Geographic Range Expansion of Ephippion Guttifer (Tetraodontidae)

Home , Ephippion, Guinean puffer, Oceanic puffer, Sphoeroides, Sphoeroides annulatus

Accepted Article (Agreement No.070401150009). data fisheries fromin Galiciafisheries-dependent monitoring program small-scale the of was agreementThis study bythe facilitated Galiciato betweenand Xuntade analyse CSIC Fundinginformation [email protected] Email: c/ CabelloCientíficas, Eduardo 6,36208Vigo, IIM-CSIC, Spain Pontevedra, R. Correspondence Pontevedra, Spain 4 s/n,36310Vigo,Spain dasAbelleiras Universidade deVigo,RúaFonte 3 2 IIM 1 Cofradía de Pescadores San José, Avda. de Bueu s/n 36940, Cangas do Morrazo, de CangasdoMorrazo, Bueus/n36940,Cofradía Pescadores SanJosé,Avda.de Departamento deBioquímica, XenéticaeInmunoloxía, FacultadedeBioloxía, Asociation AzulEcoloxía Instituto de deInvestigaciones Marinas,Investigaciones ConsejoSuperior Científicas,

B Geographic range expansion of expansionof Geographic range R. -CSIC, c/ Cabello Eduardo 6,36208Vigo, Spain Pontevedra, AÑÓN B

AÑÓN pagination andproofreading process, which version undergone full peer review but has not This article hasbeen accepted for publication intheJournalofFishBiologyand , Instituto de de Instituto Investigaciones de Marinas, Investigaciones Consejo Superior

1 , A. and the Versionof Record. Please cite this A LONSO -F Blue Ecology.– Blue28,36002 c/Herreros Pontevedra,Spain ERNÁNDEZ

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AND DE A. 761 Accepted Article Distribution KEYWORDS confirmed invalidhistorical revision for species the first time, maturemale showinga anactivelyspawning in phase. identification. caught ( waters The firstrecord ofthe in the RíadeVigo n orth capture eve capture ,

- NE west Histological examination of reproductive tissue

Atlantic,

Spain) isreported r prickly

reported reported ates . Morphometric

reproductive biology reproductive

a previous record and establishes puf in fer

the north Ephippion guttiferEphippion

based onabased ,

meristic - eastern ,

smooth puffer male specimenmm 570 of and DNAbarcod Atlantic Ocean (Tetraodontidae) (Tetraodontidae) was ,

this this as the northernmost tropicalization

carried out in thiscarried outin . e data

from Galician

confirm

total length total

A ed

the the

( L T )

Accepted Article (Carneiro (Carneiro fromrecord Bayof the (Tortonese Biscay entire Ocean including hardandmuddy ofenvironments todepths puffer ( puffer rostrata (Quéro found as the puffer fishes of South America, but approximately 30 tropical alsoTetraodontidae), are196There a depth of a depth trammel L.

1766),

west

blunthead as deep as as deep S lagocephalus Lagocephalus from Morocco and the extreme western Mediterraneanfrom andtheextremewestern Morocco S On E et alet .

and temperate

(Bloch ix phippion nets nets pac et et al c. the

10 species species of coast ofAfrica toAngola,nearBenguela (Shao .

, hygaster 5 m 5 occur in occur

of 2003 January Guinean puffer described . ,

1786 local fishermenlocal 2014).

puffer 480 in the in the mouth of RíadeVigo the

guttifer ; Bañón C entral Africa entral

), . m commonly tetraodontids shallow waters

species

2017

seas (Nelson the prickly

Sphoeroides pachygaster Sphoeroides (Shipp species species in 26oftetraodontid genera puffer fishes(

is a marine demersal marine is a fish inhabiting shallow coastal and estuarine c. substrat & , a

50 m50 (Shipp

spend theirentire life infreshwater cycles

Santás Sphoeroides marmoratus Sphoeroides live specimen of ,

1974) off called blowfishcalled

and puffer a

( have beenrecorded

Praia deBarraPraia ( L. et al (Shao , ,

2011 . but South

1758

. Ephippion , , , some species some species

2016). 1974) ): et al et 19 ), - east Asia ( the 86) and it has also been reported off Portugal 86) andithasalsobeenreportedoff Portugal the smooth . E ,

Caribbean sharpnose Caribbean

and , 2014). It 2014). It isknown inthe eastern . (Müller (Müller & T Tetraodontids are Tetraodontids are

in the in the guttifer guttifer

and globefish 4 guttifer 1 is associated with is associatedwith a of variety habitats °

Yamanoue in can be can be depthsfound atgreater 42' (Lowe southern part

puffer Atlantic (

Figure 46.021'' N et et al

(Bennett roschel 1848)roschel Sea

1838 . Lagocephalus laevigatusLagocephalus , es

2014). Thereone is 1) southward along along southward the , European waters coastal et al et

mostly that live ) and ) and

was caught - 1831 ;

puffer of 8° . , ,

in tropical regions Galicia, 2011)

51' the bluntheadthe ), family marine fishes, ,

the oceanicthe which

worldwide Canthigaster Canthigaster 24.998'' W 24.998'' . Most

in the Spain Atlantic Atlantic may be , such .

) in A

at at Accepted Article standard 5´ standard barcoding region ofthe mitochondrial specimen respect with to 2016)Matsuura, areingeneralagreementcounts descriptions of withprevious species 1974; the (Shipp, featuresthatdiagnostic morphometric andmeristic fin,and caudal theirshapefrom changing ventral surface thenear anus.Thedorsalandlateral ofthesurfaces body flaps.with two There halfway betweenthesnout anterior and margin of eye,oneside the ofsnout, oneach each Naturais 70% ethanol in theand wasdeposited fishcollection oftheMus data guttifer (Tortonese, 1986), mainly from databases biogeographic (OBIS andGBIF) andwas dot distribution based map created ongeoreferenceddata was inonlinemarine reported (Bañón C_FishF1t1 were

The body A muscle sample was collected muscleA wascollected sample ofnumberThe structure nostrils, the of dorsal et et measured atotal length ( in collected fromit al

Santiago deCompostelaSantiago - ., 2016).A651nucleotide C_FishR1t1 (Ivanova C_FishR1t1 (Ivanova the the was . M Mediterranean was including present the record

white. Numerous spots cover white inor inor differences beattributed could to the

were elongated Two andinflatable. nostrils diagnose specimens examined by

following Shipp (1974). no scales, althoughsmall spinules data

Sea are in given L

this specimen as T et et al ) of 570 mm) of570 andadditional morphometric

(Hachaichi, 1981) with the numberwith the reference - ., 2007), following procedures described., 2007),following elsewhere long sequence wasdeposited in GenBank under fromwhich elongate elongate

Table I ( Figure Shipp E. guttifer coI laterally to

to extract DNA to extract andsequence the

. The specimenwas completed

fin andcoloration rays pattern are

gene, primer usingthe cocktail and

ed 2

(1974) ). were

the the body overitspigmented area

The freshspecimen of the . were Measurements a larger were

round Iberian Peninsula Iberian

. greenish in colour, whilegreenish in colour, the with MHN

located approximately

Luis IglesiasdeCiencias present belly onthe

size of sampled the bibliograp posteriorly USC25103 then then preserved in and meristic and meristic nd meristic hic hic records, .

T E. he main

Accepted Article However, information the reproductiveof biology round gamete production (Valdez from cycles, reproductive seasonal, ortwowith one actively periods reproductive to year dynamics (Brown as (Ward sequences resulted in distance values al a within analysis species of and specimen G of sequence vouchered only with the together “Tetraodontiformes Galician waters”. from Barcod the 2013). explored numberaccession KY498013.Thetaxonomic reported enetic ., 2009). has also has also et al et Molecul The A Studies on reproductive onreproductive areStudies biology fishessential to understand population distance . The - .

species genetic divergence intherange meanof value

by NJ Arothron C e occur ., 2009). morphological diagnosisofthewas morphological bythe species corroborated

conducting conducting a omparison ofdifferences nucleotide between of (www.boldsystems.org) Life Database tree clustered in a strongly supported clade the the supportedclade astrongly in clustered tree other tetraodontid other coI

ar, morphologicalar, anddistributional data between red

cluster analysis -

Müller 1841 Peterson

DNA barcodingin succeded the validating identificationof in other tetraodontid in othertetraodontid the the neighbo coI et al et

species

to sequences was 0.31%, while while 0.31%, was sequences ., 2011). Marine pufferfish

17.59% 17.59% also - u Pineda r - joining joining available in the database the in available

like assign from of species

taxa et alet those previously documented previously those amongfish genera E. guttifer E.

ed (NJ) (NJ)

., 2014; Sánchez identification (Kaleshkumar

the analysis usingMEGA(Tamura 6 E. gutti Galician

available in public databases (KP998097). databases (KP998097). public in available by searching for

Lagocephalus coI

of this specimen distances between the Galician Galician the between distances fer E. guttifer es

sequence of the the of sequence

sample to varied between 13.21% 13.21% between varied of thesample was

et al et has not previously been previously has not have arange ofannual - s Cárdenas

found ., 2015

Swainson 1839 Swainson and tetraodontidother E. guttifer for fishfor the project ).

et al et can be foundcan be Galician Galician

molecular ., 2011). es further further

E. guttifer , showing (Ward from from specimen .

et al et in et et ., -

Accepted Article examined. no evidence tissueof hermaphroditic was found of histological sequential inany the slides members spermatozoa the in lumen ofthelobules (Brown specimen sexually mature was able andpotentially to thereproduce, since exhibited testes 3 Sertoli guttifer spermatozoa the in l Peterson ofmaturityspecimen‟s stage wasestablished asthespawningphase capable (Brown to equivalent spermatogonia development thegerminal along evident epithelium ( mass al. correspondits and dehydrated, embedded paraffin in buffered formalin. more specimens would be needed to robustly determinemore specimens towould beneeded robustly developmentofthegonads. stage ( c ,

) 2011). , eosin forhistologicaleosin examination. Thespecimen andclassifiedwas sexed ( . d Testicular tissue wasofthelobular type(TakashimaTesticular 1995), Hibiya, & - ) , with developinggermwithin germinal enclosed cells formed cysts byenveloping The Galician specimen wasamale testes andits represented 1.3% ofeviscerated Gonads cell processes. Within maturation of cyst each issynchronous germcells ( ).

et al. The most result from obtained important of analyses gonadswasthat the the of the Tetraodontidaethe of family,

Although thisfirst isthe histologicalAlthough ofreproductiveanalysis traits i

the unrestricted unrestricted the spermatogonial type(1981). by proposed Grier ing , 2011),intheactively sincethetestes spawningsubphase, exhibited were removed

maturity phase usinghistologicalcriteria (Grier

Serial Serial umen ofthelobules ( subsamples from the Galicianspecimen from the

wax

of , sectioned 3μm at and

E. guttifer the gonad( the Figure

- 3 Peterson etPeterson al.,2011). ( seems agonochor tobe b five ) ). Spermatogenesis in is cystic

in total) were extracted, and

stained withhaematoxylin immediately ,

1981; Brown Figure Common in ic

fixed in 3.6% species since species ( with The a ) n according to , - ( Peterson b E. guttifer Figure ) ) E.

et et ,

Accepted Article 2011 fish and of this of species this al guttifer thereject possibility that he location lagocephalus tetraodontid inthereported area oftheGulf Spanish of Biscay Gascogne Golfe de Dieuzeide (1955: 356) topreviousAccording his Sea identifyreliably neither previous inAtlantic Europeanoccurrence poorly watersis with sporadic intrusionsin southern observedsince 1974decade (Gómez ., 2008)orelsewhere inFr , es constitutes off ; Quéro

in the

early

It hasbeenreported It in the literature that The increaseinnumber, as aswell changes in the distribution the the exact catch location

has otherwisenever been the the

highlighted reports ( fishes Spanish coast GulfSpanish ofthe of record European AtlanticEuropean Ocean et al et

recorded in

, the northernmost eastern inthe confirmed record such E. guttifer

the Bayof Biscay . Tortonese ,

. ‟ 2008)

The present (recorded in the Bay of Biscay). by as the red as thered cornetfish ,

in SanSebastián(Lozano who only noted the the who onlynoted same information French in Tortonese Tortonese (1986) . In Galician waters . InGalician waters work (Tortonese to .

, Tortonese (1986)reportedTortonese onespecimen from incorrectly

ench waters

1986 the western Mediterranean nor the record reported areain the French ; Carneiro

could be could - Gesteira Gesteira has been relatedhas been

morphological and descriptivecharacters e placed the location

Biscay stablishes (Béarez ,

this this Fistularia petimba ,

1973), the et alet found, this is supported also et al could be

E. guttifer but - . et al et , documented Rey

2014) . a newfish species ,

the exactthe location

which raises doubtsaboutwhich raises validity the 2011)

However, at However, at ., 2017) ,

1952) original record to a

Sea .

misidentification However, these global warming

is a and on map.the ( .

of the . Hachaic Lacépède

Taking n Atlantic and there and there are No

was by numerous Atlantic Ocean

that evidence

by the the Bay of (Quèro Biscay

of hi account in

was stated not a rise of0.24° time was reported by oceanic puffer

Moreover, ,

1803 (Bañón „

tetraodontiform

Galician waters signalé dans le African species African species 1981

reports ( , but the Cantabrianthe , the only , the Bañón &Sant of only occurrences of

) of the presence the .

we cannot Its Its .

the the

two needed to provide

. C per per C & L.

á d et et s, Accepted Article Bañón Bañón References examination general Muñoz forproviding advice and during discussions his fruitful Cangas staff fordonatingthespecimen kindly and We tothank wouldlike ACKNOWLEDGEMENTS anddistributionabundance of(Rijnsdorp fish species and north tropicali grunt Sande body size - Pomadasys incisus , easte , ,

2008), the redbanded2008), the seabream

78 northnorthernmost inthe occurrence (Tetraodontiformes, Tetraodontidae) from (north Galicianwaters 10.1111/j.1439 easternin the Atlantic. (Syngnathiformes: from Fistularidae) Anorthernmost Galician waters. occurrence R R z . & . & ation of ,

rn Atlantic fish different speciesrepresenting biogeographicaffinities, habitats 1574

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ournal of

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Accepted Article Shao, K., Matsuura, K., Shao, K.,Matsuura, L., Leis, J. Liu,Hardy, G.,Jing,L.& M.(2014). Sánchez Rijnsdorp Quéro Quéro OBIS (2018). Nelson Shipp , , ,

, R Science theeffectResolving of Maritime Tetraodontiformes. Paris: 2018 Oceanographic Commission.Available at NJ the GulfCoastthe Laboratory Research Museum www.iucnredlist.org/details/193758/0 dx.doi.org/10.2305/IUCN.UK.2014 guttifer doi.org/ ofMazatlan, Sinaloa,Mexico. coast Sphoeroides annulatus & J J

J - . . . - - Cárdenas .,

, C C Ceballos L :

A S John Wiley&Sons, Inc. . . .

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Porché Delachaux &Niestlé

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, The pufferfishes oftheAtlantic (Tetradontidae) Ocean. . &

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– M

– . Vayne 832 1583 C .

Vayne Annales delaSociété desAnnales sciences naturelles delaCharente A . & , .

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climate change on fish populations. climate onfishpopulations. change (Jenyns, 1842) (Tetraodontiformes, from Tetraodontidae) the , . 0426.2011.01754.x Wilson

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Accepted Article

doi.org/ mitogenomic perspective 10.1371/journal.pone.0017410

. PLoS ONE

e17410 . Accepted Article Mass MERISTIC BIOMETRIC Galicia,near Spain T ABLE Standard lengthStandard ( Fork length ( length Total ( Gonad Eviscerated Total Body width Body depth lengthPectoral Anal Preanal length finDorsal base length Predorsal length Interorbital distance Postorbital length Eye diameter Preorbital length Head length rays rays fin pectoral finNo. ofrays caudal of No. rays No. of fin anal No. of fin rays dorsal

fin base length

mass 1

mass Measurement and

(g)

mass ( (g) L

L L

F H T

)

) )

(g) S )

meristic collectedfrom

Measurement 2504 2907 32.5 297 285 123 445 529 570 140 35 61 58 22 43 11 19 10 97 68 28 8

(mm)

specimen specimen % 66.7 64.0 13.7 13.0 27.6 21.8 31.5 15.3 7.9 4.9 9.7 6.3

of Ephippion guttiferEphippion

Shipp (1974) 12.3 14.5 29.9 14.4 106 5.1 18 8 9 11 – – – – – – – – – – – – – – – – – –

10 11

16.6 9.0 18.2 37.8 402 20 19.1

from Accepted Article Spain ( FIGURE 570

mm total

pipo guttifer Ephippion

length).

caught n the in í d Vg, oten aii, north Galicia, southern Vigo, de Ría - west

Accepted Article Typesetter exact location oftherecordedpoint theGalician specimen. ( FIGURE ● , h peet eod ( record present the ), 1 2

inset map.inset double a Insert togreenChange theredspot in insetmap.

Distribution map showing georeferenced location points of points location georeferenced showing map Distribution

● ) - , headed arrow linking the green spot on the main map with the with map main the on spot green the linking arrow headed

and

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guttifer

Accepted Article Typesetter and Spz ( of detail with testes spawning E FIGURE Spc xternal xternal 1 ); (d) );

opooy f h tse (cl i cm) in (scale testes the of morphology

the

3 histological detail of Spg, of detail histological Change

Morpholog eaoarpae centre melanomacrophage a) to etc a) (a) spermatozoa (Spz) filling the the filling (Spz) spermatozoa ical

n histological and .

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istological

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(a) )