Research Article
Research Article
Hidden species within the genus Ocys Stephens: the widespread species O. harpaloides (Audinet-Serville) and O. tachysoides (Antoine) (Coleoptera, Carabidae, Bembidiini)
expand article infoDavid R. Maddison, Roy Anderson§
‡ Oregon State University, Corvallis, United States of America
§ Unafilliated, Belfast, United Kingdom
Open Access


Beetles previously considered to be Ocys harpaloides (Audinet-Serville) from northern Africa, Spain, France, the United Kingdom, France, and Belgium belong to two species. These species can be distinguished using DNA sequences of 28S rDNA, 18S rDNA, COI, CAD, and topoisomerase I. A key, diagnoses, and images are provided to enable identification of specimens based upon characteristics of male and female genitalia, as well as microsculpture and other external structures. Through examination of the holotype of Bembidium harpaloides v. tachysoides Antoine, as well as designation of lectotypes of Bembidion harpaloides Audinet-Serville and Ocys melanocephalus Stephens, and designation of a neotype for Tachis rufescens Guérin-Ménéville, the valid names of the two species were determined to be Ocys harpaloides and Ocys tachysoides (Antoine).

Key Words

DNA taxonomy, cryptic species, Europe, ground beetle, Bembidiini, Ocys


Among European carabid beetles of the tribe Bembidiini Stephens, Ocys harpaloides (Audinet-Serville, 1821) is one of the more distinctive, looking more similar to members of other tribes (for example, the trechine Trechus Clairville, 1806 or the tachyine Porotachys Netolitzky, 1914) than to other bembidiines. As currently circumscribed, Ocys harpaloides is a common and widespread Mediterranean-Atlantic species, ranging from the Azores, Algeria and Morocco north to southern Norway, Britain and Ireland, and east to Italy, the Balkans and Greece (Lindroth 1992).

The only congener with which O. harpaloides is documented to coexist throughout much of this range is Ocys quinquestriatus (Gyllenhal), a species from which it can easily be distinguished by numerous morphological characters (Lindroth 1974; see below). There are more than 20 additional species of Ocys Stephens; almost all of these are rare, and restricted to small geographic areas (e.g., Giachino and Vailati 2012; Magrini and Degiovanni 2009; Neri et al. 2011; Toledano and Wrase 2016; Vigna Taglianti 1994). Many of these distinctive, localized endemics occur at higher elevations in mountains in Spain, northern Africa, or Italy (Magrini and Degiovanni 2009; Neri, et al. 2011; Netolitzky 1942), or on islands (Magrini et al. 1998). However, one species similar to O. harpaloides, Ocys tachysoides (Antoine, 1933), has been reported from low elevation habitats in Morocco (Antoine 1933; 1955), Spain (Toribio 2013), and a montane region of Portugal (Sciaky 1998).

In Northern Ireland, two morphologically distinct forms of “Ocys harpaloides”, correlated with habitat, have been reported (Anderson et al. 2000): specimens found among coastal rocks (Fig. 1A) are paler and larger (Fig. 2A), and those found in woodlands (Fig. 1B) are darker and smaller (Fig. 2B) (Anderson et al. 2000). Although this pattern might be a result of ecophenotypic variation, the observation suggested the possibility that two species could be hiding within “Ocys harpaloides”. However, initial morphological investigations did not reveal additional, definitive differences (Anderson et al. 2000).

Figure 1.

Habitats. A: East of Colliery Bay, Fair Head, Northern Ireland; B: Hollymount NNR near Downpatrick, Northern Ireland.

Figure 2.

Live Ocys. A: Ocys harpaloides (Benderg Bay, Co. Down, Northern Ireland); B: Ocys tachysoides (Barnett’s Demesne, Belfast, Northern Ireland).

Figure 3.

Habitus. A: Ocys harpaloides (Rinagree Point, Co. Londonderry, Northern Ireland); B: Ocys tachysoides (Annadale House, Belfast, Northern Ireland). Scale bars indicate 1 mm.

In 2011 we began an examination of the DNA sequences of these forms, and more detailed study of morphological variation. This revealed that two species were indeed present in Northern Ireland, and elsewhere in the range of “Ocys harpaloides”. As this paper was being prepared for publication, Raupach et al. (2016) reported the existence of two forms of “Ocys harpaloides” in Germany and France based upon the cytochrome oxidase I gene; these two forms correspond to the two species detailed here. In this paper, we document the differences between the two species, and through examination of types of the available names, show that the valid name of the second species is Ocys tachysoides (Antoine), which is much more widespread than previously believed.


Members of Ocys were examined from the collections listed below. Each collection’s listing begins with the coden used in the text.

BMNH The Natural History Museum, London

CTVR Luca Toledano Collection, Verona, Italy

DE Dominique Echaroux Collection, Etréchy, France

MNHN Muséum National d’Histoire Naturelle, Paris

OSAC Oregon State Arthropod Collection, Oregon State University

PB Paolo Bonavita Collection, Rome, Italy

RAC Roy Anderson Collection, Belfast, United Kingdom

Morphological methods. General methods of specimen preparation for morphological work, and terms used, follow Maddison (1993; 2008). Genitalia were prepared, after dissection from the body, by treatment in 10% KOH at 65 °C for 10 minutes followed by multi-hour baths of distilled water, 5% glacial acetic acid, distilled water, and then ethanol. Male genitalia were then mounted in Euparal between two small coverslips attached to archival-quality heavyweight watercolour paper, and, once dried, pinned beneath the specimen. For all type specimens except that of Ocys melanocephalus Stephens, genitalia were removed from Euparal through soaking in 100% ethanol, placed in small plastic vials containing glycerol, and pinned beneath the specimen.

Photographs of male genitalia were taken with a Leica Z6 and JVC KY-F75U camera using Microvision’s Cartograph software for extended depth of field (EDF) processing; the images thus potentially have some artifacts caused by the EDF algorithm.

The following measurements were made:

Length: body length, from apex of the labrum to apex of the longer elytron.

GCR : gonocoxite ratio = gonocoxite 2 length / gonocoxite 1 length (Fig. 4)

Measurements were made either using Microvision’s Cartograph software processing images from a JVC KY-F75U camera on a Leica Z6 lens, or on a Leica M3Z stereomicroscope with a Leitz graticule.

Figure 4.

Diagram of gonocoxite of a female Ocys showing measurements taken.

The density of the microsculpture lines on the left elytron was measured by counting the number of microsculpture lines that cross a 0.1 mm longitudinal line placed in the centre of the third elytral interval approximately 1/3 of the distance between the front and back of the elytron.

Taxon sampling for DNA studies. We obtained DNA sequence data for 15 Ocys specimens (Table 1), and combined these with results from the two Ocys specimens sequenced in Maddison (2012). For outgroups, we chose two or more species representing different lineages within each of the five largest genera of Bembidiina (Bembidion Latreille, Asaphidion Chaudoir, Amerizus des Gozis, Sinechostictus Motschulsky, and Lionepha Casey (Maddison 2012)). The outgroup species are listed in Table 2.

Locality details for sequenced Ocys exclusive of O. quinquestriatus. Four-digit numbers at the start of each row are D.R. Maddison DNA voucher numbers. Specimen 0569 had been previously sequenced by Maddison (2012).

Ocys harpaloides
2760 Northern Ireland: Colliery Bay, Ballycastle, Antrim, 55.2116°N 6.1910°W. 5.vii.2011. At grass roots on slope above high water mark. R. Anderson
2762 Northern Ireland: Colliery Bay, Ballycastle, Antrim, 55.2116°N 6.1910°W. 5.vii.2011. At grass roots on slope above high water mark. R. Anderson
2759 Ireland: Wexford, North Slob. 52.3546°N 6.4082°W. 20.vii.2011. Under driftwood on sandy beach. R. Anderson
0569 Belgium: Schorisse, Bos Ter Rijst. Approximately 50.78°N 3.69°E. 17.iv.1997. Temperate broadleaf forest. Konjev Desender
2937 Belgium: Schorisse, Bos Ter Rijst. Approximately 50.78°N 3.69°E. 17.iv.1997. Temperate broadleaf forest. Konjev Desender
2853 Belgium: Halve Maan, Oostende. 51.237°N 2.932°E. 23.ix.1995. Coastal salt marsh (non tidal). K. Desender.
4983 Spain: Sitges. 41.2383°N 01.8244°E. 9.ix.2006. W Maddison.
Ocys tachysoides
2758 Northern Ireland: Belfast, Belvoir Forest. 54.5544°N, 5.9404°W. 10.iii.2011. Under bark dead alder and willow. R. Anderson
2761 Northern Ireland: Belfast, Belvoir Forest. 54.5544°N, 5.9404°W. 10.iii.2011. Under bark dead alder and willow. R. Anderson
2763 Northern Ireland: Belfast, Belvoir Forest. 54.5544°N, 5.9404°W. 10.iii.2011. Under bark dead alder and willow. R. Anderson
2898 Northern Ireland: Co. Down, Mount Stewart. 54.5491°N 5.6037°W. 12.x.2011. Under bark of sycamore/cedar/pine by sea. R. Anderson
2899 Northern Ireland: Co. Down, Murlough NNR. 54.2461°N 5.8330°W. 15.xi.2011. Under bark of sycamore, sandy wood near sea. R. Anderson
2936 Belgium: Moerzeke, De Kramp. Approximately 51.06°N 4.15°E. 1.iii.1999. Tidal freshwater marsh along the River Schelde. Konjev Desender
2938 Belgium: Kastel, Gespoelde. Approximately 51.05°N 4.16°E. 11.ix.1998 Tidal freshwater marsh along the River Schelde. Konjev Desender
4606 Germany: Nordrhein-Westfalen, Landkreis Unna, Selm-Bork, Lippeaue. 51.6533°N 7.4442°E. Under bark of Salix. K. Hannig

Sampling of Bembidiina species. Four-digit numbers in the “#” column entries for Ocys are D.R. Maddison voucher numbers for sequenced specimens. Data for all non-Ocys specimens, as well as to specimens 1077 and 0569, were obtained from GenBank from previous studies (Maddison 2008; 2012; Maddison et al. 1999; Maddison and Ober 2011; Maddison and Swanson 2010; Wild and Maddison 2008), and more data about the specimens are presented in those papers. Locality details for Ocys specimens are given in Table 1 and in Maddison (2012). For each gene, GenBank accession numbers are listed.

# COI 28S 18S CAD Topo
Bembidion chalceum Dejean EF649200 EF648892 EF648647 EF649431 EU677650
Bembidion rapidum (LeConte) JN171095 EU677690 JN170224 EU677543 EU677642
Bembidion transversale Dejean GU454797 EU677688 JN170242 EU677541 EU677639
Bembidion variegatum Say JN171131 JN170458 JN170245 JN170937 JN171310
Asaphidion curtum (Heyden) JN170977 GU556078 AF002792 JN170736 JN171163
Asaphidion yukonense Wickham JN170979 JN170273 JN170139 EU677540 EU677638
Amerizus wingatei (Bland) JN170974 JN170267 JN170136 JN170732 JN171160
Amerizus (Tiruka) sp. JN170972 JN170265 JN170134 JN170730 JN171158
Sinechostictus elongatus (Dejean) JN171152 JN170479 JN170260 JN170965 JN171332
Sinechostictus (Pseudolimnaeum) sp. 3 JN171150 JN170474 JN170259 JN170960 JN171329
Lionepha erasa (LeConte) JN171141 JN170468 JN170252 JN170948 JN171320
Lionepha osculans (Casey) JN171143 JN170470 JN170254 JN170950 JN171322
Lionepha disjuncta (Lindroth) JN171142 JN170469 JN170253 JN170949 JN171321
Ocys quinquestriatus (Gyllenhal) 1077 JN171145 JN170472 JN170257 JN170954 JN171324
Ocys harpaloides (Audinet-Serville) 0569 GU556103 JN170256 JN170953
2759 KX907141 KX907154 KX907168 KX907176 KX907188
2760 KX907142 KX907155 KX907177 KX907189
2762 KX907143 KX907156 KX907178 KX907190
2853 KX907148 KX907161 KX907183 KX907195
2937 KX907152 KX907165 KX907199
4983 KX907153 KX907166
Ocys tachysoides (Antoine) 2758 KX907144 KX907157 KX907169 KX907179 KX907191
2761 KX907145 KX907158 KX907180 KX907192
2763 KX907146 KX907192 KX907181 KX907193
2898 KX907147 KX907160 KX907182 KX907194
2899 KX907151 KX907164 KX907198
2936 KX907149 KX907162 KX907184 KX907196
2938 KX907150 KX907163 KX907185 KX907197
4606 KX907187 KX907175 KX907172 KX907186 KX907167

DNA sequencing. Genes studied, and abbreviations used in this paper, are: 28S: 28S ribosomal DNA (D1-D3 domains); 18S: 18S ribosomal DNA (near full-length); COI: cytochrome oxidase I; CAD: carbamoyl phosphate synthetase domain of the rudimentary gene; Topo: topoisomerase I. In addition, sequences of the wingless gene and arginine kinase were acquired for specimens Ocys harpaloides 2759 and Ocys tachysoides 2758.

For all except specimen number 4606, DNA was extracted from specimens preserved in 95% ethanol using a Qiagen DNeasy Blood and Tissue Kit. Fragments for the seven genes were amplified using the Polymerase Chain Reaction on an Eppendorf Mastercycler Pro Thermal Cycler, using TaKaRa Ex Taq and the basic protocols recommended by the manufacturers. Primers and details of the cycling reactions used are given in Maddison (2012). The amplified products were then purified, quantified, and sequenced at the University of Arizona’s Genomic and Technology Core Facility using a 3730 XL Applied Biosystems automatic sequencer. Assembly of multiple chromatograms for each gene fragment and initial base calls were made with Phred (Green and Ewing 2002) and Phrap (Green 1999) as orchestrated by Mesquite’s Chromaseq package (Maddison and Maddison 2014; Maddison and Maddison 2015b) with subsequent modifications by Chromaseq and manual inspection. Multiple peaks at a single position in multiple reads were coded using IUPAC ambiguity codes.

For specimen 4606, RNA was extracted from an RNA-Later preserved specimen using a Qiagen RNeasy Kit and Trizol (Life Technologies). Isolation of mRNA was done with an NEBNext® Poly(A) mRNA Magnetic Isolation Module (New England BioLabs). The library was prepared using an NEBNext Ultra RNA (New England BioLabs) kit using 1000 ng of input RNA and NEBNext Multiplex Oligos for Illumina (New England BioLabs). The finished library was quantified using a Qubit Florometer (Life Technologies) and fragment distribution characterized on a 2100 Bioanalyzer (Agilent Technologies). The library was then sequenced on an Illumina 2500 at the Oregon Health Sciences University’s Massively Parallel Sequencing Shared Resource, with reads demultiplexed using CASAVA v1.8. The 100-base, paired-end run produced 36 million reads, which were assembled using CLC Genomics Workbench version 8.5, using the de novo assembly feature with default parameter values. The resulting assembly was converted to a blastable database using NCBI’s makeblastdb tool. Using the Asaphidion yukonense sequences from Maddison (2012) as query sequences, the specimen 4606 transcriptome was searched using nBLAST, yielding single contig sequences for 28S, 18S, and Topo. For CAD, three overlapping contigs were found; they were identical in the overlapping regions, and were merged into a single sequence. For COI, three contigs were found, one containing full-length COI; the other two short fragments could be eliminated from consideration because they BLASTed to non-bembidiines in GenBank or contained stop codons.

Sequences have been deposited in GenBank with accession numbers KX907141 through KX907199 (Table 2). All sequences fall into the “genseq-4” category (Chakrabarty et al. 2013).

Alignment and phylogenetic analysis. The appropriate alignment was obvious for COI, CAD, and Topo, as there were no insertion or deletions (indels) evident in the sampled sequences. The two ribosomal genes were aligned by MAFFT version 7.130b (Katoh and Standley 2013), using the L-INS-i search option and otherwise default parameter values. No data were excluded from analyses, other than trimming of ends of sequences that were notably longer than others.

Models of nucleotide evolution where chosen with the aid of jModelTest version 2.1.7 (Darriba et al. 2012; Guindon and Gascuel 2003). Among the models supported by GARLI, the models chosen by the Bayesian Information Criterion were 28S: TVMef+G, 18S: SYM+I+G, COI: TIM1+I+G, CAD: TVMef+I+G, and Topo: TrN+G. Likelihood analyses of nucleotide data were conducted using GARLI version 2.0 (Zwickl 2006), as orchestrated by the Zephyr package of Mesquite (Maddison and Maddison 2015a). Twenty-five search replicates were employed for maximum likelihood trees, and 200 replicates for bootstrap analyses.


There are two distinct clades in each of the five gene trees (Fig. 5): one clade contains all of the larger, paler specimens, and another contains all of the smaller, darker specimens. As outlined below in the taxonomic section, the valid name for the larger, paler specimens is Ocys harpaloides (Audinet-Serville), and the valid name for the smaller, darker specimens is Ocys tachysoides (Antoine). Based upon the arguments presented below in the Discussion, we consider these different species, and for simplicity’s sake we will speak of them as different species in this section, and use their valid names. Likelihood bootstrap support for these two species is strong (Table 3), being above 90% in at least two of the well-sampled genes.

Figure 5.

Phylogenetic trees of the five studied genes. Outgroups not shown. Branch length is shown proportional to relative divergence, as estimated by GARLI. The larger, paler specimens with genitalia as in Fig. 6A and B are shown in orange, and the smaller, darker specimens with genitalia as in Fig. 6C and D are shown in brown.

Support for the monophyly of each of the two species in data for each gene. Numbers shown are maximum likelihood bootstrap percentages.

Clade 28S 18S COI CAD Topo
O. harpaloides 100 100 100 95 95
O. tachysoides 100 100 84 63 100

In COI, there are 11 sites at which three to six of the O. tachysoides (all specimens except 2899) show a secondary peak in chromatograms; there are three different sites at which 1 to 3 O. harpaloides (2759, 2760, 2853) have a secondary peak. The secondary peaks suggest an alternative version of COI, which is presumably a nuclear copy, or “numt” (Thalmann et al. 2004), thus causing some uncertainty about the source of our sequenced COI. However, in spite of the uncertainty, the gene tree clearly supports two distinct species (Fig. 5).

A detailed examination of differences between and within each of these two species also reveals many sites at which they differ (Table 4). There are six fixed amino-acid differences between the two species across the three protein-coding genes.

Variation observed in the DNA sequences between and within species. The number of specimens examined per species ranged from 3 to 5, with each species represented by 2 to 3 localities. “nuc”: nucleotides; “aa”: amino acids. The * indicates that one of the fixed differences is an extra base in O. tachysoides relative to O. harpaloides.

fixed differences between species sites varying within species % nuc differences between species
gene nuc aa nuc aa
28S 22* - 1 - 2.1-2.2
18S 15 - 1 - 0.9
COI 50 2 9 0 7.2-8.4
CAD 5 1 6 0 0.50-1.0
Topo 16 3 0 0 2.2

Morphological characteristics were in general correlated with DNA sequences. All five males in the Ocys harpaloides clade in Fig. 5 had genitalia matching those shown in Fig. 6A, B, and all seven males in the Ocys tachysoides clade had genitalia matching Figs 6C, D (Table 5). Irish populations also had the colour of the elytral disc correlated with the gene tree clades, but the correlation was not as clear in Belgium (Table 5).

Figure 6.

Male aedeagi. A, B: Ocys harpaloides, both from Colliery Bay, Ballycastle, Northern Ireland (A: voucher V100671, B: voucher V100670); C, D: Ocys tachysoides, both from Belvoir Forest, Belfast, Northern Ireland (C: voucher DNA2761; D: voucher DNA2758). Scale bars 0.1 mm.

Morphological features and habitats of the sequenced specimens. “Disc colour” refers to the colour of the elytral disc relative to the apex and lateral margins. Male genitalic type A is that shown in Fig. 6A, B, and genitalic type C is that shown in Fig. 5C, D; FA refers to female genitalia type shown in Figs 8A and 9A.

Region Disc colour Genitalic type Habitat
O. harpaloides Northern Ireland paler A ocean shore
O. harpaloides Northern Ireland paler A ocean shore
O. harpaloides Republic of Ireland paler A ocean shore
O. harpaloides Belgium slightly paler FA broadleaf forest
O. harpaloides Belgium slightly paler A coastal salt marsh
O. harpaloides Belgium ± dark A broadleaf forest
O. harpaloides Spain paler A open woodland
O. tachysoides Northern Ireland dark C woodland
O. tachysoides Northern Ireland dark C woodland
O. tachysoides Northern Ireland dark C woodland
O. tachysoides Northern Ireland dark C woodland
O. tachysoides Northern Ireland dark C woodland
O. tachysoides Belgium dark C freshwater marsh
O. tachysoides Belgium dark C freshwater marsh
O. tachysoides Germany dark C woodland

On the island of Ireland, all three O. harpaloides clade males were found on the ocean shore, at two localities, whereas all five O. tachysoides clade males were found in woodland habitats, at three localities (Table 5). That distinction was not as evident on the mainland of Europe, as two of the O. harpaloides specimens from Belgium were found in a broadleaf forest, inland (Table 5).


Our data show that specimens in northern Europe referred to “Ocys harpaloides” belong to two species. The DNA data alone strongly suggests that two species live in Northern Ireland and Belgium: the gene trees for two certainly unlinked genes (nuclear 28S and mitochondrial COI) show the same clades for specimens that broadly overlap in geographic range, with specimens of both forms sampled from each region. This pattern, combined with identical patterns for 18S, CAD, and Topo, indicates that the two forms are not currently exchanging genes. This is confirmed by the correlated morphological traits, in particular the distinctive male genitalia.

Toribio (2013) recognized these two species in populations in southern Spain and Morocco, but did not recognize that O. tachysoides was as widespread as O. harpaloides.

By comparing our sequences with those presented by Raupach et al. (2016), we have confirmed that the two COI forms they documented correspond to O. harpaloides (their French specimens) and O. tachysoides (their German specimens).

Taxonomic section

The following key allows one to identify Ocys specimens from much of Europe, outside of those regions containing localized endemics (that is, outside of higher elevations in mountains in Spain, northern Africa, or Italy (Magrini and Degiovanni 2009; Neri et al. 2011; Netolitzky 1942), on islands (Magrini et al. 1998).

1 Pronotum with obtuse and rounded hind angles, with the hind margin protruding posteriorly in the middle relative to the hind angles; colour dark brown with bluish or greenish reflections Ocys quinquestriatus (Gyllenhal)
Pronotum with sharp hind angles, approximately a right angle, with the hind margin more or less straight; colour without notable metallic reflections 2
2 Elytra rufous or brown, in most specimens paler centrally with darker apical and lateral margins. Microsculpture of elytra consisting of more closely spaced, transverse lines with less of a tendency to form meshes (Fig. 7A, B). Aedeagus with ventral margin in most specimens bent downward toward the thinner apex; anterior sclerites of the internal sac more rounded; with a brush sclerite of normal size (Fig. 6A, B). Spermatheca more curved (Fig. 9A, B) Ocys harpaloides (Audinet-Serville)
Elytra dark brown to black in colour, in some specimens paler along the suture, but entire discal region not paler than margins. Microsculpture of elytra consisting of less closely spaced, transverse lines with more of a tendency to form meshes (Fig. 7C, D). Aedeagus with ventral margin straighter, and apex wider, with a blunter tip; anterior sclerites of the internal sac more angulate; with a very small brush sclerite (Fig. 6C, D). Spermatheca straighter (Fig. 9C, D) Ocys tachysoides (Antoine)
Figure 7.

Microsculpture at center of disc of left elytron, third and fourth intervals. A, B, O. harpaloides, both from Colliery Bay, Ballycastle, Northern Ireland (A: voucher V100670, B: voucher V100672). C, D. O. tachysoides (C: Murlough NNR, Co. Down, Northern Ireland, voucher DNA2899, D: Mount Stewart, Co. Down, Northern Ireland, voucher DNA2898). Scale bar 0.1 mm.

Figure 8.

Female genitalia. A: O. harpaloides, Bos Ter Rijst, Schorisse, Belgium, voucher DNA0569. B: O. tachysoides, Murlough NNR, Co. Down, Northern Ireland, voucher V100983. Scale bar 0.1 mm.

Figure 9.

Spermatheca. A, B, O. harpaloides (A: Bos Ter Rijst, Schorisse, Belgium, voucher DNA0569, B: Beuda, Girona, Spain, voucher V100984) C, D. O. tachysoides (B: Murlough NNR, Co. Down, Northern Ireland, voucher V100983, D: Mount Stewart, Co. Down, Northern Ireland, voucher V100982). A bubble of air has been digitally removed from within the spermatheca shown in C. Scale bar 0.1 mm.

Ocys harpaloides (Audinet-Serville, 1821)

Figs 2A; 3A; 6A, B; 7A, B; 8A; 9A, B; 10A, B; 11A, B

Bembidion harpaloides Audinet-Serville, 1821:78. Lectotype male, here designated, in the Dufour collection of the MNHN, examined (see Nomenclatural notes, below), with two labels: “5244” [handwritten], “LECTOTYPE Bembidion harpaloidesAudinet-Serville 1821 designated 2016 D.R. Maddison and R. Anderson” [partly handwritten, with red lines around the border]. Type locality: Clavados, Bretagne, France.

Tachis rufescens Guérin-Ménéville, 1823:123. Type specimens lost (see below). Type locality as specified in the original description: an island on the Seine River (presumably Île Seguin or Île Saint-Germain), Meudon, France. Neotype male, here designated, in the MNHN, labeled “JUVISY Aval RD 4-VI-45 M. DEWAILLY” [partly handwritten], “Ocys harpaloides Serv. M. Dewailly det.” [partly handwritten], “NEOTYPE Tachis rufescens Guérin-Ménéville designated 2016 D.R. Maddison & R. Anderson” [partly handwritten]. This specimen is from Juvisy-sur-Orge, Essonne, France, which is 18km SE of Meudon; it was formerly housed in the collection of Mr. Dominique Echaroux.

Ocys melanocephalus Stephens, 1828:10. Lectotype male, here designated, in BMNH, examined, with six labels: “BRITISH ISLES J. Stephens Coll. BM 1853- 46”, “O melanocephalum” [handwritten], “♂”, “harpaloides det Netolitzky” [handwritten], “NHMUK010363535” [with matrix barcode], “LECTOTYPE Ocys melanocephalus Stephens designated 2016 D.R. Maddison and R. Anderson” [partly handwritten, with red lines around the border]. Type locality: British Isles.

Bembidium dubium Wollaston, 1857:23. Holotype female, in BMNH, examined. Type locality: Madeira. Junior primary homonym of Bembidium dubium Heer, 1838. Synonymy tentative, based primarily on the colour of the type.

Nomenclatural notes

Interpretation of Audinet-Serville’s name rests in part on discovery of his specimens, as his description is not sufficient to distinguish between the two species. Audinet-Serville notes that his specimen or specimens of Bembidion harpaloides are from “M. de Brébisson”, presumably the botanist Louis Alphonse de Brébisson. Audinet-Serville’s Coleoptera specimens were acquired by Léon Jean Marie Dufour (Thierry Deuve, pers. comm.), whose collection was incorporated into the Muséum National d’Histoire Naturelle in Paris. In March 2016, David H. Kavanaugh searched for several hours for the type series of Bembidion harpaloides Audinet-Serville in Dufour’s collection at the MNHN, with help from Azadeh Taghavian. The following notes are in part from Kavanaugh (pers. comm.). Dufour’s collection is well-organized, and in the section containing bembidiines, there is only one box with Ocys. It contains four pins with Ocys specimens. These are housed under the heading “Bembidion (Tachys) rufescens”, a name in use around that time for this complex (e.g., Dejean 1831). In that region of the box, there are no additional pin holes, suggesting that there have been no specimens of Ocys removed from Dufour’s collection. The four pins contain a total of eight specimens: the first two pins contain one specimen each, the third pin houses five specimens, each on a separate card, and the last pin houses a single specimen. The last specimen can be excluded from consideration, as its label contains a locality (“Carcassonne”, in southern France) that differs from the locality stated by Audinet-Serville. The other three pins each have one or two labels: the first specimen has a label that apparently contains the number 249; the second the number 5244, and the remaining pin has two labels, one with a small amount of indecipherable text, the other with the number 124. All of the Ocys in Dufour’s collection belong to this species, as determined by colour pattern and microsculpture. In addition, the first two are males, and dissection revealed genitalia that match genitalic type A (i.e., as in Fig. 6A, B). The labels on the three pins are in different handwriting, and the pins themselves differ in structure. Thus, it appears as if the three came from different sources, and likely only one of them represents authentic Audinet-Serville material. According to Antoine Mantilleri (Thierry Deuve, pers. comm.), the handwriting on the first specimen is likely that of Dufour. This suggests that the first specimen is not an authentic Audinet-Serville specimen, but rather an original Dufour specimen; however, it is possible that Dufour added a label to the specimen after having acquired Audinet-Serville’s collection. The writer of the other labels is not known. We could discover no information about Audinet-Serville’s handwriting, other than a single example of his signature, and thus we do not know if any of the labels might have been written by him, or whether they may have been added by later researchers. Thus, it is not clear which specimen on the three candidate pins is Audinet-Serville’s, and it likely will never be determined with certainty. We consider the value in stabilizing the name more important than the uncertainty of the choice. As all of these specimens belong to the same species, as the specimen on the second pin is a male in sufficient condition and with undamaged genitalia, and as there is hint that the specimen on the first pin is not an Audinet-Serville specimen, we have chosen the second specimen as the lectotype of Bembidion harpaloides Audinet-Serville.

The collection of Félix Édouard Guérin-Méneville was incorporated into the Chaudoir collection at the MNHN (Cambefort 2006) in the Chaudoir magasin. David Kavanaugh and Azadeh Taghavian searched for types of Tachis rufescens Guérin-Ménéville in the Chaudoir magasin. They found numerous Ocys, but every one could be eliminated as Guérin-Ménéville specimens through consideration of the labels, as all specimens had a mechanically printed label (against other Guérin-Ménéville specimens), and the labels specified a locality, collector, or collection that did not coincide with those of the Ocys examined by Guérin-Ménéville (Kavanaugh, pers. comm.). The Guérin-Ménéville specimens of Ocys are thus lost. In the interests of stabilizing the nomenclature, we sought a neotype for Tachis rufescens. Within the MNHN, there are specimens from three localities around Paris (Vaires, Chelles, and Chalifert). All of these specimens belong to the following species, Ocys tachysoides (Thierry Deuve, pers. comm.); we have confirmed this for the specimens from Vaires by examination of male genitalia. However, the lack of Ocys harpaloides in the twentieth-century specimens in the MNHN does not necessarily mean that the species was absent from Paris in 1823: in the 123 years that passed between the description of Tachis rufescens and the collection of the Viares specimens in 1946, the habitats around Paris may have changed. The possibility that Ocys harpaloides lived in Paris in the 1820s is made more likely by the presence, in the collection of Mr. Dominique Echaroux, of two series of specimens of Ocys harpaloides from just outside of Paris: one specimen from Bouray-sur-Juine and seven from Juvisy-sur-Orge. The latter is only 18km SE of the type locality of Tachis rufescens. Thus, both species have lived in the general Paris area, and a specimen of either species could be designated as a neotype. However, designation of the Vaires specimen, for example, would lead to Ocys rufescens as the name of the following species, with O. tachysoides as a junior synonym. This would change the name used for the following species in the Iberian Peninsula and Africa, where it is known as O. tachysoides (per Toribio 2013). In the interests of stability, we choose as neotype of Tachis rufescens one of the Juvisy-sur-Orge males; with this designation, Tachis rufescens is maintained as synonym of O. harpaloides.

In the BMNH, under “melanocephalum”, are seven specimens (four males, three females) in the J. Stephens collection, all with the label “British Isles J. Stephens Coll. BM. 1853 – 46”. We consider this to be the type series of Ocys melanocephalus Stephens. Three of the males in this series were dissected, and genitalia examined; they all match genitalic type A (i.e., as in Fig. 6A, D). We have chosen one of these specimens as the lectotype.

In the Wollaston collection of the BMNH, in Insecta Maderensia, Drawer 2, is a single teneral female, without a determination label. This specimen is presumed to be the holotype of Bembidium dubium Wollaston, 1854. Its colour suggests that it belongs to this species. However, the species membership of the type is of no nomenclatural importance, as the name is a junior primary homonym of Bembidium dubium Heer, 1838, itself a junior synonym of Bemidion assimile (Gyllenhal, 1810).

An additional name (Carabus tempestivus Panzer, 1799) is listed as a synonym of O. harpaloides by some authors (e.g. Stephens, 1828). However, this name is a synonym of Trechus quadristriatus (Schrank, 1781), as documented by Erichson (1837).


Body length 4.2–6.1 mm (Toribio 2013 and our observations; average length of males 5.0 mm (n=5), of females 5.51 mm (n=5)). Head and pronotum a clear orange-brown; anterior and central part of elytra of the same colour, but sides and posterior region of elytra dark brown (Fig. 3), and with the epipleural gutter and suture yellow-brown. Microsculpture of elytral disc more transverse, with more close-set parallel lines and fewer meshes (Fig. 7A, B); density of microsculpture lines 25–26 per 0.1 mm (n=4 males). Hind margin of pronotum in most specimens straighter than in O. tachysoides, such that the hind margin is directed more or less laterally near the hind angles (Fig. 10A), occasionally with a slight emargination laterally (Fig. 10B). Elytra more parallel-sided, greatest width behind middle. Basal margin at shoulder slightly arcuate with a forward-directed concavity (Fig. 11A, B). Elytral striae 2 through 4 less marked in the apical third. Aedeagus with ventral margin bend slightly downward toward apex (Fig. 6A, B); apex more rounded. Anterior edge of central sclerite complex of internal sac more rounded; brush sclerite larger; dorsal membranes of internal sac darker. Gonocoxite relatively long (Fig. 8A), GCR=0.64–0.69 (average 0.67, n=5); spermathecal margin (opposite the efferent duct of the spermathecal gland) curved (Fig. 8A, B; n=5), with tip pointed toward duct of gland.

Figure 10.

Hind margin of pronotum. A, B, O. harpaloides (A: North Slob, Wexford, Ireland, voucher DNA2759, B: Halve Maan, Oostende, Belgium, voucher DNA2853). C, D. O. tachysoides (B: Belvoir Forest, Belfast, Northern Ireland, voucher DNA2763, D: Murlough NNR, Co. Down, Northern Ireland, voucher DNA2899). Scale bar 0.1 mm.

Figure 11.

Left shoulder. A, B, O. harpaloides (A: North Slob, Wexford, Ireland, voucher DNA2759, B: Bos Ter Rijst, Schorisse, Belgium, voucher DNA2937) C, D. O. tachysoides (B: River Schelde, De Kramp, Moerzeke, Belgium, voucher DNA2936, D: Murlough NNR, Co. Down, Northern Ireland, voucher DNA2899). Scale bar 0.1 mm.

Geographic distribution

(Fig. 12). In Africa, from Morocco, Algeria, and Tunisia. In Europe from Spain, France, Belgium, Italy, Ireland, and the United Kingdom. Examination of specimens in additional collections will likely show it to be more widely distributed.

Figure 12.

Distribution map of Ocys harpaloides and O. tachysoides. Only records of which we are confident are included. In addition to specimens we have examined, and those from Toribio (2013), Antoine (1956), and Sciaky (2009), we have included three publically available records from the Barcode of Life Database (, corresponding to BOLD records GBCOU8213-14, GBCOU8667-14, and FBCOH116-12, as well as specimens identified by Luca Toledano and Paolo Bonavita in their collections (CTVR and PB, respectively) based upon images and diagnoses we provided to them.

Specimens examined

In addition to type specimens, and those listed in Table 1, we examined specimens from Morocco: Tétouan (BMNH); Algeria (BMNH); France: Carcassone (MNHN), Bouray-sur-Juine (DE); Spain: San Roque (BMNH), Beuda, Girona (OSAC), and Sant Carles de Peralta (RAC).

Ocys tachysoides (Antoine, 1933)

Figs 2B; 3B; 6C, D; 7C, D; 8B, 9C, D; 10C, D; 11C, D

Bembidium harpaloides v. tachysoides Antoine, 1933:79. Holotype female, in the MNHN, examined, with three labels: “O. Nefifik (Maroc) Antoine I.1928” [partly handwritten], “Holotype” [handwritten on red paper], “tachysoides Antoine det. m.” [partly handwritten]. Type locality: Mouth of the Oued Nefifikh, east of Casablanca, Morocco (in the vicinity of 33.72°N, 7.34°W).

Nomenclatural notes

The type of Bembidium harpaloides v. tachysoides Antoine belongs to this species. We have examined external features of the holotype, including elytral microsculpture (which has a density of 21 lines per 0.1 mm), as well as shape of the spermatheca. Both second gonocoxites are broken in the holotype, and thus we were not able to measure their lengths.


Body length 4.0–5.8 mm (Toribio 2013 and our observations; average length of males 4.79 mm (n=7) of females 5.16 mm (n=7)). Head and pronotum a deep red-brown contrasting with the dark brown to blackish elytra (Fig. 3), only the epipleural gutters and sometimes sutures paler, light reddish-brown. Occasionally with a bluish lustre. Microsculpture of elytral disc less transverse, with more of tendency to form sculpticells (Fig. 7C, D); density of microsculpture lines 20–21 per 0.1 mm (n=4 males). Hind margin of prothorax with a slight emargination laterally, such that the hind margin at the hind angles is directed posteriorly (Fig. 10C, D). Elytra less parallel-sided, greatest width around middle. Basal margin at shoulder in most specimens more or less straight (Fig. 11C), relatively few specimens slightly arcuate with a forward-directed concavity (Fig. 11D). Elytral striae 2 through 4 more marked in the apical third. Aedeagus with ventral margin bend more or less straight (Fig. 6C, D); apex less rounded, blunt. Anterior edge of central sclerite complex of internal sac angulate; brush sclerite smaller; dorsal membranes of internal sac paler, less evident. Gonocoxite relatively short (Fig. 8B), GCR=0.55–0.60 (average 0.58, n=5); spermathecal margin opposite the efferent duct of the spermathecal gland more or less straight (Fig. 8C,D; n=5).

Geographic distribution

(Fig. 12). In Africa, known from Morocco (Antoine 1955). In Europe, from Portugal (Sciaky 1998), Spain (Toribio 2013), France, Belgium, Germany, and the United Kingdom. Examination of specimens in additional collections will likely show it to be more widely distributed.

Specimens examined

In addition to the type specimen, and those listed in Table 1, we examined specimens from France: Vaires (MNHN).


We are extremely thankful to the individuals who worked so hard on our behalf as we were attempting to locate original Audinet-Serville and Guérin-Ménéville specimens at the Muséum National d’Histoire Naturelle, Paris, and to designate primary types for the associated names. Most importantly, Thierry Deuve patiently dealt with our many queries, examined numerous specimens, and visited Dominique Echaroux. His thoughtful responses, as we puzzled through the best approach to resolving the nomenclatural issues, were invaluable. Azadeh Taghavian’s help in this regard can also not go unmentioned. In addition, David Kavanaugh spent many valuable hours while in Paris personally looking for Ocys types, and we could have not completed the project without his help; we are also thankful for his advice as we considered the best path, and for hand-carrying the Audinet-Serville material back from Paris. Several others aided these efforts as well, and we wish to thank them: Antoine Mantilleri (for his advice about Audinet-Serville material), and Paul Johnson (for providing copies of pages from Horn & Kahle). Finally, we wish to thank Mr. Dominique Echaroux, from Etréchy (Essonne, France), for providing specimens of Ocys harpaloides from his collection, and for allowing us to designate one of them as a neotype of Tachis rufescens.

We are also very thankful for Beulah Garner’s stewardship of the specimens at the Natural History Museum in London, and for sending a portion of the type series of Ocys melanocephalus Stephens so that we might designate a lectotype.

For various helpful discussions we thank Marcos Toribio, Luca Toledano, Paolo Bonavita, and Paolo Neri. For their thoughtful reviews of the manuscript, we thank Michael Raupach, Pier Mauro Giachino, and Joachim Schmidt. We also thank Luca Toledano and Paolo Bonavita for examining the specimens in their collections and providing additional localities for Fig. 12.

We thank all those who provided material preserved for DNA studies, including Joachim Schmidt, Frederik Hendrickx, and Wayne Maddison, and in particular the late Konjev Desender. We also thank John S. Sproul for extracting the RNA of specimen DNA4606, James M. Pflug for preparing the sequencing library, and Illumina Inc. for helping resolve some sequencing issues.

This work was funded in part by the Harold E. and Leona M. Rice Endowment Fund at Oregon State University.


  • Anderson R, McFerran D, Cameron A (2000) The ground beetles of Northern Ireland (Coleoptera - Carabidae). Ulster Museum, Belfast., i-x, 1–246 pp.
  • Antoine M (1933) Notes d’entomologie marocaine. XIV Carabiques nouveaux ou intéressants (Ins. Coléopt.). Bulletin de la Société des Sciences Naturelles du Maroc 13: 69–101.
  • Antoine M (1955) Coléoptères carabiques du Maroc. Première partie. Mémoires de la Société des sciences naturelles et physiques du Maroc, Nouvelle série 1: 1–77.
  • Audinet-Serville JG (1821) Faune Française ou histoire naturelle, générale et particulière des animaux qui se trouvent en France, constamment ou passagèrement, à la surface du sol, dans les eaux qui le baignent, et dans le littoral des mers qui le bornent. Coléoptères., Paris, 96 pp.
  • Cambefort Y (2006) Des coléoptères, des collections, et des hommes. Muséum national d’Histoire naturelle, Paris, 375 pp.
  • Chakrabarty P, Warren M, Page LM, Baldwin CC (2013) GenSeq: An updated nomenclature and ranking for genetic sequences from type and non-type sources. ZooKeys 346: 29–41. doi: 10.3897/zookeys.346.5753
  • Darriba D, Taboada GL, Doallo R, Posada D (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature Methods 9: 772. doi: 10.1038/nmeth.2109
  • Dejean PFMA (1831) Spécies général des Coléoptères de la collection de M. le Comte Dejean. Paris, I-VIII, 1–384 pp.
  • Erichson WF (1837) Die Käfer der Mark Brandenburg. Erster Band. Erste Abtheilung. Morin, Berlin, xiii+384 pp.
  • Giachino PM, Vailati D (2012) Ocys phoceus n. sp., a new interesting orophilous species of Central Greece (Coleoptera, Carabidae). Fragmenta Entomologica 44: 65–70. doi: 10.4081/fe.2012.32
  • Guérin-Ménéville FE (1823) Note topographie sur quelques insects coléoptères, et Description de deux espèces des genre Badister et Bembidion. Bulletin des Sciences, par la Societé Philomathique de Paris 1823: 121–124.
  • Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology 52: 696–704. doi: 10.1080/10635150390235520
  • Katoh K, Standley DM (2013) MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability. Molecular Biology and Evolution 30: 772–780. doi: 10.1093/Molbev/Mst010
  • Maddison DR (1993) Systematics of the Holarctic beetle subgenus Bracteon and related Bembidion (Coleoptera: Carabidae). Bulletin of the Museum of Comparative Zoology 153: 143–299.
  • Maddison DR (2008) Systematics of the North American beetle subgenus Pseudoperyphus (Coleoptera: Carabidae: Bembidion) based upon morphological, chromosomal, and molecular data. Annals of Carnegie Museum 77: 147–193. doi: 10.2992/0097-4463-77.1.147
  • Maddison DR (2012) Phylogeny of Bembidion and related ground beetles (Coleoptera: Carabidae: Trechinae: Bembidiini: Bembidiina). Molecular Phylogenetics and Evolution 63: 533–576. doi: 10.1016/j.ympev.2012.01.015
  • Maddison DR, Baker MD, Ober KA (1999) A preliminary phylogenetic analysis of 18S ribosomal DNA of carabid beetles (Insecta: Coleoptera). In: Ball GE, Casale A, Vigna Taglianti A (Eds) Phylogeny and Classification of Caraboidea (Coleoptera: Adephaga). Museo Regionale di Scienze Naturali, Atti, Torino, Italy, 229–250.
  • Maddison DR, Ober KA (2011) Phylogeny of minute carabid beetles and their relatives based upon DNA sequence data (Coleoptera, Carabidae, Trechitae). ZooKeys 147: 229–260. doi: 10.3897/zookeys.147.1871
  • Maddison DR, Swanson AP (2010) A preliminary characterization of Bembidion perspicuum LeConte, with a reclassification of related species (Coleoptera, Carabidae) north of México. ZooKeys 43: 15–31. doi: 10.3897/zookeys.43.390
  • Magrini P, Cecchi B, Lo Cascio P (1998) A new species of Ocys from Italy (Coleoptera Carabidae). Redia 81: 73–81.
  • Magrini P, Degiovanni A (2009) Un nuovo Ocys Stephens, 1828 della Sardegna (Insecta Coleoptera Carabidae). Quaderno di Studi e Notizie di Storia Naturale della Romagna 29: 95–103.
  • Neri P, Bonavita P, Gudenzi I, Magrini P, Toledano L (2011) Bembidiina of Italian and Corsican Fauna: identification keys. Quaderno di Studi e Notizie di Storia Naturale della Romagna 33: 1–183.
  • Netolitzky F (1942) Bestimmungstabelle der Bembidion-Arten des paläarktischen Gebietes. Koleopterologische Rundschau 28: 29–124.
  • Raupach M, Hannig K, Moriniere J, Hendrich L (2016) A DNA barcode library for ground beetles (Insecta, Coleoptera, Carabidae) of Germany: The genus Bembidion Latreille, 1802 and allied taxa. ZooKeys 592: 121–141. doi: 10.3897/zookeys.592.8316
  • Sciaky R (1998) Trechus jeannei n. sp. della Spagna settentrionale e note su altre specie di carabidi della Penisola Iberica (Coleoptera, Carabidae). Fragmenta Entomologica, Roma 30: 243–251.
  • Stephens JF (1828) Illustrations of British entomology; or, a synopsis of indigenous insects: containing their generic and specific distinctions; with an account of their metamorphoses, times of appearance, localities, food, and economy, as far as practicable. Mandibulata. Vol. II. Baldwin & Cradock, London, 112 pp.
  • Thalmann O, Hebler J, Poinar HN, Pääbo S, Vigilant L (2004) Unreliable mtDNA data due to nuclear insertions: a cautionary tale from analysis of humans and other great apes. Molecular Ecology 13: 321–335. doi: 10.1046/j.1365-294X.2003.02070.x
  • Toledano L, Wrase DW (2016) Two unexpected findings of the genus Ocys Stephens, 1828: a new species from China and another, giant, one from Iraq (Coleoptera, Carabidae, Bembidiini). Entomologische Blätter und Coleoptera 112: 387–392.
  • Toribio M (2013) Datos sobre algunos Carábidos de la Península Ibérica (Coleoptera). Revista gaditana de Entomología 4: 1–5.
  • Vigna Taglianti A (1994) Un nuovo Ocys dell’Appennino abruzzese (Coleoptera, Carabidae). Bollettino dell’Associazione Romana di Entomologia 49: 129–136.
  • Wild AL, Maddison DR (2008) Evaluating nuclear protein-coding genes for phylogenetic utility in beetles. Molecular Phylogenetics and Evolution 48: 877–891. doi: 10.1016/j.ympev.2008.05.023
  • Wollaston TV (1857) Catalogue of the Coleopterous Insects of Madeira in the collection of the British Museum. London, 234 pp.
  • Zwickl DJ (2006) Genetic algorithm approaches for the phylogenetic analysis of large biological sequence datasets under the maximum likelihood criterion. PhD Dissertation, The University of Texas at Austin, Austin, Texas.