Tardigrades in the alpine region of Northeast China with an integrative description of Crenubiotus liangshuiensis sp. nov. Author Zhang, Jing-Yu 0000-0003-4595-3211 College of Wildlife and Protected Area, Northeast Forestry University, Harbin, 150040, Heilongjiang, China. zhangjingyu@foxmail.com Author Sun, Xue-Ling 0000-0002-0135-4447 Heilongjiang Provincial Natural Resources Rights and Interests Investigation and Monitoring Institute, Harbin, 150080, Heilongjiang, China. Author Wang, Ning Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin, 150040, Heilongjiang, China. Author Hao, Li 0009-0001-8016-7331 College of Wildlife and Protected Area, Northeast Forestry University, Harbin, 150040, Heilongjiang, China. haoli958@sina.com Author Ma, Cheng-Xue 0000-0001-6100-1291 College of Wildlife and Protected Area, Northeast Forestry University, Harbin, 150040, Heilongjiang, China. Author Zhao, Na 0000-0001-9234-4908 College of Life Sciences, Northeast Forestry University, Harbin, 150040, Heilongjiang, China. Author Li, He- Ping 0000-0001-8284-0449 College of Wildlife and Protected Area, Northeast Forestry University, Harbin, 150040, Heilongjiang, China. lihepinghrb2002@nefu.edu.cn Author Zhao, Min 0000-0001-9751-454X College of Life Sciences, Northeast Forestry University, Harbin, 150040, Heilongjiang, China. 55899783@qq.com Author Yang, Sheng-Tao 0009-0005-8462-3890 Heilongjiang Provincial Natural Resources Rights and Interests Investigation and Monitoring Institute, Harbin, 150080, Heilongjiang, China. 13697385@qq.com text Zootaxa 2024 2024-08-02 5492 1 96 108 http://dx.doi.org/10.11646/zootaxa.5492.1.5 journal article 10.11646/zootaxa.5492.1.5 1175-5326 13212067 336B4DA3-553E-4EAA-9277-272D386378DB Crenubiotus liangshuiensis sp. nov. ( Figs 2–6 , Table 2 ) Material examined: 11 specimens and 1 egg mounted on microscope slides in Hoyer’s medium, 1 specimen fixed on a SEM stub, and 12 specimens processed for DNA sequencing. Type locality: 47º11′03.8″N , 128º53′45.6″E ; 416 m asl : China , Yichun , Liangshui National Nature Reserve . Substrate: moss on fallen logs . Etymology: The new species is named after the Liangshui National Nature Reserve, where it was discovered. Type depositories: The holotype , 10 paratypes (slide Nos. NMS0011–21 ) and 1 egg ( NMS0022 ) are deposited in the Physiology Lab , College of Wildlife and Protected Area , Northeast Forestry University , Harbin , China . FIGURE 2. Crenubiotus liangshuiensis sp. nov. habitus and cuticle pores under PCM: A–dorsoventral projection, holotype (NMS0011); B–the dorsal cuticle between legs II and III, the dotted ellipse indicate larger irregular holes distributed in band; C– the paratype (NMS0012) whole body, the black box correspond to the enlarged images in B. Scale bars: 50 μm (A), 10 μm (B). FIGURE 3. Crenubiotus liangshuiensis sp. nov. cuticle granulation and pores under PCM. A–Leg IV internal granulation; B–Leg IV quadrangular pores; C–bulges on Leg III; D–Dorso-caudal band of granulation. A dashed empty arrow indicates the sparse tubercles of internal side, a white full arrow indicates the bigger quadrangular pore, a black full arrow indicates the bulges; an arrowhead indicates the granulation band. A, B: Holotype (NMS0011); C, D: Paratype (NMS0012). Scale bar: 10 µm. Description of the new species. Length up to 291 µm, colourless, eyes present. Body cuticle with numerous irregular pores ( Fig. 2 ), large quadrangular pores up to 1.2×2.9 µm in diameter ( Fig. 3B ) and small, elliptical pores about 0.5 µm in diameter, all evident in PCM and NCM, as well as in SEM. The small elliptical pores are distributed randomly on the entire body cuticle ( Fig. 2 ), and the larger irregular quadrangular pores are mostly distributed in the middle of each segment, forming 9–10 transverse bands on the dorsum ( Fig. 2B ), slightly larger in size on the caudal portion of the body. Pore bands clearly distinguishable under LCM. Evident bulges are present on the external and lateral portions of the leg I–III surface ( Fig. 3C , Fig. 4A ). The granulation is present on all legs (e.g. Fig. 4E ). Small cuticular tubercles are present in a dorso-lateral caudal band just anterior to the hind legs ( Fig. 3D ) and on the dorsal cuticle of legs IV, with some sparse tubercles present on the internal side of legs IV ( Fig. 3A ). Claws are slender, of the richtersiusid type ( Fig. 4 ). Primary branches with distinct accessory points, a long, common tract with a system of internal septa, the common tract apparently longer than half of the entire claw height. The bucco-pharyngeal apparatus is of the Macrobiotus type , pharynx with triangular apophyses, two rodshaped macroplacoids (2<1), and a microplacoid close to the second macroplacoid ( Fig. 5 ). The first macroplacoid is anteriorly narrowed and constricted in the middle, whereas the second has a subterminal constriction. TABLE 2. Measurements and pt values of Crenubiotus liangshuiensis sp. nov. type series.

RANGE		MEAN		SD	Holotype
CHARACTER	N
µm	pt	µm	pt	µm	pt	µm	pt
Body length	11	225.2	-	291.3	821.8	-	956.3	261.6	892.3	19.5	40.4	258.0	861.9
Buccopharyngeal tube
Buccal tube length	11	26.2	-	31.1	29.3	1.7	29.9
Stylet support insertion point	11	20.0	-	24.0	74.1	-	78.4	22.3	76.2	1.3	1.3	23.2	77.4
Buccal tube external width	9	2.5	-	3.2	8.8	-	10.8	2.9	9.9	0.3	0.6	3.2	10.8
Buccal tube internal width	9	1.5	-	2.0	4.8	-	6.7	1.7	5.9	0.2	0.6	2.0	6.7
Placoid lengths
Macroplacoid 1	11	4.3	-	5.7	15.1	-	18.7	4.9	16.9	0.4	1.2	5.3	17.6
Macroplacoid 2	11	3.6	-	4.6	12.1	-	15.4	4.1	14.0	0.3	1.0	4.6	15.4
Microplacoid	10	0.8	-	1.2	2.6	-	4.0	1.0	3.3	0.1	0.5	1.0	3.3
Macroplacoid row	10	9.5	-	11.7	35.4	-	38.6	10.7	36.5	0.7	1.2	11.5	38.6
Placoid row	10	11.6	-	14.0	41.9	-	46.6	13.0	44.2	0.9	1.4	14.0	46.6
Claw I heights
External primary branch	9	5.4	-	6.8	20.4	-	23.5	6.5	21.9	0.5	0.9	6.8	22.6
External secondary branch	9	4.4	-	5.9	16.7	-	19.8	5.5	18.5	0.5	1.0	5.8	19.4
Internal primary branch	6	5.1	-	6.7	19.3	-	22.4	6.2	21.2	0.6	1.1	6.7	22.4
Internal secondary branch	4	5.5	-	5.7	18.0	-	19.0	5.6	18.6	0.1	0.4	5.6	18.9
Claw II heights
External primary branch	10	5.4	-	7.3	20.3	-	24.4	6.8	22.8	0.5	1.2	7.3	24.2
External secondary branch	8	4.7	-	6.5	17.6	-	21.7	5.7	19.1	0.5	1.2	6.5	21.7
Internal primary branch	2	5.5	-	7.1	20.6	-	23.6	6.3	22.1	1.1	2.1	7.1	23.6
Internal secondary branch	2	4.8	-	6.4	18.0	-	21.3	5.6	19.7	1.1	2.3	6.4	21.3
Claw III heights
External primary branch	7	5.4	-	7.2	20.2	-	24.1	6.7	22.8	0.6	1.4	7.2	24.0
External secondary branch	5	4.6	-	6.2	17.5	-	21.1	5.7	19.6	0.7	1.4	6.1	20.3
Internal primary branch	4	5.2	-	7.2	19.6	-	24.2	6.2	21.7	0.8	1.9	7.2	24.2
Internal secondary branch	4	4.2	-	6.3	15.9	-	20.9	5.2	18.2	0.8	2.1	6.3	20.9
Claw IV heights
Anterior primary branch	9	5.7	-	8.0	21.4	-	26.5	7.4	25.0	0.7	1.7	7.7	25.9
Anterior secondary branch	8	4.7	-	6.4	17.7	-	22.6	6.1	20.5	0.6	1.5	6.4	21.4
Posterior primary branch	6	5.7	-	7.9	21.6	-	26.2	7.2	24.5	0.8	1.8	7.8	26.2
Posterior secondary branch	6	4.5	-	6.6	17.0	-	23.3	5.9	20.0	0.8	2.5	6.3	21.1

The egg is white, spherical, with conical processes comprising a main body and a distal elongated and flexible portion ( Fig. 6 ). Some process apices are bifurcated and may contain internal septa forming a single distal bubble ( Fig. 6D ). The single egg found has a diameter of 73.4 µm including the processes and 52.1 µm without them; there are about 16 processes on the circumference. The processes are 12.2 µm high and have a basal diameter of 9.4 µm (average values of four processes).