taxonID	type	description	language	source
03DC776A9122FFA4FCF52E9094C7C057.taxon	description	(FIGS 2, 3; TABLE 1) Diagnosis: Body size in vivo 1100 – 1400 × 25 – 40 μm; 14 – 25 and 26 – 40 somatic kineties on head and trunk, respectively; single nuclear group composed of three or four macronuclei and two micronuclei; glabrous stripe narrow, corresponding to area occupied by two somatic kineties; cortical granules colourless and about 0.5 μm in diameter. Type locality: A mangrove wetland on Techeng island, Zhanjiang, China (21 ° 09 ′ 40 ″ N, 110 ° 25 ′ 38 ″ E), where the water temperature was 26 ° C and salinity about 25 ‰ (Fig. 1 C). Type specimens: A protargol-impregnated slide containing the holotype specimen marked with an ink circle is deposited in the Laboratory of Protozoology, Ocean University of China, China (No. YY 2012112308). One paratype slide is deposited in the Natural History Museum, London, UK, with registration number NHMUK 2015.9. 15.1. Etymology: The species-group name quadrinucleatus reflects the fact that this organism usually possesses four macronuclei. Description: Fully extended cells about 1300 × 35 μm in vivo; body flexible and contractile (Figs 2 A – C, 3 A – D). Cell distinctly tripartite, with neck, tail and trunk regions (Figs 2 A – C, 3 A – C). Head conspicuously claviform; tail wedge-shaped (Figs 2 A, D, 3 A, F). Body colour dark brown at low magnification due to multiple refractile inclusions, with several food vacuoles containing ingested algae (Figs 2 D, G, 3 E, F). Single nuclear group located in centre of trunk, containing three or four macronuclei, 7 – 10 μm in diameter, and two micronuclei, 2 – 4 μm in diameter (Figs 2 G, 3 E, H, I). Colourless cortical granules, c. 0.5 μm in diameter, scattered between ciliary rows, which are not found in glabrous stripe (Figs 2 F, 3 G). Locomotion by gliding between sand grains and organic debris. Cell surface densely ciliated with unciliated zone, glabrous stripe, about as wide as two somatic kineties (Figs 2 I, J, 3 J – M). Entire infraciliature consisting of dikinetids. About 18 and 35 somatic kineties on head and trunk, respectively, with cilia c. 13 μm long. Anterior and posterior secant system formed on left side of glabrous stripe where some kineties abut to bristle kinety (Figs 2 H, J, 3 K, M). Oral infraciliature consisting of uninterrupted circumoral kinety with cilia about 5 μm long and roundish patch of disordered dikinetids in centre of oral cavity with cilia forming conspicuous tuft (Figs 2 E, H, 3 K, L). Comparison: Since the genus Trachelolophos was established by Foissner & Dragesco (1996 b), only the following two species have been reported.	en	Yan, Ying, Xu, Yuan, Al-Farraj, Saleh A., Al-Rasheid, Khaled A. S., Song, Weibo (2016): Morphology and phylogeny of three trachelocercids (Protozoa, Ciliophora, Karyorelictea), with description of two new species and insight into the evolution of the family Trachelocercidae. Zoological Journal of the Linnean Society 177 (2): 306-319, DOI: 10.1111/zoj.12364, URL: http://dx.doi.org/10.1111/zoj.12364
03DC776A9123FFA2FCCC2B1F97FAC12D.taxon	description	(FIGS 4, 5; TABLE 1) Diagnosis: Body size in vivo 500 – 1000 × 25 – 35 μm; 9 – 19 and 17 – 26 somatic kineties on head and trunk, respectively; single nuclear group composed of two or three macronuclei and one micronucleus; narrow glabrous stripe, corresponding to area occupied by two somatic kineties; cortical granules colourless and about 0.5 μm in diameter. Type locality: The intertidal zone of a bathing beach in Qingdao, China (35 ° 55 ′ 45 ″ N, 120 ° 12 ′ 59 ″ E), where the water temperature was 16 ° C and salinity about 33 ‰ (Fig. 1 A). Type specimens: A protargol-impregnated slide containing the holotype specimen marked with an ink circle is deposited in the Laboratory of Protozoology, Ocean University of China, China (No. YY 2013052403). One paratype slide is deposited in the Natural History Museum, London, UK, with registration number NHMUK 2015.9. 15.2. Etymology: The species-group name binucleatus reflects the fact that this organism usually has two macronuclei. Description: Fully extended cells about 700 × 30 μm in vivo; body flexible and flattened ribbon-like with claviform head and pointed tail (Figs 4 A – C, 5 A – C). Body colour dark at low magnification with neck and tail portion transparent due to packed inclusions (Figs 4 A, D, 5 A, D). Single nuclear group located in centre of trunk, containing two or three macronuclei, 7 – 10 μm in diameter, and one micronucleus, 3 – 6 μm in diameter (Figs 4 A, D, I, 5 D, G, H). Colourless cortical granules, c. 0.5 μm in diameter, arranged in line between ciliary rows and scattered in glabrous stripe (Figs 4 F, 5 E, F). Locomotion by gliding between sand grains and organic debris. Cell surface densely ciliated with unciliated zone, glabrous stripe, about as wide as two somatic kineties (Figs 4 F, H, 5 J, M). Entire infraciliature consisting of dikinetids with cilia c. 10 μm long (Figs 4 H, I, 5 G). About 14 and 19 somatic kineties on head and trunk, respectively. Anterior and posterior secant system formed on left side of glabrous stripe where some kineties abut to bristle kinety (Figs 4 F, H, 5 J, M). Oral infraciliature consisting of uninterrupted circumoral kinety and conspicuous ciliary tuft located in centre of oral cavity (Figs 4 F, G, 3 I, L). Comparison: Similar to Trachelolophos quadrinucleatus sp. nov., the current new species should be compared with its known congeners. All measurements in μm. Abbreviations: CV, coefficient of variation (%); Ma, macronuclei; Mi, micronuclei; NG, nuclear groups; SD, standard deviation; SK, somatic kineties; N, number of specimens. Trachelolophos filum can be separated from the new species by having more somatic kineties on the trunk (26 – 35 vs. 17 – 26) and more macronuclei (6 – 30 forming a strand vs. two or three forming a nuclear group) (Foissner & Dragesco, 1996 b). Trachelolophos gigas differs from T. binucleatus sp. nov. in possessing a longer body length (2000 μm vs. 500 – 1000 μm), more somatic kineties on the trunk (52 – 71 vs. 17 – 26) and more macronuclei (17 – 33 macronuclei forming a strand vs. two or three forming a nuclear group) (Foissner & Dragesco, 1996 b). Trachelolophos binucleatus sp. nov. differs from T. quadrinucleatus sp. nov. (above) in having far fewer somatic kineties on the trunk (17 – 26 vs. 26 – 40) and fewer macronuclei (two or three vs. three or four).	en	Yan, Ying, Xu, Yuan, Al-Farraj, Saleh A., Al-Rasheid, Khaled A. S., Song, Weibo (2016): Morphology and phylogeny of three trachelocercids (Protozoa, Ciliophora, Karyorelictea), with description of two new species and insight into the evolution of the family Trachelocercidae. Zoological Journal of the Linnean Society 177 (2): 306-319, DOI: 10.1111/zoj.12364, URL: http://dx.doi.org/10.1111/zoj.12364
03DC776A9125FFAFFCCC28C89636C05F.taxon	description	(FIGS 6, 7; TABLE 1) This species was reported by Raikov & Kovaleva (1968) and no redescriptions have been made since then. Furthermore, in the original report it was insufficiently described and no details about the live morphology, or drawings, are available. Hence, an improved definition and a redescription, based mainly on the Chinese population, are presented here. Improved diagnosis: Body size in vivo 800 – 1500 × 20 – 40 μm; 13 – 16 and 18 – 21 somatic kineties on head and trunk, respectively; two nuclear groups, each of which is composed of about four macronuclei and two micronuclei; glabrous stripe corresponding to area occupied by five or six somatic kineties; cortical granules yellowish and c. 0.5 μm in diameter. Deposition of voucher material: A voucher slide with protargol-impregnated specimens is deposited in the Laboratory of Protozoology, Ocean University of China (No. YY 2012121301). Redescription based on the Chinese population: Fully extended cells about 1200 × 30 μm in vivo; body flexible and contractile with cross-section elliptical; cell distinctly tripartite, with neck, tail and trunk regions (Figs 6 A, 7 A). Head conspicuously claviform; tail wedgeshaped (Figs 6 A, B, 7 A, C). Endoplasm greyish and opaque due to multiple refractile inclusions (Figs 6 B, C, 7 B – D). Two nuclear groups, each of which contains about four macronuclei, 9 – 15 μm in diameter, and two micronuclei, c. 4 – 5 μm in diameter (Figs 6 A, C, H, 7 B, G, I, J). Small yellowish cortical granules, c. 0.5 μm in diameter, distributed between ciliary rows and in glabrous stripe (Figs 6 D, 7 E, F). Locomotion by gliding between sand grains and organic debris. Cell surface densely ciliated with unciliated zone, glabrous stripe, about as wide as five or six somatic kineties (Figs 6 E, G, H, 7 G, H, L). Entire infraciliature consisting of dikinetids with cilia about 10 μm long (Figs 6 G, H, 7 G). About 15 and 19 somatic kineties on head and trunk, respectively. Anterior and posterior secant system formed on left side of glabrous stripe where some kineties abut to bristle kinety (Figs 6 F, H, 7 L). Oral ciliature consisting of circumoral kinety, which is interrupted by three inserted brosse kineties (Figs 6 E – H, 7 H, L). Remarks: Based on the original description given by Raikov & Kovaleva (1968), this species has an elongated spindle-shaped body, a pointed tail that forms a slight hook and a glabrous stripe as wide as six kineties. Given these characteristics, this Guangzhou population corresponds well to the original report. Raikov & Kovaleva (1968) describe Tracheloraphis similis as colourless but provide no information on its cortical granules. Based on our study, however, this species has yellowish cortical granules but they are so small (c. 0.5 μm in diameter) that the whole cell looks colourless at low magnification. The other minor differences between these two populations are the body length in vivo (600 – 800 μm vs. 800 – 1500 μm in Guangzhou population) and the number of somatic kineties (16 vs. 13 – 16 on head, 18 – 21 on trunk in Guangzhou population). These differences are probably populationdependent because these values overlap with each other. Consequently, we identified the Guangzhou isolate as a population of T. similis. MOLECULAR PHYLOGENY BASED ON SSU RRNA GENE SEQUENCES The length (bp), GC content and GenBank accession numbers of the two species are as follows: Trachelolophos quadrinucleatus sp. nov. – 1554, 47.04 %, KT 361660; Tracheloraphis similis – 1628, 47.91 %, KT 361661. The resulting topologies generated using ML and BI are generally concordant and thus only a single topology with support values generated from both analyses is presented (Fig. 8). As described in previous studies (Yan et al., 2013, 2015), the family Trachelocercidae is a monophyletic group (88 % ML, 1.00 BI), being a sister clade to the family Kentrophoridae (87 % ML, 1.00 BI). Within Trachelocercidae, the genus Apotrachelocerca occupies a basal position. The topology then separates into two clades: Kovalevaia and Trachelolophos form one clade with low support (22 % ML, 0.52 BI), while Prototrachelocerca, Trachelocerca and Tracheloraphis form the other clade with high support (98 % ML, 1.00 BI).	en	Yan, Ying, Xu, Yuan, Al-Farraj, Saleh A., Al-Rasheid, Khaled A. S., Song, Weibo (2016): Morphology and phylogeny of three trachelocercids (Protozoa, Ciliophora, Karyorelictea), with description of two new species and insight into the evolution of the family Trachelocercidae. Zoological Journal of the Linnean Society 177 (2): 306-319, DOI: 10.1111/zoj.12364, URL: http://dx.doi.org/10.1111/zoj.12364
