Revision of the Culicoides (Avaritia) Imicola complex Khamala & Kettle (Diptera: Ceratopogonidae) from the Australasian region Author Bellis, Glenn Author Dyce, Alan Author Gopurenko, David Author Yanase, Tohru Author Garros, Claire Author Labuschagne, Karien Author Mitchell, Andrew text Zootaxa 2014 2014-02-27 3768 4 401 427 journal article 46339 10.11646/zootaxa.3768.4.1 50d9d914-e2ab-4187-89bb-5d47a7bcc9df 1175-5326 218566 2698BC2A-7B32-44A5-8856-2EB8846DEBD5 Culicoides nudipalpis Delfinado ( Figs 3, 8, 9 , 14, 17, 20 , 23, 27, 30, 33 , 40, 41, 42, 45) Culicoides nudipalpis Delfinado 1961 :655 ; Wirth & Hubert 1989 :281 (description of male and female); Meiswinkel & Baylis, 1998 (separation from C. imicola ); Lien et al., 1998 :35 ( Taiwan record, description of male and female); Yu et al. 2005 : 1018 ( China record, description of male and female); Dyce et al. , 2007 :19 (female wing illustrated). Type material examined. Philippines : Tala, Rizal, 21.May.1958 , M. Delfinado ( paratype , female, USNM ) Non-type material examined. Timor Leste : Hera, 30.Mar.2004 , Lt Tp., E. Watkins, ( 2 females , NTQIC ); Cape Tefara, lt. tp. 5.Aug.1969 , DG. Nicholls ( 5 females , 1 male , ANIC ); Suai, Cafe Cooperativa Timor, 9°23.046'S ; 125°10.675'E , Lt Tp, 13–14.May.2010 , B. Cookson ( 1 female , 2 males NTQIC ); Ainaro, Surucraic, 15.Nov.2001 , E. Watkins ( 1 female , NTQIC ), Bobonaro, Maliana, Rai Fun, 7-May-2002 , E. Watkins ( 1 female , NTQIC ); Oecusse, Samora, 24-Feb-2005 , G. Bellis ( 1 female , NTQIC ); Cova Lima, Suai, 9.31°S , 125.26°E , 5.May.2002 , G. Bellis ( 1 female , NTQIC ); Oecusse, Pante Makassar, 7-Dec-2002 , G. Bellis ( 1 female , NTQIC ); Ermera, Letefoho Eraulo, 22.Jul.2004 , E. Watkins ( 1 female , NTQIC ); Viqueque, 19.Jul.2000 , A. Moss ( 1 female , NTQIC ); Dili, Cristo Rei, Hera, 19.Jul.2000 , A. Moss ( 1 female , NTQIC ). Indonesia , Irian Jaya, Merauke, Kumbe, VLi378, 5.Aug.1980 ( 1 male , ANIC ). Diagnosis. Female: The only species in the Imicola complex with the combination of wing with proximal dark marking on costa of similar length to stigmatic dark spot, apical half of cell r2 included in post-stigmatic pale spot, posterior margin of apical pale marking in cell m1 curving posteriorly to reach vein M2 well before the wing margin, often crossing vein M2 and joining the pale spot in cell m2 and with PH ratio less than 0.75. Male : there are currently no reliable characters for separating males of this species from those of C. imicola . These two species are the only species in the Imicola complex with the posterior margin of apical pale marking in cell m1 curving posteriorly to reach vein M2 well before the wing margin, often crossing vein M2 and joining the pale spot in cell m2 Description. Adult . In addition to characters listed in the diagnosis, palpus ( Fig. 14 ) pale brown with 3rd segment slightly swollen, broadest about mid way along, sensory area a superficial rounded patch of aggregated sensoria in a shallow depression spanning nearly the full width of segment; legs ( Fig. 20 ) pale brown, fore & mid femora with pale apical band, all tibiae with pale basal band and weak, broad apical pale band; haltere pale. Male hypopygium (Fig. 40) with ventral membrane of ninth sternite bearing a sparse covering of spicules. Aedeagus (Fig. 42) with peg broad at base and noticeably paler than distal process. Immatures. Unknown. Distribution. ( Fig. 45 ) Philippines ( Delfinado 1961 ; Wirth & Hubert 1989 ), Indonesia : Bali, West Timor, Papua , Flores , Sumbawa, Lombok and Maluku ( Wirth & Hubert 1989 ; Sukarsih et al. 1993; Dyce et al. 2007 ), Timor Leste ( Wirth & Hubert 1989 ), Taiwan ( Lien et al. 1998 ) and China , Hainan ( Yu et al. 2005 ). Biology. Largely unknown. Bellis et al. (2005) investigated some of the aspects of biology pertinent to vector capacity and found this species to feed on mammals and have a relatively high survival rate. Sukarish et al. (1993) reported collecting this species in association with cattle. Remarks. Dyce (1979) reported some confusion in the type series of this species. He could not locate the holotype female and the specimen illustrated as C. nudipalpis by Delfinado (1961) was apparently labeled as an allotype of C. radicitus . Wirth & Hubert (1989) designated a neotype which is now lodged in the USNM. Meiswinkel & Baylis (1998) correctly noted that C. nudipalpis is morphologically most similar to C. imicola and reported on characters used to distinguish these two species. They were unable to observe differences between males of these two species and although differences are apparent in the ratio of the length between the interocular seta and the base of the cibarium to the length between the base of the cibarium and the apex of the labium (Bellis, Garros & Labuschagne unpublished data), these differences require further study to confirm their validity. Results from the present study confirm the observations of Meiswinkel & Baylis (1998) that the PH ratio of female C. nudipalpis is less than that reported for C. imicola and this remains the most reliable means of separating these species. Genetic analyses reported herein support the validity of C. nudipalpis as a taxon and analyses of CAD and concatenated CAD +COI data place it as sister to C. imicola as morphological similarity might suggest although COI analysis did not agree with this close relationship. Wirth & Hubert (1989) reported that these two species do not occur sympatrically although Yu et al. (2005) have subsequently reported both species occurring on Hainan in China and Lien et al. (1998) reported C. nudipalpis in nearby Taiwan so these species are not as geographically separate as previously thought and workers in this region are encouraged to check material to ensure accurate identification. TABLE 6. Ratio of length:width of antennal flagellar segments of females of Culicoides bolitinos , C. brevitarsis and C. asiatica . Antennal flagellomere Antennal flagellomere Gene: COI

1	2	3	4	5	6	7
C. brevitarsis mean	1.27	1.03	1.08	1.28	1.39	1.38	1.41
range	1.09–1.47	0.88–1.14	0.86–1.23	1.08–1.50	1.08–1.58	1.17–1.54	1.17–1.67
n	9	9	9	9	9	9	9
C. asiatica mean	1.21	0.91	1.08	1.20	1.37	1.35	1.35
range	1.09–1.36	0.78–1.00	1–1.43	1–1.40	1.21–1.64	1.29–1.5	1.23–1.64
n	7	8	8	8	8	8	8
C. bolitinos mean	1.36	1.12	1.28	1.43	1.6	1.52	1.67
(ex Meiswinkel 1995) range	1.25–1.50	1.03–1.25	1.17–1.48	1.29–1.56	1.38–1.77	1.33–1.70	1.48–1.83
n	25	25	25	25	25	25	25

TABLE 6. (Continued).

8	9	10	11	12	13
C. brevitarsis mean	1.53	2.51	2.53	2.55	2.54	3.99
range	1.07–1.83	1.69–3.00	2.08–2.83	2.15–2.85	2.08–3.25	3.57–4.46
n	9	9	9	8	9	8
C. asiatica mean	1.49	2.40	2.47	2.61	2.36	3.97
range	1.36–1.79	2.14–2.92	2.31–2.64	2.29–2.92	2–2.71	3.57–4.46
n	8	8	8	7	7	7
C. bolitinos mean	1.81	2.63	2.6	2.54	2.5	4.04
(ex Meiswinkel 1995) range	1.62–1.92	2.31–2.92	2.44–2.79	2.22–2.85	2.07–2.88	3.69–4.62
n	25	25	25	25	25	25

FIGURE 46. Maximum likelihood tree inferred from concatenated CAD & COI sequences at Culicoides species in the Imicola complex and other representative Avaritia. Terminal labels indicate specimen ID and species (Table 7). Tree rooted at C. orientalis . Scale bar = 0.05 substitutions per site. Node support values generated from 1200 replicate ML searches (values <60% not shown). Bayesian phylogenetic analysis pp supports estimated from 15000 sampled trees are indicated in the tree as circles at nodes, where closed circles represent> 98% pp and open circles represent 70–98 % pp. FIGURE 47. Maximum likelihood tree inferred from COI sequences at Culicoides species in the Imicola complex and other representative Avaritia. Terminal labels indicate specimen ID and species (Table 7). Tree rooted at C. orientalis . Scale bar = 0.05 substitutions per site. Node support values generated from 1200 replicate ML searches (values <60% not shown). Bayesian phylogenetic analysis pp supports estimated from 15000 sampled trees are indicated in the tree as circles at nodes, where closed circles represent> 98% pp and open circles represent 70 – 98 % pp. Specimen AY286329 Culicoides pseudopallidipennis accessed from GenBank. FIGURE 48. Maximum likelihood tree inferred from concatenated CAD sequences at Culicoides species in the Imicola complex and other representative Avaritia. Terminal labels indicate specimen ID and species (Table 7). Tree rooted at C. orientalis . Scale bar = 0.05 substitutions per site. Node support values generated from 1200 replicate ML searches (values <60% not shown). Bayesian phylogenetic analysis pp supports estimated from 15000 sampled trees are indicated in the tree as circles at nodes, where closed circles represent> 98% pp and open circles represent 70–98 % pp. TABLE 8. Sequence statistics for each gene (COI, CAD) and at each codon position. codon 1st 2nd 3rd all Notes: # informative = number of parsimony informative sites; CI = consistency index; CI-ex = consistency index excluding invariant sites; RI = retention index; RC = rescaled consistency index (CI * RI).