Biodiversity Data Journal :
Taxonomic Paper
|
Corresponding author:
Academic editor: Vladimir Blagoderov
Received: 05 Aug 2015 | Accepted: 21 Nov 2015 | Published: 26 Nov 2015
© 2015 Søren Nielsen, Michael Kristensen, Thomas Pape
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:
Nielsen S, Kristensen M, Pape T (2015) Three new Scandinavian species of Culicoides (Culicoides): C.boyi sp. nov., C.selandicus sp. nov. and C.kalix sp. nov. (Diptera: Ceratopogonidae). Biodiversity Data Journal 3: e5823. https://doi.org/10.3897/BDJ.3.e5823
|
In the context of a major monitoring program of Culicoides in Denmark and Sweden due to the appearance of bluetongue disease in 2007–2008, a large number of specimens were collected by light traps and sorted morphologically, with COI barcodes generated for selected specimens.
Three species are described as new to science based on both morphological and molecular data: Culicoides (Culicoides) boyi sp. nov. (Denmark: Jutland), C. (C.) selandicus sp. nov. (Denmark: Zealand) and C. (C.) kalix sp. nov. (Sweden: Norrbotten). All are diagnosed morphologically as well as by molecular barcoding. A key to slide-mounted females of all Scandinavian species of Culicoides (Culicoides) is presented.
Biting midges; new species; Scandinavia; morphology; molecular barcoding
Following the outbreak of bluetongue disease in 2007–2008, a major entomological monitoring program of Culicoides was implemented in Denmark and Sweden to collect a large material of Culicoides (
Biting midges were collected in 2007–2008 in Sweden (
For documentation of the identification, the head, wings and the posterior abdominal segments were removed from the female individuals and slide mounted, and the remaining parts of the animal were processed for DNA analysis as described by
The holotypes and paratypes of the three new species are deposited in the collection of the Natural History Museum of Denmark, University of Copenhagen (ZMUC).
Culicoides boyi:
Culicoides boyi:
Culicoides dk1:
Culicoides boyi:
Female: Eyes bare and contiguous dorsally (Fig.
Culicoides (Culicoides) boyi sp. nov., female. Details of adult morphology.
Wing length 1,641 ± 10 μm (Fig.
Two normal-sized spermathecae, ovoid, of almost equal size, with a short neck and a third rudimentary one (Fig.
Male: Unknown.
Named as a tribute to Boy Overgaard Nielsen, an outstanding Danish entomologist and current emeritus at Aarhus University.
Palaearctic – Denmark (northern Jutland).
The length divided by the width of the first flagellomere is significantly higher in C. boyi compared to all other Scandinavian species of this subgenus.
Culicoides dk3:
Culicoides selandicus:
Female: Eyes bare and contiguous dorsally (Fig.
Culicoides (Culicoides) selandicus sp. nov., female. Details of adult morphology.
Wing length 1,339 ± 33 μm (Fig.
Spermathecae not observed.
Male: Unknown.
The species epithet refers to the name of the major Danish island Sjælland (Latin = Selandia; English = "Zealand" or more rarely "Sealand"), where the type series was collected.
Palaearctic – Denmark (Zealand).
Culicoides selandicus may be confused with C. kalix but differs habitually by the extensive dark areas on the wings. The head/proboscis ratio of C. selandicus (1.16 ± 0.06) is smaller than in C. kalix (1.29 ± 0.07), although a small overlap should be expected when more specimens are measured. The average number of antennal sensilla coeloconica (12.29±0.95) is higher than in C. kalix (10.70 ± 0.82), and the first flagellomere has a higher average of sensilla (4.1) compared to C. kalix (3.0). The P3/P2 ratio (0.96 ± 0.06) differs from that of C. kalix (0.87 ± 0.08), but with a large overlap. Second maxillary palp segment is about as long as the third, while in C. kalix the second palp segment is longer than the third. Third maxillary palp segment is more slender (PR = 3.2 ± 0.3) than that of C. kalix (PR = 2.9 ± 0.2). Maxillary palp segments four and five of equal length provides a difference from C. kalix, where the fifth palp segment is longer than the fourth. The ratio of mandibular vs. maxillary teeth is significantly higher in C. selandicus (1.31 ± 0.12) than in C. kalix (1.17 ± 0.12).
Culicoides Kalix:
Culicoides kalix:
Female: Eyes bare and contiguous dorsally (Fig.
Culicoides (Culicoides) kalix sp. nov., female. Details of adult morphology.
Wing length 1,423 ± 39 μm (Fig.
Two functional spermathecae, ovoid, of almost equal size, with a short neck; a third rudimentary one present (Fig.
Male: Unknown.
The species epithet refers to the municipality Kalix in northern Sweden, in which the type series was collected.
Palaearctic – Sweden (Norrbotten).
Culicoides kalix could be confused with C. selandicus but may be separated from this as already discussed above under the description of the latter species and as outlined in the key.
Key to females of Scandinavian species of Culicoides (Culicoides) The problems in constructing reliable keys to adults of the European species of Culicoides (Culicoides) due to overlapping morphometric measures are well known (e.g., Note that we are following |
||
1 | Wing with a spot in cell cu | 2 |
– | Wing without a spot in cell cu | 9 |
2 | Third segment of maxillary palp about as long as or longer than the second segment (P3/P2 ≥ 0.94). Wing with the dark hour-glass mark in cell r3 broadest above the longitudinal fold above vein M1 | 3 |
– | Third segment of maxillary palp shorter than the second segment (P3/P2 ≤ 0.93). Wing with the dark hour-glass mark in cell r3 broadest above the longitudinal fold above vein M1 or broadest at or below the fold above M1 | 6 |
3 | Wing darkened around the entire length of vein M1 | 4 |
– | Wing darkened for some part of vein M1 | 5 |
4 | Wing with two dark marks in cell m1. The dark hour-glass mark in cell r3 is broad and roughly square in outline. Third segment of maxillary palp longer than second segment (P3/P2 > 1.01); PR (palp ratio, i.e., length/width of third segment) < 2.8. Fronto-vertex/ocellus ratio < 0.78 | C. newsteadi Austen & C. halophilus Kieffer (separation based on morphological characters currently not possible) |
– | Wing with only one dark mark in cell m1. The dark hour-glass mark in cell r3 is not broad and square in outline. Third segment of maxillary palp at most as long as second segment (P3/P2 ≤ 1.0); PR > 3.0. Fronto-vertex/ocellus ratio > 0.87 | C. selandicus Nielsen, Kristensen & Pape, sp. nov. |
5 | Wing with small pale spots at the tip of veins M1, M2, and Cu1. AR (antennal ratio, i.e., length of flagellomeres 9–13 divided by the length of flagellomeres 1–8) > 1.08. Ratio length/width of first flagellomere < 1.6 | C. punctatus (Meigen) |
– | Wing without spots at the tip of veins M1, M2 and Cu1. AR < 1.08. Ratio length/width of first flagellomere > 1.7 | C. boyi Nielsen, Kristensen & Pape, sp. nov. (part) |
6 | Wing with the dark hour-glass mark in cell r3 broadest above the longitudinal fold above M1 or hour-glass mark with continuous outline and of equal width above and at the longitudinal fold above vein M1 | 7 |
– | Wing with the dark hour-glass mark in cell r3 broadest at or below the longitudinal fold above vein M1 | 8 |
7 | Wing with the dark hour-glass mark in r3 broadest above the longitudinal fold above vein M1. The dark areas in wings surround the entire length of the veins M1 and M2. Mandibular teeth 12–14, maxillary teeth 12–17. Number of antennal sensilla coeloconica 9–12 | C. kalix Nielsen, Kristensen & Pape, sp. nov. |
– | Wing with the dark hour-glass mark in cell r3 with continuous outline and equal widths above the longitudinal fold and at the fold above vein M1. The dark areas in wings do not surround veins M1 and M2. Mandibular teeth 16–20, maxillary teeth 19–21. Number of antennal sensilla coeloconica 12–19 | C. pulicaris (Linnaeus) |
8 | Small species (wing length < 1400 µm). Wing markings are faint but sharply defined. Third segment of maxillary palp rhomboid. AR (antennal ratio, i.e., length of flagellomeres 9–13 divided by the length of flagellomeres 1–8) < 1.09. Head/proboscis ratio > 1.26. Number of antennal sensilla coeloconica 7–12 | C. impunctatus Goetghebuer (part) |
– | Large species (wing length > 1600 µm). Wing with extensive and vaguely defined dark markings. Third segment of maxillary palp cigar-shaped. AR > 1.09. Head/proboscis ratio < 1.24. Number of antennal sensilla coeloconica 12–20 | C. deltus Downes and Kettle (part) |
9 | Wing with distinct dark markings; the dark hour-glass mark in the middle of cell r3 broadest above the longitudinal fold above the longitudinal fold above cell M1 | C. boyi Nielsen, Kristensen & Pape, sp. nov. (part) |
– | Wing with vaguely defined markings; the dark hour-glass mark in cell r3 broadest at or below the fold above M1 | 10 |
10 | PR (palp ratio, i.e., length/width of third segment) < 3.5. Fronto-vertex/ocellus ratio > 1.2 | 11 |
– | PR > 3.5. Fronto-vertex/ocellus ratio < 1.0 | C. grisescens Edwards |
11 | Small species (wing length < 1400 µm). Third segment of maxillary palp rhomboid. AR (antennal ratio, i.e., length of flagellomeres 9–13 divided by the length of flagellomeres 1–8) < 1.09. Head/proboscis ratio > 1.26. Number of antennal sensilla coeloconica 7–12 | C. impunctatus Goetghebuer (part) |
– | Large species (wing length > 1600 µm). Third segment of maxillary palp cigar-shaped. AR > 1.09. Head/proboscis ratio < 1.24. Number of antennal sensilla coeloconica 12–20 | C. deltus Edwards (part) |
Relevant comparisons for separating the three new species from their morphologically most similar Scandinavian congeners are given in Table
Pairwise morphometric comparisons between three new species of Culicoides (Culicoides) and the morphologically most similar Scandinavian species, including divergence in COI sequences. The significance of differences between measurements was determined by multiple comparison test after Kruskal-Wallis (PK-w< 0.05) followed by a Conover-Inman test for all pairwise comparisons (
1 - Species; 2 - Flagellum, length (μm); 3 - Antennal ratio (AR: length of flagellomeres 9–13 divided by length of flagellomeres 1–8); 4 - First flagellomere, length/width; 5 - Maxillary palp, length (μm); 6 - Maxillary palp ratio PR (length/width of third palp segment); 7 - Maxillary palp ratio P3/P2 (length of third maxillary palp segment divided by length of second); 8 - Length of wing (μm); 9 - Spermatheca ratio S/R; 10 - Head/proboscis ratio; 11 - Mandibular teeth; 12 - Maxillary teeth; 13 - Ratio M/M; 14 - Fronto-vertex /ocellus; 15 - Antennal sensilla coeloconica.
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
15 |
C. pulicaris |
742± 26 |
1.09± 0,03 |
1.52± 0.07 |
255.6± 14.3 |
2.9± 0.2 |
0.84± 0.05 |
1626± 67 |
121± 0.08 |
1.19± 0.04 |
16.7± 1.2 |
19.5± 1.1 |
1.17± 0.08 |
1.2± 0.3 |
15.47± 1.97 |
C. boyi |
746± 44 |
1.03± 0.04 |
1.78± 0.07 |
245.1± 17.7 |
2.9± 0.3 |
1.04± 0.11 |
1641± 10 |
1.05± 0.3 |
1.29± 0.07 |
15.1± 1.2 |
17.1± 1.4 |
1.13± 0.10 |
1.5± 0.2 |
14.71± 1.26 |
Divergence of COI sequences = 16.5% |
||||||||||||||
C. newsteadi |
591± 45 |
1.04± 0.05 |
1.53± 0.15 |
193.6± 18.7 |
2.6± 0.2 |
1.11± 0.10 |
1291± 12 |
1.04± 0.02 |
1.32± 0.13 |
13.4± 1.4 |
15.9± 1.5 |
1.18± 0.20 |
0.3± 0.3 |
7.70± 0.67 |
C. selandicus |
616± 10 |
1.12± 0.04 |
1.56± 0.11 |
216.8± 8.6 |
3.2± 0.3 |
0.96± 0.06 |
1339± 33 |
ND |
1.16± 0.06 |
15.0± 1.0 |
19.6± 1.5 |
1.31± 0.12 |
1.5± 0.8 |
12.29± 0.95 |
Divergence of COI sequences = 16.2% |
||||||||||||||
C. newsteadi |
591± 45 |
1.04± 0.05 |
1.53± 0.15 |
193.6± 18.7 |
2.6± 0.2 |
1.11± 0.10 |
1291± 12 |
1.04± 0.02 |
1.32± 0.13 |
13.4± 1.4 |
15.9± 1.5 |
1.18± 0.20 |
0.3± 0.3 |
7.70± 0.67 |
C. kalix |
646± 14 |
1.13± 0.04 |
1.46± 0.06 |
212.4± 4.0 |
2.9± 0.2 |
0.87± 0.08 |
1423± 39 |
1.17± 0.08 |
1.29± 0.07 |
12.8± 0.6 |
14.9± 1.5 |
1.17± 0.12 |
1.2± 0.3 |
10.70± 0.82 |
Divergence of COI sequences = 15.6% |
||||||||||||||
C. selandicus |
616± 10 |
1.12± 0.04 |
1.56± 0.11 |
216.8± 8.6 |
3.2± 0.3 |
0.96± 0.06 |
1339± 33 |
ND |
1.16± 0.06 |
15.0± 1.0 |
19.6± 1.5 |
1.31± 0.12 |
1.5± 0.8 |
12.29± 0.95 |
C. kalix |
646± 14 |
1.13± 0.04 |
1.46± 0.06 |
212.4± 4.0 |
2.9± 0.2 |
0.87± 0.08 |
1423± 39 |
1.17± 0.08 |
1.29± 0.07 |
12.8± 0.6 |
14.9± 1.5 |
1.17± 0.12 |
1.2± 0.3 |
10.70± 0.82 |
Divergence of COI sequences = 5.9% |
The recent arrival of bluetongue virus in northern and western Europe (
The European fauna of Culicoides (Culicoides) is in need of a thorough taxonomic revision based on both morphological and molecular data, but this will be a massive undertaking that reaches far beyond the scope of the present paper. The Scandinavian fauna of Culicoides (Culicoides) has been sampled extensively as a result of the recent bluetongue epidemic, and the current study is based on thousands of specimens examined [by SAN] from many localities widely distributed through Denmark and Sweden. The three species here described as new do not match any of the European species as keyed by
Revisionary taxonomy of European Ceratopogonidae suffers from the well-known constraints of old names and insufficient or missing type material. A number of nominal species currently listed in synonymy under C. newsteadi and C. pulicaris (see
The molecular data from the three species named in the present paper were analysed by
Cladogram based on COI-data showing clusters of species of Culicoides (s.str.) from western Europe. GenBank numbers with a concluding country code to show geographic origin. Numbers on branches are bootstrap values, and branch lengths are equivalent to computed evolutionary distance (scale at bottom). Redrawn from
It is noteworthy that the three new species were collected at single locations or from a few locations in close proximity in spite of a very large sampling. These species are most likely more widely distributed, as are the majority of the well-known biting midge species, and what may look like a restricted geographical occurrence may be due to either a patchy distribution or a very short adult flying period.
We are very grateful for constructive comments from Dr. Art Borkent, Research Associate of the Royal British Columbia Museum, American Museum of Natural History, and Instituto Nacional de Biodiversidad as well as from Dr. Bruno Mathieu, Institut de Parasitologie et de Pathologie Tropicale, Faculté de Médecine de Strasbourg. The project was supported by the Danish Ministry of Food, Agriculture and Fisheries by grant no. 3304FVFP077 to MK.
SAN and MK conceived the study. SAN identified Culicoides specimens. SAN and MK analysed the data. SAN, MK and TP interpreted the results. SAN and TP drafted the manuscript. All authors read and approved the final manuscript.