taxonID	type	description	language	source
070387C8D63E050B4E4F23A5D9F5FB55.taxon	diagnosis	Diagnosis Body flexible; 125 – 155 9 25 – 35 lm in vivo; citrine-yellow cortical granules about 1 lm in diameter; about 35 adoral membranelles; one buccal cirrus, one anterior and one posterior frontal row with a frontoventral cirrus to the left of them; four or five postoral ventral cirri; four or five ventral rows; two pretransverse ventral cirri; c. seven transverse cirri; on average 37 left and 38 right marginal cirri; five dorsal kineties, dorsal kinety 4 starting at level of buccal vertex; about eight macronuclear segments; two micronuclei; brackish water habitat. Type locality and ecology Zhanjiang Mangrove National Nature Reserve (21 ° 55 0 N, 109 ° 75 0 E), Guangdong Province, China. Water temperature 23 ° C, salinity 15 psu. Type specimens A protargol slide containing the holotype specimen (see Figs 1 B, C, 2 J, K; registration no. LXT 2013120701 / 1) and one paratype slide (registration no. LXT 2013120701 / 2) were deposited in the Laboratory of Protozoology, Ocean University of China (OUC). A second paratype slide was deposited in the Natural History Museum, London (registration number NHMUK 2015.7.10.1). Etymology The species-group name monilata is a composite of the Latin noun ‘ monile’ (necklace) and the suffix ‘ ata’ (having a feature) and refers to the moniliform macronucleus. Description Body 125 – 155 9 25 – 35 lm in vivo, flexible but not contractile, sometimes slightly twisted, elongate, generally elliptical in outline with both ends rounded and head region slightly bent to left. Ratio of length to width about 5: 1, dorsoventrally flattened about 2: 1 with ventral side flat and dorsal side bulging evenly when viewed laterally (Figs 1 A, 2 A – E). Anterior end with a conspicuous transparent, thin collar. Adoral zone occupying about 28 % of body length (Figs 1 A, 2 A – D). Buccal cavity wide, right margin of cavity in mid-cell region. Pellicle thin and flexible. Cortical granules citrine-yellow, spherical, approximately 1 lm in diameter, rendering cell citrine-brownish at low magnification (Fig. 2 A – E, H). Several cortical granules grouped together around dikinetids, arranged in short rows between dorsal kineties and cirral rows (Figs 1 E, F, 2 G). Cytoplasm colourless, usually packed with granules (1 – 5 lm in diameter), and food vacuoles usually containing diatoms and sometimes small ciliates (Fig. 2 I, L, T). Contractile vacuole not observed. Locomotion by slowly crawling on substrate and amongst debris, or occasionally by swimming in water. Cilia of anterior membranelles approximately 15 lm long in vivo. All cirri on ventral side about 10 – 12 lm long except the stronger frontal cirri, which have cilia about 15 lm long, and the transverse cirri, which are about 15 – 20 lm long and project beyond posterior cell margin. Infraciliature as shown in Figures 1 B – D, G, 2 J – T. Adoral zone composed of 27 – 41 membranelles, distal end of adoral zone extending far onto right-ventral side of cell (Distal End of adoral zone-value c. 0.4; Berger, 1999, 2006). Bases of membranelles unequal in length, those in distal part comprise three short, equal-length rows of kinetosomes, those in proximal part with four rows, one short and three long (Figs 1 B, G, 2 J, L). Undulating membranes more or less in Oxytricha - pattern; paroral and endoral membranes almost equal in length, slightly curved and optically intersect in posterior third; endoral membrane with single-rowed kinetosomes, starting at midpoint of buccal field; paroral with multiple-rowed kinetosomes, commencing ahead of endoral (Fig. 1 B, G). Constantly three enlarged frontal cirri, rightmost one just behind distal end of adoral zone; single buccal cirrus right of paroral membrane (Figs 1 B, G, 2 F, J, L). Two frontal rows; anterior row composed of five to eight cirri, posterior row four to six cirri. One frontoventral cirrus located left of posterior end of anterior frontal row (Figs 1 B, G, 2 J). Four or five short ventral cirri rows, each composed of three to eight cirri. Four or five sparsely distributed postoral ventral cirri located left of ventral cirral rows, anterior-most one just behind buccal vertex (Figs 1 B, D, G, 2 R – T). Six to eight transverse cirri arranged in J-shaped row; two fine pretransverse ventral cirri located close to transverse cirri (Figs 1 B, D, G, 2 P, Q). Two marginal rows, almost confluent posteriorly. Right marginal row composed of 32 – 44 cirri, commencing near anterior end of anterior frontal row; left marginal row composed of 32 – 41 cirri, starting about level with posterior end of paroral membrane (Figs 1 B, G, 2 J). Five dorsal kineties; dorsal kinety 4 commences at about level of buccal vertex, the other four extend almost entire body length; dorsal bristles about 4 lm long in vivo (Figs 1 B, C, 2 K, M – O). Strong fibres associated with cirri are visible in protargol-stained specimens; pharyngeal fibres extremely strong, extending posteriorly and occupying about one-third of body length (Figs 1 G, 2 T). Macronucleus left of cell median, moniliform, usually composed of eight globular to ellipsoidal segments; consistently two ellipsoidal micronuclei (Figs 1 C, G, 2 J, L, M, P).	en	Luo, Xiaotian, Gao, Feng, Yi, Zhenzhen, Pan, Ying, Al-Farraj, Saleh A., Warren, Alan (2017): Taxonomy and molecular phylogeny of two new brackish hypotrichous ciliates, with the establishment of a new genus (Ciliophora, Spirotrichea). Zoological Journal of the Linnean Society 179 (3): 475-491, DOI: 10.1111/zoj.12451, URL: https://www.mendeley.com/catalogue/48f35782-3ad5-3367-86d9-31aa846460b9/
070387C8D63805054DDC27F4D957FBA4.taxon	description	(FIGS 3 A – I, 4 A – H; TABLE 2) Diagnosis Body 70 – 110 9 40 – 60 lm in vivo; generally oval in outline, colourless to slightly grey-greenish; about 22 adoral membranelles; three frontal cirri; one buccal cirrus; amphisiellid median cirral row comprises about 11 cirri and terminates ahead of mid-body; three to seven frontoventral cirri; two pretransverse ventral cirri; five transverse cirri; 19 left and 24 right marginal cirri on average; three dorsal kineties; two macronuclear segments; two micronuclei; brackish water habitat. Type locality and ecology A small puddle near Zhan Qiao Pier in Qingdao (36 ° 06 0 N, 120 ° 31 0 E), Shandong Province, China. Water temperature 21 ° C, salinity 20 psu. Type specimens The protargol slide containing the holotype specimen (see Fig. 3 B, C, G; registration no. PY 2010092801 / 1) was deposited in the Laboratory of Protozoology, Ocean University of China (OUC). A paratype slide was deposited in the Natural History Museum, London (registration number NHMUK 2015.7.10.2). Etymology The species-group name ovalis (Latin adjective; eggshaped, oval) refers to the oval body outline. Description Cell about 70 – 110 9 40 – 60 lm in vivo, oval in outline (Figs 3 A, 4 A – F), anterior end slightly narrowed and bends leftwards when cell changes direction during locomotion, posterior end more or less broadly rounded. Ratio of length to width about 5: 3 – 2: 1; dorsoventrally flattened about 2: 1. Pellicle very flexible. Buccal field narrow, transparent, and conspicuous in vivo, occupying about 1 / 3 of body length. Cytoplasm colourless; whole body except buccal field usually packed with numerous dark-greenish algae (3 – 10 lm across; we are unsure whether they are endosymbionts or not) and several food vacuoles containing diatoms and small ciliates, rendering cell slightly grey-greenish (Figs 3 I, 4 A – G). Several bacterial plaques of varying size on dorsal surface, each composed of c. 1 – 2 9 0.5 lm bacilli (Fig. 4 H). Cortical granules absent. Contractile vacuole about 10 lm across, positioned left of midline in mid-body region (Fig. 3 A). Locomotion by slowly crawling on substrate; swimming cells not observed. Cilia of anterior membranelles approximately 12 lm long in vivo. All ventral cirri about 8 – 10 lm long except frontal and transverse cirri, which have cilia about 12 lm long; transverse cirri do not project beyond posterior body margin (Fig. 3 A). Dorsal bristles about 3 lm long in vivo. Infraciliature as shown in Figure 3 B – I. Adoral zone composed of 19 – 25 membranelles with about four membranelles in distal portion located apically. Bases of distal membranelles comprise three short equal-length rows of kinetosomes; bases of proximal membranelles comprise one short and three long rows of kinetosomes (Fig. 3 B, D, E). Paroral and endoral membranes equal in length, arranged in parallel, occupying only about half length of buccal field (Fig. 3 B, D, I). Consistently three enlarged frontal cirri located near anterior end of cell, arranged in a slightly oblique pseudorow (Fig. 3 B, D, G). Single enlarged buccal cirrus located right of anterior end of paroral membrane (Fig. 3 B, D, G – I). Amphisiellid median cirral row (ACR) usually composed of eight to 11 cirri, commencing behind rightmost frontal cirrus and terminating at about level of buccal vertex (Fig. 3 B, D, I). Three to seven (usually four) frontoventral cirri left of anterior portion of ACR (Fig. 3 B, D, G, H). Usually five transverse cirri arranged in J-shaped or U-shaped row. One to three, usually two, pretransverse ventral cirri positioned ahead of transverse cirri. Two marginal rows not reaching posterior end of body and thus separated posteriorly. Right marginal row commencing near anterior part of ACR, composed of 19 – 31 cirri; left marginal row starting about level with buccal vertex, composed of 16 – 24 cirri (Fig. 3 B, G – I). Invariably three dorsal kineties more or less shortened anteriorly; number of bristles per kinety increases from left to right (Fig. 3 C, F). Two ellipsoidal macronuclear segments slightly left of cell midline, transparent in vivo, conspicuously separated but connected to each other via a thread (Figs 3 A, C, E, 4 A, E); rear end of anterior macronuclear segment a little behind level of buccal vertex, rear end of posterior segment slightly anterior of transverse cirri when observed in vivo. Two spherical micronuclei, c. 2 lm in diameter, each closely associated with a macronuclear segment (Fig. 3 C, H). CV, coefficient of variation in %; Max., maximum; Mean, arithmetic mean; Med., median value; Min., minimum; N, number of specimens examined; SD, standard deviation. SSU RRNA GENE SEQUENCE AND PHYLOGENETIC ANALYSES The SSU rRNA gene sequence of Pol. monilata sp. nov. (GenBank accession number KT 192639) is 1698 bp long and has a G + C content of 45.11 %; that of L. ovalis sp. nov. (GenBank accession number KP 266625) is 1729 bp long and has a G + C content of 45.58 %. The topologies of the BI and ML trees inferred from SSU rRNA gene sequences were basically congruent with variable support values; therefore, only the ML topology (with nodal support from both methods) is shown (Fig. 5). The phylogenetic analyses showed that neither order Stichotrichida nor Sporadotrichida is monophyletic. Within these two orders, three families out of seven (i. e. Trachelostylidae, Gonostomatidae, and Spirofilidae) and one TWO NEW BRACKISH HYPOTRICHOUS CILIATES 483 G A E B C D F H ACR, amphisiellid median cirral row; CV, coefficient of variation in %; Max., maximum; Mean, arithmetic mean; Med., median value; Min., minimum; N, number of specimens examined; SD, standard deviation. subfamily (i. e. Stylonychinae) are monophyletic. Protogastrostyla pulchra and Protogastrostyla sterkii cluster in a clade that branches off at the base of the ingroup. Lamtostyla ovalis sp. nov. clusters with five stichotrichid species, i. e. Uroleptoides magnigranulosus, Bistichella cystiformans, Bistichella variabilis, Parabistichella variabilis, and Orthoamphisiella breviseries, near the base of the SSU rRNA gene trees forming a group that is sister to a large assemblage comprising most of the stichotrichids, sporadotrichids, and (core) urostyloids for which SSU rRNA gene sequence data are available. Polystichothrix monilata sp. nov. is sister to Pseudogastrostyla flava with high support (96 % ML, 1.00 BI), and is next most closely related to Rubrioxytricha ferruginea, Rubrioxytricha haematoplasma, Ponturostyla enigmatica, and Pseudocyrtohymena koreana. Except for Pseudogastrostyla flava, all of these species have 18 frontal-ventral-transverse cirri and are members of the subfamily Oxytrichinae. In order to further investigate evolutionary relationships within the family Oxytrichidae, we mapped morphological characters onto the phylogenetic tree (Fig. 5). Based on cirral patterns, Oxytrichidae is divided into three groups, viz. oxytrichids with 18 frontal-ventral-transverse cirri (blue stars), oxytrichids with more than 18 cirri (red stars), and oxytrichids with fewer than 18 cirri (green stars). The Oxytrichidae can also be separated into two groups based on the presence or absence of dorsal kinety fragmentation, viz. those with dorsomarginal kineties but lacking dorsal kinety fragmentation (orange triangles), and those both with dorsomarginal kineties and dorsal kinety fragmentation (black triangles). However, grouping patterns based on either of these two morphological character states are not possible in the topology of the SSU rRNA gene trees (Fig. 5).	en	Luo, Xiaotian, Gao, Feng, Yi, Zhenzhen, Pan, Ying, Al-Farraj, Saleh A., Warren, Alan (2017): Taxonomy and molecular phylogeny of two new brackish hypotrichous ciliates, with the establishment of a new genus (Ciliophora, Spirotrichea). Zoological Journal of the Linnean Society 179 (3): 475-491, DOI: 10.1111/zoj.12451, URL: https://www.mendeley.com/catalogue/48f35782-3ad5-3367-86d9-31aa846460b9/
