ZooKeys 866: 65-83 (20 | 9) A peer-reviewed open-access journal doi: 10.3897/zookeys.866.32708 RESEARCH ARTICLE #Zookey S http:/ /Z00 keys -pen soft.net Launched to accelerate biodiversity research The genus Macrobrachium (Crustacea, Caridea, Palaemonidae) with the description of a new species from the Zaomu Mountain Forest Park, Guangdong Province, China Xiao-Zhuang Zheng', Wen-Jian Chen', Zhao-Liang Guo! | Department of Animal Science, School of Life Science and Engineering, Foshan University, Nanhai 528231, Guangdong Province, China Corresponding author: Zhao-Liang Guo (zlguo@fosu.edu.cn) Academic editor: 1, Wehrtmann | Received 28 December 2018 | Accepted 29 April 2019 | Published 24 July 2019 Attp://zoobank. ore/C44329A4-2293-45E 1-91 FO-DO8BE935788C Citation: Zheng X-Z, Chen W-J, Guo Z-L (2019) The genus Macrobrachium (Crustacea, Caridea, Palaemonidae) with the description of a new species from the Zaomu Mountain Forest Park, Guangdong Province, China. ZooKeys 866: 65-83. https://doi.org/10.3897/zookeys.866.32708 Abstract Evidence-based information is the foundation for addressing urgent global challenges in conservation and sustainable management of the freshwater biodiversity. The present study expands current knowledge of the genus Macrobrachium in Zaomu Mountain Forest Park, Guangdong Province based on the morphol- ogy, colouration, distribution, and molecular characteristics of Macrobrachium maculatum, M. inflatum, M. nipponense, and an undescribed new species, M. laevis. Macrobrachium laevis sp. nov. can be distin- guished from its congeners by a combination of characters, which includes the shape of rostrum, smooth carapace, and male second pereiopod. Macrobrachium laevis sp. nov. displays striking colour pattern, which could help to distinguish this species from other congeneric species in living specimen. Further- more, the molecular characteristics of mitochondrial cytochrome c oxidase subunit I (COI) showed that this species has a sufficient interspecific divergence from its congeners. Keywords biodiversity, freshwater prawn, molecular phylogeny, morphology, oriental region, taxonomy Copyright Xiao-Zhuang Zheng et al. 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. 66 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) Introduction The genus Macrobrachium Spence Bate, 1868 comprises 242 species and subspecies inhabiting fresh to brackish environments (De Grave and Fransen 2011). Macrobra- chium species are native to all continents except for Europe (Holthuis and Ng 2010). Prawns of the genus Macrobrachium are widely distributed in China. They can be found in various water bodies, including lakes, reservoirs, rivers, ponds, streams, ditch- es, swamps, and subterranean waters. Interest in Macrobrachium as a food has emerged throughout the world because of their delicious and unique flavor, and large size. Macrobrachium species have become an attractive food source, with good economic potential and high commercial interest in China. In addition, some colourful members of the genus Macrobrachium have attract- ed attention as ornamental pet prawn, and are traded in the ornamental fish market. Li et al. (2007) confirmed the existence of 33 species of the genus Macrobrachium in China. Recently, Guo and He (2008), He et al. (2009) and Chen et al. (2018) reported three new species from Guangdong Province, southern China. Furthermore, three new troglobitic species were reported in Guangxi Zhuang Autonomous Region, southwest China (Li and Luo 2001; Li et al. 2006; Pan et al. 2010; Lan et al. 2017; Cai and Ng 2018). A total of 39 species of the genus Macrobrachium have been recorded from China. The continued description of new species within Macrobrachium is a strong indication that there is still undiscovered species richness across the full taxonomic spectrum of Macrobrachium in China. The taxonomy of the genus Macrobrachium is mainly based on morphological characters, such as the relative length of the articles of the second pereiopods in fully developed males, rostrum shape and indentation, and colouration (Holthuis 1950; Chace and Bruce 1993). Some of these morpho- logical characteristics have been proven highly variable within the species (e.g., rostrum shape and colouration). Furthermore, the second chelipeds in particular show a very high level of developmental and sexual variation, including allometric growth in males (Short 2004). This makes it a challenge to identify and distinguish different species, and almost impossible to identify juvenile, immature and adult female specimens. Thus, comprehensive molecular characterisations have become a crucial step towards resolv- ing these longstanding taxonomic issues (Liu et al. 2007; Jose and Harikrishnan 2019). The Zaomu Mountain Forest Park (22°43'—22°45'N, 112°45'-112°47'E) was rat- ed as national 4A scenic area in 2012 (Zhang 2014). In recent years, tourism in the Zaomu Mountain Forest Park has been fast growing. The park has been reconstructed with newly climbing trestles, streams for drifting, and amusement facilities. However, the increasing exploitation of tourist resources has failed to recognise the conservation needs of different species that are found in this ecosystem. These may have negatively affected the species biodiversity of the prawn fauna in the scenic area. So far, the Macrobrachium fauna of the Zaomu Mountain Forest Park has not been accurately surveyed. To understand the diversi- ty of the prawn fauna in the scenic area, intensive field surveys were carried out in the period from 2017 to 2018. The results of these field surveys have shown that there are four species of Macrobrachium; one of them is considered as a new species to science, M. /aevis sp. nov. The genus Macrobrachium with the description of a new species... 67 Materials and methods Study area The Zaomu Mountain Forest Park (112°45'—112°47'E, 22°43'—22°45'N) is located in the Youngmei Town, Gaoming District, 38 km West Foshan, Guangdong Province. The area stretches approximately 11 km from north to south and is 5 km wide. The total area is 55 square km. The main mountain peak is 804.5 meters above sea level, and is known as the highest peak of Foshan City (Zhang 2014). The Zaomu Moun- tain Forest Park has a subtropical maritime monsoon climate, which is warm and humid throughout the year. The mean annual temperature and precipitation are 22.5 °C and 1681.2 mm, respectively (Lu et al. 2003). There are many streams, ponds and reservoirs spread out the Zaomu Mountain Forest Park, and the Yangmei River runs through near the west. The locations of the sampling stations are shown in Figure 1. Collecting specimens Samples were collected by a hand net with a mesh size of 0.8 mm. All specimens ob- tained were fixed in 95 % ethanol. Ethanol was replaced after 24 h with fresh 75 % ethanol. The drawings were made with the aid of a drawing tube mounted on an Olympus BX-41 compound microscope. Genetic analyses Genomic DNA was isolated from the muscle tissue of the abdomen by using the Universal Genomic DNA Kit (Beijing, China). A fragment of the COI (619bp) gene was amplified with conventional polymerase chain reaction (PCR) using two primers LCO1490 (5’-GGTCAACAAATCATAAAGATATTGG-3’) and HCO2198 (5°-TAAACTTCAGGGTGACCAAAAAATCA-3’) (Folmer et al. 1994). PCR cycling conditions consisted of a 3 min denaturation at 94 °C, followed by 35 cycles of denaturation at 94 °C for 30 s, annealing at 45-47 °C for 60 s, extension at 72 °C for 60 s, and a final extension at 72 °C for 5 min. PCR amplification se- quences were obtained by sanger dideoxy sequencing (Applied Biosystems 3730), after verification with the complementary strand. The sequenced fragments were edited and aligned using Codon Code Aligner v. 8.0.2 (Codon Code Corporation, Dedham, MA, USA) and corrected by the naked eye. All sequences of this study have been deposited in GenBank Nucleotide Sequence Database (see Table 1 for accession numbers). Six specimens of Macrobrachium laevis sp. nov. and 21 specimens of nine described species, namely Macrobrachium maculatum, M. formosense, M. meridionalis, M. nipponense, M. inflatum, M. dongaoensis, M. asperulum, M. fukienense, and Palaemon modestus were analysed in the prensent study. Sequences of M. asperulum 68 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) 112°45'0"E 112°50'0"E 22°50'0"N Guangdong Province, China Gaoming 5 e Zaomu Mountain Forest Pagx Sibao Reservoir Heshan 6,000 Meters —————— | 112°45'0"E 112°50'0"E Figure |. A schematic map of Guangdong Province, China. The expanded map shows locations of the Zaomu Mountain Forest Park and the 12 sampling sites. and M. fukienense were obtained from GenBank for comparative and phylogenetic analyses. Two phylogenetic methods, maximum likelihood (ML) and neighbor- joining (NJ) were implemented. The best-fitting model for sequence evolution was determined by Modelgenerator (Goss et al. 2014) and selected by the Akaike Information Criterion (AIC). Pairwise genetic distances were calculated using the Kimura 2-parameter model with the pairwise deletion option in the MEGA 5 program. The phylogenetic tree was estimated using a NJ and ML method by MEGA 5 (Tamura et al. 2011), and the confidence level in the generated tree was obtained by using 1,000 bootstraps. Table I. List of locality, geographical coordinates, and GenBank accession numbers of eight palaemonid The genus Macrobrachium with the description of a new species... species used in the present study. Species Locality Geographical coordinates GenBank accession numbers M. dongaoensis Dong’ao Island, Zhuhai 22°01'39"N, 113°42'54"E MK412789 M. formosense Chancheng, Foshan 22°56'39"N, 112°53'41"E MK412773 Chancheng, Foshan 22°56'39"N, 112°53'41"E MK412780 M. inflatum Dongfang, Hainan 18°52'50"N, 108°59'29"E MK412787 Dongfang, Hainan 18°52'50"N, 108°59'29"E MK412788 M. laevis sp. nov. Gaoming, Foshan 22°43'60"N, 112°47'10"E MK412774 Gaoming, Foshan 22°43'60"N, 112°47'10"E MK412775 Heshan, Jiangmen 22°41'06"N, 112°44'59"E MK412776 Heshan, Jiangmen 22°41'06"N, 112°44'59"E MK412777 Gaoming, Foshan 22°43'60"N, 112°47'10"E MK412781 Gaoming, Foshan 22°43'60"N, 112°47'10"E MK412782 M. maculatum Gaoming, Foshan 22°45'18"N, 112°46'04"E MK412770 Gaoming, Foshan 22°45'18"N, 112°46'04"E MK412771 Gaoming, Foshan 22°45'18"N, 112°46'04"E MK412785 Gaoming, Foshan 22°45'18"N, 112°46'04"E MK412786 M. meridionalis Chancheng, Foshan 22°56'39"N, 112°53'41"E MK412778 Chancheng, Foshan 22°56'39"N, 112°53'41"E MK412779 M. nipponense Gaoming, Foshan 22°39'54"N, 112°41'45"E MK412772 Gaoming, Foshan 22°45'18"N, 112°46'04"E MK412783 Gaoming, Foshan 22°44'49"N, 112°47'04"E MK412784 Outgroup P modestus Wulanhaote, Neimenggu 46°19'20"N, 121°54'45"E MK412768 Wulanhaote, Neimenggu 46°19'20"N, 121°54'45"E MK412769 Abbreviations The following abbreviations are used throughout the text: alt altitude, m merus, breadth, p palm, Cc carpus, rl rostral length, measured from cl carapace length, measured from the rostral tip to the postorbital the postorbital margin to the pos- margin, terior margin of the carapace, stn —_ sampling station, f finger, tl total length, measured from the i ischium, rostral tip to the posterior margin I length, of the telson. All measurements are in millimetres. Specimens were deposited in the Department of Animal Science, School of Life Science and Engineering, Foshan University (FU). 70 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65—83 (2019) Systematic accounts Palaemonidae Rafinesque, 1815 Genus Macrobrachium Spence Bate, 1868 Macrobrachium laevis sp. nov. http://zoobank.org/E4A945BD-0988-40FB-B8E7-5199396E62D 1 Figs 2,3 Material examined. Holotype: Adult male (FU, 2018-01-15-01), tl 66.2 mm, cl 18.8 mm, rl 9.4 mm; a stream near the bamboo park, the Zaomu Mountain Forest Park, Guangdong Province China (22°43'60"N, 112°47'10"E, alt. 182 m, stn. 7), 15 January 2018. Paratypes: 7 males (FU, 2018-01-15-02) tl 45.0-61.1 mm, cl 11.8- 16.4 mm, rl 7.2-9.3 mm. 14 females, 2 ovigerous females (FU, 2018-01-15-03), tl 39.8-61.5 mm, cl 9.9-17.3 mm, rl 6.5—9.3 mm, same data as for holotype. Paratypes: 2 males (FU, 2018-01-15-04), tl 32.1-48.8 mm, cl 8.0—-14.2 mm, rl 5.0—8.0 mm. 1 fe- male, tl 40.0 mm, cl 11.0 mm, rl 5.9 mm, a small stream near the Luohan hill, Heshan, Jiangmen City, Guangdong Province China (22°41'10"N, 112°43'33"E, alt. 140 m, stn.11), 12 May 2018. Paratypes: 8 males (FU, 2018-01-15-05), tl 43.3-51.2 mm, cl 11.8-14.4 mm, rl 8.5—10.1 mm. 14 female, 10 ovigerous females, tl 39.2—-60.1mm, cl 10.1-16.5 mm, rl 6.5—9.6 mm, Longquan Gorge near Heshan, Jiangmen City, Guang- dong Province China (22°41'6"N, 112°44'59"E, alt. 180 m, stn. 10), 12 May 2018. Diagnosis. Rostrum 0.51—0.71 of cl, tip slightly bent downwards, reaching to end of third segment of antennular peduncle. Rostral formula: 3-4+5-8/2-3 (usually 3), teeth equally spaced. Cephalothorax, abdomen, and second pereiopods smooth, with- out microspinules. Second pereiopods shorter than tl in both sexes; merus 1.0—1.2 times as long as the ischium; carpus 4.5—5.2 times as long as width, 1.2—1.4 times as long as merus and 0.8—1.0 times as long as palm. Palm not inflated, 4.8—5.3 times as long as wide. Movable finger 0.66—0.86 times as long as palm, without any gape when crossed. Fixed finger with one proximal tooth; moveable finger with two proximal teeth. All segments smooth, with only a small amount of spines along the lateral sur- faces of the palm. Eggs large; size 1.1-1.4 x 1.5—-1.8 mm diameter. Description. Rostrum (Fig. 2A, B) rl 0.51—0.71 of cl, high, reaching downward to end of third segment of antennular peduncle. Dorsal margin with 8—12 teeth, three or four equally spaced teeth behind orbit; ventral margin with two or three teeth (usually three). Carapace (Fig. 2A) smooth; antennal spine well developed, situated below lower orbital angle. Hepatic spine slightly larger than antennal spine, and slightly above level of antennal spine. Antennule (Fig. 2A, B) bearing sharp stylocerite, reaching end of eye; anterior mar- gin of basal segment distinctly convex, second segment 0.46 times as long as basal seg- ment, 0.83 time as long as distal segment. All segments with submarginal plumose setae. Antenna (Fig. 2A, B) bearing scaphocerite large, rectangular, 2.4—2.6 times as long as wide. Outer margin almost straight, ending with a strong spine, overreached by lamella. The genus Macrobrachium with the description of a new species... Fal Figure 2. Macrobrachium laevis sp. nov., holotype male (FU, 2018-01-15-01), cl 18.8 mm. A Entire animal, lateral view B cephalothorax and cephalic appendages, dorsal view C first pereiopod D second pereiopod E fingers of second pereiopod F third pereiopod G fifth pereiopod H telson. Mandibles, maxillulae, maxillae, first maxillipeds, second maxillipeds and branchial formula typical for genus. Third maxillipeds with robust endopod and ischiomerus slightly bow-shaped, with rows of long simple setae on distal inner and outer mar- gins. Carpus 0.70 times length of ischiomerus, with row of long simple setae on inner margin and sparse row of simple setae on outer margin; distal segment 0.83 times of penultimate segment, with long simple setae on inner margin. Exopod reaching distal end of ischiomerus, with plumose setae distally, basal with well-developed oval lateral plate; two arthrobranchs, one rudimentary, obscured by the larger one. 72, Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) First pereiopods (Fig. 2C) slender, overreaching antennal scale by carpus; carpus 1.6— 2.0 times as long as chela; fingers shorter than palm, 0.80—0.90 times as long as palm. Second pereiopods (Fig. 2D, E) shorter than tl. Shape and segment ratios of left and right second pereiopods similar in both sexes, extending beyond the antennal scale by 1/2 of carpus; merus 1.0—1.2 times as long as ischium; carpus 4.5—5.2 times as long as wide, 1.2—1.4 times as long as merus, 0.80—1.0 times as long as palm; palm not inflated, 4.85.3 times as long as wide, movable finger 0.66—0.86 times as long as palm; fingers not gaping when crossed; fixed finger with one tooth at proximal, move- able finger with two proximal teeth; all segments smooth, only small amount of spines along lateral surfaces of palm. Third pereiopods (Fig. 2F) extending to end of third segment of antennular pedun- cle by distal propodus; propodus 2.5—3.3 times as long as dactylus, with 5—7 spines on posterior margin; dactylus 5.5 times as long as wide, terminating in small claw. Fourth pereiopods (Fig. 2A) extending to end of third antennular peduncle segment by distal propodus, somewhat similar to third pereiopods. Fifth pereiopods (Fig. 2G) extending to end of third segment of antennular pedun- cle; propodus 3.4—6.5 times as long as dactylus, with 5—7 spines on posterior margin; dactylus 3.5 times as long as wide, terminating in small claw. First pleopods of male with endopod of approximately half-length of exopod, slight- ly concave at inner margin, tip rounded, without appendix interna. Second pleopods with well-developed appendix masculina, reaching middle of en- dopod, approximately twice as long as appendix interna with numerous stiff setae. Abdomen (Fig. 2A) glabrous, smooth, pleura of first to third somites broadly rounded; pleura of fourth and fifth somites also rounded, but with almost rectangular posterolateral angle; sixth somite 1.2—1.4 times as long as fifth somite, 0.59-0.67 times as long as telson. Telson (Fig. 2H) smooth, 0.46—0.61 times of cl, longer than sixth abdominal seg- ment; dorsal surface furnished with two pairs of stout, movable, spine; posterior mar- gin tapering regularly to a sharp point with two pairs of posterior a spine; numerous plumose setae present between inner pair of spine. Uropodal diaeresis with a spine shorter than lateral angle. Eggs large, size 1.1-1.4 x 1.5-1.8 mm. Live colour patterns. The juvenile was yellowish and semi-transparent (Fig. 3E); the adult male had a few indistinct longitudinal yellow stripes on the carapace, with one transverse yellow band on the first abdominal somite. All segments of the second pereiopods were golden (Fig. 3A, B). The ovigerous females had a pale yellow longi- tudinal stripe on the mid-dorsal surface from the rostrum to the tail, which extended to both sides of the abdominal somites. The palms of the third to fifth pereiopods had black and white rings (Fig. 3D). The eggs were brown (Fig. 3C). Molecular phylogenetic results. Neighbour-joining (NJ) and maximum likelihood (ML) trees inferred from partial COI sequences (619 bp) from ten species of Palaemonidae, including the new species, are shown in Figure 4. Macrobrachium laevis sp. nov. is clustering The genus Macrobrachium with the description of a new species... i) Figure 3. The colour of living Macrobrachium laevis sp. nov. A Lateral view of adult male B dorsal view of adult male C lateral view of ovigerous female D dorsal view of the fresh moulting female E lateral view of the immature male. 74 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) M L / 54/ 4s|Macrobrachium laevis sp.m. MK 412781 N J Macrobrachium laevis sp.n. MK412782 Macrobrachium laevis sp.n. MK412774 100/100 Macrobrachium laevis spn. MK412775 Macrobrachium laevis spn. MK412776 Macrobrachium laevis spn. MK412777 Macrobrachium maculatum MK412786 78/92 Macrobrachium maculatum MK412771 63/83 100/100 65/67, |Macrobrachium maculatum MK412770 82/82L_ Vacrobrachium maculatum MK412785 Macrobrachium asperulum AB250520 98/100 Macrobrachium asperulum AB250551 ee 100/100 Macrobrachium asperulum AB250549 Macrobrachium asperulum AB250550 Macrobrachium fukienense FM958065 Macrobrachium formosense MK412773 68/70] Vacrobrachium meridionalis MK412778 Tate esc Macrobrachium meridionalis MK412779 Macrobrachium formosense MK412780 100/100 Macrobrachium nipponense MK412772 F/T1 Macrobrachium nipponense MK412783 67/94 Macrobrachium nipponense MK412784 98/100 |Macrobrachium inflatum MK412787 Macrobrachium inflatum MK412788 Macrobrachium dongaoensis MK412789 96/* Palaemon modestus MK412768 100/100 L Palaemon modestus MK412769 —<—=. ned 0.05 Figure 4. Phylogenetic relationships among Macrobrachium laevis sp. nov. and the other nine species, an- alysed by maximum likelihood (ML) and neighbour-joining (NJ) methods with Palaemon modestus as the out-group taxa. Bootstrap values of ML (left) and NJ (right) are indicated above the branches of the clades. with M. maculatum with high bootstrap support (96 % in ML tree and 96 % in NJ tree). Interspecific genetic divergence (K2P) among these ten species is summarised in Table 2. The pairwise distance was 0.15—0.62%. The new species was closest to M. asperulum (0.1475—-0.1496) and M. maculatum (0.1095—0.1154), and the morphological characters supported this relationship. Moreover, the genetic divergence between M. /aevis sp. nov., M. inflatum, M. nipponense and M. dongaoensis were 0.1558—0.1598, 0.1576—0.1617 and 0.2154—0.2218, respectively, supporting the morphological differentiation of the three species. Of the species analysed, Z. dongaoensis was most genetically divergent from the new species (0.2154—0.2218). The genus Macrobrachium with the description of a new species... 75 Table 2. Pairwise genetic distance among nine Marobrachium prawn species based on the COI gene. 1 2 3 7 8 9 (1) M. maculatum 0.138 | 0.169 ao ss a 0.192 | 0.171 | 0.103 (2) M. asperulum 0.138 0.162 0.171 | 0.203 | 0.151 | 0.135 (3) M. inflatum 0.169 | 0.162 "0.014 | 0.053 | 0.197 | 0.146 | 0.148 (4) M. nipponense 0.166 | 0.160 | 0.014 0.050 | 0.050 | 0.193 | 0.144 | 0.145 (5) M. formosense 0.167 | 0.171 | 0.053 aaa 0.001 | 0.208 | 0.147 | 0.137 (6) M. meridionalis 0.167 | 0.171 | 0.053 | 0.050 | 0.001 0.208 | 0.146 | 0.138 (7) M. dongaoensis 0.192 | 0.203 | 0.197 0.208 0.207 | 0.182 (8) M. fukienense 0.171 | 0.151 | 0.146 0.146 | 0.207 0.155 (9) M. laevis sp. nov. 0.103 | 0.135 | 0.148 0.139 | 0.192 | 0.155 Etymology. Species name is derived from /aevis (Latin) in reference to the smooth- ness of the segments of the second pereiopod, carapace, and abdomen. Remarks. Macrobrachium laevis sp. nov. shows close similarity with VM. maculatum Liang and Yan 1980 regarding the ratios of various segments of the second pereiopods and in the rostral shape. Macrobrachium laevis sp. nov. can be distinguished from M. maculatum by the smooth second pereiopod whose margin of the palm has scattered microspinules (versus second pereiopod with microspinules on its whole surface); the second tooth of the movable finger placed on the proximal one-quarter (versus on the proximal one-fifth); the lack of papillae along the cutting edges (versus numerous papil- lae along the cutting edges); the finger slightly longer than the merus (versus the finger distinctly shorter than merus); the wider scaphocerite (2.4—2.6 times as long as wide) (versus 3.5 times); and ovigerous females carrying smaller eggs (1.1—1.4 x 1.5—1.8 mm) (versus larger egg sizes, 1.60—1.68 x 2.12—2.36 mm). Macrobrachium laevis sp. nov. is morphologically close to M. asperulum von Martens 1868 regarding the form of the rostrum and egg size. Macrobrachium laevis sp. nov. can be distinguished from M. aspe- rulum by its smooth carapace and second chelipeds and lack of denticles on the cutting edges (versus with rough carapace and chelipeds, and the presence of approximately ten denticles on the cutting edges), and the second tooth of the movable finger at about proximal one-quarter (versus second tooth of the movable finger on the proximal two- fifths). Macrobrachium laevis sp. nov. superficially resembles M. inflatum Liang & Yan, 1985; however, Macrobrachium laevis sp. nov. can be distinguished from M. inflatum by its shorter rostrum with fewer dorsal teeth and reaching beyond the end of the third an- tennular peduncle segment, with 8—12 dorsal teeth (versus rostrum reaching beyond the scaphocerite, with 12-17 dorsal teeth); the palm of male second pereiopod being not inflated (versus inflated) and 4.8—5.3 times as long as wide (versus 3.5—3.6 times); the finger distinctly longer than merus (versus the finger as along as the merus); the ischium shorter than the merus (versus the ischium distinctly longer than the merus); and the ovigerous females bearing larger-sized eggs (1.1—1.4 x 1.5—1.8 mm) (versus 0.53-0.59 x 0.62-0.69 mm). Macrobrachium laevis sp. nov. is closely related to M. fukienense Liang & Yan, 1980. It is possible to distinguish Macrobrachium laevis sp. nov. from M. Jukienense by the presence of more dorsal and postorbital teeth (8-12 dorsal and 3—4 postorbital teeth) (versus 7-8 dorsal and 1—2 postorbital teeth); the second tooth of the Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) 76 ye vedo [| ove 8ST arse Rages by 0-LE0 98°0-CZL'0 Piadeans hare COLE 9L1-06'T Vag AA a a} bare 39 Xx ThO-€6-0 X 89°0-F5'0 69°0-79'0 * 6S°0-€S'0 x 9'[-C'] x 9Z*I-80'I YBN NRCS Se Comes Ba A ee al eco i a | (UU) ozZIS Sood C/I C/I T/T C/T yewurxoid C/I Ja3uy I]qeaaour JO y1003 jewrxoid oy] uD YIM siusurses [Ty | YIM sjusursas [Ty] jo urszeur ynod ydaoxa Z/{ Tewrxoid oy1 ud jewrxoid oy] ud jewrxoid oy1 uEG 941 UC LeplaN sluaursas IV ale yum QnoyUM siusuTBas [TV syusuId—as [TV jewrxosd oy] ud YIM syuauTs—s [TV P/] [eurxoid oy uEG saqnutdsosorur poreos yim wiyed jo surdeur idaoxa “yOOUIS puoosas ay} Jo uOoFNGINsIq quawisas AIaA9 UO sapnuTdsossIy Ts ke is [>> w/j Jo oney 0'I-€6'°0 a a L8°0-9£L'0 780-620 d/o Jo oney ST ajo ORF 6P-EY OLLY OCS & eV q/T wyed jo oney podotsizad puosag £0-S'0 8°0—-9°0 Ol 9°0 L0-9°0 £'0-S'0 TOFTH J? ORF a a a a a a a oe a ae ee ee ee ea cy ae y-€ (Cull C6 yj901 1gso Jeisod Jo oquINN S1suaovsuop "Ay ‘s1audsU09 sy) pur ‘ou “ds szaav] WNIGIvAGoLIVPY JO SOVISTIAIDeIeYD [edISOjoYydIoypy *¢E VjIqeL UNIVINIVUL "AT ‘sou ‘ds szaavy ‘Jy WINJISOYY The genus Macrobrachium with the description of a new species... Fel. movable finger of the male second pereiopods on the proximal one-quarter (versus on the proximal half). Macrobrachium laevis sp. nov. is also closely related to M. nipponense De Haan, 1849. Macrobrachium laevis sp. nov. can be distinguished from M. nipponense by morphological characters of the male second pereiopods. The second pereiopods of Macrobrachium laevis sp. nov. are distinctly shorter than those of M. nipponense; the finger are distinctly longer than the merus (versus the finger shorter than the merus), and without setae on the cutting edge (versus the cutting edge with the long dense setae) (Fig. 3A, B versus Fig. 6A). It is possible to distinguish living Macrobrachium laevis sp. noy. from other congeners by its striking colour pattern (Fig. 3). Morphological differ- ences between these congeneric species are presented in Table 3. Habitat. Specimens of Macrobrachium laevis sp. nov. were collected from two streams and a river. The stream was near bamboo park, the Zaomu Mountain For- est Park, Foshan City (22°43'60"N, 112°47'10"E, alt. 182 m, stn. 7) (Fig. 5A). This stream runs through land covered with a secondary forest, with beds of rock and patch- es of gravel. The stream width and depth were 2.0—3.5 m and 0.6—0.9 m, respectively, with fast flowing water. The water parameters of the stream at the time of collection (15 January 2018) were: temperature 13.8 °C, pH 7.0, dissolved ammonia nitrogen 0.2 mg/I, and dissolved oxygen 4.0 mg/l. The prawns were found at the bottom of the streams together with an atyid shrimp, Caridina cantonensis Yu 1938. The specimens were also collected from another small stream near the Luohan hill, Heshan, Jiang- men City, Guangdong Province (22°41'10"N, 112°43'33"E, atl. 140 m, stn. 11). The environmental conditions were very similar to the first stream. The water parameters of the stream at the time of collection (12 May 2018) were: temperature 25.6 °C, pH 6.5, dissolved ammonia nitrogen 0.2 mg/l, and dissolved oxygen 4.5 mg/l. Additional specimens were collected from the Longquan Gorge, near Heshan, Jiangmen City (22°41'6"N, 112°44'59"E, atl. 180 m, stn. 10) (Fig. 5B). It is a small river, with a total length of 6 km. The total drop of the river is 108 meters. The river resembles a jade belt and is deeply embedded at the bottom of the Zaomu Mountain. ‘The river has flowing water, with rocks interspersed with sand patches at its bottom. The water parameters of the river at the time of collection (12 May 2018) were temperature of 26.1 °C, pH 7.0, dissolved ammonia nitrogen 0.2 mg/I, and dissolved oxygen 6.0 mg/l. Distribution. So far only known from the type locality and nearby localities in the Guangdong Province, southern China. Macrobrachium nipponense (De Haan, 1849) Fig. 6A Material examined. Five females, tl 48.5—52.8 mm, cl 14.2—16.3 mm, 4 males, tl 51.3— 65.9 mm, cl 18.0-25.6mm, Sibao Reservoir, Heshan, Jiangmen City (22°44'14"N, 112°50'17"E, alt. 84 m, stn. 1), 3 September 2017; 2 females, tl 48.5—50.4 mm, cl 15.2— 16.4 mm, | male, tl 51.3 mm, cl 17.3mm, Lingshan Garden, Gaoming, Foshan City (22°45'42"N, 112°44'39"E, alt. 44.9 m, stn. 2), 17 May 2018; 3 females, tl 46.3-49.2 78 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) [ “or Si i ag Figure 5. Habitats of Macrobrachium laevis sp. nov. A Stream near bamboo park, Zaomu Mountain For- est Park, Foshan City (type locality) B the Longquan Gorge, near Heshan, Jiangmen City. Both localities are situated in the Guangdong Province, southern China. The genus Macrobrachium with the description of a new species... os) inlugshuliu uly Li Figure 6. Photographs of Macrobrachium species. A M. nipponense, living specimen, male BM. maculatum, living specimen, male C M. inflatum, living specimen, female. mm, cl 14.1-15.3 mm, 2 males, tl 51.3-61.4 mm, cl 18.2—23.4 mm, Xikong Reservoir, Gaoming, Foshan City (22°42'35"N, 112°43'25"E, alt. 22.4 m, stn. 3), 17 May 2018; 2 females, tl 42.5-44.1 mm, cl 13.2—14.6 mm, 4 males, tl 48.3—-59.4 mm, cl 17.4-22.9 mm, Yangmei River, Gaoming, Foshan City (22°45'18"N, 112°46'04"E, alt. 49 m, stn. 5), 9 September 2017; 4 females, tl 41.4-50.3 mm, cl 12.3-16.3 mm, 3 males, tl 47.2— 65.5 mm, cl 17.5-24.5 mm, stream near Hengkong Village, Gaoming, Foshan City (22°44'49"N, 112°47'04"E, alt. 72 m, stn. 6), 9 September 2017; 1 female, tl 42.5 mm, cl 13.2, 1 male, tl 48.3 mm, cl 16.7 mm, a stream near Datian Village, Gaoming, Foshan City (22°44'22"N, 112°46'36"E, alt. 56 m stn. 9), 17 May 2018. Remarks. Macrobrachium nipponense were found in reservoirs, streams, rivers, and ponds of the Zaomu Mountain Forest Park. The species is native and broadly dis- tributed throughout East Asia (ie. China, Japan, Korea, Vietnam, and Myanmar), 80 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) (Cai and Ng 2002; Li et al. 2007). Macrobrachium nipponense was introduced into Singapore, Philippines, Uzbekistan, Iraq, Russia, Belarus, Moldova, and Iran (Chong et al. 1987; Alekhnovich and Kulesh 2001; Mirabdullaev and Niyazov 2005; Cai and Shokita 2006; De Grave and Ghane 2006; Salman et al. 2006). Macrobrachium nip- ponense is commercially important in Guangdong Province where it is sold live in local fish markets, and is locally consumed at home and in restaurants as a special dish. Colouration. ‘The body has a lighter green and transparent colour, and the cara- pace has an M-shaped mark on the side (Fig. 6A). Distribution. China, Japan, Korea, Myanmar, and Vietnam. Macrobrachium maculatum Liang & Yan, 1980 Fig. 6B Material examined. Three females, tl 45.8—54.0 mm, cl 12.0-18.3 mm, 4 males, tl 35.6-75.8 mm, cl 9.6-19.8 mm, Yangmei River, Gaoming, Foshan City (22°45'18"N, 112°46'04"E, alt. 49 m, stn. 5), 9 September 2017. Remarks. ‘The present specimens are consistent with the original description and illustration by Liang and Yan (1980) and Liu et al. (1990). This species is widely dis- tributed in the southeastern China. Macrobrachium maculatum has an economic im- portance and is usually found in the same habitat with M. nipponense. Macrobrachium maculatum inhabits freshwater and has been found in rivers, reservoirs, and streams. This species seeks shelter among aquatic vegetation. Colouration. The body is very dark brown, the cephalothorax has diagonal yellow stripes, and the abdomen has large spots (Fig. 6B). Distribution. Southeastern China (Anhui, Hunan, Fujian, and Guangdong Provinces). Macrobrachium inflatum Liang & Yan, 1985 Fig. 6C Material examined. Two females, tl 46.8—50.2 mm, cl 13.8—15.3 mm, 1 male, tl 52.1 mm, cl 14.2 mm, Qianlonggu, Gaoming, Foshan City (22°42'48"N, 112°44'54"E, alt. 124 m, stn. 4), 9 September 2017; 2 females, tl 45.5-51.0 mm, cl 13.4-16.1 mm, 3 males, tl 46.5-60.1 mm, cl 15.2—21.3 mm, Yangmei River, Gaoming, Foshan City (22°45'18"N, 112°46'04"E, alt. 49 m, stn. 5), 9 September 2017; 3 females, tl 40.5—54.3 mm, cl 12.5—20.6 mm, 2 males, tl 41.5—65.2 mm, cl 16.2—23.7 mm, Sibao Reservoir, Heshan, Jaingmen City (22°44'14"N, 112°50'17"E, alt. 84 m, stn.1), 17 August 2017. Remarks. Specimens were confidently assigned to the present species due to their inflated palm, the upturned rostrum and the rostral formula, as well as the ratio of the segments in the male second pereiopods. Macrobrachium inflatum is usually found together with M. nipponense. The genus Macrobrachium with the description of a new species... 81 Colouration. The body is translucent and light green. The rostrum is transparent to almost colourless. The cephalothorax has blue-black diagonal strips, and the abdo- men shows blue-black transverse strips. The second pereiopods have transversal yellow bands on the merus and carpus. All joints of third to fifth pereiopods have transversal yellow bands. ‘The eggs are yellow (Fig. 6C). Distribution. Southeastern China (Jiangsu, Anhui, Hunan, Guangdong, and Yunnan Provinces). Acknowledgements This study was supported by the project “Survey of marine and coastal biodiver- sity protection in prioritised region of national environmental protection project” (PM-zx097-201812-244). Part of this work was funded by Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Life Science and Engineering, Foshan University, Foshan, 528231, Guangdong, China (2018A02), and by a special fund for the cultivation of science and technology innovation of the university students in Guangdong in 2019 (climbing plan) (pd- jh2019b0510). Additional support for this project was provided by Guangdong College students’ innovation and entrepreneurship (XJ2018247 and XJ2018222). Special thanks to Dr. El-Ashram Saeed and Werner Klotz for correcting the manu- script. We are grateful to Célio Magalhaes and another anonymous reviewer who provided very useful suggestions, which improved of our work. We also thank Xi- aoxue Bao, Chunjun Zhong, Guocai Guo, Dengxu Li, and Zilong Huang, students of the Foshan University, for helping us to collect the studied specimens used for this study. References Alekhnovich AV, Kulesh VF (2001) Variation in the parameters of the life cycle in prawns of the genus Macrobrachium Bate (Crustacea, Palaemonidae). Russian Journal of Ecology 32: 454-459. https://doi.org/10.1023/A:1012586218096 Cai YX, Ng PKL (2002) The freshwater palaemonid prawns (Crustacea: Decapoda: Caridea) of Myanmar. Hydrobiologia 487: 59-83. https://doi.org/10.1023/A:1022991224381 Cai YX, Ng PKL (2018) Freshwater shrimps from karst caves of southern China, with descrip- tions of seven new species and the identity of Typhlocaridina linyunensis Li and Luo, 2001 (Crustacea: Decapoda: Caridea). Zoological Studies 57: 1-33. Cai YX, Shokita S (2006) Report on a collection of freshwater shrimps (Crustacea: Decapoda: Caridea) from the Philippines, with descriptions of four new species. Raffles Bulletin of Zoology 54: 245-270. Chace FA, Bruce AJ (1993) The caridean shrimps (Crustacea: Decapoda) of the ‘Albatross’ Philippine Expedition 1907-1910, part 6: Superfamily Palaemonoidea. Smithsonian Con- tributions to Zoology 543(543): 1-152. https://doi.org/10.5479/si.00810282.543 82 Xiao-Zhuang Zheng et al. / ZooKeys 866: 65-83 (2019) Chen QH, Chen WJ, Guo ZL (2018) Caridean prawn (Crustacea, Decapoda) from Dong’ao Island, Guangdong, China. Zootaxa 4399(3): 315-328. https://doi.org/10.11646/ zootaxa.4399.3.2 Chong SSC, Khoo HW, Ng PKL (1987) Presence of the Japanese freshwater prawn Macro- brachium nipponense (De Haan, 1849) (Decapoda: Caridea: Palaemonidae) in Singapore. Zoologische Mededelingen Leiden 61: 313-317. De Grave S, Fransen CHJM (2011) Carideorum catalogus: the recent species of the dend- robranchiate, stenopodidean, procarididean and caridean shrimp (Crustacea: Decapoda). Zoologische Mededelingen Leiden 89(5): 195-589. De Grave S, Ghane A (2006) The establishment of the Oriental River Prawn, Macrobrachium nipponense (de Haan, 1849) in Anzali Lagoon (Iran). Aquatic Invasions 1(4): 204-208. https://doi.org/10.339 1/ai.2006.1.4.2 De Haan W (1833-1850) Crustacea. In: von Siebold PF (Ed.) Fauna Japonica sive descriptio animalium, quae in itenere per Japoniam, jussu et auspiciis superiorum, qui summum in India Batava imperium tenent, suscepto, annis 1823-1830 collegit, notis, observationibus et admumbrationibus illustravit. Lugduni-Batavorum, 243 pp. [I—xxxi, plates A-J, L-Q, 1-55] Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit i from diverse metazoan invertebrates. Mo- lecular Marine Biology and Biotechnology 3(5): 294-299. Goss EM, Tabima JE Cooke DEL, Restrepo S, Fry WE, Forbes GA (2014) The Irish potato famine pathogen Phytophthora infestans originated in central Mexico rather than the Andes. Proceedings of the National Academy of Sciences of the United States of America 111(24): 8791-8796. https://doi.org/10.1073/pnas. 1401884111 Guo ZL, He SL (2008) One new and four newly recorded species of the genus Macrobrachium (Decapoda: Caridea: Palaemoindae) from Guangdong Province, southern China. Zootaxa 196 = 25; He SL, Gao J, Guo ZL (2009) Macrobrachium pentazona, a new freshwater palaemonid prawn (Decapoda: Caridea: Palaemoindae) from Guangdong Province, China. Zootaxa 2140: 38-44. Holthuis LB (1950) The Decapoda of the Siboga expedition. Part 10. The Palaemonidae col- lected by the Siboga and Snellius Expeditions with remarks on other species. I. Subfamily Palaemonidae. Siboga-Expeditie Leiden 39a(9): 1-268. Holthuis LB, Ng PKL (2010) Nomenclature and taxonomy. In: New MB, Valenti WC, Tidwell JH, D’Abramo LR, Kutty MN (Eds) Freshwater Prawns: Biology and Farming (1* edn). Wiley-Blackwell, Oxford, 12-17. https://doi.org/10.1002/978 1444314649 .ch2 Jose D, Harikrishnan M (2019) Evolutionary history of genus Macrobrachium inferred from mitochondrial markers: a molecular clock approach. Mitochondrial DNA Part A 30: 92— 100. https://doi.org/10.1080/24701394.2018.1462347 Lan C, Wu Z, Li W (2017) A new troglobitic shrimp from Guangxi Zhuang Autonomous Region: Macrobrachium duanensis sp. Journal of Jishou University 38(2): 61-62. Li JC, Cai YX, Clarke A (2006) A new species of troglobitic freshwater prawn of the genus Macrobrachium from southern China (Crustacea: Decapoda: Palaemonidae). Raffles Bul- letin of Zoology 54: 277-282. The genus Macrobrachium with the description of a new species... 83 Li W, Luo Z (2001) A new troglobitic shrimp from Guangxi. Journal of Guangxi Normal Uni- versity 19(12): 72-74. [in Chinese with English abstract] Li XZ, Liu RY, Liang XQ, Chen GX (2007) Fauna Sinica. Invertebrata. Vol. 44. Crustacea De- capoda Palaemonoidea. Science Press, Beijing, 381 pp. [in Chinese with English abstract] Liang XQ, Yan SL (1980) Description of two new species of Macrobrachium (Decapoda Caridea) from Fujian, China. Acta Zootaxonomica Sinica 5(1): 30-34. [in Chinese with English abstract] Liang XQ, Yan SL (1985) New species and new record of Palaemoninae from China (Crustacea Decapoda). Acta Zootaxonomica Sinica 10(3): 253-258. [in Chinese with English abstract] Liu MY, Cai YX, Tzeng CS (2007) Molecular systematics of the freshwater prawn genus Mac- robrachium Bate, 1868 (Crustacea: Decapoda: Palaemonidae) inferred from mtDNA se- quences, with emphasis on east Asian species. Zoological Studies 46(3): 272-289. Liu RY, Liang XQ, Yan SL (1990) A study of the Palaemoninae (Crustacea Decapoda) from China I. Macrobrachium, Leander and Leandrites. Transactions of The Chinese Crustacean Society No. 2: 102—134. [in Chinese with English abstract] Lu YD, Xue KN, Li ZK (2003) Investigation on the wild ornamental plants in Gaoming City, Guangdong. Guangdong Forestry Science and Technology 19(3): 35-37. Mirabdullaev IM, Niyazov DS (2005) Alien decapods (Crustacea) in Uzbekistan. Abstracts of the II International Symposium Invasion of alien species in Holarctic (BOROK-2), Borok, Russia, 27 September 27—1 October, 113-114. Pan YT, Hou Z, Li SQ (2010) Description of a new Macrobrachium species (Crustacea: De- capoda: Caridea: Palaemonidae) from a cave in Guangxi, with a synopsis of the stygo- biotic Decapoda in China. Journal of Cave and Karst Studies 72: 86-93. https://doi. org/10.4311/jcks20091sc0087 Rafinesque CS (1815) Analyse de la Nature, ou Tableau de l'Univers et des Corps Organisés. UImprimerie de Jean Barravecchia, Palermo, 224 pp. https://doi.org/10.5962/bhl title. 106607 Salman SD, Page TJ, Naser MD, Yasser AG (2006) ‘The invasion of Macrobrachium nipponense (De Haan, 1849) (Caridea: Palaemonidae) into the Southern Iraqi marshes. Aquatic Inva- sions 1: 109-115. https://doi.org/10.3391/ai.2006.1.3.2 Short JW (2004) A revision of Australian river prawn, Macrobrachium (Crustacea: De- capoda: Palaemonidae). Hydrobiologia 525: 1-100. https://doi.org/10.1023/ B:HYDR.000003887 1.50730.95 Spence Bate CS (1868) On a new genus, with four new species of freshwater prawns. Proceed- ings of the Zoological Society of London 1868: 363-368. Tamura K, Dudley J, Nei M, Kumar S (2011) MEGAS: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mo- lecular Biology & Evolution 28(10): 2731-2739. https://doi.org/10.1093/molbev/msr121 Von Martens E (1868) Uber einige ostasiatische Siisswasserthiere. Archiv fiir Naturgeschichte Berlin 34(1): 1-67. https://doi.org/10.5962/bhl.part.20475 Yu SC (1938) Studies on Chinese Caridina with descriptions of five new species. Bulletin of the Fan Memorial Institute of Biology, Zoology 8: 275-310. Zhang YL (2014) Charming Yang He: Building the town in the scenery. Insight China 9: 60-62.