53"N, 125°35'03"E; leg. Freitag & Pangantihon 05. Jul. 2018 “(890)L”; GenBank: OK316814; BOLD: COLPH067-21.
Byrrhinus convexus (Blackburn, 1896): Luzon, Australia
Byrrhinus ferax Wooldridge, 1993: Mindoro, Mindanao (Cotabato, Davao); Borneo (Malaysia: Sabah)
Byrrhinus negrosensis sp. nov.: Negros
Byrrhinus punctatus (Pic, 1922): Luzon, Mindoro
Byrrhinus villarini sp. nov.: Negros
Byrrhinus subtestaceus Pic, 1923: Luzon
Byrrhinus tarawakanus Deléve, 1973: Palawan, Tawi-Tawi
The two new Byrrhinus species described here, B. negrosensis sp. nov. and B. villarini sp. nov., increase the total number of Philippine Byrrhinus from five to seven. This contribution using the integrative taxonomic approach provides the first additional record on Philippine Byrrhinus species in the last 28 years and provides the first records from the Visayas (Fig.
Characters used to differentiate limnichid species, such as pubescence and punctation patterns and aedeagal structure (
Phylogenetic analysis retrieved strongly supported clades (BS = 100) corresponding to the two newly-described species (Fig.
The clustering of conspecific sequences in the gene tree is also reflected in the statistical parsimony network generated for the two new species (Fig.
Entomofaunal diversity has been experiencing a tremendous decline in both local (
We would like to thank the Bureau of Fisheries and Aquatic Resources (BFAR) for their continuous assistance in issuing Gratuitous Permit (GP 0133-17 and renewals) which allows sampling and collecting specimens from aquatic habitats. We also would like to thank local government units of Murcia and Valencia and their Environment and Natural Resources Office for their Prior Informed Consents and thus granting permission to conduct sampling in their locality and for providing logistical assistance in the field.
We also thank the members of the Biodiversity Laboratory, Marc Sabordo, Dr Jhoana Garces, Clister Pangantihon and Arthien Pelingen, for their valuable contribution in sampling, sorting and safekeeping the specimens.
Additionally, we thank Dr Thomas von Rintelen and Mr Robert Scheiber at Museum für Naturkunde, Berlin, Germany (MfN) for supervising the molecular laboratory work training during the internship of the first author under the Biodiversity Teaching in a Philippine-Cambodian-German Network (BIO-PHIL) project. We would like to express our sincerest gratitude to Dr Manfred A. Jäch (
The study was partly funded by the School of Science and Engineering Industry 4.0 Research Fund (SI4-013) of the Ateneo de Manila University. Inspection of type specimens at European collections and taxonomy training of the first author were made possible through the Biodiversity Teaching in a Philippine-Cambodian-German Network (BIO-PHIL) project funded by the German Academic Exchange Service (DAAD project BIO-PHIL 57393541).
Data type: Genetic distance
Table S1
Explanation note: Table S1. Kimura 2-parameter (K2P) genetic distance of the samples of Byrrhinus specimens, based on their aligned partial COI-3’ sequences of 723 bp length (in %).