Research Article |
Corresponding author: William E. Moser ( moserw@si.edu ) Academic editor: Fredric Govedich
© 2022 William E. Moser, Dennis J. Richardson, Charlotte I. Hammond, Lourdes Rojas, Eric Lazo-Wasem, Anna J. Phillips.
This is an open access article distributed under the terms of the CC0 Public Domain Dedication.
Citation:
Moser WE, Richardson DJ, Hammond CI, Rojas L, Lazo-Wasem E, Phillips AJ (2022) Resurrection and redescription of Clepsine pallida Verrill, 1872 (Hirudinida, Glossiphoniidae) with a phylogeny of the genus Alboglossiphonia. ZooKeys 1127: 135-154. https://doi.org/10.3897/zookeys.1127.86004
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Alboglossiphonia pallida (Verrill, 1872) comb. nov. is resurrected and redescribed based on morphological and molecular data from specimens of the type locality (New Haven County, Connecticut, USA) that demonstrate it is distinct from North American Alboglossiphonia heteroclita, European Alboglossiphonia heteroclita, and Alboglossiphonia papillosa. Alboglossiphonia pallida is characterized by having dark chromatophores on the dorsal surface arranged lateral to patrilaterally and medially as a thin line or interrupted thin line along with three pairs of eye spots (with the first pair closest together), six pairs of crop ceca, and a united gonopore. Additional sampling of specimens of the genus Alboglossiphonia is needed to understand its phylogeny especially as many species have not been collected since their description.
Alboglossiphonia heteroclita, Clitellata, Glossiphonia, Glossiphonia swampina, Glossiphoniiformes, leech, Rhynchobdellida
The species concept of Alboglossiphonia heteroclita (Linnaeus, 1761) has become very heterogenous over time through a combination of formal synonymy and informal accumulation of diagnostic morphological characters. Other species of the genus Alboglossiphonia exhibit similar taxonomic confusion, including Alboglossiphonia hyalina (O.F. Müller, 1774), Alboglossiphonia inflexa (Goddard, 1908), Alboglossiphonia novaecaledoniae (Johansson, 1918), Alboglossiphonia papillosa (Braun, 1805), and Alboglossiphonia striata (Apáthy, 1888) (
Hirudo heteroclita was originally described from Europe by
In North America,
In describing Glossiphonia concolor from Europe,
Hirudo swampina was described by
Ignoring the similarities of H. swampina to the genus Placobdella,
In recent phylogenetic studies,
The convoluted history of this assemblage is given in Fig.
During the course of a survey of the leech fauna of south-central Connecticut, individuals matching the description of Clepsine pallida Verrill, 1872 were collected by hand from submerged substrate in the West River, New Haven, New Haven County, the type locality of C. pallida. Specifically, collections were made from the West River at Konolds Pond (41°20'52.1"N, 72°58'41.6"W) and Whalley Avenue Bridge (41°19'30.13"N, 72°57'26.76"W) south to the “Duck Pond” (41°18'51.30"N, 72°57'21.75"W) as illustrated on page 12 of
Molecular analyses were conducted on newly collected material according to
Polymerase chain reactions (PCR) were prepared using the Illustra PuRe Taq Ready-To-Go PCR beads from GE Health Care (cat. no. 27-9559-01). Primers were purchased from Invitrogen and were comprised of two primers each for cytochrome c oxidase subunit I (COI) as specified by
Purified PCR products were sequenced using the HCO2198 and LCO1490 primers for the COI products by the W.M. Keck Foundation Biotechnology Resource Laboratory at Yale University. DNA sequences were edited and assembled using Geneious Prime (v. 2020.1.2, Biomatters Ltd.). Novel sequences were deposited in GenBank (
Species, collection locality, museum catalog number, and Genbank accession information or BOLD for sequences included in this study.
Species | State/province | Country | Location | Catalog number /citation | GenBank or BOLD# |
---|---|---|---|---|---|
Alboglossiphonia iberica | Huelva | Spain | 8789, |
N/A | |
Alboglossiphonia quadrata | Namibia |
|
AY962455 | ||
Alboglossiphonia heteroclita | Germany | 9195, |
N/A | ||
Alboglossiphonia heteroclita | Michigan | USA |
|
AF116016 | |
Alboglossiphonia heteroclita | Michigan | USA | BSC-160.1, SUNY Buffalo State | ANNMO802-20 | |
Alboglossiphonia heteroclita | Michigan | USA | BSC-160.2, SUNY Buffalo State | ANNMO803-20 | |
Alboglossiphonia heteroclita | Michigan | USA | BSC-160.6, SUNY Buffalo State | ANNMO807-20 | |
Alboglossiphonia heteroclita | Wisconsin | USA | BSC-160.3, SUNY Buffalo State | ANNMO804-20 | |
Alboglossiphonia heteroclita | Wisconsin | USA | BSC-160.4, SUNY Buffalo State | ANNMO805-20 | |
Alboglossiphonia pallida | Connecticut | USA | Konolds Pond, West River |
|
ON738431 |
Alboglossiphonia pallida | Connecticut | USA | Konolds Pond, West River |
|
ON738432 |
Alboglossiphonia pallida | Connecticut | USA | Konolds Pond, West River |
|
ON738433 |
Alboglossiphonia pallida | Connecticut | USA | Konolds Pond, West River |
|
ON738434 |
Alboglossiphonia pallida | Connecticut | USA | Clarks Pond |
|
ON738435 |
Alboglossiphonia papillosa | Siberia | Russia | Lake Gusinoe |
|
KM095100 |
Alboglossiphonia papillosa | Siberia | Russia | Lake Gusinoe |
|
KM095101 |
Alboglossiphonia papillosa | Siberia | Russia | Lena River basin | RMBH Hir13/3, |
MH286269 |
Alboglossiphonia papillosa | Siberia | Russia | Lena River basin | RMBH Hir13/4, |
MH286270 |
Alboglossiphonia papillosa | Siberia | Russia | Lena River basin | RMBH Hir13/5, |
MH286271 |
Alboglossiphonia papillosa | Siberia | Russia | Lena River basin | RMBH Hir13/2, |
MH286268 |
Alboglossiphonia papillosa | Siberia | Russia | Lena River basin | RMBH Hir13/1, |
MH286267 |
Alboglossiphonia sp. 2 | Myanmar | RMBH HIR58/2 |
MN295404 | ||
Alboglossiphonia sp. | Victoria | Australia | Melbourne | MRD16Gloss2, |
MG976199 |
Alboglossiphonia lata | Primorsky Krai | Russia | RMBH HIR58/1, |
MN295414 | |
Alboglossiphonia lata | South Korea | RMBH HIR113/4, |
MN393286 | ||
Alboglossiphonia lata | South Korea | RMBH HIR103/5, |
MN393275 | ||
Alboglossiphonia lata | South Korea | RMBH HIR110/5, |
MN393279 | ||
Alboglossiphonia lata | South Korea | RMBH HIR113/3, |
MN393284 | ||
Alboglossiphonia lata | South Korea | RMBH HIR112/1, |
MN393281 | ||
Alboglossiphonia lata | South Korea | RMBH HIR114/12, |
MN393288 | ||
Alboglossiphonia lata | South Korea | RMBH HIR111/22, |
MN393280 | ||
Alboglossiphonia lata | South Korea | RMBH HIR114/1, |
MN393287 | ||
Alboglossiphonia lata | South Korea | RMBH HIR109/1, |
MN393276 | ||
Alboglossiphonia lata | South Korea | RMBH HIR110/32, |
MN393277 | ||
Alboglossiphonia lata | South Korea | RMBH HIR113/32, |
MN393285 | ||
Alboglossiphonia weberi | Hawaii | USA |
|
AY962453 | |
Alboglossiphonia sp. | South Korea | HJK-2020, |
MN503262 | ||
Glossiphonia complanata | United Kingdom |
|
AY047321 | ||
Glossiphonia complanata | Mecklenburg-Vorpommern | Germany |
|
HM246608 | |
Glossiphonia elegans | Connecticut | USA | West River |
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JQ073858 |
Glossiphonia elegans | Connecticut | USA | West River |
|
JQ073860 |
The best partitioning scheme was tested using ModelFinder within IQ-TREE (
Examination of the type series of Clepsine pallida (
In further examination of the
The second lot (now
The following redescription of the new combination Alboglossiphonia pallida (Verrill, 1872) is based upon the holotype of Clepsine pallida (
Dark chromatophores on the dorsal surface arranged lateral to patrilaterally and medially as a thin line or interrupted thin line along with three pair of eye spots (where the first pair are closest together), six pair of crop ceca, and a united gonopore.
Body narrowly ovoid to narrowly pyriform. Rounded anterior region. Dorsum buff to translucent, smooth (without papillae), and with small, black chromatophores that form thin lines with scattered areas; thin, interrupted mid-dorsal line with larger chromatophore patches (typically on sensory annuli); black chromatophores in a lateral pattern on the sensory annulus of the lateral to paralateral region (Figs
Cylindrical, blunt-tipped protrusible proboscis (approximate length of 14 annuli), opening at the center of the oral sucker. Short esophagus and diffuse salivary glands that are distributed in the anterior third of the body (Fig.
Male atrium opening into paired narrowly ovoid atrial cornua that extends laterally and narrows abruptly at junction with ejaculatory ducts and extends posteriad (Fig.
Uncorrected p-distances between COI sequences of each species are given in Table
COI uncorrected pairwise sequence differences among specimens of Alboglossiphonia included in this study. Values presented are range followed by average in parentheses ().
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | |
---|---|---|---|---|---|---|---|---|---|---|
A. iberica (1) | — | — | — | — | — | — | — | — | — | — |
A. quadrata (2) | 17.05 | — | — | — | — | — | — | — | — | — |
A. heteroclita Germany (3) | 16.47 | 16.13 | — | — | — | — | — | — | — | — |
Alboglossiphonia sp. S. Korea HJK (4) | 15.28 | 14.92 | 11.71 | — | — | — | — | — | — | — |
A. lata/A. weberi (5) | 15.28–17.05 (16.26) | 15.21–16.96 (16.03) | 13.27–14.38 (13.68) | 13.18–14.44 (13.43) | 0–3.96 (2.23) | — | — | — | — | — |
Alboglossiphonia sp. 2 Myanmar (6) | 14.74 | 16.48 | 14.22 | 13.65 | 6.18–7.45 (6.84) | — | — | — | — | — |
Alboglossiphonia sp. Australia (7) | 16.16 | 14.9 | 14.54 | 13.65 | 7.45–9.19 (8.19) | 7.92 | — | — | — | — |
A. papillosa (8) | 15.10–15.95 (15.22) | 16.64–17.56 (16.77) | 12.16–12.87 (12.26) | 13.97–14.83 (13.97) | 9.83–12.05 (10.81) | 10.3–11.12 (10.3) | 11.89–12.40 (11.89) | 0–0.86 (0.25) | — | — |
A. heteroclita USA (9) | 16.70–17.70 (17.08) | 16.32–17.71 (16.92) | 11.69–12.91 (12.16) | 13.38–14.75 (13.40) | 10.14–13.94 (11.61) | 10.94–13.11 (11.81) | 12.38–14.14 (13.18) | 8.40–9.77 (9.04) | 0.33–3.07 (1.89) | — |
A. pallida (10) | 17.36–17.58 (17.43) | 16.84–17.10 (16.95) | 12.72–12.94 (12.80) | 15.16–15.54 (15.31) | 10.86–13.29 (11.80) | 11.17–11.55 (11.35) | 12.72–13.14 (12.87) | 9.07–9.70 (9.24) | 5.78–8.35 (6.74) | 0.24–1.05 (0.73) |
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
The molecular dataset included COI sequences of 41 specimens (37 members of Alboglossiphonia and two specimens each of Glossiphonia complanata and Glossiphonia elegans that served as outgroups; Table
Maximum likelihood phylogeny (lnL = −2868.139) of Alboglossiphonia pallida and congeners based on mitochondrial COI sequence data partitioned by codon. Maximum likelihood bootstrap values above 70 are shown at the internodes. GenBank accession number and locality following each species name is provided for each terminal. Branches are drawn proportional to the amount of change.
The genus Alboglossiphonia is well supported as monophyletic (bs = 100). Alboglossiphonia pallida and A. papillosa were represented by more than one individual in our analysis and each of these species was monophyletic with strong support (A. pallida bs = 100, A. papillosa bs = 98). The clade of A. pallida specimens (bs = 100) was adjacent to two sequences of Alboglossiphonia sp. from Wisconsin (bs = 54). Individuals of A. heteroclita were not each other’s closest relatives. Alboglossiphonia heteroclita (GenBank: AF116016) from Michigan placed with three sequences of A. heteroclita from Michigan (BOLD:ANNMO802, BOLD:ANNMO803) and two sequences of A. heteroclita from Wisconsin (BOLD:ANNMO804, BOLD:ANNMO805) in a series of branches with short internodes and moderate support values (bs = 74–81). Alboglossiphonia heteroclita (9195) from Germany placed sister to a sequence of Alboglossiphonia from South Korea (GenBank: MN503262), albeit with low support (bs = 63), suggesting these are separate species, the latter not otherwise represented in GenBank. The clade of A. heteroclita from Germany + Alboglossiphonia sp. from South Korea (MN503262) placed adjacent to clades including A. heteroclita from Michigan and Wisconsin, A. pallida, A. papillosa, A. lata, A. weberi, Alboglossophonia sp. from Myanmar, and Alboglossiphonia sp. from Australia (bs = 98). Alboglossiphonia pallida + A. heteroclita from Michigan and Wisconsin was sister to A. papillosa (bs = 75). Alboglossiphonia lata specimens from South Korea and the specimen from Russia (MN295414) placed within a strongly supported clade (bs = 100) that included the specimen of A. weberi (GenBank: AY962453) from Hawaii, USA. Sequences of two unidentified specimens of Alboglossiphonia (MN295404 from Myanmar and MG976199 from Australia) placed sister to one another with strong support (bs = 100), within the A. lata/weberi clade, and sister to the A. weberi specimen from Hawaii (bs = 75). The sequences of A. iberica (8739) from Spain and A. quadrata (GenBank: AY962455) from Nambia were sister to one another (bs = 100), and that clade was well supported as sister to all other specimens of Alboglossiphonia in the tree (bs = 100).
Alboglossiphonia pallida was strongly supported by morphological and molecular evidence as a species within the genus Alboglossiphonia and distinct from North American A. heteroclita, European A. heteroclita, A. lata, A. weberi, A. iberica, and A. papillosa. Alboglossiphonia pallida is characterized by having dark chromatophores on the dorsal surface arranged lateral to patrilaterally and medially as a thin line or interrupted thin line along with three pair of eye spots (where the first pair are closest together, the defining characteristic of the genus Alboglossiphonia), six pair of crop ceca, and a united gonopore. The non-monophyly of A. heteroclita continues to pose a challenge. The A. heteroclita specimens from North America were 11.69–12.91% different from A. heteroclita from Europe, indicating that the North American specimens are not A. heteroclita and most likely represent an undescribed species. The A. heteroclita specimens from North America were 0.33–3.07% different from one another. These specimens form a strongly supported clade with A. pallida (bs = 100), although the North American A. heteroclita specimens as a group were 5.78–8.35% different from the A. pallida specimens. North American specimens assigned to A. heteroclita are typically characterized by a lack of pigmentation on translucent bodies (
In Europe, A. heteroclita had been a heterogenous concept and known more by the infraspecific varieties. These synonymies have since been elevated to the species rank with A. hyalina (O.F. Müller, 1774) having yellow chromatophores and no dark chromatophores and A. striata (Apáthy, 1888) with dark, transverse pigmentation (
Sequences of specimens from Asia, Australia, and Hawaii form a strongly supported clade, except for a single sequence from South Korea (MN503262). Sequences of A. lata form a clade with short internodes that were poorly supported for the most part. The clade predominantly consisted of sequences from South Korea, yet also included a single sequence from Primorsky Krai, Russia (MN295414) and the sequence of A. weberi from Hawaii, USA (AY962453). Alboglossiphonia lata is a widely distributed species that is considered invasive and spread via the aquatic plant trade. In particular, the specimen of A. weberi from Hawaii should be reexamined to determine if this might be an occurrence record of the invasive A. lata, which would be concerning for the Hawaiian island ecosystem. The sequences of Alboglossiphonia from Myanmar and Australia are supported as members of the genus and likely represent species distinct from one another and not otherwise represented in this analysis or publicly available databases (e.g., GenBank, BOLD), yet the specimens need to be examined to determine the species identification as there have been seven described species from Australasia and Oceania.
The sequence of Alboglossiphonia quadrata (AY962455) from Namibia has likely been assigned the incorrect name.
In this study, COI was largely successful at distinguishing congeners of Alboglossiphonia, but it had limited utility in resolving the relationships between species. Combining COI data with other loci, especially nuclear loci, is needed to determine relationships between glossiphonid species with confidence. The addition of sequences of more Alboglossiphonia species will improve our understanding of relationships within the genus. This study included all publicly available Alboglossiphonia sequences, although this represents only about one-third of the diversity of the genus.
Alboglossiphonia pallida (Verrill, 1872) is resurrected and redescribed based on morphological and molecular data that demonstrate it is distinct from the specimen assigned to A. heteroclita from Michigan and Wisconsin and A. heteroclita from Europe, as well as other species of Alboglossiphonia. Additional sampling of Alboglossiphonia is needed to understand its phylogeny especially as many species have not been collected since their original description.
Kristen E. Richardson and Krystal A. Lazos, Quinnipiac University assisted in field collection. Katie Ahlfeld, Department of Invertebrate Zoology, National Museum of Natural History, Smithsonian Institution, provided valuable assistance in specimen management. Harry Coyle, New Haven Parks and Recreation provided valuable logistic insight. We are grateful to Dr Christian Albrecht, Justus-Liebig University, Giessen, Germany, and to Uwe Jueg, Ludwigslust, Germany, for providing the sequence data of Alboglossiphonia heteroclita from Germany and A. iberica. We thank Dr Milan Bull, Connecticut Audubon Society, for permission to collect at Sturges Pond, Larsen Sanctuary, Fairfield, Connecticut. We greatly appreciate Uwe Jueg and Clemens Grosser, Elstertrebnitz, Germany, for their insightful comments on the genus Alboglossiphonia. The State of Connecticut, Department of Environmental Protection provided permission to collect leeches. Funding for this project was provided in part by the Department of Biological Sciences and Schools of Arts and Sciences and Health Sciences at Quinnipiac University, Hamden, Connecticut.