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1 TITLE: Atlantic expansion of the African caridean shrimp uncicornis Holthuis 2 and Maurin, 1952 (: ), a potential amphi-Atlantic : 3 implications of global climate change in marine fauna conservation 4 5 Atlantic expansion of the African caridean shrimp Lysmata uncicornis Holthuis and 6 Maurin, 1952 (Caridea: Lysmatidae). 7 8 RUNNING TITLE: Lysmata uncicornis across Atlantic waters 9 10 E. GONZÁLEZ-ORTEGÓN1*, J. E. GARCÍA-RASO2, R. CALADO3, I. LÓPEZ DE 11 LA ROSA1, M. GUERRERO4 and J.A. CUESTA1 12 13 1Instituto de Ciencias Marinas de Andalucía, CSIC, Avda. República Saharaui, 2, 11519 14 Puerto Real, Cádiz, Spain 15 2Universidad de Málaga, Departamento de Biología , Facultad de Ciencias, 16 Campus de Teatinos s/n, Málaga, Spain 17 3ECOMARE, Centre for Environmental and Marine Studies (CESAM), Department of 18 Biology, University of Aveiro, Santiago University Campus, 3810-193 Aveiro, Portugal 19 4Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, Puerto Real, 20 Cádiz, Spain 21 *Corresponding Author: Enrique González-Ortegón 22 [email protected] 23 24 Tel.: +34 956832612; fax: +34 956 83 47 01 25 26 May 2019 27 Abstract 28 29 The present study reports the occurrence of several specimens of the African caridean 30 shrimp Lysmata uncicornis Holthuis and Maurin, 1952 in the NE Atlantic coast (Gulf of 31 Cadiz, Spain and the Algarve, Portugal). Lysmata uncicornis is a poorly studied species 32 that has been originally described from the Atlantic waters of Morocco, where it was 33 first collected inside the port of Casablanca in a rocky bottom at 4-5 m depth. While no 34 scientific publication has previously reported this species outside the waters of 35 Morocco, several specimens have been collected in the coastal waters of the Gulf of 36 Cadiz and the Algarve. This species may be able to expand successfully northward 37 along European waters, probably favoured by global warming. It is possible that this 38 expansion may also be enhanced through the marine aquarium trade. Recently, Lysmata 39 arvoredensis Giraldes, Macedo, Brandão, Baeza and Freire, 2018 a new species of 40 shrimp from the south coast of Brazil was described. However, morphological and 41 genetic comparisons revealed no differences between L. uncicornis and L. arvoredensis. 42 Therefore, L. arvoredensis should be considered as a junior synonym of L. uncicornis. 43 While L. uncicornis may well be an amphi-Atlantic species, such as L. grabhami 44 (Gordon 1935), for now, the introduction of L. uncicornis in Brazilian waters cannot be 45 ruled out. 46 47 Keywords: , Atlantic waters, Lysmata arvoredensis, amphi-Atlantic, 48 marine ornamental shrimp, introduced species, exotic species, tropicalization, 49 ornamental species, marine aquarium trade 50 51

1 52 Introduction 53 It is commonly reported that more non-native species occur rather at higher than lower 54 latitudes than that of their native home ranges (Guo et al. 2012). Several African species 55 are currently moving northward and arriving to Southwest European coasts, likely due 56 to global warming (e.g. Perez-Miguel et al. 2019). This natural expansion may 57 eventually be accelerated by other vectors, such as ballast water or human transactions. 58 As an example, one can refer the global trade of marine species for domestic and public 59 aquaria (Padilla and Williams 2004; Calado and Chapman 2006). Caridean shrimps 60 within genus Lysmata Risso, 1816 are commonly ranked among the top 10 most heavily 61 collected and traded marine ornamental invertebrates (Wabnitz et al. 2003). Coloration 62 is probably the most common feature used to recruit shrimp species to the marine 63 aquarium trade (see Calado et al. 2003 and Calado 2008 for details) and are likely one 64 of those invertebrate groups experiencing an increase in demand among hobbyists 65 (Calado et al. 2017). Despite the fact that biological invasions often contribute to native 66 biodiversity decline, and global warming and anthropogenic activities are expected to 67 elevate the potential of introducing non-native species worldwide, they continue to be 68 overlooked in marine conservation plans (Giakoumi et al., 2016). 69

70 Lysmata species, popularly termed as “peppermint shrimps”, occur worldwide in 71 tropical, warm to cold-temperate seas, usually on rocky boulder slopes and reef habitats, 72 being currently represented by 48 valid species (WoRMS, 2018), 24 of these being 73 present in the Atlantic Ocean (Western Atlantic: WA, Eastern Atlantic: EA) and 74 Mediterranean Sea (M). Chace (1997), Fransen (1991), d´Udekem d´Acoz (1999, 2000) 75 and Wirtz et al. (2016) have cited the following species to these geographic regions: 76 WA: L. anchisteus Chace, 1972, L. intermedia (Kingsley, 1878), L. rathbunae Chace, 77 1970 and L. wurdemanni (Gibbes, 1850); WA and EA: L. grabhami (Gordon, 1935) and 78 L. moorei (Rathbun, 1901); EA: L. olavoi Fransen, 1991, L. stenolepis Crosnier and 79 Forest, 1973 and L. uncicornis Holthuis and Maurin, 1952; EA and M: L. nilita Dohrn 80 and Holthuis, 1950 and L. seticaudata (Risso, 1816). Rhyne and Lin (2006) analysed 81 the complex L. wurdemanni and L. rathbunae for the WA and described 4 new species 82 for this region: L. ankeri Rhyne and Lin, 2006, L. bahia Rhyne and Lin, 2006, L. 83 pederseni Rhyne and Lin, 2006 and L. boggessi Rhyne and Lin, 2006, and redescribed 84 the species Lysmata rathbunae Chace, 1970 and L. wurdemanni. It is worth highlighting 85 that the presence of L. wurdemanni in WA is valid, but further studies are still needed 86 (Rhyne et al. 2009). More recently new Atlantic species have been described, namely L. 87 rafa Rhyne and Anker, 2007 from Florida (WA) (Rhyne and Anker 2007), L. hochi 88 Baeza and Anker 2008 from the Caribbean Sea (WA) (Baeza and Anker 2008), L. 89 jundalini Rhyne, Calado and Dos Santos, 2012 from Puerto Rico (WA) (Rhyne et al. 90 2012), L. udoi Baeza et al. 2009 from Venezuela (WA) (Baeza et al. 2009), L. baueri 91 Prakash and Baeza 2017 from Mexico (WA) (Prakash and Baeza 2017), Lysmata 92 napoleoni De Grave and Anker, 2018, from St. Helena (CA) (De Grave and Anker 93 2018) and Lysmata arvoredensis Girardes, Macedo, Brandão, Baeza and Freire, 2018 94 from Brazil (WA) (Giraldes et al. 2018).

95 In addition to the five species reported from the EA and the M, two alien 96 Lysmata species originating from the Indian Ocean have been reported for the M: L. 97 kempi Chace, 1997 (Froglia and Deval 2014) and L. vittata (Stimpson, 1860) 98 (Abdelsalam 2018). It is also worth highlighting that in the WA, namely in Brazil, the

2 99 alien species L. vittata has also been recorded and it was, at first, erroneously reported 100 as a new species: L. rauli Laubenheimer and Rhyne, 2010 (Soledade et al. 2013).

101 On what concerns the Atlantic waters of the Iberian Peninsula, only one species of 102 Lysmata has ever been officially documented: L. seticaudata (see Zariquiey Álvarez 103 1968; d’Udekem d’Acoz 1999). Another species that is also known to occur in this 104 region is L. nilita. This species has been recorded and collected in underwater caves at 105 Sagres in the Algarve (Ilhotes do Martinhal, 37º00'56"N 8º55'01"W) (southern 106 Portugal), but this finding has never been reported in the scientific literature (P. 107 Chevaldonné and R. Calado, personal observations). In the present study, we report the 108 occurrence of a third species of Lysmata in Atlantic Iberian waters: L. uncicornis. This 109 species that, was formerly solely known to occur in the Atlantic coasts of Morocco, has 110 expanded its range northward, from Morocco to the Gulf of Cadiz and the Algarve (SW, 111 Iberian Peninsula), likely as a consequence of ongoing ocean global warming with 112 potential implication on marine fauna conservation. 113 114 115 Material and Methods 116 Two specimens of L. uncicornis were collected in the coastal waters of Cadiz 117 (Spain) by traps for shrimp fishery at the breakwater pier close to the Naval Station 118 Rota (36°37'28.1"N 6°20'44.7"W) and one in Valdelagrana beach (36°33'24.5"N 119 6°13'45.8"W) in November 1997, five in the coastal waters of Portimão in the inner part 120 of the eastern breakwater pier at the entrance to Portimão harbour (37°06'33.30", N 121 8°31'26.50"W) in June 2017, and five at the breakwater pier at the entrance to Cadiz 122 harbour (36°32'33.05" N, 6°17'15.54"W) in January 2018 (Figure 1). 123 The specimens of L. uncicornis and L. seticaudata from Cadiz (Spain) and 124 Portimão (Portugal) examined in the present study were identified according to Holthuis 125 and Maurin (1952), Lagardère (1971), Chace (1997), Baeza and Anker (2008), Froglia 126 and Deval (2014), Anker and Cox (2011), Giraldes et al. (2018) and Abdelsalam (2018), 127 and photographed (Figure 3). Two specimens were deposited in the and 128 Stomatopoda Collection (Spanish Institute of Oceanography in Cadiz, 129 Spain) under catalogue numbers IEO-CD-CAD18/2463-2464 (see Table 1). 130 Two paratypes of L. uncicornis from Naturalis Biodiversity Centre, Leiden 131 (RMNH.CRUS.D.7812) collected in Casablanca, Morocco by Maurin in 1951, were 132 comparatively studied with Iberian L. uncicornis and sequenced (Table 1). 133 Total genomic DNA was extracted from pereiopod and/or pleon muscle tissue of 134 two specimens of Lysmata uncicornis and one of L. seticaudata, collected in Cadiz 135 harbour, and paratypes of L. uncirconis, following a modified Chelex 10% protocol by 136 Estoup et al. (1996). Partial sequences of the mitochondrial 16S and Cox1 genes were 137 amplified. Cycling conditions of the polymerase chain reaction (PCR), length of the 138 sequences obtained, and primers used for each gene are the same that in previous works 139 (see Perez-Miguel et al. 2019). PCR products were sent to Stab-Vida laboratories to be 140 purified and then bidirectionally sequenced. 141 Sequences were edited using the software Chromas v. 2.0. The final DNA 142 sequences obtained were compared with those of several Lysmata species retrieved 143 from the Genbank database. New sequences of 16S and Cox1 are deposited in Genbank 144 under the accession numbers MN294750 to MN294752, MN296350, MT002801, and 145 MT002837. 146 An evolutionary distances analysis was carried out in MEGA6 (Tamura et al. 147 2013), for concatenate 16S and Cox1 sequences obtained in the present study from

3 148 specimens of L. uncicornis and L. seticaudata, as well as the sequences of other selected 149 group of Lysmata species downloaded from GenBank (http://www.ncbi.nlm.nih.gov), 150 that according to the phylogeny by Giraldes et al. (2018) cluster with L. arvoredensis 151 and as outgroup. The phylogenetic reconstruction analyses were inferred from 152 neighbour-joining using the p-distance method. The nodal confidence of the obtained 153 topologies was assessed via 2000 bootstrap replicates. 154 155 156 Results 157 158 Local fishermen in the harbours of Cadiz (Spain) and Portimão (Algarve, 159 Portugal) had observed that non-native shrimps similar in appearance to L. uncicornis 160 had been collected before in traps prior to the present study, but were likely mistaken as 161 L. seticaudata. Since 2006, Portuguese companies trading ornamental species for 162 marine aquariums have been regularly collecting L. uncicornis in the piers and rocky 163 shores around the Portimão area, along with L. seticaudata, as both species occur in 164 sympatry (Figure 2). These companies have been supplying both Lysmata species to 165 buyers all over Europe and the USA, with L. uncicornis being popularly known as 166 candy-cane cleaner shrimp. The average proportion of L. seticaudata/L. uncicornis 167 recorded in baited traps used to capture them from the wild has varied from 100:1 to 168 less than 1000:1. 169 170 The most relevant features for assessing the identity of L. uncicornis compared 171 to native Lysmata spp. species present in the EA and M are: (1) the semi-translucent 172 bodies with conspicuous transverse red bands (Figure 2, 3a, 3b); (2) the dorsal 173 antennular flagellum with the presence of the antennule accessory branch with a single 174 segment (Figure 3d); the absence of a pterygostomian spine in the anterior end of the 175 carapace (Figure 3c); (3) the antennular peduncle with stylocerite reaching nearly to or 176 beyond distal end of basal segment; (4) the antennal scale 3 times as long as wide; (5) 177 two teeth of dorsal rostral series situated on the carapace posterior to orbital margin 178 (Figure 3c) and (6) the second pereiopod with 19-28 carpal articles. 179 180 DNA sequences of 16S generated from the specimens collected in the present 181 study fit 99.4-99.8% with those of Lymata arvoredensis and Lysmata uncicornis 182 paratype, only varying on 1 or 2 mutations in 532 bp. Although the authors describing 183 L. arvoredensis did not include any sequences of gene Cox1 in their publication, they 184 did upload an unpublished sequence of this gene belonging to Lysmata arvoredensis to 185 Genbank (MF380416) named as Lysmata sp.. When we compare the Cox1 sequence of 186 L. uncircornis from Cadiz with that downloaded from Genbank, they fit 99.8%, with 187 only one mutation in 626 bp. Moreover, when we compare both of these sequences 188 from Cox 1 with those obtained from the paratype of L. uncicornis there is also only one 189 mutation when contrasted with Lysmata sp. (= L. arvoredensis) and two mutations for 190 L. uncicornis originating from Cadiz in 626 bp. In the final tree (assembled for 16S and 191 Cox1), all sequences of L. uncicornis (from Cadiz and the paratype from Casablanca) 192 cluster together within a clade with L. arvoredensis, well separated from the rest of all 193 other Lysmata species included in the analysis (Figure 4). 194 195 Discussion 196 197

4 198 Among the species of Lysmata present in the EA with a reduced or poorly developed 199 accessory branch on the lateral antennular flagellum (Holthuis and Maurin 1952; 200 Crosnier and Forest 1973; Fransen 1991; Chace 1997) we can find L. stenolepis, L. 201 olavoi and the amphi-Atlantic L. grabhami, in addition to L. uncicornis. However, as 202 Baeza and Anker (2008) mentioned, both L. stenolepis and L. olavoi are deep-water 203 forms (occurring at more than 120 m deep), while L. uncicornis inhabits shallow rocky 204 waters. Besides, L. uncicornis differs from L. olavoi in the absence of a pterygostomial 205 tooth and a postero-lateral tooth on the fourth abdominal pleura (Fransen 1991). In L. 206 stenolepis the rostrum bears only one small ventral tooth, while in L. uncicornis there 207 are a variable number of ventral teeth (1-3); these species also differ in the proportions 208 of their pereiopods and the armature of the dactylus of their third to fifth pereiopods 209 (Crosnier and Forest 1973; Fransen 1991). The differences to L. grabhami, namely on 210 live specimens, are too conspicuous, as this species is yellow with a continuous 211 middorsal white stripe from the tip of the rostrum to the end of the telson and, at both 212 sides of this one, present a dorsal longitudinal red band (see Kassuga et al 2015). 213 214 Giraldes et al. (2018) mentioned that the stylocerite of L. uncicornis exhibits a 215 series of denticules in its outer margin, while that of L. arvoredensis exhibited none. 216 However, the original description of L. uncicornis mentioned “In some specimens the 217 lateral edges and more especially the outer margin of the stylocerite are denticles near 218 the end of the latter (“Chez quelques specimens les bords latéraux et plus spécialement 219 le bord externe du stylocérite sont lenticules près de l'extrémité de celui-ci.” see 220 Holthuis and Maurin 1952, page 199). In the same sense, the two paratypes of L. 221 uncicornis and the five specimens collected in Cadiz showed a variation with respect to 222 the presence or not of denticles in both margins, with some specimens even showing a 223 bifid stylocerite tip (Figure 3e, f). In addition, specimens from Cadiz exhibited mesial 224 setae in the inner margin of this appendage, as described for L. arvoredensis. 225 The same authors referred that the second pereiopod of L. arvoredensis shows 226 11–16 subsegments in the merus and 22–24 subsegments in the carpus, in the range of 227 L. uncicornis 11-14 and 19-28 articles, respectively, (Holthuis and Maurin 1952), and in 228 specimens collected in Cadiz show 13 and 21-23 respectively. Giraldes et al. (2018) 229 also refer that the accessory branch of the antennular flagellum cannot be 230 distinguishable before the unguiform free segment in L. uncicornis (see Fig. 1d in 231 Holthuis and Maurin 1952), while it is distinguishable in L. arvoredensis (Giraldes et al. 232 2018). However, this is an artefact created by the orientation of the drawing, as in the 233 outer face it is indeed distinguishable, but not in its inner face. It has also been referred 234 by the authors describing L. arvoredensis that the number of setae in the ventral margin 235 of the propodus in pereiopods 3, 4 and 5 are different for both species (6-8, 6-8, 5 in L. 236 uncicornis, while these are 10–12, 9–12, and 9–12 in L. arvoredensis). In the specimens 237 surveyed in the present work, as well as in the paratype of L. uncicornis, the number of 238 setae in the ventral margin of the propodus in pereiopods 3, 4 and 5 is within this range 239 (the most common sequence from Cadiz specimens being 11, 9-12, 9 and 12, 11-12, and 240 8-11 in the paratypes of L. uncicornis) to that of L. arvoredensis. Thus, the discrepancy 241 between the specimens from Brasil and those early recorded from Morocco are likely 242 due to the fact of Holthuis and Maurin (1952) having recorded their observations using 243 an observation plane (posterior border, page 200 in Holthuis and Maurin 1952) that did 244 not allow them to see the spines of a second row that are practically transparent and 245 somehow masked by a pereiopod. Finally, Giraldes et al. (2018) also refer that the 246 length of the first pereiopod, exceeds the scaphocerite by nearly the length of the 247 dactylus in L. uncicornis (according Holthuis and Maurin description), while this does

5 248 not occur in L. arvoredensis. When analyzing L. uncicornis specimens collected at 249 Cadiz, it was possible to see that the length of this appendage exceeds lightly (i.e Cadiz 250 specimens), or was at least similar, to that cited for L. arvoredensis (i.e Rota specimen). 251 Thus, it seems that this feature may likely be a size‐ dependent morphological trait. 252 Baeza and Anker (2008) mentioned that L. uncicornis appears to be 253 morphologically close to L. hochi. Lysmata uncicornis exhibits the merus of the third 254 pereiopod with five spines, three spines on the flexor margin of the dactylus of the third 255 to fifth pereiopod (Figure 3g) and a lesser number of segments (11) in the merus of the 256 second pereiopod. By contrast, the merus of the third pereiopod of L. hochi has two to 257 four spines, two spines on the flexor margin of the dactylus of the third to fifth 258 pereiopod and a higher number of segments (15) in the carpus of the second pereiopod. 259 In addition to these features pointed by Baeza and Anker (2008), the dorsal rostral 260 formula of L. hochi is 2+3, while that of L. uncicornis is 2+4–5. We would like to 261 highlight, that the flexor margin of the dactylus of the third to fifth pereiopod in L. 262 uncicornis varies between three and four spines (Figure 3g,h), even in the two analyzed 263 paratypes (Figure 3i); in addition, while the bifid stylocerite tip has been suggested to be 264 a key diagnosing feature for L. hochi, it has also been recorded in some specimens of L. 265 uncicornis. As such, this feature should be used with caution when aiming to confirm 266 the morphological identification of L. hochi. 267 268 The DNA and morphology of L. uncicornis paratypes and Cadiz specimens, as 269 well as the live coloration of Cadiz specimens, do not exhibit any differences to L. 270 arvoredensis. Therefore, based on all these evidences, we recommend L. arvoredensis 271 to be considered a junior synonym of L. uncicornis. At this point there are two 272 possibilities to explain the occurrence of L. uncicornis in Brazilian waters: 1) a new 273 amphi-Atlantic Lysmata species, much like L. grabhami and several other decapod 274 species (Wirtz 2004; Almeida et al. 2013); or 2) a recent introduction of this species in 275 Brazil, as the two specimens of L. arvoredensis reported were found in a region that is 276 commonly monitored (e.g. Bouzon and Freire 2007; Teschima et al. 2012) and were 277 present in an artificial structure deployed for a few months at the collection site 278 (Giraldes et al. 2018). This misidentification would be similar to that reported for L. 279 vittata, which was incorrectly described as a new species (L. rauli) by Laubenheimer 280 and Rhyne (2010). The erroneous naming of that “new” Lysmata species was based on a 281 unique specimen collected in Salvador, Bahía (Brazil), when in fact it should have been 282 reported as an introduction of a congeneric species from the Indo-Pacific (Soledade et 283 al. 2013). 284 285 Distribution 286 Lysmata uncicornis is distributed from Morocco to Congo, in the west Atlantic African 287 coast (Holthuis and Maurin 1952; Lagardère 1971). The specimens collected in the Gulf 288 of Cadiz (between Cadiz and Portimão) and those ones collected in the south coast of 289 Brazil (as L. arvoredensis by Giraldes et al. 2018) represent the first occurrences of L. 290 uncicornis, outside of its original distribution, Atlantic African waters. As referred 291 above, earlier observations by fishermen suggest that this species has been present for 292 several years in the SW Atlantic waters of the Iberian Peninsula. Other decapod 293 crustacean species such as Ogyrides rarispina Holthuis, 1951, Brachynotus atlanticus 294 Forest, 1957,Xaiva mcleayi (Barnard, 1947) and Afropinnotheres monodi Manning, 295 1993, support the known African influence in this geographic region (Hothuis 1977; 296 García Raso 1985; García Raso and Manjón-Cabeza 1996; Subida et al. 2011). 297 Moreover, the distribution of several marine organisms is currently shifting in latitude,

6 298 likely as a response to a changing climate, with the present finding likely being an 299 example of an ongoing process of tropicalization (Cuesta et al. 2016; Encarnação et al. 300 2019). However, the recent record in the south coast of Brazil (as L. arvoredensis by 301 Giraldes et al. 2018) could also be related with human-mediated introduction of non- 302 native species. Shrimps of genus Lysmata are certainly a trademark of the marine 303 aquarium trade industry (Calado 2008) and the novelty of the attractive colours of L. 304 uncicornis, along with its limited offer and high retail value may foster the trade of this 305 species worldwide. Thus, as already described for many other taxa (Padilla and 306 Williams 2004), the introduction of non-native species via the global marine aquarium 307 trade should not be overlooked. 308 309 The present study signalling the presence of L. uncicornis in the SW of Iberian 310 coasts increases the number of species of this genus occurring in European waters. The 311 habitat of L. uncicornis is likely to be the same of L. seticaudata, as both species occur 312 in sympatry, but their feeding habits may differ, and additional studies are required to 313 see if any type of interspecific competition occurs. Lysmata uncicornis is less prone to 314 being stocked in captivity than L. seticaudata, but it can be easily induced to spawn and 315 cultured through its larval stages using available protocols (e.g., Calado et al. 2005; 316 Calado et al. 2007). Future studies should compare the morphology of L. uncicornis 317 larvae raised in the laboratory with earlier studies that sampled Lysmata larvae from the 318 plankton in adjacent waters to the study region (Caroli 1918; Kurian 1956; Bourdillon- 319 Casanova 1960; Barnich, 1996), in order to determine for how long may have this 320 species potentially remained unnoticed. 321 322 Conservation 323 As already reported for other marine invertebrates (e.g. Perez-Miguel et al. 324 2019), it is expected that Atlantic African species, such as L. uncicornis, spread their 325 distribution along the west Atlantic coasts of Europe. These new distribution patterns 326 are prompted by ongoing climate change, as it expands the extent of suitable habitat for 327 tropical species in European waters. Although the tropicalization of temperate marine 328 ecosystems is unavoidable, the spread of non-native species is now a hot issue at a 329 global scale (Dawson et al. 2017) and is at the forefront of research in conservation 330 (Bonanno and Orlando-Bonaca 2019). An unprecedented level of human activities in 331 coastal waters, including the marine aquarium trade, may increase the risks of invasion 332 (Calado 2008). Preventing the occurrence of non-native species is regarded as the 333 cornerstone of non-native species management, and management efforts, which control 334 non-native species in the marine realm, are needed to prevent new introductions and the 335 establishment of these non-native species (see Chan et al. 2019). 336 337 Acknowledgments 338 339 MG wish to thank to his uncle José Lazaro for the teaching in shrimp fishing and 340 dedicate this paper to his memory. We also thank Charles Fransen, Bram van der Bijl 341 and Wendy van Bohemen from Naturalis Biodiversity Centre for the loan specimens 342 and the editors Dr. Sammy De Grave and Lena Menzel for their useful comments. 343 344 Funding information 345 346 Financial support was given by CSIC through Intramural Research program 2018 under 347 grant number 201830I081. Thanks are also due for the financial support to CESAM

7 348 (UID/AMB/50017/2019), to FCT/MEC through national funds, and the co-funding by 349 the FEDER, within the PT2020 Partnership Agreement and Compete 2020. 350 351 Compliance with ethical standards 352 353 Conflict of interest The authors declare that they have no conflict of interest. 354 355 Ethical approval This article does not contain any studies with performed by 356 any of the authors. 357 358 Sampling and field studies The necessary permit for sampling has been obtained by 359 the authors from the competent authorities as mentioned in the acknowledgements. 360 361 Data availability All data generated or analysed during this study are included in this 362 published article 363 364 References 365 Abdelsalam K (2018) First record of exotic Lysmatid shrimp Lysmata vittata 366 (Stimpson, 1860) (Decapoda: Caridea: Lysmatidae) from the Egyptian 367 Mediterranean coast. Mediterr Mar Sci 0: 124-131. 368 Almeida AO, Terossi M, Araujo-Silva CL, Mantelatto FL (2013). Description of 369 Alpheus buckupi spec. nov., a new amphi-Atlantic snapping shrimp (Caridea: 370 Alpheidae), based on morphological and molecular data. Zootaxa 3652(4):437-452. 371 Anker A, Cox D (2011) A new species of the shrimp genus Lysmata Risso, 1816 372 (Crustacea, Decapoda) from Guam. Micronesica 41:197-214. 373 Baeza JA, Anker A (2008) Lysmata hochi n. sp., a new hermaphroditic shrimp from the 374 southwestern Caribbean Sea (Caridea: Hippolytidae). J Crustacean Biol 28:148-155. 375 Baeza JA, Bolaños JA, Hernandez JE, López R (2009) A new species of Lysmata 376 (Crustacea, Decapoda, Hippolytidae) from Venezuela, southeastern Caribbean Sea. 377 Zootaxa 2240:60-6. 378 Barnich R (1996) The Larvae of the Crustacea Decapoda (excl. Brachyura) in the 379 Plankton of the French Mediterranean Coast (Identification Keys and Systematic 380 Review). PhD dissertation, Cuvillier Verlag, Götingen, Germany, 189 pp. 381 Bourdillon-Casanova L (1960) Le meroplancton du Golfe de Marseille: les larves des 382 crustace´s decapodes. Rec Trav St Mar Endoume 30:1–286. 383 Bonanno G, Orlando-Bonaca M (2019) Non-indigenous marine species in the 384 Mediterranean Sea—Myth and reality. Environ Sci Technol 96:123-31. 385 Bouzon JL, Freire AS (2007) The Brachyura and Anomura fauna (Decapoda; 386 Crustacea) in the Arvoredo Marine Biological Reserve on the southern brazilian 387 coast. Braz J Biol 67:321-325. 388 Calado R (2008) Marine ornamental shrimp: biology, aquaculture and conservation. 389 Wiley-Blackwell, Hoboken, p 262 390 Calado R, Chapman CM (2006) Aquarium species: deadly invaders. Mar Pollut Bull 391 52:599–601 392 Calado R, Lin J, Rhyne AL, Araujo R, Narciso L (2003) Marine ornamental decapods 393 — popular, pricey, and poorly studied. J Crustacean Biol 23:963–973 394 Calado R, Figueiredo J, Rosa R, Nunes ML, Narciso L (2005) Larval culture of Monaco 395 shrimp Lysmata seticaudata (Decapoda: Hippolytidae): effect of temperature, 396 rearing density and larval diet. Aquaculture 245:221-237.

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11 Figure captions

Figure 1. Worldwide distribution of Lysmata uncicornis in the Atlantic coasts. Data from Holthuis and Maurin (1952), Lagardère (1971), Giraldes et al. (2018), and present study (inset).

Figure 2. Lysmata uncicornis (top and right) and L. seticaudata (left) specimens collected in Portimão (Algarve, Portugal). Scale bar = 1 cm.

Figure 3. Lysmata uncicornis, a-h, specimens collected in the Gulf of Cadiz (Spain), i, paratype RMNH.CRUS.D.7812 (Holthuis and Maurin, 1951). a, Dorsal view: specimen showing semi-translucent body with transverse red bands; b, Lateral view; c, Rostrum details; d, Dorsal antennular flagellum with trace of accessory branch; e,f stylocerite, dorsal view; g,h, dactyls of pereiopod 3, lateral view;i, dactyls of pereiopod 3 from paratype, lateral view. Scale bars, (a–c) = 1 cm; (d, e, g–i) = 1 mm; (f, h) = 2 mm. Photographs by E. González-Ortegón.

Figure 4. Topology of neighbour-joining tree based on 16S and Cox1 genes concatenated sequences, showing inferred phylogenetic relationships within a selected group of Lysmata species. Numbers close to nodes indicate bootstrap support (only values above 70% shown). After the name of each species the accession number of sequences in Genbank is included (16S/Cox1).

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