Research Article |
Corresponding author: Javier Ponce-Saavedra ( ponce.javier0691@gmail.com ) Academic editor: José Antonio Ochoa
© 2019 Ana F. Quijano-Ravell, Luis F. de Armas, Oscar F. Francke, Javier Ponce-Saavedra.
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:
Quijano-Ravell AF, de Armas LF, Francke OF, Ponce-Saavedra J (2019) A new species of the genus Centruroides Marx (Scorpiones, Buthidae) from western Michoacán State, México using molecular and morphological evidence. ZooKeys 859: 31-48. https://doi.org/10.3897/zookeys.859.33069
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A new species of scorpion belonging to the genus Centruroides Marx, 1890 is described from the Coalcomán mountain range, western Michoacán State, Mexico. Its general aspect resembles Centruroides ruana Quijano-Ravell & Ponce-Saavedra, 2016, and C. infamatus (C. L. Koch, 1844), but it is a smaller species having lower pectinal tooth counts; also, males of C. ruana have the pedipalp chelae slightly thicker, whereas C. infamatus has a subaculear tubercle nearer to the base of the aculeus. Another species with similar aspect is Centruroides ornatus Pocock, 1902; however, a preliminary molecular analysis of the mitochondrial gene mRNA 16S showed genetic divergence (measured as p-distance) near to 10% between these species, and lower differences between the new species with respect to C. infamatus (4.63%) and C. ruana (5.07%). The molecular evidence together with the morphological characters (integrative taxonomy) are sufficient for recognizing the Coalcomán population as a separate and valid species.
bark scorpions, Coalcomán Range, North America, striped scorpions, taxonomy
Buthid scorpions of the genus Centruroides Marx, 1890 (Buthidae) are widely distributed in Mexican territory, from which 45 nominal species and two subspecies have been recognized (
From Michoacán State, eight species belonging to this genus have been described or recorded (
Centruroides elegans (Thorell, 1876) and C. pallidiceps Pocock, 1902 were mentioned from Michoacán by
The Coalcomán Range is located in the west of Michoacán and forms part of the western-most region of the Sierra Madre del Sur. Its highest elevations reach almost 2900 m a.s.l. and contain well conserved areas with high levels of endemism for animals and plants (
In the present contribution we describe a new species of the genus Centruroides from the Coalcomán Range, based on several specimens of both sexes under an integrative taxonomic perspective, using morphological and molecular evidence.
The examined specimens are deposited in 75% ethanol in the following institutions: CAFBUM: Colección Aracnológica del Laboratorio de Entomología “Biol. Sócrates Cisneros Paz”, Facultad de Biología, Universidad Michoacana de San Nicolás de Hidalgo, Morelia, Michoacán, México; CNAN: Colección Nacional de Arácnidos, Instituto de Biología, Universidad Nacional Autónoma de México, D.F; and IESC: Instituto de Ecología y Sistemática, La Habana, Cuba.
The specimens were examined and measured with a Zeiss Stemi DV4 stereomicroscope, equipped with a 0.1 mm ocular micrometer. Photographs were obtained with a microscope eyepiece camera 3.1mp AmScope MU300. Digital images obtained were processed and edited with Adobe Photoshop CS5.The distribution map was generated with ESRI ArcGIS online. We obtained two hemispermatophores from one male of the new species as a complementary structure for the description.
Nomenclature and measurements follow
In addition to the morphological diagnostic characters, a molecular analysis using sequences of mitochondrial gene RNAm 16S was carried out with specimens of four populations of C. ornatus, one of C. balsasensis and two localities of C. infamatus. Also, included was one sequence of the type population of C. ruana (Quijano-Ravell & Ponce-Saavedra 2016). All sequences were obtained from specimens captured at different dates by several collectors that were found in the CNAN, CAFBUM and IESC collections, except the one of C. infamatus from Uruapan which was downloaded from GenBank (AF439753).
For the genetic analyses, DNA was extracted from muscle tissue preserved at 96% ethanol (pedipalps and legs fragments) using the FitzSimmons protocol (
DNA sequences were aligned with MEGA X: Molecular Evolutionary Genetics Analysis software (
The analysis involved 11 nucleotide sequences (Table
Male holotype (CNAN-T01315), Michoacán: Coalcomán de Vázquez Pallares municipality: La Nieve (18°49.070'N, 103°02.653'W, 2230‒2260 m a.s.l.), 07-VIII-2002, O. Francke, E. González S. y S. Reynaud colls, determined as Centruroides infamatus ornatus by R. J. Moreno B., 02-VII-2004. Paratypes: 17 ♂♂, 38 ♀♀ Michoacán: Coalcomán de Vázquez Pallares municipality: La Nieve, 10.VII.2006, 2246 m, O. Francke, J. Ponce, M. Córdova, A. Jaimes, G. O. Francke & V. Capovilla, colls: 1 male (peines 22-21) 1 female (18-18). (CNAN-T01316), 3 ♂♂, 5 ♀♀ (CAFBUM S0150), 3 ♂♂ adult, 1 ♂ juvenile, 3 ♀♀ (IESC-3.3796 to. 3.3802). Michoacán: Coalcomán de Vázquez Pallares municipality: La Nieve, 7. VIII. 2002, 2265 m, O. Francke, E. Gonzalez-Santillán & S. Reynaud colls.
The proposed name is a patronym honoring Biol. Mario Manuel Romero Tinoco, who has dedicated his life to increasing our knowledge of the “hot land” in Michoacán State, and for his relevant and continued contributions to the people that inhabit those beautiful places.
A medium-sized species belonging to the Centruroides infamatus subgroup (as defined by
Centruroides romeroi sp. nov. closely resembles C. ruana and C. infamatus, (
(Figs
Measurements (in mm) of the holotype and four paratype males of Centruroides romeroi sp. nov. Abbreviations: L=Length; W=Width; D= Depth; Ca = Carapace; MeS= Mesosomal segment; MS= Metasomal segment; Ves=Vesicle; Fmr=Femur; Ptla= Patella; Hand= hand of chelae of pedipalp; Fix F= Fixed finger; MovF= Movable finger; BP= Basal plate of pectines; P.C.= Pectinal tooth count.
Measurement | Holotype | Paratypes | ||||
---|---|---|---|---|---|---|
Male 1 | Male 2 | Male 3 | Male 4 | Male 5 | ||
L Ca | 4.60 | 4.40 | 4.40 | 4.40 | 4.60 | 4.60 |
LMeSVII | 4.00 | 3.80 | 3.20 | 4.00 | 4.40 | 3.80 |
W MeSVII | 4.10 | 4.20 | 4.00 | 4.20 | 4.20 | 4.20 |
L MSI | 4.00 | 3.80 | 3.60 | 3.80 | 4.20 | 3.80 |
L MSII | 4.60 | 4.60 | 4.20 | 4.40 | 5.00 | 4.60 |
L MSIII | 5.20 | 5.00 | 4.80 | 5.00 | 5.80 | 5.20 |
L MSIV | 5.80 | 5.80 | 5.40 | 5.60 | 6.40 | 5.60 |
L MSV | 6.60 | 6.60 | 6.20 | 6.20 | 7.20 | 6.20 |
W MSI | 2.20 | 2.20 | 2.20 | 2.20 | 2.20 | 2.20 |
W MSII | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 |
W MSIII | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 |
W MSIV | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 |
W MSV | 2.20 | 2.20 | 2.20 | 2.20 | 2.20 | 2.00 |
D MSI | 1.80 | 2.00 | 1.80 | 1.80 | 1.80 | 1.80 |
D MSII | 1.80 | 2.00 | 1.80 | 1.80 | 1.80 | 1.80 |
D MSIII | 1.80 | 2.00 | 1.80 | 1.80 | 1.80 | 1.80 |
D MSIV | 1.80 | 2.00 | 1.80 | 1.80 | 1.80 | 1.80 |
D MSV | 2.00 | 2.10 | 2.00 | 2.00 | 2.00 | 2.00 |
L Ves | 3.00 | 2.90 | 3.00 | 2.80 | 3.00 | 3.00 |
W Ves | 1.60 | 1.60 | 1.60 | 1.60 | 1.60 | 1.60 |
D Ves | 1.60 | 1.40 | 1.60 | 1.60 | 1.60 | 1.60 |
L Fmr | 4.80 | 4.80 | 4.60 | 4.60 | 4.90 | 4.60 |
W Fmr | 1.20 | 1.20 | 1.20 | 1.20 | 1.20 | 1.20 |
L Ptla | 5.00 | 5.00 | 4.80 | 4.80 | 5.20 | 4.80 |
W Ptla | 1.60 | 1.60 | 1.60 | 1.60 | 1.60 | 1.60 |
L Hand | 3.40 | 3.60 | 3.40 | 3.60 | 3.80 | 3.60 |
W Hand | 1.60 | 1.60 | 1.60 | 1.80 | 1.80 | 1.80 |
D Hand | 1.60 | 1.60 | 1.60 | 1.60 | 1.60 | 1.60 |
L Fix F | 4.20 | 4.40 | 4.00 | 4.00 | 4.40 | 4.10 |
L Mov F | 5.00 | 5.00 | 4.60 | 4.80 | 5.00 | 5.00 |
L BP | 0.50 | 0.60 | 0.60 | 0.50 | 0.50 | 0.60 |
W BP | 1.00 | 1.10 | 1.10 | 1.00 | 1.00 | 1.00 |
P.C. | 21-21 | 21-22 | 21-22 | 22-23 | 20-20 | 22-22 |
Carapace. Anterior margin with median notch broadly “V” shaped, reaching the level of the posterior margin of the first pair of lateral eyes, weakly crenulate and scarcely setose; three pairs of lateral eyes subequal in size (Fig.
Mesosoma. Tergites with moderate median longitudinal carina (Figs
Sternum type 1, triangular, very finely granular, with two long, median subdistal macrosetae; posterior depression long, wide and deep (Fig.
Genital operculum (Fig.
Pectines. Tooth count 21/22. Basal plate rectangular, anterior margin almost straight, with small median V-shaped notch, posterior margin straight (Fig.
Metasoma. Moderately elongated and not incrassate distally (Figs
Chelicerae with dentition typical for the genus (Fig.
Pedipalps orthobothriotaxic A-α; moderately elongated (length/width ratio of femur and patella = 4.8 and 3.6, respectively). Femur with dorsal, external and ventral intercarinal spaces finely and densely granulose (Fig.
Legs. Slender, with carinae granulose to subserrate and intercarinal tegument coriaceous to minutely granulose. Prolateral and retrolateral pedal spurs strong and somewhat curved in all legs. Ventral surface of tarsomere II densely covered by long macrosetae irregularly arranged into two longitudinal, broad, dense rows converging basally. Claws rather short and curved.
Differs from males as follows: color pattern somewhat darker. Metasoma and pedipalps shorter and robust (Figs
Measurements (in mm) of six female paratypes of Centruroides romeroi sp. nov. Abbreviations: L=Length; W=Width; D= Depth; Ca = Carapace; MeS= Mesosomal segment; MS= Metasomal segment; Ves=Vesicle; Fmr=Femur; Ptla= Patella; Hand= hand of chelae of pedipalp; Fix F= Fixed finger; MovF= Movable finger; BP= Basal plate of pectines; P.C.= Pectinal tooth count.
Measurement | Paratypes | |||||
---|---|---|---|---|---|---|
Female 1 | Female 2 | Female 3 | Female 4 | Female 5 | Female 6 | |
L Ca | 4.60 | 4.80 | 4.40 | 4.20 | 4.20 | 4.60 |
LMeSVII | 3.00 | 3.20 | 3.20 | 3.40 | 3.20 | 3.60 |
W MeSVII | 5.00 | 4.60 | 4.60 | 5.00 | 4.60 | 5.00 |
L MSI | 3.20 | 3.20 | 3.20 | 2.80 | 2.60 | 3.20 |
L MSII | 3.60 | 3.60 | 3.80 | 3.40 | 3.20 | 3.80 |
L MSIII | 4.00 | 4.00 | 4.00 | 3.80 | 3.40 | 4.20 |
L MSIV | 4.60 | 4.40 | 4.60 | 4.20 | 3.80 | 4.60 |
L MSV | 5.40 | 5.20 | 5.20 | 5.00 | 4.60 | 5.20 |
W MSI | 2.60 | 2.60 | 2.60 | 2.40 | 2.40 | 2.60 |
W MSII | 2.40 | 2.40 | 2.40 | 2.20 | 2.20 | 2.40 |
W MSIII | 2.40 | 2.40 | 2.40 | 2.20 | 2.20 | 2.40 |
W MSIV | 2.40 | 2.40 | 2.20 | 2.20 | 2.00 | 2.40 |
W MSV | 2.20 | 2.40 | 2.20 | 2.00 | 2.00 | 2.40 |
D MSI | 2.00 | 2.20 | 2.00 | 2.00 | 2.00 | 2.20 |
D MSII | 2.00 | 2.20 | 2.00 | 2.00 | 2.00 | 2.20 |
D MSIII | 2.00 | 2.20 | 2.00 | 2.00 | 2.00 | 2.20 |
D MSIV | 1.80 | 2.00 | 1.80 | 1.80 | 1.80 | 2.00 |
D MSV | 1.80 | 2.00 | 1.80 | 1.80 | 1.80 | 2.00 |
L Ves | 2.40 | 2.40 | 2.40 | 2.20 | 2.30 | 2.40 |
W Ves | 1.60 | 1.60 | 1.60 | 1.40 | 1.40 | 1.60 |
D Ves | 1.40 | 1.40 | 1.40 | 1.40 | 1.40 | 1.60 |
L Fmr | 4.40 | 4.20 | 4.40 | 4.20 | 4.00 | 4.60 |
W Fmr | 1.20 | 1.20 | 1.20 | 1.20 | 1.20 | 1.20 |
L Ptla | 4.80 | 4.80 | 4.80 | 4.60 | 4.20 | 5.00 |
W Ptla | 1.60 | 1.60 | 1.60 | 1.40 | 1.60 | 1.60 |
L Hand | 3.40 | 3.20 | 3.40 | 3.00 | 3.00 | 3.40 |
W Hand | 1.80 | 1.60 | 1.80 | 1.40 | 1.40 | 1.80 |
D Hand | 1.60 | 1.60 | 1.60 | 1.40 | 1.40 | 1.60 |
L Fix F | 4.40 | 4.40 | 4.20 | 4.00 | 3.80 | 4.40 |
L Mov F | 5.00 | 5.00 | 4.80 | 4.80 | 4.40 | 5.20 |
L BP | 0.60 | 0.60 | 0.60 | 0.60 | 0.60 | 0.60 |
W BP | 1.20 | 1.00 | 1.20 | 1.20 | 1.10 | 1.00 |
P.C. | 18-16 | 18-18 | 18-18 | 17-17 | 18-18 | 19-18 |
Variation of the pectinal tooth counts in Centruroides romeroi sp. nov. Abbreviations: n, sample by sex; N, total of examined combs; * Mode.
Sex | n | Pectinal tooth counts | Average | Standard Deviation | ||||||
---|---|---|---|---|---|---|---|---|---|---|
16 | 17 | 18 | 19 | 20 | 21 | 22 | ||||
Female | 90 | 2 | 22 | 47* | 16 | 2 | 1 | 17.96 | 0.84 | |
Male | 46 | 3 | 3 | 13 | 21* | 8 | 20.60 | 1.05 | ||
N | 136 |
Pectinal tooth count varies among both sexes (Table
La Nieve (2030 to 2260 m a.s.l.) belongs to the Coalcomán Range, the predominant vegetation is pine forest, and the climate is temperate sub humid (Cw). The scorpions were collected at night, with portable U.V. lights, under stones and fallen rotten trees, under bark of Pinus sp., in the yards and walls of the local school and houses of the village. During the collection the temperature and relative humidity of the air were 10‒12 °C and 90%, respectively. Centruroides romeroi sp. nov. is sympatric with Vaejovis coalcoman Contreras-Félix & Francke, 2014.
The 16S mitochondrial marker was used successfully by several authors for delimiting several species in the genus Centruroides such as the cryptic species C. exilicauda (Wood) and C. sculpturatus (Ewing) (
The results showed stronger genetic divergence (measured as p-distance and the Jukes-Cantor model) between the population of C. romeroi sp. nov. and populations of C. ornatus at two localities of the municipality of Morelia, Michoacán (p-distance = 0.076–0.079), and with two populations at Chapala, Jalisco (p-distance = 0.093–0.098), with one population of C. balsasensis (p-distance = 0.128) rather than to C. infamatus from two localities of Michoacán (p-distance = 0.463) and with the type population of C. ruana (p-distance = 0.049). These differences were consistent both using p-distance and the Jukes-Cantor model (Table
Genetic distances among different populations of four species of Centruroides from Michoacán and Jalisco state including Centruroides romeroi sp. nov. The outgroup was Centruroides fulvipes from Puerto Ángel, Oaxaca. The evolutionary distances were computed using the Jukes-Cantor method [2] and are in the units of the number of base substitutions per site. p-distance (Nucleotide). This distance is the proportion (p) of nucleotide sites at which two sequences being compared are different. Distances with C. infamatus in blue, distances with C. ornatus in green.
Especie localidad | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 |
---|---|---|---|---|---|---|---|---|---|---|---|
1. C fulvipes Oaxaca** | 0.8883 | 0.9837 | 0.9492 | 0.8840 | 0.8973 | 0.8706 | 0.8973 | 0.8536 | 0.8533 | 0.8619 | |
2. C infamatus Uruapan | 0.5205 | 0.1259 | 0.1231 | 0.0000 | 0.1008 | 0.0977 | 0.0647 | 0.0478 | 0.0736 | 0.0825 | |
3. C balsasensis Tepalcatepec | 0.5479 | 0.1159 | 0.1690 | 0.1263 | 0.1753 | 0.1652 | 0.1652 | 0.1404 | 0.1453 | 0.1552 | |
4. C infamatus SEscalante1 | 0.5205 | 0.0000 | 0.1159 | 0.0000 | 0.1008 | 0.0977 | 0.0647 | 0.0478 | 0.0736 | 0.0825 | |
5. C infamatus SEscalante2 | 0.5192 | 0.0000 | 0.1162 | 0.0000 | 0.1011 | 0.0950 | 0.0619 | 0.0478 | 0.0738 | 0.0828 | |
6. C ornatus Chapala | 0.5233 | 0.0943 | 0.1563 | 0.0943 | 0.0946 | 0.0191 | 0.1133 | 0.1051 | 0.0275 | 0.0359 | |
7. C ornatus Isla Alacranes | 0.5151 | 0.0916 | 0.1482 | 0.0916 | 0.0892 | 0.0189 | 0.1070 | 0.0989 | 0.0247 | 0.0331 | |
8. C ruana Buenavista | 0.5233 | 0.0620 | 0.1482 | 0.0620 | 0.0595 | 0.1051 | 0.0997 | 0.0507 | 0.1008 | 0.1039 | |
9. C romeroi sp. nov. Coalcomán | 0.5097 | 0.0463 | 0.1281 | 0.0463 | 0.0463 | 0.0981 | 0.0926 | 0.0490 | 0.0805 | 0.0835 | |
10. C ornatus Morelia | 0.5096 | 0.0701 | 0.1321 | 0.0701 | 0.0703 | 0.0270 | 0.0243 | 0.0943 | 0.0763 | 0.0081 | |
11. C ornatus Tiripetío | 0.5123 | 0.0782 | 0.1402 | 0.0782 | 0.0784 | 0.0350 | 0.0323 | 0.0970 | 0.0790 | 0.0081 |
Centruroides romeroi sp. nov., ventral aspect of hemispermatophores of adult ♂ from “La Nieve” of Coalcomán municipality, Michoacán, Mexico. a Left side b Right side. Abbreviations: il, internal lobe. el, external lobe. ml, medial lobe. pf, pedal flexure. Tf, trunk flexure. Scale bar: 0.5 mm.
Phylogenetic trees of the Centruroides infamatus subgroup obtained by A maximum parsimony and B maximum likelihood analysis from 500 replicates bootstrap consensus. The specimens used as terminal belong to populations of the species that inhabit localities near the Coalcomán Range. Population identifications are represented by different colors.
Localities and sequences used to obtain genetic distances and bootstrap consensus tree for five species and one outgroup including C. romeroi sp. nov.
Species | Locality | Code | GenBank registration | Author | Museum where is deposited the voucher specimen |
---|---|---|---|---|---|
Centruroides fulvipes (Pocock, 1898) | Puerto Ángel, Oaxaca, Mex. | C. fulvipes Oaxaca | MK876846 | Quijano-Ravell, et al. | CAFBUM |
Centruroides infamatus (C. L. Koch, 1844) | Zumpimito, Uruapan del Progreso, Michoacán, Mex. | C. infamatus Uruapan | AF439753 |
|
CAFBUM |
Centruroides balsasensis Ponce-Saavedra & Francke, 2004 | Tepalcatepec, Michoacán, Mex. | C. balsasensis Tepalcatepec | MK787193 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
Centruroides infamatus (C. L. Koch, 1844) | Salvador Escalante, Michoacán, Mex. | C. infamatus SEscalante1 | MK877232 | Quijano-Ravell, et al. | CAFBUM |
Centruroides infamatus (C. L. Koch, 1844) | Salvador Escalante, Michoacán, Mex. | C. infamatus SEscalante2 | MK877233 | Quijano-Ravell, et al. | CAFBUM |
Centruroides ornatus Pocock, 1902 | Chapala, Jalisco, Mex. | C. ornatus Chapala | MK774709 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
Centruroides ornatus Pocock, 1902 | Isla de Los Alacranes, Chapala, Jalisco, Mex. | C. ornatus Isla Alacranes | MK774710 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
Centruroides ruana Quijano-Ravell & Ponce-Saavedra | Felipe Carrillo Puerto, Buenavista, Michoacán, Mex. | C. ruana Buenavista | MK789720 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
Centruroides romeroi sp. nov. | La Nieve, Coalcomán de Vázquez Pallares, Michoacán, Mex. | C. romeroi sp. nov. Coalcomán | MK789721 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
Centruroides ornatus Pocock, 1902 | Morelia, Michoacán, Mex. | C. ornatus Morelia | MK774718 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
Centruroides ornatus Pocock, 1902 | Tiripetío, Morelia, Michoacán, Mex. | C. ornatus Tiripetío | MK774721 | Quijano-Ravell and Ponce-Saavedra. | CAFBUM |
These molecular results support our decision of considering the Coalcomán population as an isolated and taxonomically valid species.
The authors thank Dr Omar Chassin Noria for his support with the DNA extraction.