# Phylogenetic analysis of Chirocephalus (Branchiopoda: Anostraca), with the description of a new species from Algeria

Lylia Boumendjel, Mounia Amarouayache, Celine Bonillo, Laurent Sorba, Thibaut Bagni, Nicolas Rabet

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Lylia Boumendjel, Mounia Amarouayache, Celine Bonillo, Laurent Sorba, Thibaut Bagni, et al. Phylogenetic analysis of Chirocephalus (Branchiopoda: Anostraca), with the description of a new species from Algeria. Biologia, 2023, 79 (2), pp.495-518. 10.1007/s11756-023-01544-x. hal-04585106

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# Phylogenetic analysis of the genus Chirocephalus, with the description of a new species Chirocephalus algeriensis sp.nov. (Branchiopoda: Anostraca) from Algeria

Lylia BOUMENDJEL (✉) ly.boumendjel@gmail.com)

MNHN: Museum National d'Histoire Naturelle https://orcid.org/0009-0004-8040-9507

MOUNIA AMAROUAYACHE

University of Annaba: Universite Badji Mokhtar Annaba

CELINE BONILLO

MNHN: Museum National d'Histoire Naturelle

LAURENT SORBA

Office de l'environnement de Corse

THIBAUT BAGNI

Sorbonne Université: Sorbonne Université

NICOLAS RABET

Sorbonne Université: Sorbonne Université

## Research Article

**Keywords:** Phylogeny, Molecular taxonomy, Fairy shrimp, Chirocephalidae, North-Africa, Europe

**Posted Date:** May 16th, 2023

**DOI:** https://doi.org/10.21203/rs.3.rs-2833817/v1

**License:** © This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License

**Version of Record:** A version of this preprint was published at Biologia on November 1st, 2023. See the published version at https://doi.org/10.1007/s11756-023-01544-x.
Abstract

Molecular analyses based on the comparison of *Chirocephalus* species mtDNA cytochrome oxidase I (COI) sequences obtained from Algeria, Morocco and France with other available sequences were performed. The analysis clarified the status of the genus *Chirocephalus* Prévost, 1803 in Algeria and its phylogenetic links with other populations. Three lineages occur in Algeria representing three different species: *Chirocephalus salinus* Daday, 1910, C. sanhadjaensis Boumendjel, Rabet and Amarouayache, 2018 and a new species sampled in Reghaïa (Alger, Algeria) near Algiers. All these species, belonging to the "diaphanus" group, seem to be endemic to Algeria except *C. salinus* also present in France (Corsica).

The new species *C. algeriensis* sp. nov. is described and its affinities with other species of the genus are presented. This species is until now restricted to Algiers region and shares several characters with *C. diaphanus* Prévost, 1803. It is separated from all other species by the shape and size of the second antennal appendages of the male and the female. The resting eggs of *C. algeriensis* sp. nov. are morphologically similar to those of many *Chirocephalus* but a variation of size of eggs can be used at a specific level.

Introduction

Molecular phylogenetic studies on fairy shrimp species have increased in the last decades (Ketmaier et al. 2005, 2008, 2012; Aguilar 2011; Reniers et al. 2013; Zarattini et al. 2013; Lindholm et al. 2016; Aguilar et al. 2017; Kappas et al. 2017; Rodríguez-Flores et al. 2017; Mioduchowska et al. 2018; Lukić et al. 2019; Paula et al. 2019). They present an important source of information that can help to further resolve taxonomy problems. In this group, morphological characters traditionally delimit the species but, in some cases, molecular phylogeny challenges the specific delimitation and improves traditional morphological characters.

To date, fifty-four species of *Chirocephalus* Prévost, 1803 have been recorded worldwide. This genus is distributed throughout the Palearctic under several types of climates (Rogers 2013). New species of *Chirocephalus* are still discovered so far (Rogers and Soufi 2013; Cottarelli et al. 2017; Boumendjel et al. 2018) and work on molecular phylogeny gradually begins to shed light on the evolution of this genus (Reniers et al. 2013).

*Chirocephalus diaphanus* Prévost, 1803 is one of the most widespread European species of fairy shrimp (Brtek and Thiéry 1995; Mura 2001). Its distribution in Western Europe extends almost continuously from Great Britain to the Iberian Peninsula, and as far east as the Rhine in Germany (Brtek and Thiéry 1995). A single occurrence of *C. diaphanus* is known from the Benelux countries, in pools in South Limburg, Netherlands Paulssen (2000). Further east, it was recorded south in the Apennine and Balkan peninsulas, reaching the Black Sea in Romania; an isolated population occurs at the mouth of the Vistula River in Poland (Brtek and Thiéry 1995). In the Mediterranean area, populations were recorded in Sicily, Sardinia and Crete (Brtek and Thiéry 1995), North Africa in Morocco (Thiéry 1991), Tunisia (Guerney 1909; Daday 1910; Seurat 1922; Massal 1951; Turki and Turki 2010) and Algeria (Simon 1886; Daday 1910; Gauthier 1928; Beladjal et al. 1995; Samraoui et al. 2002). This species can colonize a wide variety of freshwater habitats (flooded ditches, fallow fields, peat bogs), both in the lowlands and highlands areas (Brtek and Thiéry 1995; Mura et al. 2003; Fonseca et al. 2008) and has a wide range of ecological tolerances (Brtek and Thiéry 1995; Mura 1999), in terms of temperature, dissolved oxygen and potential hydrogen. Part of this diversity could be linked to a strong genetic diversity (Reniers et al. 2013).

Indeed, this species can be considered as a species complex, and it will be necessary to gradually improve the taxonomy of this genus with an integrated analysis combining morphology and genetics. In this context, we will designate as *C. diaphanus sensu stricto*, the populations forming a clade with the French populations collected around Montauban from where the species was originally described. *C. diaphanus sensu lato* refers to all populations that are morphologically identified but not necessary in this clade.

Some authors recognize several subspecies of *C. diaphanus* based on morphological differences (Daday 1913; Pesta 1921; Stoicescu 1992). However, no consensus has been reached yet about the taxonomic status of these taxa due to substantial intraspecific morphological variability and a lack of DNA sequence data from populations across subspecies ranges (Brtek and Thiéry 1995; Marincek and Petrov 1995; Brtek 1997; Mura 2001; Mura et al. 2002).

In Algeria *C. diaphanus*, was mentioned by several authors (Simon 1886; Daday 1910; Gauthier 1928; Beladjal et al. 1995; Samraoui and Dumont 2002) in humid and sub-humid areas of the Northern region. The species *C. salinus* Daday, 1913 was also identified in some localities where the alleged *C. diaphanus* had previously been recorded (Samraoui 2002). To clarify the systematic position of Algerian *Chirocephalus* species, we have sequenced the COI gene of Algerian, Moroccan, and French populations to start a comparative analysis based on genetic and morphological characters.

A new species *Chirocephalus algeriensis* sp. nov. from Reghaïa, is described and compared with the populations of *C. diaphanus* from mainland France, which allows us to detect morphological variability in Algeria to better understand diversity elsewhere.

Materials and methods

Material examined

Current populations of *Chirocephalus* were obtained from 12 different locations in Algeria, 2 in Morocco and 14 in France. Specimens were collected in the field, using a net of 400 µm mesh size, between December 2013 and March 2018 and by culturing animals in the laboratory from pond sediments. These were collected after a complete drying of the temporary pools. Breeding was performed between 12 and 16°C under a natural day/night light cycle, with continuous aeration, using dry sediment (200 g) from pool in 20 L container full of distilled water. The specimens were fed on algae and bacteria grown from the sediment. All specimens were preserved on 96% ethanol to make it available for both morphological studies and molecular analyses. Details of sampling locations are provided in (Table 1) and (Fig. 1).
<table><caption>Table 1
Populations codes, GenBank accession number, locations, corresponding latitude and longitude, detailed sampling information (sampling date, collector/reference, wild/breeding) for populations of *Chirocephalus* included in the study</caption><thead><tr><th>Taxon</th><th>Code Haplotype</th><th>N° accession</th><th>Locality</th><th>Latitude</th><th>Longitude</th><th>Sampling date</th><th>Collector/Reference</th><th>Wild/ Breeding</th></tr></thead><tbody><tr><td rowspan="2">C. sanhadjaensis<br>Boumendjel et al, 2018</td><td>M1</td><td>OP596278</td><td>Ain-Magroun, Skikda, Algeria</td><td>36°50'21"N</td><td>07°17'01"E</td><td>16/12/2016</td><td>L. Boumendjel</td><td>B</td></tr><tr><td>M2</td><td>OP596279</td><td></td><td></td><td></td><td>02/03/2017</td><td></td><td></td></tr><tr><td rowspan="15">C. salinus<br>Daday, 1910</td><td>Sa1</td><td>OP595366</td><td>Berrihene, El Tarf, Algeria</td><td>36°50'02"N</td><td>08°06'44"E</td><td>11/02/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa2</td><td>OP595367</td><td>Malahmar, El Tarf, Algeria</td><td>36°25'43"N</td><td>08°13'40"E</td><td>09/05/2016</td><td>G. Gouasmia</td><td>B</td></tr><tr><td></td><td></td><td></td><td></td><td></td><td>18/03/2017</td><td>A. Harket and S. Litim</td><td>W</td></tr><tr><td>Sa3</td><td>OP595368</td><td>Souk Rguibet, El Tarf, Algeria</td><td>36°53'42"N</td><td>08°17'30"E</td><td>17/03/2017</td><td>M. Amarouayache</td><td>B</td></tr><tr><td></td><td></td><td>Bonifacio, Corsica-Sud, France</td><td>41°27'55"N</td><td>9°14'37"E</td><td>13/09/2016</td><td>L. Sorba</td><td>B</td></tr><tr><td></td><td></td><td>Berrihene, El Tarf, Algeria</td><td>36°50'02"N</td><td>08°06'44"E</td><td>11/02/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa4</td><td>OP595369</td><td>Les Salines, Annaba, Algeria</td><td>36°49'60"N</td><td>7°47'56"E</td><td>02/03/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td></td><td></td><td>Airport, Annaba, Algeria</td><td>36°48'58"N</td><td>7°48'25"E</td><td></td><td></td><td></td></tr><tr><td>Sa5</td><td>OP595370</td><td>Ben Mhidi, El Tarf, Algeria</td><td>36°46'22"N</td><td>07°53'41"E</td><td>02/03/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa6</td><td>OQ588789</td><td>Tahouna, Souk Ahras, Algeria</td><td>36°19'53"N</td><td>7°54'0"E</td><td>18/03/2017</td><td>A. Harket and S. Litim</td><td>W</td></tr><tr><td></td><td></td><td>Ain Seynour, Souk Ahras, Algeria</td><td>36°25'43"N</td><td>08°13'41"E</td><td>08/03/2017</td><td></td><td></td></tr><tr><td>Sa7</td><td>OQ588790</td><td>Bonifacio, Corsica-Sud, France</td><td>41°27'55"N</td><td>9°14'37"E</td><td>13/09/2016</td><td>L. Sorba</td><td>B</td></tr><tr><td>Sa8</td><td>OP595371</td><td>Ben Mhidi, El Tarf, Algeria</td><td>36°46'22"N</td><td>07°53'41"E</td><td>02/03/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa9</td><td>OP595372</td><td>Laalalig, Annaba, Algeria</td><td>36°45'50"N</td><td>07°40'42"E</td><td>03/02/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa10</td><td>OP595373</td><td>Ben Mhidi, El Tarf, Algeria</td><td>36°46'22"N</td><td>07°53'41"E</td><td>02/03/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa11</td><td>OP595374</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>Sa12</td><td>OP595375</td><td>Bonifacio, Corsica-Sud, France</td><td>41°27'55"N</td><td>9°14'37"E</td><td>13/09/2016</td><td>L. Sorba</td><td>B</td></tr><tr><td>Sa13</td><td>OP595376</td><td>Airport, Annaba, Algeria</td><td>36°48'58"N</td><td>7°48'25"E</td><td>02/03/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td>Sa14</td><td>OP595377</td><td>Sidi Salem, Annaba, Algeria</td><td>36°50'30"N</td><td>7°49'35"E</td><td>17/03/2017</td><td>L. Boumendjel</td><td>B</td></tr><tr><td>Sa15</td><td>OP595378</td><td>Airport, Annaba, Algeria</td><td>36°48'58"N</td><td>7°48'25"E</td><td>02/03/2017</td><td>M. Amarouayache</td><td>W</td></tr><tr><td rowspan="3">C. diaphanus<br>Prévost, 1803</td><td>D1</td><td>OP597701</td><td>Marcillé-la-Ville, Mayenne, France</td><td>48°18'34"N</td><td>0°30'49"W</td><td>21/04/2014</td><td>N. Rabet</td><td>W</td></tr><tr><td></td><td></td><td>Chambord, Loir-et-Cher, France</td><td>47°38'17"N</td><td>1°30'10"E</td><td>04/05/2015</td><td></td><td></td></tr><tr><td>D2</td><td>OP597702</td><td>Villers-lès-Nancy, Meurthe-et-Moselle, France</td><td>48°39'52"N</td><td>6°6'19"E</td><td>03/04/2013</td><td></td><td>W</td></tr></tbody></table>
<table><thead><tr><th>Taxon</th><th>Code Haplotype</th><th>N° accession</th><th>Locality</th><th>Latitude</th><th>Longitude</th><th>Sampling date</th><th>Collector/Reference</th><th>Wild/ Breeding</th></tr></thead><tbody><tr><td></td><td></td><td></td><td>Bouconville sur Madt, Meuse, France</td><td>48°50'35"N</td><td>5°43'53"E</td><td>13/10/2013</td><td></td><td>B</td></tr><tr><td></td><td>D3</td><td>OP597703</td><td>Mennecy, Essonne, France</td><td>48°32'58"N</td><td>2°26'51"E</td><td>12/04/2014</td><td></td><td>B</td></tr><tr><td></td><td>D4</td><td>OP597704</td><td>Signes, Var, France</td><td>43°16'7"N</td><td>5°46'39"E</td><td>05/03/2013</td><td>J.C. Tempier</td><td>W</td></tr><tr><td></td><td>D5</td><td>OP597705</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td></td><td>D6</td><td>OP597706</td><td>Ampus, Var, France</td><td>43°35'52"N</td><td>6°24'54"E</td><td>09/03/2013</td><td>N. Rabet</td><td>W</td></tr><tr><td></td><td>D7</td><td>OP597707</td><td>Gonfaron, Var, France</td><td>43°20'36"N</td><td>6°15'35"E</td><td>09/03/2013</td><td></td><td>W</td></tr><tr><td></td><td>D8</td><td>-</td><td>Chamarande, Essonne, France</td><td>48°31'22"N</td><td>2°13'28"E</td><td>05/2013</td><td>N. Rabet</td><td>W</td></tr><tr><td></td><td></td><td></td><td>Regnéville-sur-Mer, Manche, France</td><td>49°1'37"N</td><td>1°32'52"W</td><td>16/03/2016</td><td></td><td>W</td></tr><tr><td></td><td></td><td></td><td>Aurseulles, Calvados, France</td><td>49°6'43"N</td><td>0°42'8"W</td><td>2009</td><td></td><td>B</td></tr><tr><td></td><td></td><td></td><td>Guérande, Loire Atlantique, France</td><td>47°18'2"N</td><td>-2°26'45"O</td><td>03/2017</td><td>A. Soileux</td><td>W</td></tr><tr><td></td><td>D9</td><td>-</td><td>Gonfaron, Var, France</td><td>43°20'36"N</td><td>6°15'35"E</td><td>09/03/2013</td><td>N. Rabet</td><td>W</td></tr><tr><td></td><td>D10</td><td>-</td><td>Méjannes le Cap, Gard, France</td><td>44°13'30"N</td><td>4°22'4"E</td><td>22/07/2013</td><td>N. Rabet</td><td>W</td></tr><tr><td></td><td>4FChaiM2</td><td>JX990358</td><td>Chaillevette, Charente-Maritime, France</td><td>45°43'30"N</td><td>1°3'30"E</td><td>-</td><td>Reniers et al. (2013)</td><td>-</td></tr><tr><td></td><td>FLarivM1</td><td>JX990355</td><td>Lac des Rives, Hérault, France</td><td>43°51'42"N</td><td>3°15'52"E</td><td>-</td><td></td><td>-</td></tr><tr><td></td><td>FRoquehV1</td><td>JX990350</td><td>Roque Haute, Hérault, France</td><td>43°18'20"N</td><td>3°21'58"E</td><td>-</td><td></td><td>-</td></tr><tr><td></td><td>4FLeuV1</td><td>JX990353</td><td>Leucate, Aude, France</td><td>42°54'01"N</td><td>3°1'13N</td><td>-</td><td></td><td>-</td></tr><tr><td></td><td>5VijlM1</td><td>JX990339</td><td>Vijlen, Netherlands</td><td>50°47'19"N</td><td>5°57'50"N</td><td>-</td><td></td><td>-</td></tr><tr><td></td><td>FVarloV1</td><td>JX990345</td><td>Flassans sur Issole, Var, France</td><td>43°20'59"N</td><td>6°14'21"E</td><td>-</td><td></td><td>-</td></tr><tr><td></td><td>FValligV2</td><td>JX990347</td><td>Valliguières, Gard, France</td><td>44°0'42"N</td><td>4°34'42"E</td><td>-</td><td></td><td>-</td></tr><tr><td></td><td>P_RCL2_01</td><td>MF458642</td><td>South-East of France</td><td>-</td><td>-</td><td>-</td><td>Corse et al. (2017)</td><td>-</td></tr><tr><td>Chirocephalus sp1</td><td>Z</td><td>OP599900</td><td>Khénifra, Morocco</td><td>32°56'31"N</td><td>5°28'49"W</td><td>11/04/2006</td><td>E. Quéinnec and A. Lecot</td><td>W</td></tr><tr><td>Chirocephalus sp2</td><td>BRilV2</td><td>JX990360</td><td>Rila, Bulgaria</td><td>42°11'27"N</td><td>23°35'2"E</td><td>-</td><td>Reniers et al. (2013)</td><td>-</td></tr><tr><td>Chirocephalus sp3</td><td>COX1</td><td>AY189000</td><td>Ozieri, Sardinia</td><td>-</td><td>-</td><td>-</td><td>Ketmaier et al. (2003)</td><td>-</td></tr><tr><td>Chirocephalus sp4</td><td>COX1</td><td>AY188996</td><td>Italy</td><td>-</td><td>-</td><td>-</td><td>Ketmaier et al. (2003)</td><td>-</td></tr><tr><td>Chirocephalus sp5</td><td>SDonM3</td><td>JX990335</td><td>Doñana, Spain</td><td>36°56'52"N</td><td>6°24'57.9168"W</td><td>-</td><td>Reniers et al. (2013)</td><td>-</td></tr><tr><td></td><td>MBensV1</td><td>JX990344</td><td>Benslimane, Morocco</td><td>33°37"N</td><td>7°7'13"W</td><td>-</td><td></td><td>-</td></tr></tbody></table>
<table><thead><tr><th>Taxon</th><th>Code Haplotype</th><th>N° accession</th><th>Locality</th><th>Latitude</th><th>Longitude</th><th>Sampling date</th><th>Collector/Reference</th><th>Wild/ Breeding</th></tr></thead><tbody><tr><td rowspan="2">Chirocephalus sp6</td><td>PsagV1</td><td>JX990337</td><td>Sagres, Portugal</td><td>37°3'24"N</td><td>8°57'2"W</td><td>-</td><td rowspan="2">Reniers et al. (2013)</td><td>-</td></tr><tr><td>PSagM1</td><td>JX990338</td><td></td><td></td><td></td><td>-</td><td>-</td></tr><tr><td>C. algeriensis sp.nov.</td><td>A</td><td>OP596277</td><td>Reghaïa, Algeria</td><td>36°44'30"N</td><td>3°22'7"E</td><td>09/03/2017</td><td>L. Boumendjel</td><td>B</td></tr><tr><td>C. spinicaudatus Simon, 1886</td><td>P2</td><td>OP596276</td><td>Saron-sur-Aube, Marne, France</td><td>48°35'11"N</td><td>3°43'52"E</td><td>12/2012</td><td>N. Rabet</td><td>B</td></tr><tr><td>Eubranchipus grubii Dybowski, 1860</td><td>G1</td><td>OP596274</td><td>Contrevoz, Ain, France</td><td>5°38'38"E</td><td>45°46'41"N</td><td>22/03/2015</td><td>L. Augettant</td><td></td></tr><tr><td>C. ruffoi Cottarelli et Mura, 1984</td><td>COX1</td><td>AY188999</td><td>Pollino National Park, Calabria, Italy</td><td>-</td><td>-</td><td>-</td><td>Ketmaier et al. (2003)</td><td>-</td></tr><tr><td>C. marchesoni Ruffo et Vesentini, 1957</td><td>H28</td><td>JN246530</td><td>Lake Pilate, Marches, Italy</td><td>-</td><td>-</td><td>-</td><td></td><td>-</td></tr><tr><td>C. sibyllae Cottarelli et Mura, 1975</td><td>COX1</td><td>AY189001</td><td>Palazzo Borghese, Marches, Italy</td><td>-</td><td>-</td><td>-</td><td></td><td>-</td></tr><tr><td rowspan="12">C. kerkyrensis Ketmaier et al., 2012</td><td>H1</td><td>JN246503</td><td>Merhojes Pond, Cepa, Dumre, Albania</td><td>40°56'39"N</td><td>19°51'58"E</td><td>-</td><td></td><td>W</td></tr><tr><td>H3</td><td>JN246505</td><td>Avena Pond, Basilicata, Italy</td><td>40° 2'60"N</td><td>16°14'42"E</td><td>-</td><td></td><td>W</td></tr><tr><td>H6</td><td>JN246508</td><td></td><td></td><td></td><td>-</td><td></td><td>W</td></tr><tr><td>H9</td><td>JN246511</td><td>Pond T3, Circeo National Park, Latina, Italy</td><td>41°20'53"N</td><td>13° 2'49"E</td><td>-</td><td></td><td>W</td></tr><tr><td>H10</td><td>JN246512</td><td></td><td></td><td></td><td>-</td><td></td><td></td></tr><tr><td>H13</td><td>JN246515</td><td></td><td></td><td></td><td>-</td><td></td><td></td></tr><tr><td>H14</td><td>JN246516</td><td>Pond A1, Castel Porziano, Rome, Italy</td><td>41°41'7"N</td><td>41°43'11"E</td><td>-</td><td></td><td>W</td></tr><tr><td>H17</td><td>JN246519</td><td>Bosco di Foglino, Nettuno, Rome, Italy</td><td>41°26'28"N</td><td>12°43'55"E</td><td>-</td><td></td><td>W</td></tr><tr><td>H19</td><td>JN246521</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>H24</td><td>JN246526</td><td>Pond KO3, Kokkini, Corfu Is, Greece</td><td>39°36'58"N</td><td>19°49'12"E</td><td>-</td><td></td><td>W</td></tr><tr><td>H27</td><td>JN246529</td><td>Pond KO4, Kokkini, Corfu Is, Greece</td><td>39°37'41"N</td><td>19°49'52"E</td><td>-</td><td></td><td>W</td></tr></tbody></table>

## DNA extraction, PCR amplification and sequencing

Molecular analyses were performed at the MNHN "Service de Systématique Moléculaire" (SSM) in Paris. For this study, single individuals of *Chirocephalus salinus* (Algeria and France), *C. sanhadjaensis* from Ain-Magroun (Algeria), *C. algeriensis* sp. nov. from Reghaïa (Algeria), *C. diaphanus sensu strecto* (France), *C. diaphanus sensu lato* (Morocco), and *C. spinicaudatus* (France) were processed to be included in the molecular analyses.

From one up to 10 adult individuals (male and female) from each population of *Chirocephalus* were processed to be included in the molecular analyses. They were identified using determination keys to the species of Daday (1910) and Boumendjel et al. (2018). Further *Chirocephalus* COI sequences representative of all the available *Chirocephalus* species and major evolutionary lineages covering the range of the genus in Europe and North Africa (Ketmaier et al. 2003; Ketmaier et al. 2012; Reniers et al. 2013; Zarattini et al. 2013) were downloaded from GenBank to be included in the analyses (see GenBank accession numbers in Table 1). We also re-sequenced populations from Auseulles (Normandy) already sequenced in the paper of Reniers et al. 2013. A new sequence of *Eubranchipus grubii* from France, was used as outgroup.
Genomic DNA for each single individual of *Chirocephalus* was extracted from thoracic segments using the NucleoSpin® 96 Tissue Kit. After homogenization, tissues were digested overnight at 65° in a volume of 2.5 ml of BP (Proteinase Buffer) Proteinase K, and 18 ml of T1, 200 µL for each sample. To complete the lysis, buffers (BQ1, BW, B5 and BE) were prepared in a gutter and placed in the robot Eppendorf epMotion 5075 along with the DNA sample. A fragment of the mtDNA gene encoding for cytochrome oxidase subunit I (COI mtDNA) was amplified using the universal LCO1490 and HCO2198 primers described by Folmer (Folmer et al. 1994). The PCR reaction was performed out on a Bio-Rad C1000 Touch Thermal Cycler for a total volume of 20 µl. Each reaction contained the following composition: 12.44 µl (H2O), 2 µl (Taq10x buffer coralLoad), 1 µl of DMSO (DiMethyl Sulfoxide), 1 µl of BSA (Bovine Albumin Serum), 0.8 µl of dNTPs (DeoxyNucleotide TriPhosphate), 0.32 µl (HCO2198), 0.32 µl (LCO1490), 0.12 µl of Taq (recombinant DNA polymerase, Quiagen) and 2 µl of the DNA.

The amplification consisted of an initial denaturation step of 94°C for 5 min followed by 40 cycles of 94°C for 30 s, 50°C for 30 s, and 72°C for 90 s, followed by a final extension at 72°C for 8 min. After PCR, 3 µl of each PCR product was separated by electrophoresis on a 2% agarose gel stained with Ethidium Bromide at 100 V for 20 min and visualised with a UV Transilluminator. The PCR products, which showed a clear and single band of the correct expected length, were sent for sequencing by the method of Sanger et al. (1977) to the "Eurofins" sequencing service company MWG Operon (Germany). They were sequenced in both directions using the forward and reverse primers (LCO1490 and HCO2198) that we have provided to the company. Chromatograms were imported and edited with Chromas version 2.6.4.

## Phylogenetic analyses

Nucleotide sequences obtained were aligned with multiple sequences alignment programs by CLUSTALW (Thompson et al. 1994) in Geneious 11 and collapsed into haplotypes prior to genetic analysis. A refinement alignment was tested using the Gblocks 0.91b program to eliminate poorly aligned positions and divergent regions, but this did not change the alignment. We chose not to have any missing data by reducing the number of usable sequences in Genbank.

Best-fitting DNA substitution models were assessed for each codon without partition or for each codon position using Partition Finder following the Bayesian Information criterion (BIC) (Lanfear et al. 2017).

A phylogenetic tree was constructed on the alignment of haplotypes by applying the Neighbor joining (NJ) implemented in the Geneious 11.1.5 Tree Builder software, with a 1000 replicas bootstraps and HKY Genetic distance model and using the Maximum likelihood (ML) analyses with RAxML version 8.2.10 were run under the GTR model and a 1000 replicas bootstraps. A Bayesian tree was constructed using MrBayes package version 3.1.2 by running the General Time-Reversible with HKY + G, SYM + G and F81 models for each codon positions and 10 000 000 generations. For a general overview of nucleotide differences among species, percentual pair-wise nucleotide difference per site for COI data was also estimated. Pair-wise distances in % were calculated using Geneious distance (Table 2).
<caption>Table 2</caption>

Pairwise values between *Chirocephalus* species (*Chirocephalus* sp1: new specie from Morocco; *Chirocephalus* sp2: *C. diaphanus* from Bulgaria; *Chirocephalus* sp3: *C. salinus* from Italy; *Chirocephalus* sp4: *C. diaphanus* from Italy; *Chirocephalus* sp 5: *C. diaphanus* Spain and Morocco; *Chirocephalus* sp6: *C. diaphanus* from Portugal)

<table><thead><tr><th></th><th>1</th><th>2</th><th>3</th><th>4</th><th>5</th><th>6</th><th>7</th><th>8</th><th>9</th><th>10</th><th>11</th><th>12</th><th>13</th><th>14</th></tr></thead><tbody><tr><td>1. C. salinus</td><td>0.2-5.9</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>2. C. sanhadjaensis</td><td>13.1-15.5</td><td>0</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>3. C. diaphanus ss</td><td>9.4-12.2</td><td>13.4-14.5</td><td>0.2-4</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>4. Chirocephalus sp1</td><td>15.6-16.6</td><td>13.3</td><td>13.1-14</td><td>0</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>5. Chirocephalus sp2</td><td>10.5-12</td><td>13.8-14</td><td>8.8-9.6</td><td>13.6</td><td>0</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>6. Chirocephalus sp3</td><td>15.1-15.8</td><td>15.8-16</td><td>12.1-13.3</td><td>16.6</td><td>13.3</td><td>0</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>7. Chirocephalus sp4</td><td>7.7-8</td><td>10.5-12.5</td><td>7.9-9</td><td>13.4</td><td>9.4</td><td>4.6</td><td>0</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>8. C. algeriensis sp.nov.</td><td>10.3-11.6</td><td>12.1-12.3</td><td>7.2-8.3</td><td>12.9</td><td>9.6</td><td>11.6</td><td>8.1</td><td>0</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>9. Chirocephalus sp.5</td><td>9.6-11.2</td><td>13.4-13.8</td><td>6.8-7.9</td><td>13.3-14.5</td><td>8.8-10.3</td><td>11</td><td>7</td><td>9-9.4</td><td>2.9</td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>10. Chirocephalus sp6</td><td>9.9-11.6</td><td>12.1-12.3</td><td>5.7-7.4</td><td>12.5</td><td>9.4</td><td>11.4</td><td>7.7-8</td><td>8.1-8.5</td><td>6.6-7.2</td><td>0.4</td><td></td><td></td><td></td><td></td></tr><tr><td>11. C. marchesonii</td><td>14.9-16.4</td><td>15.5</td><td>15.1-16</td><td>15.6</td><td>16.2</td><td>19.7</td><td>16.2</td><td>16.2</td><td>15.6-16</td><td>15.5-15.8</td><td>0</td><td></td><td></td><td></td></tr><tr><td>12. C. ruffoi</td><td>11-12.3</td><td>13.4-13.6</td><td>8.3-9.6</td><td>14.5</td><td>9.6</td><td>4.6-6.6</td><td>2</td><td>7.7</td><td>7.7-8</td><td>9.2-8.8</td><td>16.2</td><td>0</td><td></td><td></td></tr><tr><td>13. C. kerkyrensis</td><td>12.1-14</td><td>10.5-12.1</td><td>9.6-11.8</td><td>13.3-14.4</td><td>11.8-12.9</td><td>13.3-14.4</td><td>10.1-11.2</td><td>10.5-11.4</td><td>9.6-10.7</td><td>10.9-10.9</td><td>13.1-14.7</td><td>10.9-11.4</td><td>0.2-5.5</td><td></td></tr><tr><td>14. C. sibyllae</td><td>12-14.4</td><td>13.4-13.6</td><td>10.7-11.8</td><td>14.7</td><td>10.9</td><td>13.3</td><td>9.6</td><td>11.8</td><td>9.4</td><td>10.3</td><td>16.9</td><td>10.3</td><td>11.4-12.3</td><td>/</td></tr></tbody></table>

## Samplings and morphological analysis

Thirteen adult individuals, 6 males and 7 females of Reghaïa population were examined. They were measured from the head anterior margin to the furca posterior margin under a stereomicroscope (Nikon SMZ-1) equipped with an ocular micrometer. Seven specimens (4 males and 3 females) had the antennal appendages, second antennae, genitalia and thoracic appendages dissected and mounted on glass microscope slides for examination under a microscope (WILD HEERBRUGG SWITZERLAND M20-35442) equipped with a camera lucida. The animals are deposited at the Muséum national d'Histoire naturelle (MNHN) of Paris, France. Specimens from different populations of *Chirocephalus diaphanus* from France were also sorted and dissected for examination of the male antennal appendages under a microscope.

We also observed the antennal appendages and the second antennae of male and female species of *Chirocephalus* from other regions of Algeria and France stored at MNHN, Paris to determine the morphological variations between these species. Locality names are provided according to labels and other information in the species description and bibliography (Table 3). A new population of *C. algeriensis* sp.nov. was found in Bab Ezzouar (Algiers) by L. Beladjal. These specimens are deposited at the MNHN of Paris.
<caption>Table 3</caption>

List of *Chirocephalus* species from Algeria and France present at the Paris MNHN (France), including MNHN accession number, sampling locality, sampling date, Collector, wild/breeding, and study

<table><thead><tr><th>N° accession MNHN</th><th>Locality</th><th>Collection date</th><th>Taxon</th><th>Collector</th><th>Wild/ Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2021-8503</td><td>Sidi Salem, Annaba, Algeria</td><td>29/09/2015</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8504</td><td>Sidi Salem, Annaba, Algeria</td><td>12/04/2015</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8505</td><td>Chambord Park, Loir-et-Cher, France</td><td>04/05/2015</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8506</td><td>Episy pond Fontainebleau, Essonne, France</td><td>20/04/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8507</td><td>Marcillé-la-Ville, Mayenne, France</td><td>29/05/2014</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8508</td><td>Marcillé-la-Ville, Mayenne, France</td><td>29/05/2014</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8509</td><td>Mennecy, Essonne, France</td><td>12/04/2014</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8510</td><td>Mennecy, Essonne, France</td><td>12/04/2014</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8511</td><td>Khénifra, Morocco</td><td>11/04/2006</td><td>Chirocephalus sp</td><td>E. Quéinnec and A. Lecot</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8512</td><td>Khénifra, Morocco</td><td>11/04/2006</td><td>Chirocephalus sp</td><td>E. Quéinnec and A. Lecot</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8513</td><td>Bouconville sur Madt, Meuse, France</td><td>13/10/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8514</td><td>Méjannes-le-Clap, Gard, France</td><td>22/07/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8515</td><td>Méjannes-le-Clap, Gard, France</td><td>22/07/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8516</td><td>Chamarande, Essonne, France</td><td>05/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8517</td><td>Chamarande, Essonne, France</td><td>05/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8518</td><td>Bonifacio, Corsica-Sud, France</td><td>29/11/2013</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8519</td><td>Bonifacio, Corsica-Sud, France</td><td>29/11/2013</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8520</td><td>Regnéville-sur-Mer, Manche, France</td><td>16/03/2016</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8521</td><td>Regnéville-sur-Mer, Manche, France</td><td>16/03/2016</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8522</td><td>Sidi Salem, Annaba, Algeria</td><td>05/12/2013</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8523</td><td>Sidi Salem, Annaba, Algeria</td><td>06/12/2013</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8524</td><td>Souk Rguibet, El Tarf, Algeria</td><td>13/03/2015</td><td>C. salinus</td><td>S. Ghaouaci</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8525</td><td>Souk Rguibet, El Tarf, Algeria</td><td>13/03/2015</td><td>C. salinus</td><td>S. Ghaouaci</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8526</td><td>Aurseulles, Calvados, France</td><td>2009</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8527</td><td>Aurseulles, Calvados, France</td><td>2009</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8528</td><td>Malahmar, Souk Ahras, Algeria</td><td>09/05/2016</td><td>C. salinus</td><td>G. Gouasmia</td><td>B</td><td>Genetics and morphological</td></tr></tbody></table>
<table><thead><tr><th>N° accession<br>MNHN</th><th>Locality</th><th>Collection<br>date</th><th>Taxon</th><th>Collector</th><th>Wild/<br>Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2021-8529</td><td>Malahmar, Souk Ahras, Algeria</td><td>09/05/2016</td><td>C. salinus</td><td>G. Gouasmia</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8530</td><td>Ampus, Var, France</td><td>09/03/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8531</td><td>Ampus, Var, France</td><td>09/03/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8532</td><td>Signes, Var, France</td><td>05/03/2013</td><td>C. diaphanus</td><td>J.C Tempier</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8533</td><td>Signes, Var, France</td><td>05/03/2013</td><td>C. diaphanus</td><td>J.C Tempier</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8534</td><td>Gonfaron, Var, France</td><td>09/03/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8535</td><td>Gonfaron, Var, France</td><td>09/03/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8536</td><td>Khénifra, Morocco</td><td>11/04/2006</td><td>Chirocephalus<br>sp</td><td>E. Quéinnec and A.<br>Lecot</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8537</td><td>Etendard-St Maximin, Var, France</td><td>08/03/2013</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8538</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8539</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8540</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8541</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8542</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8543</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8544</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8545</td><td>Souk Rguibet, El Tarf, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8546</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8547</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8548</td><td>Ain-Magroun, Skikda, Algeria</td><td>10/06/2016</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8549</td><td>Ain-Magroun, Skikda, Algeria</td><td>10/06/2016</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8550</td><td>Ain-Magroun, Skikda, Algeria</td><td>10/06/2016</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8551</td><td>Ain-Magroun, Skikda, Algeria</td><td>06/12/2016</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8552</td><td>Ain-Magroun, Skikda, Algeria</td><td>16/12/2016</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8553</td><td>Ain-Magroun, Skikda, Algeria</td><td>16/12/2016</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8554</td><td>Ain-Magroun, Skikda, Algeria</td><td>05/01/2017</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8555</td><td>Ain-Magroun, Skikda, Algeria</td><td>05/01/2017</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8556</td><td>Ain-Magroun, Skikda, Algeria</td><td>02/03/2017</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr></tbody></table>
<table><thead><tr><th>N° accession<br>MNHN</th><th>Locality</th><th>Collection<br>date</th><th>Taxon</th><th>Collector</th><th>Wild/<br>Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2021-8557</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8558</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8559</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8560</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8561</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8562</td><td>Berrihene, El Tarf, Algeria</td><td>11/02/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8563</td><td>Malahmar, Souk Ahras, Algeria</td><td>09/05/2016</td><td>C. salinus</td><td>G. Gouasmia</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8564</td><td>Malahmar, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8565</td><td>Malahmar, Souk Ahras, Algeria</td><td>09/05/2016</td><td>C. salinus</td><td>G. Gouasmia</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8566</td><td>Malahmar, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8567</td><td>Bonifacio, Corsica-Sud, France</td><td>27/02/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8568</td><td>Bonifacio, Corsica-Sud, France</td><td>27/02/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8569</td><td>Bonifacio, Corsica-Sud, France</td><td>27/02/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8570</td><td>Bonifacio, Corsica-Sud, France</td><td>27/02/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8571</td><td>Bonifacio, Corsica-Sud, France</td><td>03/05/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8572</td><td>Bonifacio, Corsica-Sud, France</td><td>03/05/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8573</td><td>Bonifacio, Corsica-Sud, France</td><td>03/05/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8574</td><td>Bonifacio, Corsica-Sud, France</td><td>03/05/2017</td><td>C. salinus</td><td>L. Sorba</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8575</td><td>Guérande, Loire Atlantique, France</td><td>03/2017</td><td>C. diaphanus</td><td>A. Soileux</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8576</td><td>Guérande, Loire Atlantique, France</td><td>03/2017</td><td>C. diaphanus</td><td>A. Soileux</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8577</td><td>Malahmar, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8578</td><td>Malahmar, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8579</td><td>Malahmar, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8580</td><td>Malahmar, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8581</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8582</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8583</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8584</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S.<br>Litim</td><td>W</td><td>Genetics and<br>morphological</td></tr></tbody></table>
<table><thead><tr><th>N° accession<br>MNHN</th><th>Locality</th><th>Collection<br>date</th><th>Taxon</th><th>Collector</th><th>Wild/<br>Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2021-8585</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8586</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8587</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8588</td><td>Tahouna, Souk Ahras, Algeria</td><td>18/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8589</td><td>Sidi Salem, Annaba, Algeria</td><td>26/01/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8590</td><td>Sidi Salem, Annaba, Algeria</td><td>26/01/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8591</td><td>Sidi Salem, Annaba, Algeria</td><td>05/01/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8592</td><td>Sidi Salem, Annaba, Algeria</td><td>05/01/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8593</td><td>Sidi Salem, Annaba, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8594</td><td>Sidi Salem, Annaba, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8595</td><td>Sidi Salem, Annaba, Algeria</td><td>17/03/2017</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8596</td><td>Sidi Salem, Annaba, Algeria</td><td>10/05/2016</td><td>C. salinus</td><td>L. Boumendjel</td><td>B</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8597</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8598</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8599</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8600</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8601</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8602</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8603</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8604</td><td>Ain Seynour, Souk Ahras, Algeria</td><td>08/03/2017</td><td>C. salinus</td><td>A. Market and S. Litim</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8605</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8606</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8607</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8608</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8609</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8610</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8611</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr><tr><td>MNHN-IU-2021-8612</td><td>Laalalig, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and morphological</td></tr></tbody></table>
<table><thead><tr><th>N° accession<br>MNHN</th><th>Locality</th><th>Collection<br>date</th><th>Taxon</th><th>Collector</th><th>Wild/<br>Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2021-8613</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8614</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8615</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8616</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8617</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8618</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8619</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8620</td><td>Les Salines, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8621</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8622</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8623</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8624</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8625</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8626</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8627</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8628</td><td>Airport, Annaba, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8629</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8630</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8631</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8632</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8633</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8634</td><td>Reghaïa, Algeria</td><td>29/03/2017</td><td>Chirocephalus<br>sp</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8635</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8636</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8637</td><td>Ben Mhidi, El Tarf, Algeria</td><td>02/03/2017</td><td>C. salinus</td><td>M. Amarouayache</td><td>W</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8638</td><td>Reghaïa, Algeria</td><td>29/03/2017</td><td>Chirocephalus<br>sp</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8639</td><td>Ain-Magroun, Skikda, Algeria</td><td>02/03/2017</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8640</td><td>Ain-Magroun, Skikda, Algeria</td><td>02/03/2017</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr></tbody></table>
<table><thead><tr><th>N° accession<br>MNHN</th><th>Locality</th><th>Collection<br>date</th><th>Taxon</th><th>Collector</th><th>Wild/<br>Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2021-8641</td><td>Ain-Magroun, Skikda, Algeria</td><td>02/03/2017</td><td>C. sanhadjaensis</td><td>L. Boumendjel</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8642</td><td>Souk Rguibet, El Tarf, Algeria</td><td>09/06/2016</td><td>C. salinus</td><td>S. Ghaouaci</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8643</td><td>Souk Rguibet, El Tarf, Algeria</td><td>09/06/2016</td><td>C. salinus</td><td>S. Ghaouaci</td><td>B</td><td>Genetics and<br>morphological</td></tr><tr><td>MNHN-IU-2021-8648</td><td>Le Chênes, Aube, France</td><td>09/05/2021</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8649</td><td>Chambord, Loir et Cher, France</td><td>08/05/2021</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8650</td><td>Leudeville, Essonne, France</td><td>15/03/2020</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8651</td><td>Chamarande, Essonne, France</td><td>31/03/2021</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8652</td><td>Plaisance du Touch, Haute Garonne, France</td><td>09/06/2020</td><td>C. diaphanus</td><td>N. Rabet</td><td>B</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8653</td><td>Chambord, Loir et Cher, France</td><td>18/07/2021</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8654</td><td>Saint Oulph, Aube, France</td><td>18/04/2018</td><td>C. diaphanus</td><td>N. Rabet and J.F<br>Cart</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8655</td><td>Marolles, Essonne, France</td><td>01/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8656</td><td>Marolles-En- Hurepoix, Essonne, France</td><td>01/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8657</td><td>Cheptainville, Essonne, France</td><td>01<br>/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8658</td><td>Cheptainville, Essonne, France</td><td>01/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8659</td><td>Cheptainville, Essonne, France</td><td>02/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8660</td><td>Cheptainville, Essonne, France</td><td>02/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8661</td><td>Cheptainville, Essonne, France</td><td>02/04/2018</td><td>C. diaphanus</td><td>N. Rabet</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2021-8666</td><td>Alger, Algeria</td><td>04/2020</td><td>Chirocephalus<br>sp</td><td>L. Belajdal</td><td>B</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-419</td><td>Le Croisic, Pays de Loire, France</td><td>1908</td><td>C. diaphanus</td><td>Chevreux</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-422</td><td>Mare Lakraf, Alger, Algeria</td><td>1/30/1923</td><td>C. diaphanus</td><td>H. Gauthier</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-430</td><td>Kef El Akdar, Alger, Algeria</td><td>1887</td><td>C. diaphanus</td><td>E.Simon</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-457</td><td>Frais vallon, Alger, Algeria</td><td>1880</td><td>C. diaphanus</td><td>Lataste</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-416</td><td>Sucy en Brie, Val-de-Marne, France</td><td>3/23/1924</td><td>C. diaphanus</td><td>M. André</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-829</td><td>Argenteuil, Val-d'Oise, France</td><td>-</td><td>C. diaphanus</td><td>E.Simon</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-967</td><td>Morbihan, Bretagne, France</td><td>4/30/1998</td><td>C. diaphanus</td><td>S. Buard</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-425</td><td>Lectoure, Occitanie, France</td><td>3/11/05</td><td>C. diaphanus</td><td>Kopperhorn</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-426</td><td>Mantes la jolie, Yvelines, France</td><td>-</td><td>C. diaphanus</td><td>Kopperhorn</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-428</td><td>Meudon, Île de France, France</td><td>3/15/05</td><td>C. diaphanus</td><td>A.Dollfus</td><td>W</td><td>morphological</td></tr></tbody></table>
<table><thead><tr><th>N° accession<br>MNHN</th><th>Locality</th><th>Collection<br>date</th><th>Taxon</th><th>Collector</th><th>Wild/<br>Breeding</th><th>Study</th></tr></thead><tbody><tr><td>MNHN-IU-2007-429</td><td>Villiers sur mer, Normandy, France</td><td>-</td><td>C. diaphanus</td><td>A. Dollfus</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-436</td><td>Saône et Loire, Bourgogne-Franche-Comté, France</td><td>2/13/05</td><td>C. diaphanus</td><td>Dr. Jullien</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-438</td><td>Le Vernet, Auvergne-Rhône-Alpes, France</td><td>-</td><td>C. diaphanus</td><td>E.Simon</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-442</td><td>Angers, Pays de la Loire, France</td><td>3/1/05</td><td>C. diaphanus</td><td>Raimbault</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-446</td><td>St Quai Portrieux, Bretagne, France</td><td>-</td><td>C. diaphanus</td><td>Leprieur</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-450</td><td>Île de Noirmoutier, Pays de la Loire, France</td><td>-</td><td>C. diaphanus</td><td>A. Dollfus Lemire</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-451</td><td>Douai, Hauts de France, France</td><td>-</td><td>C. diaphanus</td><td>Dr. Moniez</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-452</td><td>Evreux, Normandy, France</td><td>-</td><td>C. diaphanus</td><td>E. Simon</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-459</td><td>Lanouaille, Nouvelle-Aquitaine, France</td><td>-</td><td>C. diaphanus</td><td>Ch. Allaud</td><td>W</td><td>morphological</td></tr><tr><td>MNHN-IU-2007-464</td><td>Verdon, Grand Est, France</td><td>12/10/09</td><td>C. diaphanus</td><td>Kadidez Vatiluk</td><td>W</td><td>morphological</td></tr></tbody></table>

Eggs for SEM microscopy were collected directly from female's brood pouch of the three Algerian populations representing three species: *C. salinus*, *C. sanhadjaensis* and *C. algeriensis* sp.nov. They were dried and sputter coated with Platinum (LEICA EM ACE600). For micrographs we used a Scanning Electron Microscope (HITACHI SU3500).

## Results

A total of 57 haplotypes aligned (543 pb), including 28 new sequences, and three which are identical to those from Genbank. All haplotypes are listed in (Table 1). A phylogenetic tree was constructed using these sequences (Fig. 2). The results of different methods of phylogenetic inference (NJ, ML, BA) recognize 12 distinct evolutionary lineages with a high support in all methods, embedded in a large polytomy except for haplotypes of *Chirocephalus marchesonii* from Italy; *Chirocephalus* sp1 from Morocco, and *C. sanhadjaensis* from Algeria. These lineages represent both single haplotypes *C. marchesonii*, *Chirocephalus* sp1 from Morocco, *Chirocephalus* sp2 from Bulgaria, *C. sibyllae* and *C. algeriensis* sp.nov. from Reghaïa as well as clades up to 15 haplotypes whose relationships are well resolved and supported. The different populations of *C. salinus* in northeastern Algeria and Corsica (France) emerge as a single clade. Another clade includes haplotypes from Italy (*Chirocephalus* sp3, *Chirocephalus* sp4 and *C. ruffoi*). Another clade included populations from Ben Slimane in Morocco with populations from Doñana in Southern Spain. A similar clade that groups Portuguese populations, that is a sister group of the France-Netherlands clade. The different populations of *C. diaphanus* from different geographical areas of continental France and northern population from Netherlands converge towards the division of the clade into three well-supported subclades. The first clade grouped haplotypes from south-eastern France (D4, D5, D6, D7, FValligV2, FVarloV1, P_RCL2_01). The second clade consists of a single population from the north of the Netherlands (5VijlM1). The third clade is shared between populations from northern France (D2, D3, D4), populations from southern France (FRoquehV1, FLarivM1, 4FLeuV1) and population from Southwest France (4FChaiM2),

## Genetic divergence

The pairwise sequence divergence between the different populations of *Chirocephalus salinus* (Algeria-Corsica) and the *Chirocephalus* sp3 haplotype from Sardinia is quite significant 15.1 to 15.8%. On the other hand, this Italian taxon shows a slight genetic divergence between 4.6 and 6.6% with *C. ruffoi* and *Chirocephalus* sp4. The pairwise sequence divergence between *Chirocephalus* populations of the new species from Algeria *C. algeriensis* sp.nov. and *C. diaphanus* sensu stricto (France-Netherlands) is 7.2 to 8.3%. *C. algeriensis* sp.nov. a maximum divergence of 9.6% was observed with population of Bulgaria (*Chirocephalus* sp2) and a minimum divergence of 8.1% was noted with populations of Italy (*Chirocephalus* sp4) and Portugal (*Chirocephalus* sp6). The population of Portugal (*Chirocephalus* sp6) and *C. diaphanus* sensu stricto France-Netherlands) shows a slight pairwise sequence divergence ranging from 5.7 to 7.4%. *Chirocephalus* populations that appear as single clades (*C. marchesonii*, *Chirocephalus* sp1, *Chirocephalus* sp2, *C. sibyllae* and *C. algeriensis* sp.nov) show pairwise divergence between other clades ranging from 7.2 to 16.6%.

## Taxonomy

*Chirocephalus algeriensis* sp. nov.

(Figs. 4, 5, 6, 7, 8, 9)
**Etymology.** The specific epithet "algeriensis" refers to Algiers, the main region where the species were sampled.

**Type locality.** Temporary freshwater pond is located in a meadow about 35 Km East of Algiers, close to a railway, between commune of Reghaïa and that of Boudouaou (city El Kerrouche) (36° 44'29.80"N, 3° 22' 6.73"E). The pool has an oval shape and an area of about 1200 m², with a maximum depth of 0.5 m (Fig. 3).

## Type material

**Holotype:** Mature male of 9 mm total length; Algeria: Alger Province: Reghaïa temporary pond; Specimens were obtained by breeding; 2019; L. Boumendjel leg; stored in 96% ethanol (MNHN-IU-2021-8662).

**Allotype:** Mature female of 12.6 mm total length; Algeria: Alger Province: Reghaïa temporary pond; Specimens were obtained by breeding; 2019; L. Boumendjel leg; stored in 96% ethanol (MNHN-IU-2021-8663).

**Paratypes:** Mature male of 8.8 mm total length: (MNHN-IU-2021-8664); same data as Allotype. One female of 12.6 mm total length (MNHN-IU-2021-8665); same data as Allotype.

## Description

### Male

Average length 12 ± 1.6 mm (9.6–15.2 mm, N = 6).

*First antennae* filiform, with apical setae, with a length 0.6x of the second antenna proximal antennomere.

*Second antennae* (Fig. 4a) proximal antennomere curved medially, with a length 1.2x distal antennomere. Bearing an apophysis subconic, digitiform, apex with denticles, total length ~ 0.40x of the proximal antennomere. Second antenna distal antennomere thin, curving nearly at 90° in its proximal part, then straight to the apex. Proximedial projection lobiform, with curved apex, distal surface bearing apically truncated spinulae; a few spinulae occur similar near the proximedial projection.

*Antennal appendage* (Fig. 4b) total length 0.7x second antenna. Basal lamella small and triangular bearing a dorsal carina, with a length ~ 0.25x of the basal lamella, and apex elongate, total length ~ 0.20x of the basal lamella, with marginal tubercles. Basal lamella margins bearing a basal medial lobe decreasing towards the apex, Lateral margin provided with one row of papillae each with an apical denticle (Fig. 4c). Antennal appendage dorsal lamella length ~ 2x basal lamella, serrate in the distal half with medial and lateral margins bearing subtriangular papillae (larger medially) diminishing in size distally, apex acute. Outer margin of serrate, bearing four digitations equipped with a spines apical and tubercles on the surface (Fig. 4d).

*Labrum.* Distal lobe rounded; 0.5 x of length exceeds the distal margin of the labrum.

*Mandibles* asymmetrical, right mandible molar surface with two posterio-apical acute spines, equal in length, followed by a single teeth, molar surface of left mandible bears by a single robust teeth followed by eight small teeth on the lateral outer margin.

### Maxilla I

Typical for the genus (Cottarelli et al. 2010, 2017). Setae aciculate and straight; there is also a posterior ventral acute spine.

*Thoracic appendages*, typical of the genus. Thoracopods (I to X), with endopod (Fig. 5c) broad, forming right angle to limb, bearing short, plumose acute marginal spines. Exopod oval, margined with a few small plumose spines and a long plumose setae apically. Epipodite subcylindrical, length 3x width. Praepipodite completely divided in two, broadly rounded distally, with margin serrate and inerm (Fig. 5b).

*Thoracopods I* (Fig. 5a), with the first three endites (I to III) arcuate, rounded, with long plumose setae. Endites IV to VI rounded as long as broad, bearing respectively 4, 5 and 6 short plumose acute spines and some long plumose setae (Fig. 5d).

*Thoracopods VI* (Fig. 5g), with endite IV, V and VI lamellar as long as broad, bearing respectively 2, 2 and 6 short plumose acute spines and some long plumose setae (Fig. 5h).

*Thoracopods XI* (Fig. 5e) without praepipodite and with a conical epipodite four times longer than its width, with acute apex and without spine or setae. Endite IV, V and VI rounded, as long as broad, bearing respectively 5, 4 and 4 short plumose acute spines and some long plumose setae (Fig. 5f).

*Gonopod* (Fig. 6a) typical for the genus, with a lateral conical lobe. Gonopod rigid basal portion subtriangular, medially margined in dense denticules. Eversible portion of the gonopods (Fig. 6b) represented by conical and curved apex (Fig. 6c).

*Abdominal-somites* smooth, without projections.

*Cercopod* length does not exceed the length of last three abdominal segments; cercopods bordered with long plumose setae.
## Female

Adults with an average length of 12.8 ± 0.73 mm (12-14.4 mm, N = 7).

*First antennae* with apical setae.

*Second antennae*, slightly shorter by 20% than the first pair of antennae, flattened, subconical apex acute. Second antenna margin bearing a basal medial lobe with acute spine (Fig. 6d).

*Thoracopods* I-XI (Fig. 5i, j, k) as in the male.

*First genital* somite reaches half the length of the second one (Fig. 6e).

*Brood pouch* conical, extending to the fourth abdominal somite with gonopore typical for genus.

*Abdominal somites and cercopods* similar to those of males.

*Eggs* (Fig. 7a, b) Shell with high and thin palisaded structure delimiting more or less regular polygons. Diameter of eggs: 344.03 ± 0.01 µm, n = 30.

SEM analysis of resting eggs of *C. salinus* and *C. sanhadjaensis* from Algeria shows the same egg structure of *C. algeriensis*; however, they differ in egg diameter size: *C. salinus* from Sidi Salem with 326.5 ± 19.4 µm (n = 134) (Fig. 7c, d) and *C. sanhadjaensis* from Ain-Magroun with 448.24 ± 30.93 µm (n = 90) diameter (Fig. 7e, f).

## Distribution

*Chirocephalus algeriensis* sp.nov. has a distribution restricted to the vicinity of Algiers region, not far from the coast.

## Differential diagnosis

*Chirocephalus algeriensis* sp.nov. closely resemble to *C. diaphanus sensu stricto* Prévost, 1803. *C. algeriensis* sp.nov. differs from *C. diaphanus* Prévost, 1803 by antennal appendage dorsal lamella length ~ 2x basal lamella, with a small dorsal carina and by second antenna margin of female bearing a basal medial lobe with acute spine.

In *C. algeriensis*. sp. nov. the male second antennal distal antennomere as shorter as proximal antennomare and slightly curved in its proximal portion, a characters shared by *C. cupreus* Cottarelli, Mura and Özkütük, 2007; *C. sibyllae* Cottarelli and Mura, 1975; *C. salinus* Daday, 1910; *C. hardingi* Brtek, 1965; *C. pelagonicus* Petkowski, 1986; *C. reiseri* Marcus, 1913 but not in *C. diaphanus* sensu stricto and *C. ruffoi* the distal antennomere is more curved in its proximal portion and its longer than proximal antennomere in *C. ruffoi* while in *C. diaphanus* sensu stricto distal and proximal antennomere are equal.

In *C. algeriensis* sp. nov. the basal process of the distal antennomere of the antennae is quite developed, bearing apically truncated spinulae. In *C. diaphanus* ss this process is rather developed, straight, bearing spinules and papilla with acute spines. In *C. sibyllea*, *C. cupreus*, *C. weisigi* Smirnov, 1933; *C. hardingi* Brtek, 1965 this process is quite elongated and narrow, straight, bearing a few sharp tubercles.

The dorsal lamella of *C. algeriensis* sp. nov. ~ 2x long than the basal lamella, bearing four long basolateral digitiform expansions are present in many species of the genus *Chirocephalus*: *C. diaphanus*, *C. salinus*, *C. ruffoi*. In *C. diaphanus* the dorsal lamella is slightly longer than the basal lamella. In *C. algeriensis* sp.nov. the shape of basal lamella resembles that of *C. diaphanus*, *C. diaphanus carinatus*, *C. ruffoi* Cottarelli and Mura, 1984; *C. sibyllae* and *C. cupreus* with margins bearing basal medial lobes. Basal lamella of *C. algeriensis* sp. nov. exhibits a small carina. This last resembles that of *C. sanhadjaensis*, *C. marchesonii*, *C. algidus*, where the size of carina differs between species (~ 0.25x of the basal lamella). Dorsal carina is proportionally much larger in *C. weisigi* and *C. diaphanus sensu stricto*.

In female *C. algeriensis* sp.nov. the second antenna margin bearing a basal medial lobe (tubercule) with acute spine absent in all other species of the "*diaphanus*" group.

## Morphological analysis of *C. diaphanus sensu stricto* from France

Examination of the male's second antennae of different *C. diaphanus* populations sampled from several localities in France (Gonfaron, Bouconville sur Madt, Saint Maximin, Marolles en Hurepoix, Guérande) reveals that a basal process of the distal antennomere is rather developed, straight, apically bearing spinules and papilla with acute spines, on the medio-basal margin (Fig. 8). Dorsal lamella of the antennal process is slightly longer 1.2x than the basal lamella and exhibits a carina proportionally much larger than the basal lamella ~ 0.70x (Fig. 9).

## Chirocephalus collection

Examination of the *Chirocephalus* species collection from Algeria and different populations of *C. diaphanus* ss from France present at the MNHN from Paris (Table 3) confirmed that the morphology of specimens collected in the Algiers region (Frais Vallon, Kef el Akhdar, Lekral Pond and Bab Ezzouar) are identical to the specimens of the new species *C. algeriensis* sp.nov.
Discussion

For the first time, we present a phylogenetic analysis integrating the Algerian *Chirocephalus* populations, identified so far solely by their morphology. We also
performed a phylogenetic analysis of different French populations of *Chirocephalus diaphanus* and *C. salinus* which have been described from France and
reported as also present in Algeria (Simon 1886; Gauthier 1928; Beladjal et al. 1995; Samraoui and Dumont 2002; Samraoui et al. 2006).

The phylogenetic analysis indicates that we have three distinct clades in Algeria: one clade representing *C. sanhadjensis* whose specific status is clearly confirmed here and two clades in the *diaphanus* / *salinus* species complex. None of the Algerian populations is included in the *diaphanus* subclade sensu stricto. In contrast, the populations of Corsica morphologically identified as *C. salinus* are associated with many populations from the Northeast of the country suggesting that *C. salinus* is indeed present in Algeria as already recorded by Samraoui et al. (2006). The third clade includes a new species that we have described in this paper as *C. algeriensis* sp.nov.

In addition, our study confirms the existence of several cryptic lineages related to the "*diaphanus*" group, especially in the Mediterranean basin, which confirms the results of Reniers et al. (2013). Our phylogenetics results demonstrate that some populations of *Chirocephalus*, originally assigned as *C. diaphanus* (*Chirocephalus* sp2, *Chirocephalus* sp4, *C. algeriensis* sp.nov, *Chirocephalus* sp5, *Chirocephalus* sp6) do not constitute a single monophyletic assemblage with *C. diaphanus* sensu stricto populations from France-Netherlands.

The molecular thresholds indicative for species level differentiation at COI in branchiopod crustaceans (average sequence divergence of 7–10%: Cox and
Hebert 2001; Zofkova and Timms 2009) would allow assignment of a new species. This study suggests that a detailed morphological analysis, in particular
of the antennae and the basal lamella, should make it possible to resolve taxonomy of the *C. diaphanus* sensu lato clade.

The population from Portugal (*Chirocephalus* sp6) is placed as the closest relative of *C. diaphanus* sensu stricto (France- Netherlands) with relatively high statistical support for the different phylogenetic methods used. This population could be integrated into *C. diaphanus* sensu stricto but a verification of its morphology is necessary.

It seems that species with a narrower geographical distribution should present a lower genetic diversity (Doorenweerd et al. 2020; Nekola and White 1999).
Overall, it seems according to this genetic analysis that the population groups of *Chirocephalus* (*Chirocephalus* sp2, *Chirocephalus* sp4, *C. algeriensis*
sp.nov., *Chirocephalus* sp5) are quite genetically isolated from other populations of *C. diaphanus* sensu stricto (from France-Netherlands). According to
Reniers et al. (2013), the highly divergent haplotypes found in different Mediterranean areas hint at the long-term presence of suitable habitat in these areas
throughout the Pliocene and Pleistocene. Our results confirm the hypothesis proposed by De Meester et al. (2002) to explain the apparent paradox shown by
most freshwater organisms which produce resting stages (and therefore have a high potential for dispersal) but exhibit low gene flow among their
populations, as indicated by molecular analysis.

Three main mitochondrial lineages of *C. diaphanus* ss from France and the Netherlands are grouped into a large well-supported clade. Our results are quite consistent with previous studies on *C. diaphanus* fairy shrimp species (Reniers et al. 2013). In this study we did not exactly find the distribution of the haplotypes identified by Reniers et al. (2013) since we found the P_RCL2_01 haplotype and not FNor (J (2), H (2)) in the same locality in Aurseulles, called Normandy in Reniers et al. (2013). In the Var, only the FVar (K), FVal (L), FVal (M (9)) and Fcam (M (10)) haplotype group was found. The Nordic haplotype FNor (J (2), H (2)) was therefore not confirmed in this study in France.

Knowledge of *Chirocephalus* taxonomy has improved in Algeria; three species are now clearly separated. *C. salinus* Daday, 1910 which occupies the eastern regions and *C. sanhadjensis* Boumendjel et al., 2018 restricted to Guerbes-Sanhadja-Complex, while *C. algeriensis* sp. nov. which has a centered distribution around Algiers region. This species was found cohabiting with *Tanymastix stagnalis* Linnaeus, 1758 near ponds visited and sampled by Gauthier in 1928 and where he collected *Chirocephalus* with *T. stagnalis* and *Lepidurus apus* Linnaeus, 1758. Unfortunately, part of this forest has disappeared today and replaced by urban constructions. There is a need to protect the rest of this locality.

This study is a continuation of work on non-malacostracan crustacean in Algeria (Ghaouaci et al. 2017, 2018; Menail et al. 2023). Its objective is to update
knowledge on biodiversity and to obtain a database necessary for the management and protection of wetlands.

Declarations

Acknowledgements

The authors thank the Muséum national d'Histoire naturelle Paris (France) and the laboratory UMR BOREA for their help in advancing this research. We are grateful to Souad Ghaouaci, Amel Market, Sara Litim, Ghouzala Gouasmia, Aude Soileux, Jean-Claude Tempier, Eric Queinnec, Adeline Lecot and Jean-François Cart for their sampling efforts. The authors also thank Laure Corbari, Paula Martin-Lefevre for their assistance and access to the collection. We also thank Géraldine Toutirais for her technical help (Plateau technique de Microscopie Electronique (PTME) du Muséum national d'Histoire naturelle). The authors are grateful to Dr. Lynda Beladjal for her helpful comments on the manuscript.

Funding

This study was part of a national research project (CNEPRU) financially supported by the Algerian Minister of Higher Education and Scientific Research funds (General Directorate for Scientific Research and Technology Development, GDRSDT).

Consent for publication
All authors have read and approved the final manuscript.

**Conflict of interest**

The authors declare no conflict of interest.

**Author information**

Marine Bioresources Laboratory, Marine Sciences Department, Badji-Mokhtar Annaba University, Algeria

LYLIA BOUMENDJEL; https://orcid.org/0009-0004-8040-9507; ly.boumendjel@gmail.com

MOUNIA AMAROUAYACHE; https://orcid.org/0000-0002-1244-0156; m.derbal@yahoo.fr

Sorbonne Universités, Muséum national d'Histoire naturelle, UCN, UA, CNRS, IRD, Biologie des organismes et écosystèmes aquatiques, BOREA, UMR 8067,
CP26 75231, 43 rue Cuvier, 75005, Paris Cedex 05, France

CELINE BONILLO ; https://orcid.org/0000-0002-9892-8901 ; celine.bonillo@mnhn.fr

NICOLAS RABET ; https://orcid.org/0000-0002-3578-7574; nicolas.rabet@mnhn.fr

Office de l'environnement de Corse, France

LAURENT SORBA ; laurent.sorba@oec.fr

Sorbonne Université, CNRS, INRAe, IRD, Institut d'Ecologie et des Sciences de L'Environnement de Paris, IEES-Paris, 75005, Paris, France

THIBAUT BAGNI; https://orcid.org/0000-0002-3307-5717; thibaut.bagni@sorbonne-universite.fr

Corresponding author

LYLIA BOUMENDJEL

Tel (+33) 09 52 38 66 72 ; ly.boumendjel@gmail.com

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## Figures
<figure><img src="image_2.png" /><figcaption>Figure 1</figcaption></figure>

Geographic distribution of *Chirocephalus* populations included in the study

**Black triangle**, *C. salinus* populations from France and Algeria included for genetic analysis; **black star**, *C. sanhadjaensis* populations from Ain Magroun and Belkroun included for genetic and morphological analysis; **black diamond**, *C. diaphanus* populations from France and Netherlands included for genetic analysis; **black dots**, different populations of *Chirocephalus* species included for genetic analysis; **black square**, *C. algeriensis* populations from Reghaïa included for genetic and morphological analysis; **white pentagone**, *C. diaphanus* populations from France included for morphological study; **white square**, *C. algeriensis* populations from Alger (Algeria) conserved in the MNHN collection in Paris; **white diamond**, *C. diaphanus* populations from France conserved in the MNHN collection in Paris
<figure><img src="image_3.png" /><figcaption>Figure 2</figcaption></figure>

Phylogenetic tree of *Chirocephalus* species based on COI nucleotide sequence data. The tree was constructed using the Bayesian inference (BI), neighbor joining (NJ), and Maximum likelihood (ML) algorithms, separated by a slash, only nodes with a statistical support greater than 60% labelled 0.9 posterior probability. Populations names and accession number N°. correspond to the identified populations in Table 1

<caption>Figure 3</caption>

Biotope illustration of *Chirocephalus algeriensis* sp.nov. (Reghaïa pond)
<figure><img src="image_4.png" /><figcaption>Figure 4</figcaption></figure>

*Chirocephalus algeriensis* sp.nov. a-e, male. a, second antenna. b, head, right side, anterior view. c, basal lamella. d, antennal appendage dorsal lamella. e, enlarged detail of dorsal lamella
<figure><img src="image_5.png" /><figcaption>Figure 5</figcaption></figure>

*Chirocephalus algeriensis* sp. nov. — h, male. i-k, female. a, first pair of thoracic limbs. b, enlarged detail of the praepipodite P1. c, enlarged detail of the endopod P1. d, enlarged detail of the endites P1. e, eleventh pair of thoracic limbs. f, enlarged detail of the endites P11. g, sixth pair of thoracic limbs. h, enlarged detail of the endites de the sixth pair of thoracic limbs. i, first pair of thoracic limbs. j, eleventh pair of thoracic limbs. k, sixth pair of thoracic limbs

<caption>Figure 6</caption>

*Chirocephalus algeriensis* sp. nov.. a-c, male. d-e, female. a, genital segments. b, basal process of hemipenis. c, eversible tip of hemipenis left. d, second antennae. e, abdominal segments and ovisac
**Figure 7**

SEM micrographs of eggs. a-b, eggs of *C. algeriensis* sp.nov. c-d, eggs of *C. salinus* Daday, 1910. e-f, eggs of *C. sanhadjaensis*. Scale bars: a, c, e: 200 µm; b, d and f: 100 µm

**Figure 8**

Enlarged detail of the apophysis of the distal antennomere. a, population of *C. algeriensis* sp. nov. from Reghaïa. b-f, populations of *C. diaphanus* from France. b, population from Var (Bonne Cougne). c, population from Meuse (Bouconville sur Madt). d, population from Var (L'Etendard at St Maximin).e, population from Essonne (Marolles en Hurepoix). f, population from Loire Atlantique (Guérande)
<figure><img src="image_6.png" /><figcaption>Figure 9</figcaption></figure>

Dorsal carina of the basal lamella. a, population of *C. algeriensis* sp. nov. from Reghaïa. b-g, populations of *C. diaphanus* from France. b, population of Var (Bonne Cougne). c, population of Meuse (Bouconville sur Madt). d, population of Var (L'Etendard at St Maximin). e, population of Essonne (Marolles en Hurepoix), f, population of Loire Atlantique (Guérande). g, population of Var (Signe)