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Research Article
Eusociality unveiled: discovery and documentation of two new eusocial shrimp species (Caridea: Alpheidae) from the Western Indian Ocean
expand article infoHossein Ashrafi, Kristin M. Hultgren§
‡ University of Ostrava, Ostrava, Czech Republic
§ Seattle University, Seattle, United States of America
Open Access

Abstract

The alpheid snapping shrimp genus Synalpheus Spence Bate, 1888, is a prominent component of arthropod diversity found in coral reefs. Notably, Synalpheus is the only genus of marine organisms known to exhibit eusocial behavior. Although eusociality has evolved at least four times independently in Synalpheus, all described eusocial species are from the West Atlantic, with only a single study documenting possibly eusocial species from Indonesia. In 2008, during an expedition to Madagascar organized by the Florida Museum of Natural History (FLMNH), a diverse array of sponge-dwelling species was collected, including two species of Synalpheus exhibiting colonial behavior. Through detailed examination of these specimens, we have confirmed that these two species are eusocial and represent new eusocial species of Synalpheus outside of the West Atlantic. Consequently, we provide the first official documentation of eusocial species from the Western Indian Ocean and present their taxonomic descriptions and their phylogenetic relationships with other species of the genus in this study.

Keywords

Synalpheus, phylogeny, sponge-dwelling, Madagascar, new species, S. gustavi sp. nov., S. sponjy sp. nov.

1. Introduction

The snapping shrimp in the genus Synalpheus Spence Bate, 1888, comprise a major component of the crustacean diversity within coral reefs (Coutière 1909; Banner and Banner 1975, Chace 1988, Anker et al. 2012; Anker and De Grave 2016). Remarkably, it stands as the sole genus among all marine animals to exhibit eusocial living systems (Duffy 1996, 2007). Eusociality was first discovered by Duffy in 1996, and since then, multiple occurrences of eusociality have been documented for several species in the Western Atlantic (Ríos and Duffy 2007; Hultgren et al. 2011; Hultgren and Brandt 2015; Ashrafi and Hultgren 2022). Duffy (2007) identified several key traits that define eusocial Synalpheus shrimps. These include the presence of colonies consisting of multiple generations with pronounced reproductive skew and division of labor, such as cooperative defence and cooperative care of offspring. All eusocial species (in which larvae have been characterized) also have exclusively direct development. That is, eggs hatch directly into crawling juveniles who tend to stay in the sponge, instead of planktonic larvae, allowing for the establishment of multi-generational colonies within the same host sponge. Eusocial species typically have a reduced number of eggs produced by the queen and larger egg size (Duffy 1996, 2007; Hultgren et al. 2021), relative to species with planktonic larvae who tend to have smaller and more numerous eggs.

Extensive research has delved into various aspects of this unique lifestyle, encompassing colony composition (Tóth and Bauer 2007; Duffy and Macdonald 2009; Chak et al. 2015), interactions between shrimps and sponges (Duffy et al. 2002; Tóth and Duffy 2008; Hultgren and Duffy 2010), phylogenetic relationships (Hultgren and Duffy 2011, for instance), and documentation of larger genomes in eusocial species (Jeffery et al. 2016; Chak et al. 2021; Hultgren et al. 2021). However, eusociality has been primarily studied in West-Atlantic species of Synalpheus, while little attention has been given to eusociality in other regions, such as the Indian Ocean and Pacific Ocean, with the exception of a single study conducted by Didderen et al. in 2006. This lack of exploration is primarily attributed to the fact that taxonomic studies on Indo-Pacific Synalpheus species were conducted before Duffy’s groundbreaking work in 1996 (instances of important taxonomic work in the Indo-West Pacific: Coutière 1905, 1909, 1921; De Man 1910, 1911; AH Banner 1959; Banner and Banner 1975, 1981, 1983). Nonetheless, traces of eusociality can be found in earlier studies, where skewed sex ratios were noted (Banner and Banner 1975; Banner and Banner 1981, 1983) and species were even named based on this distinct feature (e.g. S. paradoxus Banner and Banner, 1981).

Didderen et al. (2006) documented extremely large colonies (40-372 individuals) of males (non-ovigerous) individuals, with often only 1 or 0 obviously ovigerous females, in Synalpheus neptunus neptunus (Dana, 1852), living in sponges in Indonesia. They also documented large colonies of S. fossor (Paulson, 1875), S. hastilicrassus Coutière, 1905, and S. aff. neomeris (De Man, 1897), living in sponges from Indonesia. However, these species are also often collected as heterosexual pairs from other localities (i.e., Moorea, Guam, Papua New Guinea, New Caledonia, Australia, Madagascar, and Red Sea), and genetic data suggests many of these species represent cryptic species complexes (Hultgren et al. 2014; Hultgren, Ashrafi, unpublished data). Interestingly, Banner and Banner (1975) reported a colony of S. neptunus (Dana, 1852) consisting of a colony of 44 individuals with no females, which they designated as the subspecies S. neptunus germanus Banner and Banner, 1975. Genetic evidence suggests S. neptunus is a cryptic species complex with several different related species (Hultgren et al. 2014), and several specimens of S. neptunus are found alone or in heterosexual pairs, suggesting at least some of the cryptic species may exhibit a pair-living lifestyle. A thorough taxonomic revision of these cryptic species complexes—S. hastilicrassus, S. fossor, S. aff. neomeris, and S. neptunus—is needed to properly document the extent of eusociality in these Indo-West Pacific species.

In 2008, a comprehensive expedition to Madagascar was organized by the Florida Museum of Natural History (FLMNH: Gainesville, Florida, USA). The primary objective of this expedition was to study the biodiversity of the region. During this expedition, a substantial number of Synalpheus specimens were collected. In 2022, HA visited the collection and carefully studied the specimens belonging to the genus Synalpheus.

Notably, the expedition to Madagascar yielded a significant number of cryptic sponges, which were frequently sampled. This fortuitous circumstance provided an ample opportunity to study the Synalpheus species associated with these sponges. Among the inhabitants of the sponges, two distinct species of Synalpheus stood out, with each species represented by a substantial number of specimens collected from a single sponge. Interestingly, within each sponge, only a single ovigerous female was found, if any.

By conducting a meticulous examination of both species, it became evident that they had developed a eusocial living system, marking a remarkable discovery previously undocumented in the Western Indian Ocean. These two previously unknown species are now being formally described and have been incorporated into a multi-gene phylogenetic tree. This inclusion allows for the reconstruction of how many times eusociality has independently evolved outside of the Western Atlantic Synalpheus species.

2. Material and Methods

2.1. Phylogenetic Tree and taxon sampling

The materials for the present study were collected during several expeditions conducted by the Florida Museum of Natural History (FLMNH), Gainesville, Florida. These expeditions include Madagascar 2008, French Polynesia/Moorea 2008, Western Australia 2009, Guam 2010, and Saudi Arabia 2013.

The most recent comprehensive worldwide tree of Synalpheus (Hultgren et al. 2014) featured S. gustavi sp. nov. as an undescribed species (referred to as S. aff. brevifrons). However, S. sponjy sp. nov. was not included. To augment the existing tree, we sequenced multiple representatives of these species, and used existing data, from four genes: mitochondrial COI DNA (cytochrome oxidase I), mitochondrial 16S rRNA, 18S rRNA, and the nuclear PEPCK gene (phosphoenolpyruvate carboxykinase). Additionally, we extracted and sequenced DNA from museum specimens of several Synalpheus species (e.g. S. theano, S. sladeni, and S. anceps) that exhibit several morphological similarities to the new species, but were not included in the Hultgren et al. (2014) tree.

Table 1.

All species sequenced for phylogenetic analyses used in this study, including the genetics # (referenced in phylogenetic trees), locality, collection number, and GenBank accession numbers. For the complete list of the abbreviations used in the voucher numbers, see Supplementary data 1 in Hultgren et al. (2014). GenBank numbers in bold indicate sequences used in consensus tree.

Species Genetics# Locality Collection# COI PEPCK 16S 18S
Synalpheus africanus 592 São Tomé AA--06-215 KJ595031 KJ625140 KJ625020
Synalpheus cf. africanus B 595 Sao Tome OUMNH.ZC.2012-07-130 KJ595053 KJ625142 KJ595185 KJ595255
Synalpheus agelas 459 Belize CBC09-1508 KJ595032 HQ435419 KJ595245
Synalpheus anceps 2182 Guam FLMNH-UF 37335 OR482981 OR483005 OR483030
Synalpheus androsi 467 Belize CBC09-6502 KJ625033 HQ435421 KJ595246
Synalpheus ankeri 477 Panama P09-6903 KJ625041 KJ625069 HQ435473 KJ595242
Synalpheus apioceros 593 Panama RMNH.CRUST.D.54889 KJ595035 KJ625141 KJ595177 KJ595247
Synalpheus bannerorum 563, 634 Panama OUMNH.ZC.2014-04-027 KJ595036 KJ595178
Synalpheus belizensis 250 Barbados BR08-6901 KJ595038 KU682627
Synalpheus bituberculatus 1290 Singapore OUMNH.ZC.2014-11-190 OR482983 OR483007 OR483032
Synalpheus bituberculatus 1291 Singapore OUMNH.ZC.2014-11-190 OR482982 OR483006 OR483031
Synalpheus bocas 16 Jamaica JAM08-7402 KJ595041 KJ625107 HQ435426 KJ595249
Synalpheus bousfieldi 483 Belize CBC09-3605 KJ595042 KJ625073 KJ595250
Synalpheus brevifrons 481 Belize CBC09-2704 KJ625034 KJ625072 HQ435435 KJ595251
Synalpheus brooksi 476 Panama P09-3102 KJ595049 KJ625068 HQ435437 KJ595253
Synalpheus brooksi 479 Panama P09-4911 KJ625035 KJ625070 HQ435438 KJ595252
Synalpheus carpenteri 421 Jamaica JAM08-3008 KJ595052 HQ435439 KU682628
Synalpheus cayoneptunus 910 Florida VIMS 13FK2901, 2903 KM204166 KM204180 KU682629
Synalpheus chacei 457 Belize CBC09-501 KJ595059 KJ625062 HQ435440 KJ595265
Synalpheus chaki 2014_1228 Martinique MNHN-IU-2014-1228 MZ323456 MZ329362
Synalpheus charon A 565 Egypt OUMNH.ZC.2014-02-022 KJ595060 KJ625128 KJ595191 KJ595266
Synalpheus charon C 566 Taiwan FLMNH-UF 19914 KJ595064 KJ595192 KJ595269
Synalpheus corallinus 826 Jamaica JAM12-9902 KU980212 HQ435441 KU682630
Synalpheus corbariae 2014-1238 Martinique MNHN-IU-2014-1238 MZ323450 MZ329367
Synalpheus coutierei B 2164 French Polynesia FLMNH-UF 16178 OR482984 OR483008 OR483033
Synalpheus dardeaui 461 Belize CBC09-3105 KJ625036 KJ625110 HQ435442 KJ595271
Synalpheus dominicensis 482 Belize CBC09-2906 KJ477702 KJ494390 KJ477695 KU682632
Synalpheus duffyi 26 Jamaica JAM08-7403 (26), VIMS 08JAM7401-2 KJ595078 HQ435444 KU682633
Synalpheus elizabethae 944, 137 Panama P08-12504 (137) KU980213 HQ435446 KU682634
Synalpheus filidigitus 470 Belize CBC09-7603 KJ595079 KJ625066 HQ435447 KJ595275
Synalpheus fossor 570 Thailand OUMNH.ZC.2011-03-096 KJ595080 KJ595199 KJ595276
Synalpheus goodei 464 Belize CBC09-5404 KJ477698 KJ625065 HQ435448 KJ595279
Synalpheus guerini A 527, 541 Florida FLMNH-UF 9334 KJ595082 KJ595200 KJ595282
Synalpheus gustavi n. sp. 707 Madagascar FLMNH-UF 14255-1 KJ494394
Synalpheus gustavi n. sp. 2172 Madagascar FLMNH-UF 14231 OR482985 OR483010
Synalpheus gustavi n. sp. 2173 Madagascar FLMNH-UF 14259 OR482986 OR483011
Synalpheus gustavi n. sp. 2175 Madagascar FLMNH-UF 14255 OR482987 OR483012
Synalpheus gustavi n. sp. 2184 Madagascar FLMNH-UF 14257 OR483009
Synalpheus gustavi n. sp. 2180 Madagascar FLMNH-UF 14236 OR483013
Synalpheus hastilicrassus A Syn2119 Queensland FLMNH-UF 24728 OR482988 OR483015
Synalpheus hastilicrassus A 678 Queensland FLMNH-UF 17136 KJ625055 KJ625098 KJ595284
Synalpheus hastilicrassus B 596 Queensland FLMNH-UF 18204 KJ595089 KJ625143 KJ595206 KJ595287
Synalpheus hastilicrassus C Syn2111 Hong Kong FLMNH-UF 54147 OR482989 KJ625134 OR483016 KJ595288
Synalpheus hastilicrassus_D Syn2109 Guam FLMNH-UF 37332 OR482990 OR483017
Synalpheus hastilicrassus_F Syn2118 Queensland FLMNH-UF 18256 KJ625061 KJ625134 OR483014 KJ595288
Synalpheus hemphilli 571 Florida FLMNH-UF 19640 KJ595092 KJ595208 KJ595290
Synalpheus herricki 157 Curacao CU08-3202 KJ595095 HQ435449 KU682635
Synalpheus hoetjesi 202 Curacao CU08-2901 KJ625037 KJ625076 HQ435452 KJ595293
Synalpheus idios 474 Belize CBC09-8803 KJ625038 KJ625115 HQ435455
Synalpheus iocasta 1284 Singapore OUMNH.ZC.2014-11-219 OR482991 OR483018 OR483034
Synalpheus irie 38, 493 Jamaica JAM08-3601 KJ595106 KJ625117 HQ435457 KJ595294
Synalpheus kensleyi 504 Panama P07-1204 KJ625039 KJ625119 HQ435458 KJ595295
Synalpheus kuadramanus 843 Jamaica VIMS 12JAM9501 KU980214 MZ329378 KU682637
Synalpheus lani 599 Panama OUMNH.ZC.2014-04-030 KJ595107 KJ625145 KJ595210 KJ595296
Synalpheus aff. longicarpus 488 Panama P09-9101 KJ595025 KJ625075 HQ435459 KJ595240
Synalpheus macdonaldi 2014_1227 Martinique MNHN-IU-2014-1227 MZ323448 MZ329364
Synalpheus mcclendoni 237 Barbados BR08-1413 KJ595109 HQ435462 KU682639
Synalpheus microneptunus 247 Barbados BR08-6001 KJ595110 KJ625108 HQ435463 KU682640
Synalpheus aff. sanctithomae 471 Belize CBC09-7804 KJ625043 KJ625114 HQ435464 KJ595243
Synalpheus aff. sanctithomae 486 Belize CBC09-8701 KJ595029 KJ625074 HQ435465 KJ595244
Synalpheus neomeris_A 1133 Saudi Arabia FLMNH-UF 36916 OR482992 OR483019 OR483035
Synalpheus neomeris_A 573 Madagascar FLMNH-UF 12540 KJ595056 KJ595188 KJ595260
Synalpheus neptunus A 560 Queensland FLMNH-UF 17566 KJ595119 KJ625126 KJ595217
Synalpheus neptunus B 1119 Queensland FLMNH-UF 23841 OR482994 OR483021
Synalpheus cf. neptunus germanus 1117 Western Australia FLMNH-UF 28020-7 OR482993 OR483020 OR483036
Synalpheus nobilii 576 Panama OUMNH.ZC.2013-03-009 KJ595122 KJ625132 KJ595219 KJ595303
Synalpheus obtusifrons 465 Belize CBC09-6303 KJ477703 KJ494389 HQ435466 KJ494396
Synalpheus pandionis 472 Belize CBC09-8403 KJ595126 KJ625067 HQ435468 KJ595305
Synalpheus parfaiti 609 Sao Tome MNHN-IU-2010-4150 KJ595127 KJ595223
Synalpheus pectiniger 500 Jamaica JAM08--8801 KJ595129 KJ625118 HQ435470 KJ595307
Synalpheus peruvianus 601 Panama OUMNH.ZC.2013-03-056 KJ595132 KJ625147 KJ595224 KJ595310
Synalpheus plumosetosus 231 Jamaica JAM08-2704 KU980220 HQ435471
Synalpheus rathbunae 941 Panama P08--3501-2 KU980221 AY344767 KU682644
Synalpheus regalis 469 Belize CBC09-7002 KJ625042 KJ625113 HQ435474
Synalpheus ruetzleri 466 Belize CBC09-6201 KJ595136 KJ625112 HQ435475 KJ595313
Synalpheus sanctithomae 235 Barbados BR08-1201 KJ595139 AY344768 KU682647
Synalpheus sanlucasi 654 Panama OUMNH.ZC.2013-03-015 KJ625049 KJ625089 KJ595225 KJ595317
Synalpheus scaphoceris 683 Panama MNHN-IU-2010-4152 KJ625058 KJ595226 KJ595318
Synalpheus sladeni 1137 Madagascar FLMNH-UF 14474 OR482995 OR483022 OR483037
Synalpheus somalia aff. 2186 Saudi Arabia FLMNH-UF 36964 OR482996 OR483023 OR483038
Synalpheus spinifrons 611 Chile AA--07-317 KJ595145 KJ595228 KJ595321
Synalpheus sponjy n. sp. 2178 Madagascar FLMNH-UF 14228 OR482998 OR483025 OR483039
Synalpheus sponjy n. sp. 2176 Madagascar FLMNH-UF 14256 OR482997 OR483024
Synalpheus sponjy n. sp. 2179 Madagascar FLMNH-UF 14316 OR482999 OR483026
Synalpheus stimpsonii_A 553 Queensland FLMNH-UF 16622 KJ595147 KJ595229 KJ595323
Synalpheus stimpsonii_B 554 Queensland FLMNH-UF 16982 KJ595148 KJ595230 KJ595325
Synalpheus stimpsonii_C 1109 Queensland FLMNH-UF 16788 OR483000 OR483027 KJ595326
Synalpheus streptodactylus A 584 Madagascar FLMNH-UF 13933 KJ595151 KJ595234 KJ595327
Synalpheus thai 1281 Singapore OUMNH.ZC.2014-11-263 OR483001 OR483028 OR483040
Synalpheus theano 1286 Singapore OUMNH.ZC.2014-11-266 OR483002 OR483029 OR483041
Synalpheus theano 1285 Singapore OUMNH.ZC.2014-11-264 OR483003
Synalpheus theano 2183 Western Australia FLMNH-UF 21825 OR483004
Synalpheus thele 113 Jamaica JAM08-8924 KJ595156 KJ625106 KJ595335
Synalpheus ul 253 Barbados BR08-8703 KJ625044 KJ625109 HQ435482 KU682648
Synalpheus williamsi 462 Belize CBC09-5102 KU980224 KJ625064 HQ435484 KJ595338
Synalpheus yano 484 Belize CBC09-3802 KJ595161 KJ625116 HQ435485 KJ595339
Alpheus percyi (outgroup) 0 FLMNH-UF 12372 KJ477697 KJ494392 KJ477694 KJ494395

2.2. DNA extraction, amplification, and sequencing

Tissue from the gills, eggs, and/or pleopods 3-5 were used for genomic DNA extraction using the QIAGEN DNeasy Blood and Tissue kit, following standard manufacturer protocols. We amplified the COI and 16S loci using PCR and thermocycler conditions described in Ashrafi and Hultgren (2022), and 18S described in Hultgren et al. (2014). PEPCK sequences were acquired from GenBank from a previous study (Hultgren et al. 2014). For COI, we used the primer pairs jgLCO/jgHCO (Geller et al. 2013), or polyLCO/polyHCO from Brasier et al. (2016). For 16S and 18S, we used the primers, PCR conditions, and thermocycler settings described in Hultgren et al. (2014). PCR products were cleaned up using EXO-SAP and sequenced for forward and reverse sequences at MCLab (South San Francisco, CA).

2.3. Tree Construction

Forward and reverse sequences were cleaned, aligned, and translated to amino acids to check for stop codons using Sequencher (Gene Codes, Ann Arbor, MI, USA) and MEGA v. 11 (Stecher et al. 2020). All new sequences were aligned with the existing sequences for the four loci (PEPCK, 18S, 16S, and COI) from Hultgren et al. (2014) and Chak et al. (2021) using MUSCLE (Edgar et al. 2004) implemented in MEGA X (Stecher et al. 2020). For the 18S alignment, we used GBLOCKS v. 0.91.1 (Castresana, 2000; Talavera and Castresana 2007) to exclude ambiguous areas of the alignment using relaxed criteria (allowed gap positions = with half, minimum length of a block = 5), yielding an alignment of 253 bp. For COI, we trimmed the alignment to ~660 bp at the 5’ end of the COI gene (corresponding to the standard barcoding region). For the final tree, we included all taxa with at least three out of the four loci sequenced (all taxa were sequenced for COI), to optimize taxon sampling. We used JModelTest ver. 2.1.10 (Darriba et al. 2012) to get the best model of nucleotide substitution using AIC, and implemented these models in a concatenated Bayesian analysis using MrBayes v.3.27 (Nylander et al. 2004; Ronquist et al. 2012) run on the CIPRES server (Miller et al. 2010). The Bayesian tree was run for 1×108 generations with four chains, and we discarded the first 25% of the samples as burn-in. As we generated many additional sequences of COI for new specimens that were not included in the consensus tree, we also ran a single-gene Bayesian tree using COI data for 3×108 generations (again discarding the first 25% of the samples as burn-in).

2.4. Abbreviations

CL – Carapace length; FLMNHFlorida Museum of Natural History, Gainesville, Florida.

3. Results

3.1. Phylogenetic trees

The consensus phylogenetic tree constructed in this study (Fig. 1) is based on a concatenation of 16S, CO1, 18S, and PEPCK sequences. We incorporated major clades or representative species from previous studies (Hultgren et al., 2014; Chak et al. 2021) and included new species as well as morphologically similar species to gain a better understanding of the phylogenetic relationships among eusocial species in the Indo-West Pacific region. However, due to the unavailability of fresh material, we were unable to include the other known eusocial species from the Western Indian Ocean, S. crosnieri Banner and Banner, 1983, and S. paradoxus Banner and Banner, 1981, in the present analysis. Additionally, we added Synalpheus cf. neptunus germanus, a potential eusocial species from the Eastern Indian and western Pacific oceans, to the phylogenetic analysis.

Figure 1. 

Phylogenetic tree constructed for the members of the genus Synalpheus using the Bayesian Inference approach based on a concatenation of four genes: 16S, COI, PEPCK, and 18S. Bayesian posterior probabilities are provided above each branch. Taxa are shown as Synalpheus species plus unique identifier (see supplementary table S1). Eusocial species are indicated with an arrow (red, WA= West Atlantic; blue, IWP = Indo-West Pacific).

The results of the phylogenetic study using Bayesian inference (Fig. 1) indicate that eusociality has independently evolved in each eusocial species from the Indo-Pacific region included in this analysis. Eusociality arose once in the S. neptunus clade, which comprises S. neptunus neptunus (Dana 1852), S. neptunus germanus Banner and Banner, 1975, and S. theano De Man, 1910. It also arose once in S. sponjy sp. nov., which is positioned as a sister to a clade consisting of S. sladeni Coutière, 1908, S. thai Banner and Banner, 1966, and S. aff. somalia Banner and Banner, 1979. Furthermore, eusociality emerged in S. gustavi sp. nov., which is not closely related to other species in the tree. Notably, the latter species holds an isolated position relative to all other Synalpheus species in the Indo-West Pacific.

The previously recognized S. gambarelloides group is no longer a monophyletic clade (possibly due to the lack of morphological characters). Several of its members are now positioned outside the traditional S. gambarelloides clade. Synalpheus rathbunae Coutière, 1909, group; S. paraneptunus Coutière, 1909, group; S. herricki Coutière, 1909; S. pectiniger Coutière, 1907; S. androsi Coutière, 1909; S. kensleyi (Ríos and Duffy 2007); S. parfaiti (Coutière, 1898); S. agelas Pequegnat and Heard, 1979; and S. mcclendoni Coutière, 1910 are the species situated outside the main body of the S. gambarelloides clade.

In the current phylogenetic study, we identify seven distinct lineages of eusociality: the entire S. rathbunae clade; appearing multiple times within the S. paraneptunus clade; within S. brooksi Coutière, 1909, species complex; as well as involving S. chacei; S. gustavi sp. nov.; S. sponjy sp. nov.; and at least two occurrences within the S. neptunus species complex. While S. crosnieri is not encompassed within the phylogenetic analysis, it is undoubtedly closely related to S. sponjy sp. nov. owing to numerous morphological similarities between the two species. Moreover, it is noteworthy that S. paradoxus might potentially introduce a new lineage of eusociality within the genus.

The COI Bayesian gene tree (Fig. S1) indicated that multiple sequenced specimens of the two new species are reciprocally monophyletic, with Bayesian posterior probabilities = 1.

3.2. Taxonomy

Infraorder Caridea Dana, 1852

Family Alpheidae Rafinesque, 1815

Genus Synalpheus Spence Bate, 1888

Synalpheus sponjy sp. nov.

Figures 2, 3, 4

Etymology

The new species derives its name from the Malagasy word ‘sponjy’, which translates to ‘sponge’. The term is used as a noun in opposition.

Material examined

Holotype : MADAGASCAR • 1 male (CL 3.6 mm); Nosy Be; 13°25’9.7”S, 48°15’37.94”E; 17 May 2008; Bakary G, Bruggermann H, Michonneau F, Paulay G, Werner T leg.; 6-9 m, from cryptic sponge; FLMNH-UF-14409. — Paratypes: MADAGASCAR • 19 males (CL ranging from 1.99 to 3.6 mm); Nosy Vorona; 15 May 2008; Paulay G leg.; 4 m, from cryptic sponge; FLMNH-UF-14256 • 2 males (CL 3.1-3.2 mm); same data as for preceding; FLMNH-UF-14228 • 1 male (CL 3.35 mm); same data as for preceding; Anker A, Bakary G, Boissin E, Bruggemann H, Horeau T, Michonneau F, Paulay G, Werner T leg.; FLMNH-UF-14313 • 1 male (CL 3.23 mm); same data as for preceding; FLMNH-UF-14316.

Description

Small-sized species of Synalpheus. Carapace (Fig. 2A, B) glabrous, unarmed; frontal margin between rostrum and orbital teeth relatively deep, U-shaped, with moderately developed rostrum; rostrum triangular in dorsal view, subacute at tip, about twice as long as wide, reaching to about proximal third of visible part of first antennular article, slightly shorter than orbital teeth, orbitorostral process present; orbital teeth relatively stout, subtriangular, distally subacute, as long as wide, about three times as broad as rostrum at base; pterygostomial angle bluntly protruding; cardiac notch deep.

Figure 2. 

Synalpheus sponjy sp. nov., holotype (FLMNH-UF-14409). A anterior region, dorsal view B same, lateral view, C pleon, D telson and uropods, E third maxilliped.

Male pleon (Fig. 2C) with first pleuron bearing small ventrally-pointing hook-shaped projection posteroventrally; third pleuron rounded anteroventrally and angled posteroventrally; fourth pleuron rounded anteroventrally and subrounded posteroventrally; fifth pleuron rounded antero- and posteroventrally; sixth pleuron with blunt projection on posterolateral margin.

Telson (Fig. 2D) subrectangular, tapering distally, slightly shorter than endopod, flat, without depression on mid-line; dorsal margin with two pairs of relatively strong spiniform setae located at 0.3 and 0.6 telson length, respectively; posterolateral angle with no projection; posterior margin broad, slightly convex, furnished with about 10 setae, with two pairs of spiniform setae, mesial spiniform setae slightly longer than lateral ones.

Antennula (Fig. 2A, B) with peduncle relatively stout; first antennular article almost twice as long as broad; stylocerite relatively short, subacute, falling short of distal margin of first antennular article; second antennular article slightly shorter than first one, slightly longer than broad; third antennular article slightly longer than second one, subequal to first one, slightly longer than broad; lateral flagellum biramous, fused portion consisting of five subdivisions, secondary ramus with three subdivisions, aesthetascs extending from third subdivision of fused portion to end of secondary ramus.

Antenna (Fig. 2A, B) with inferior tooth of basicerite subacute, slightly overreaching first antennular article, superior tooth small, subacute; scaphocerite relatively slender, lateral tooth overreaching end of second antennular article, lateral margin straight, blade fully absent; carpocerite somewhat slender, extending beyond antennular peduncle.

Mouthparts not dissected. Third maxilliped (Fig. 2E) pediform; lateral plate with small acute hooked-shaped projection pointing laterally; antepenultimate article about 4 times as long as widest part, mesial margin furnished with setae; penultimate article short, about a sixth of antepenultimate length; ultimate article relatively stout, about 0.6 times as long as antepenultimate article, slightly tapering distally, with crown of six spiniform setae at tip; exopod slender, reaching to about 0.8 of antepenultimate article length.

Major cheliped (Fig. 3A–D) with coxa bearing small spiniform seta on dorsodistal margin; basis and ischium short; merus robust, dorsal margin convex, about twice as long as broadest part, dorsodistal margin with blunt projection; carpus short, cup-shaped; palm swollen, ovoid, about 1.5 times as long as wide, dorsodistal margin with subrounded, prominent tubercle; fingers 0.4 times as long as palm, pollex subequal to dactylus. Minor cheliped (Fig. 3E, F) with coxa bearing small spiniform seta on dorsodistal margin; basis and ischium short; merus relatively robust, ventral margin concave, about 3 times as long as broadest part; carpus vase-shaped; palm ovoid, nearly 0.7 times as long as merus, about 0.7 times as long as wide; fingers subequal, about 0.8 times as long as palm; pollex with several tufts of setae situated on lateral side, simple at tip; dactylus bearing densely-situated tufts of distally-curved setae (known as gambarelloides setae), simple at tip.

Figure 3. 

Synalpheus sponjy sp. nov., holotype (FLMNH-UF-14409). A major cheliped, B same, chela, C lateral view same, D mesial view same, E fingers minor cheliped, F mesial view same, lateral view.

Second pereiopod (Fig. 4A) with coxa bearing small spiniform seta dorso-distally; basis short; ischium relatively robust, approximately 3.5 times as long as broad; merus relatively robust, 3.5 times as long as broad, about 1.3 times as long as ischium; carpus slightly longer than merus, composed of five articles, proximal article longest, as long as sum of three medial articles, second to fourth articles subequal, distal article twice as long as fourth one; palm subequal to fingers.

Figure 4. 

Synalpheus sponjy sp. nov., holotype (FLMNH-UF-14409). A second pereiopod, B third pereiopod, C lateral view same, mesial view, D fourth pereiopod, mesial view, E fifth pereiopod, lateral view.

Third pereiopod (Fig. 4B, C) robust, coxa with small spiniform setae on dorsodistal margin; ischium vase-shaped, 1.5 times as long as broadest part; merus robust, about 3 times as long as broad, about 2.8 times as long as ischium, distal half of ventral margin concave, mesial margin of concavity furnished with thick setae and lateral margin widened; carpus slightly widening distally, slightly longer than ischium, with spiniform seta on distoventral margin; propodus with five spiniform setae on ventral margin in addition to distal pair, about 1.7 times as long as carpus; dactylus biunguiculate, inferior unguis slightly wider at base than superior one, latter one slightly longer than former. Fourth pereiopod (Fig. 4D) generally similar to third pereiopod, less robust; coxa lacking spiniform setae on dorso-distal margin; merus concavity less prominent and mesial margin lacking thick setae. Fifth pereiopod (Fig. 4E) generally similar to third pereiopod, less robust; merus without concavity; carpus about 0.7 times as long as merus, without spiniform setae on distoventral margin; propodus subequal to merus, bearing four transverse rows of microserrulate setae on distal half.

Uropods (Fig. 2D) with protopods bearing two subacute teeth; exopod with small distolateral tooth; diaeresis inconspicuous except for lateral margin terminating to small tooth; distolateral spiniform setae next to diaresis stout, not overreaching posterior margin of exopod.

Type locality

North of Madagascar: Nosy Be and Nosy Vorona.

Ecology

All the specimens were collected from cryptic sponges situated among dead coral and coral rubbles.

Remarks

The new species exhibits similarities to several rare species found in the Indo-West Pacific region, characterized by a dense brush of setae on the dactylus of the minor cheliped, known as gambarelloides setae. These species include S. sladeni Coutière, 1908; S. spongicola Banner and Banner, 1981; S. crosnieri Banner and Banner, 1983; and S. gambarelloides (Nardo, 1847) [as reported in Banner and Banner (1983)]. Originally described from Cargados Carajos Shoals (Mauritius) and reported in the Gulf of Aqaba [Banner and Banner (1981)], S. sladeni, each time based on a single specimen, can be distinguished from the new species by several notable characteristics: slender orbital teeth, presence of a scaphocerite blade, less prominent inferior tooth and more prominent superior tooth of the basicerite, highly robust exopods of the second and third maxillipeds, a subacute tooth on the dorso-distal margin of the major chela palm, and complete presence of diaeresis.

According to Banner and Banner (1983), 45 males and 6 females, suggesting a potential communal lifestyle for the species, from Tuléar, Madagascar were reported and identified as S. gambarelloides, (for further details, see Banner and Banner 1983: page 97). However, no descriptions or illustrations were provided for these specimens, except for one distinguishing characteristic of S. gambarelloides, namely the absence of an orbitorostral process. This particular trait can be utilized to differentiate the new species from S. gambarelloides.

Synalpheus sponjy sp. nov. is closely related to the other two species, S. crosnieri and S. spongicola, both of which have not been reported since their original descriptions. The new species can be distinguished from S. crosnieri (found northwest of Madagascar) by the overall shape of the major cheliped fingers. In S. sponjy sp. nov., the major cheliped is normal and straight, whereas in S. crosnieri, it appears twisted when viewed dorsally. Additionally, the shape of the meri in the third and fourth pereiopods sets the new species apart. In the new species, these meri are concave on the distal half, and the third pereiopod merus bears a row of stiff setae along the mesial margin of the concavity. However, caution should be exercised when using the latter characteristic as the drawings provided by Banner and Banner (1983) are schematic, and the description lacks precise information regarding the exact shape of the meri in the third and fourth pereiopods.

One of the authors (HA) had the opportunity to examine the type series of S. spongicola deposited in the Naturalis Biodiversity Center, Leiden, Netherlands. As mentioned by Banner and Banner (1981), S. spongicola is a relatively large species of Synalpheus. In addition to differences in size, S. sponjy sp. nov. can be distinguished from S. spongicola based on the following characters: 1) the stylocerite falls short of reaching the end of the first antennular article in the new species, whereas in S. spongicola, it extends to the end; 2) the blade of the scaphocerite is absent in the new species, while it is complete in S. spongicola; 3) the palm of the major cheliped terminates in an obtuse, robust tubercle that points forward on the dorso-distal margin in S. sponjy sp. nov., whereas in S. spongicola, it ends in an upturned subacute tooth; 4) the last three pereiopods are more robust in the new species, and the meri of the third and fourth pereiopods are ventrally concave (lacking concavity in S. spongicola); and 5) in the new species, the diaeresis is confined to the lateral tooth, whereas in S. spongicola, the diaeresis is complete. Unfortunately, as these types were very old (date), they could not be included in the phylogenetic tree. Limited information is available regarding the ecology of S. spongicola, as it has been recorded only once with 9 females and 1 male from a single collection, suggesting a potential communal lifestyle for the species.

In the phylogenetic tree (Fig. 1), S. sponjy sp. nov. forms a clade with S. aff. somalia (Banner and Banner, 1979) and S. thai (Banner and Banner, 1966), along with S. sladeni. The new species can be easily distinguished from both S. somalia and S. thai by the densely-located (gambarelloides) setae on the dactylus of the minor cheliped. Additionally, S. thai stands out within the entire genus due to its distinctive shape of the telson (see Banner and Banner 1966: fig. 19L). Synalpheus somalia can be further differentiated from the new species by the dactylus of the major cheliped being much longer than the pollex (Banner and Banner 1979: fig. 3c, e).

Synalpheus gustavi sp. nov.

Figures 5, 6, 7

Synalpheus aff. brevifrons Hultgren, Hurt and Anker, 2014; Ashrafi and Hultgren, 2022.

Etymology

The new species is named after Gustav Paulay (FLMNH), who collected the majority of the specimens that contributed to the present study, and in recognition of his generous help and warm hospitality during the authors’ visit to the FLMNH collection.

Material examined

Holotype : MADAGASCAR • 1 male (CL 3.4 mm); Nosy Vorona; 15 May 2008; Paulay G leg.; 4m, in cryptic sponge; FLMNH-UF-71394. — Allotype: MADAGASCAR • 1 female (CL 3.6 mm); same data as for holotype; FLMNH-UF-71395. — Paratypes: MADAGASCAR • 15 males (CL ranging from 1.99 to 2.98 mm); from the allotype colony; FLMNH-UF-14259 • 4 males (CL ranging from 1.73 to 2.98 mm); from the holotype colony; FLMNH UF-14255 • 19 males and 0 females (CL ranging from 1.37 to 3.35 mm); same data as for holotype; FLMNH-UF-14257 • 1 female and 3 males (CL ranging from 1.73 to 2.85 mm); same data as for holotype; FLMNH-UF-14231 • 1 female and 3 males (CL ranging from 2.86 to 3.35 mm); same data as for holotype; FLMNH-UF-14230 • 1 male (CL 2.23 mm); Nosy Vorona; 15 May 2008; Anker A leg.; cryptic sponge; FLMNH-UF-14915 • 2 males (CL 2.85-2.98 mm); same data as for holotype; FLMNH-UF-14229 • 5 males (CL ranging from 2.36-3.48); same data as for holotype; FLMNH-UF-14236.

Description

Small-sized species of Synalpheus. Carapace (Fig. 5A, B) glabrous, smooth; frontal margin between rostrum and orbital teeth shallow, broadly U-shaped, with very small rostrum; rostrum triangular in dorsal view, subacute at tip, approximately as long as wide, slightly surpassing orbital teeth, orbitorostral process present; orbital teeth shallow, broad, distally subrounded, about four times as broad as long, about three times as broad as rostrum at base; pterygostomial angle bluntly protruding; cardiac notch relatively deep.

Figure 5. 

Synalpheus gustavi sp. nov., holotype (FLMNH-UF-71394). A anterior region, B dorsal view same, C lateral view pleon, D same, allotype, E telson and uropods, F third maxilliped.

Pleon only showing sexual dimorphism in first pleonite. Male pleon (Fig. 5C) with first pleuron angulate or with tiny projection posteroventrally; third to fifth pleuron rounded anteroventrally and posteroventrally. Female pleon (Fig. 5D) with first pleuron rounded posteroventrally.

Telson (Fig. 5E) subrectangular, tapering posteriorly, remarkably shorter than endopod, approximately as long as broadest part, dorsal margin with two pairs of relatively strong spiniform setae at 0.5 and 0.8 telson length (right pairs in holotype), respectively, without depression on mid-line; distolateral angle with no projection; posterior margin broad, straight, furnished with 8 setae, with two pairs of spiniform setae, mesial spiniform setae about twice as long as lateral ones and subequal to dorsal spiniform setae.

Antennula (Fig. 5A, B) with peduncle relatively stout; first antennular article about 1.7 times as long as broad; stylocerite relatively short, subacute at tip, falling short of reaching distal margin of first antennular article; second antennular article subequal to first one, slightly longer than broad; third antennular article shortest, about 0.8 times as long as first antennular article, slightly longer than broad; lateral flagellum biramous, fused portion consisting of five subdivisions, aesthetascs extending from third subdivision of fused portion to end of secondary ramus.

Antenna (Fig. 5A, B) with inferior tooth of basicerite subacute, reaching to about middle of second antennular article, superior tooth small, subacute; scaphocerite relatively slender, lateral tooth reaching to midlength of third antennular article, lateral margin straight, blade remarkably reduced; carpocerite slender, extending beyond antennular peduncle.

Mouthparts not dissected. Third maxilliped (Fig. 5F) pediform; lateral plate with small hooked-shaped projection pointing laterally; antepenultimate article about 3 times as long as widest part; penultimate article short, about a fourth of antepenultimate article length; ultimate article very stout, not tapering distally, slightly shorter than antepenultimate article, tip widened, with 13 spiniform setae; exopod slender, reaching to about 0.8 of antepenultimate article length.

Major cheliped (Fig. 6A–C) with coxa unarmed; basis and ischium short; merus robust, dorsal margin convex, about twice as long as broadest part, dorsodistal margin blunt; carpus short, cup-shaped; palm swollen, ovoid, about 1.9 times as long as broad, dorsodistal margin with small projection; fingers about 0.6 times as long as palm, pollex subequal to dactylus; plunger slightly reduced. Minor cheliped (Fig. 6D–F) with coxa unarmed; basis and ischium short; merus relatively robust, about 2.5 times as long as broadest part; carpus vase-shaped; palm ovoid, nearly 0.7 times as long as merus, about 0.7 times as long as broad; fingers subequal, about 0.8 times as long as palm, broadened, with minute accessory projection at tip; dactylus spoon-shaped.

Figure 6. 

Synalpheus gustavi sp. nov., holotype (FLMNH-UF-71394). A major cheliped, B same, chela, C lateral view same, D mesial view minor cheliped, E same, fingers, dorsomesial view, F same, ventromesial view.

Second pereiopod (Fig. 7A, B) with basis short; ischium relatively slender, approximately three times as long as broad; merus slender, five times as long as wide, about 1.5 times as long as ischium; carpus slender, about 1.4 times as long as merus, with four subdivisions, proximal article longest, slightly longer than sum of other articles, second article slightly longer than third one, distal article about 2.7 times as long as third article; palm slightly longer than fingers; fingers very slender.

Figure 7. 

Synalpheus gustavi sp. nov., holotype (FLMNH-UF-71394). A second pereiopod, B same, distal segments, C third pereiopod, lateral view, D fourth pereiopod, lateral view, E fifth pereiopod, lateral view.

Three last pereiopods (Fig. 7C–E) shorter and more robust compared to general shape of walking legs in Synalpheus. Third pereiopod (Fig. 7C) robust, coxa armed with small spiniform setae on dorsodistal margin; ischium vase-shaped, slightly longer than widest part; merus robust, about 2.2 times as long as wide, about 2.6 times as long as ischium; carpus slightly widening distally, 1.5 times as long as ischium, with single spiniform seta on distoventral margin; propodus relatively robust, about 0.7 times as long as merus, with five spiniform setae on ventral margin in addition to distal pair; dactylus biunguiculate, superior unguis remarkably longer (around 3 times) than inferior one. Fourth pereiopod (Fig. 7D) generally similar to third pereiopod, slightly less robust; coxa lacking spiniform setae on dorsodistal margin. Fifth pereiopod (Fig. 7E) generally similar to third pereiopod, less robust; carpus about 0.7 times as long as merus, without spiniform setae on distoventral margin; propodus subequal to merus, bearing three spiniform setae on ventral margin and one pair distally, furnished with five transverse rows of microserrulate setae on distal half.

Uropods (Fig. 5E) with mesial tooth of protopod blunt, lateral tooth subrounded with subdistal blunt projection on mesial side; exopod broad, with small distolateral tooth next to slender spiniform seta; diaeresis lacking except for lateral margin terminating to small tooth; endopod slightly longer than exopod.

Type locality

North of Madagascar: Nosy Vorona.

Ecology

All the specimens were collected from cryptic sponges situated among dead coral and coral rubble.

Remarks

The new species, S. gustavi sp. nov., shares a distinctive spoon-shaped feature of the minor cheliped, along with the presence of an orbitorostral process, with several species found in the Indo-Pacific region. These species include S. anceps AH Banner, 1956; S. dorae Bruce 1988; S. harpagatrus Banner and Banner, 1975; S. laticeps Coutière, 1905; S. neptunus neptunus (Dana, 1852); S. neptunus germanus Banner and Banner, 1975; S. paradoxus Banner and Banner, 1981; S. paralaticeps Banner and Banner, 1982; and S. theano De Man, 1910. Among these morphologically similar species, we included S. neptunus neptunus, S. anceps, and S. theano in the phylogenetic tree to explore their relationship with the new species. However, the results of the phylogenetic analysis reveal that the spoon-shaped minor chela has evolved independently in S. gustavi sp. nov. compared to other Indo-Pacific species exhibiting similar characteristics in the minor chela.

The new species can be distinguished from the aforementioned species based on four distinct characteristics, allowing for a quick and accurate differentiation. These traits are as follows: 1) very shallow frontal margin of the carapace, both the rostrum and orbital teeth, 2) unique shape of the ultimate segment of the third maxilliped, 3) presence of a four-articled carpus in the second pereiopod, and 4) exceptionally narrow and slender fingers in the second pereiopod.

Synalpheus anceps, which was reported from Saipan in the Northern Mariana Islands (AH Banner 1956), is the only species, along with the new species, that possesses a very shallow frontal margin. However, there are notable differences between the two species. In S. anceps, the front (including the rostrum and orbital teeth) is less pronounced, the blade of the scaphocerite is complete, the ultimate segment of the third maxilliped is normal Synalpheus like, the second pereiopod carpus has five articles with relatively normal-shaped fingers, and the third pereiopod is generally less robust compared to that of the new species. Additionally, in S. anceps, the superior unguis of the last three pereiopods is slightly longer than the inferior unguis. On the other hand, the new species can be distinguished from S. anceps by having a squame reduced to a small projection, a distally broad ultimate segment of the third maxilliped, a four-articled carpus in the second pereiopod with very slender fingers, and the three last pereiopods with superior ungui of the dactylus significantly longer than the inferior ones.

Synalpheus dorae shares one of the four distinctive characteristics of S. gustavi sp. nov., i.e. the second pereiopod with four-articled carpus. However, there are several other notable differences between the new species and S. dorae: the less developed superior tooth of the basicerite in the new species compared to that of S. dorae, the more developed scaphocerite blade in S. dorae, the major chela palm terminating to a small projection dorso-distally in S. gustavi sp. nov. while bearing a prominent tooth in S. dorae, more robust three last pereiopods with dorsal ungui about three times as long as ventral ungui in S. gustavi sp. nov. compared to slightly longer superior ungui in S. dorae, very strong dorsal spiniform setae of telson, and very narrow posterior margin of telson in S. dorae with mesial spiniform setae being juxtaposed.

Three species, namely S. harpagatrus, S. laticeps, and S. paralaticeps, are distinguished by the specific shape of the fingers on the minor cheliped, which possess 3 or 4 teeth at the tip. In contrast, the new species has a single tooth and a tiny accessory tooth on the fingers. Additionally, the following differences, in addition to the four main distinctions, can be observed between these three species and the new species: the longer stylocerite (overreaching the first antennular article) and more developed blade of the scaphocerite in the three of them compared to S. gustavi sp. nov.; typical ultimate segment of the third maxilliped in S. laticeps and S. paralaticeps compared to broad distally and armed with 13 slender spiniform setae in S. gustavi sp. nov., and narrow distally armed with 10-12 short heavy spiniform setae in S. harpagatrus; stouter general shape of the minor cheliped in S. harpagatrus; armed merus of the third pereiopod in S. harpagatrus and S. paralaticeps but unarmed in S. laticeps and S. gustavi sp. nov.; proportion of the superior unguis of the third pereiopod dactylus to the inferior one being approximately 3 in S. gustavi sp. nov., 2 in S. paralaticeps and 1 in S. harpagatrus and S. latirostris; fully developed diaeresis in S. harpagatrus and S. paralaticeps compared to the reduced one to a lateral tooth in S. gustavi sp. nov.; straight posterior margin of the telson in S. gustavi sp. nov. compared to the convex one in the other three species.

The Red Sea-inhabiting species, S. paradoxus, can be further distinguished from S. gustavi sp. nov. by the following characteristics, in addition to the four previously mentioned differences: its very strong superior tooth of the basicerite; half-developed blade of the scaphocerite; major cheliped merus armed with a strong tooth dorso-distally, palm with strong tubercle on the dorso-distal margin, and the dactylus longer than the pollex; minor cheliped dactylus with two definite rows of setae situated dorsally and mesially; third pereiopod dactylus with the superior unguis about twice as long as the inferior unguis; complete diaeresis; and triangular distolateral angles of the telson.

The three remaining species S. neptunus neptunus, S. neptunus germanus and S. theano, are separable from the new species by various characters, in addition to the four mentioned differences (De Man 1911; Banner and Banner 1972, 1975). Regarding the frontal appendages, S. theano, S. neptunus neptunus, and S. gustavi sp. nov. have a shorter stylocerite falling short of distal margin of the first antennular article while it overreaches the first antennular article in S. neptunus germanus; S. theano and S. gustavi sp. nov. possess a longer superior tooth of the basicerite reaching to about midlength of the second antennular article or end of it, while in the other two species it fails to reach the midlength of the second antennular article; the scaphocerite blade is absent in S. neptunus germanus, reduced to a small bump in S. gustavi sp. nov., partially reduced in S. neptunus neptunus, and with no reduction in S. theano. Regarding the thoracic appendages, S. neptunus neptunus and S. neptunus germanus possess a typical shape of the ultimate segment of the third maxilliped, S. theano lacks the crown of spiniform setae replaced by setae, and that of S. gustavi sp. nov. is broadened distally with 13 spiniform setae; the major cheliped palm terminates to a tiny projection in S. gustavi sp. nov. while the other three to prominent tubercles; the minor cheliped dactylus is furnished with two rows of setae, one dorso-mesially and one mesially in S. neptunus germanus, while there is only one mesially located row of setae in S. theano and S. neptunus neptunus, and S. gustavi sp. nov. has no row of setae; the superior unguis of the third pereiopod is remarkably longer in S. gustavi sp. nov. compared to the inferior unguis, whereas it is only slightly longer in the other three species. Regarding the pleonal appendages, three differences are notable. First, the diaeresis is complete in S. theano and S. neptunus neptunus, while it is limited to its lateral tooth in S. neptunus germanus and S. gustavi sp. nov. Second, the position of dorsal pairs of spiniform setae on telson are more mesially (especially the posterior pair) in S. gustavi sp. nov. (for detailed comparison see De Man 1911: fig. 61a for S. theano, Banner and Banner 1972: fig. 3M for S. neptunus neptunus, Banner and Banner 1975: fig. 12i for S. neptunus germanus, and Fig. 5E for S. gustavi sp. nov.). Third, the posterior margin of the telson is straight in S. gustavi sp. nov., almost straight in S. neptunus germanus, and concave in S. theano and S. neptunus neptunus.

Phylogenetically speaking, S. gustavi sp. nov. was initially included in the first and only worldwide phylogenetic study of the genus Synalpheus (Hultgren et al., 2014) as S. aff. brevifrons. The new species is broadly grouped with the S. gambarelloides species group, consistent with the findings of the present study.

4. Discussion

Since the initial documentation of eusociality in Synalpheus shrimps, prerequisites for the evolution of eusociality, as well as numerous distinctive characteristics unique to eusocial species have been reported and extensively discussed (for instance, Duffy 2007; Tóth and Bauer 2007; Tóth and Duffy 2008). According to the ‘Fortress Defence Hypothesis’ proposed by Crespi (1994), eusociality can arise under three sufficient conditions. As discussed by Duffy (2007), the host sponges inhabited by Synalpheus shrimps fulfil the first condition by providing both food (long-lived resources) and shelter, making them valuable resources for defence and facilitating specialization in the protection of kin. The shrimps’ lifelong association with sponges has led to the development of various adaptations for feeding. Among the diverse shapes observed in the minor cheliped, a specialized feeding appendage, only two forms are found in eusocial species. The first form is the characteristic shape of the minor cheliped in the S. gambarelloides group, wherein the dactylus is adorned with densely located rows of setae (commonly known as gambarelloides setae). This form predominates in species of the S. gambarelloides group. The second form is characterized by spoon-shaped fingers in the minor cheliped and is observed in species of the S. neptunus group, S. paraneptunus group, and several other species in the Indo-Pacific region. When compared to other forms of minor chelipeds, these two forms can be regarded as the most advanced evolutionary adaptations among sponge-dwelling Synalpheus. It should be noted that while the first form (S. gambarelloides form) is exclusive to sponge-dwellers, the second form is not limited to them. For instance, certain types of pair-living S. neptunus are observed to inhabit coral burrows and crevices, displaying this particular form of minor cheliped. Although the gambarelloides type of minor cheliped is the predominant form in Atlantic Synalpheus species, it is relatively rare in the Indian and Pacific oceans (see the ‘remarks’ section under S. sponjy sp. nov.).

One of the most remarkable examples of division of labor was observed by Duffy (1999) in larger females of S. filidigitus Armstrong, 1949. While smaller females maintain a morphology similar to other colony members, the larger females were reported to lack the massive major cheliped (their defensive tool) and instead rely on two minor chelipeds (their feeding tools), representing a striking transition towards division of labor. Interestingly, the first observation of females with two minor chelipeds in the Indo-Pacific species of Synalpheus was documented by Banner and Banner (1983) in the colony of S. crosnieri. In this colony, a total of 147 specimens were reported, with only three females (two of them ovigerous) possessing exclusively minor chelipeds. Two of the females had two minor chelipeds present, while the third female had one of the first pereiopods missing. Another observation made by Banner and Banner (1983) regarding S. crosnieri was the relatively large size of the eggs, measuring approximately 0.6 mm in diameter. Considering all the aforementioned characteristics of S. crosnieri, this species is undeniably a eusocial species.

Synalpheus sponjy sp. nov. not only represents the species that is morphologically closest to S. crosnieri but also exhibits a similar life style. While no females were found in the collected colonies of the new species, the assemblages of the collected specimens within a sponge varied between 1 and 19 males. Like other eusocial species of Synalpheus, this species inhabits cryptic sponges situated among dead corals, making it challenging to sample the entire sponge (and the entire colony) during a typical survey of coral rubble. However, considering the number of collected specimens (24 males), and the fact that 19 of them were collected from a single sponge, the most likely conclusion is that S. sponjy sp. nov., like its sister taxon, is a eusocial species.

In contrast to the rarity of the gambarelloides small chela type in Indo-Pacific species, the prevalence of the spoon-shaped type of chela is notable in this region, with several species showing indications of evolving a eusocial system. One such species is S. neptunus germanus, described by Banner and Banner in 1975 based on 44 specimens collected from three different sites. Although all specimens were reported as juveniles, there are indications that the species is eusocial based on personal observations of the authors. Furthermore, eusociality has been observed in at least one additional species within the S. neptunus group. Didderen et al. (2006) reported four colonies of S. neptunus from Indonesia, varying in size, with the largest colony consisting of up to 388 individuals within a single sponge. The morphology and ecology of the S. neptunus complex will be further discussed in forthcoming studies (Ashrafi et al.). Another eusocial species with the characteristic spoon-shaped fingers of the minor cheliped is Synalpheus paradoxus, described from the Red Sea (Banner and Banner 1981). The species name “paradoxus” emphasizes the highly skewed sex ratio observed in this species. Colonies of S. paradoxus were found in three different sponges, with colony sizes ranging from 113 to 132 individuals, each containing only two ovigerous females.

Lastly, the newly discovered species, S. gustavi sp. nov., represents another eusocial species characterized by the spoon-shaped fingers of the small chela. Alongside the number and size of eggs, S. gustavi sp. nov. exhibits one of the most significant hallmarks of eusociality—a skewed sex ratio. Colony sizes ranged from 1 to 19 individuals, although it is possible that some records of solitary individuals originated from individuals separated from the rest of their colonies within the rubble. Among colonies with females, the sex ratio varied from 1/3 (UF 14231) to 1/19 (UF 14259, including the allotype ovigerous queen). Interestingly, colonies of S. gustavi sp. nov. showed a tendency for high rates of parasitism (primarily in the pleonal area) by bopyrid isopod parasites. The rates of individual lot parasitism ranged from 0% to 50% (mean = 14.4%), comparable to the parasitism rates reported in some eusocial Synalpheus colonies by Duffy (1992) (30% in S. brooksi) and (McGrew and Hultgren, 2011) (25% in S. elizabethae).

In summary, the two new species, S. sponjy sp. nov. and S. gustavi sp. nov., exhibit assemblages indicative of a eusocial colony structure. These include the presence of large numbers of male (non-ovigerous) individuals residing together in a single sponge, with one or zero females present. Additionally, the two new species share ecological and morphological characteristics with other described eusocial Synalpheus species. Firstly, both species inhabit sponges, a symbiotic lifestyle believed to be a prerequisite for the evolution of eusociality. Moreover, the presence of multiple size classes, including juveniles, within the same sponge suggests overlapping generations, further supporting their eusocial nature. Secondly, similar to eusocial species in the West Atlantic, the two new species are relatively small-bodied (mean CL of S. gustavi sp. nov. = 2.62, mean CL of S. sponjy sp. nov. = 2.93), comparable to the body size of other described eusocial species (mean = 3.2 mm CL), but smaller than the average body size of communal species (mean CL = 5.32) and pair-living species (mean CL = 5.57) (Hultgren et al. 2021).

5. Conclusion

Eusociality has been extensively documented and studied in several Atlantic species of Synalpheus. However, numerous Indo-West Pacific species of Synalpheus with a high likelihood of possessing a eusocial living style have remained unnoticed, as they were described and/or reported prior to 1996 when eusociality was first reported in Synalpheus by Duffy in 1996.

The recent descriptions of two new species and the presence of at least four other eusocial species in the Indo-West Pacific region emphasize the likelihood of multiple independent evolutions of eusociality within the genus Synalpheus. These findings highlight the importance of conducting taxonomic and ecological studies on species within this region.

It is important to acknowledge the significant contributions made by Coutière, De Man, and Banner and Banner in their work before 1986, which greatly enhanced our understanding of Synalpheus species in the Indo-West Pacific. However, it is evident that earlier studies did not emphasize eusocial behavior and overall living styles, primarily due to the lack of recognition of eusociality before Duffy’s publication in 1996. The limited taxonomic and ecological studies on Indo-West Pacific species of the genus indicate a noticeable gap in our understanding. The species richness of Indo-West Pacific Synalpheus, particularly in terms of eusocial species, is currently underestimated. Therefore, it is crucial to conduct further research and observations to deepen our understanding of the taxonomy, morphology, and ecology of this fascinating genus. Continued research efforts will undoubtedly illuminate the complex behaviors, social structures, and ecological interactions of Synalpheus species not only in the Indo-West Pacific region but also in other regions, especially poorly-studied ones such as the East Pacific. Ultimately, this will lead to a more comprehensive understanding of this remarkable group of organisms.

6. Author’s contributions

HA: Conceptualization, Methodology, Original draft.

KH: Conceptualization, Methodology, Review and Editing.

7. Funding

The study was financially supported by the student grant project SGS01/PřF/2023 (to HA).

8. Competing interests

The author has declared that no competing interests exist.

9. Availability of Data

All the sequences used in the study are submitted to the GenBank with the numbers provided in the Table 1.

10. Acknowledgements

We are sincerely grateful for the invaluable insights, comments, and recommendations generously provided by our esteemed reviewers, Charles H.J.M. Fransen and Sammy De Grave. The specimens used in this study were collected during the FLMNH expedition to Madagascar in 2008 under the leadership of Gustav Paulay. We would like to express our sincere appreciation to Gustav Paulay for providing us with the opportunity to study the material deposited at FLMNH. HA extends his deepest gratitude to Gustav Paulay, Arthur Anker, Amanda Bemis, John Slapcinski, and Robert Lasley for their invaluable assistance and hospitality during his visit to the FLMNH collection. This study received financial support from the student grant project SGS01/PřF/2023 (to HA).

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Supplementary material

Supplementary material 1 

Fig. S1

Orlandin E, Piovesan M, Herbin D, Carneiro E (2023)

Data type: .tif

Explanation note: Bayesian gene tree based on COI sequences. Numbers above clades indicate Bayesian posterior probabilities. Taxon labels are given as Synalpheus species, locality, and genetics identifier (given in Table S1). New species described in this study are figured in red.

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.
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