Ma ual of dissecio tech i ues fo the lio ish, Pterois volitans, f o Southe Gulf of Mexico Alfonso Aguilar-Perera, Luis Quijano-Puerto, Evelyn Carrillo-Flota
Fundac on
Arrecife Alacranes
parque nacional
UADY
CAMPUS DE CIENCIAS BIOLÓGICAS Y AGROPECUARIAS “Luz, Ciencia y Verdad”
FACULTAD DE MEDICINA
VETERINARIA Y ZOOTECNIA
*
Citaio : Aguila -Pe e a, A.1 , Quija o-Pue to, L.1, Ca illo-Flota, E.1 . Ma ual of disse io te h i ues fo the lio ish, Pterois volitans, f o Southe Gulf of Me i o . Fu da ió UADY-CONANP-CCBA. U i e sidad Autó o a de Yu atá pp. . Depa ta e to de Re u sos Ma i os T opi ales, Ca pus de Cie ias Biológi as Ag ope ua ias, U i e sidad Autó o a de Yu atá . * alfo so.aguila @uad .
Desig : L.D.G. C isia Re é Aguila Pe e a. E ail: fooi _us@hot ail. o
Acknowledgements We tha k the follo i g stude ts ho o t i uted i p oessi g lio ish sa ples: Jua Chí-Bla o, He á No elo-Pe h, Ka la Pe aza-Az o a, Sa a tha Villa ue a-Moe o u de g aduates i Ma i e Biolog UADY , Dia a A el Co o ado-Ri e a a d Ja ie Mé dez-A guia o u de g aduates i Biolog , I situto Te ológi o de Alta ia, Ta aulipas . D A i Tuz-Sulu UADY p o ided helpful a s to o tai sa ples. The olla o aio of lo ste di e -ishe s f o the Pa ue Na io al A e ife Ala a es PNAA as ital to get lio ish sa ples. We tha k the autho it of the PNAA, M.e C. C istó al Cá e es-G. Ca tó a d M. e C. Y i Ra í ez-He á dez. D . Ma k S. Pete so U i e sit of Southe Mississippi ki dl p oof ead the te t.
Index
Introduction 4 5 Objective Problematic situation with 6 the lionfish Fish collection 8 Taxonomic identification 9 General procedure 10 Considerations for sampling lionfish 11
Morphometrics and meristics Internal sampling What can be done with the recorded data? References Appendix
13 22 31 32 34
Introduction Wo ld ide, the i t odu io of spe ies to the a i e e i o e t is p o oted e tai a i iies elated to ishe i p o e e t, a ua ultu e, a d ha itat esto aio . Ho ee , the a ua iu i dust is e e o e espo si le of su h i t odu io s e ause a a ua ists a elease pet spe ies to the e io e t a eless ess he ha dli g the . The esulted i t odu ed spe ies, i a ases, ha e e o e i asi e a d a th eat to the e os ste Se e s et al. .
f o the Southe Gulf of Me i o, ith e phasis i the Pa ue Na io al A e ife Ala a es, this Ma ual is i te ded to e a p oto ol i disse io te h i ues useful fo a age s a d stude ts to o tai asi iologi al data to u de sta d the iolog of the lio ish, a d uli atel u de sta d its i asio .
As a a ua iu spe ies, the lio ish Pterois volitans – ai e to the o al eefs of the Pa ii a d I dia O ea - is i po ted US he e it osts up to USD $7 Mo is . A out ea s ago, this a ua iu ish as released into coral reefs of Florida and its i t odu io has e o e a iologi al i asio . Its high o a it , high fe u dit , a d lo a u da e of p edato s ha e allo ed its populaio to i ease a d disse i ate all o e the Atla i O ea . Its p ese e puts at isk the fu io al a d st u tu al i teg it of the a i e e os ste i this egio Cote et al. .
Bo . 1. The lio ish Pterois voli-
tans – ai e to the I do-Pa ii O ea - as i trodu ed to the Atla i O ea here it has e o e a i asi e spe ies that threats the fu io i g of the ari e e os ste .
As a st ateg to sta da dize the p o edu es fo e o di g iologi al sa ples of lio ish
4
Objective Des i e disse io te h i ues a d ho to e- ha e a p oto ol fo s ste ai all e o di g ioo d iologi al data fo p o essi g lio ish Pte- logi al sa ples of this ish a d ge e ai g a darois volitans) a d k o its iologi al a d e olo- ta ase to do u e t i fo aio o this spe ies. gi al aspe ts f o the Southe Gulf of Me i o. This a ual ill allo esou e a age s to
5
Problematic situation with the lionfish The lio ish is a ai e d elli g f o the I do-Pa ii that, due to its at a i e aspe t, is p i ed high i the a ua iu i dust Bal oa . This i dust is espo si le fo its i po taio to No th A e i a he e so e a ua ists eleased it a eless ess i to o al eefs of Flo ida i Mo is . Due to iologi al ep odu io , feedi g , e ologi al p edaio , a d o ea og aphi a i e u e ts, te pe atu e fa to s, a d to a o -p ope a age e t la k of lea legislaio i a ua iu t ade , this ish st i ed a d its populaio has g o to i asi e p opo io s Cote et al. . Fi st ge e alized sighi gs of lio ish sho i g high de siies ega i The Baha as i , a d the it as p og essi el dete ted i a othe a eas i ludi g Cu a, Do i i a Repu li , Pue to Ri o, Ve ezuela, Ho du as, Belize, a d Me i o S hoield . B the e d of , the ish as dete ted the i st i e of the Southe Gulf of Me i o ea A e ife Ala a es Aguila -Pe e a Tuz-Sulu . I this late a ea, its a u da e has al ead ea hed le els of i asio L贸pez-G贸 ez et al. .
6
The lio ish has eu oto i e o i its i spi es hi h ep ese ts a isk, ot o l to a ua ists ut also to o i e di e s ho atte pt to aptu e it Cohe a d Olek . Besides, the lio ish is feedi g o ish ju e iles a d usta ea s of o e ial i po ta e. Also, it o su es a p e s that a othe ai e ish also o su e.
Atlani O ean Gulf of Mexi o Cari
ean Sea
Bo . . The lio ish i asio has o ered al ost all Atla i O ea . The real threat of lio ish relies ot o l o its de ographi e plosio , ut also i its euroto i e o deposited i its i spi es. The lio ish i asio is o prised t o spe ies, al ost ide i al i orpholog a d oloraio : Pterois volitans y Pterois miles.
7
Fish collection The ost e o e ded a d se u e ethod fo aptu i g lio ish at least of s all od sizes less tha 5 is ha d ets. Ho e e , fo la ge ish g eate tha 5 the o l p ope ethod is eithe spea gu o Ha aiia sli g Aki s . It is e t e el e o e ded to use ha d glo es fo a oidi g a ide tal spi e pu tu es. If a o e io al spea gu is used, it is e o e ded ot to spea the ish i the head o i the sto a h to a oid otolith dest u io o sto a h o te t loses, espe i el .
Bo . . The ost o o ethod to apture lio ish is spear gu . Ho e er, this pro edure is da gerous a d glo es ust e or to a oid a ide tal pu tures ith the e o ous spi es.
8
Taxonomic identification Spe i e s of lio ish a e ide iied ta o o i all a ordi g to orpho etri size a d eight a d erisi o po e ts u er of spi es, sot ra s i i s esta lished S hultz .T o spe ies ha e ee ide iied as i trodued to the Atla i O ea : o e of the is Pterois volitans - hi h is the lio ish per se- a d the other o e is the De il´s ire ish Pterois miles . These spe ies are al ost ide i al i orpholog a d oloraio . The o l orphologi al difere e oted s ie ists is that P. volitans has o e ore sot ra i oth the dorsal a d a al i s, hile P. miles has relai el shorter pe toral i s o pared to P. volitans. The i asio i the Atla i O ea is o prised ai l P. volitans % a d i less le el % P. miles Ha er et al. .
Box. . T o spe ies of lio ish ha e i aded the Atla i O ea : the lio ish Pterois volitans) a d the De il´s ire ish Pterois miles); ut the lio ish is ost a u da t 9 % .
Red lio ish Pterois volitans)
De il´s ire ish (Pterois miles
93% Pterois volitans
7% Pterois miles
9
General procedure This a ual has t o ai pro edures for reordi g iologi al sa ples of lio ish: Morpho etri s a d erisi s a d I ter al sa pli g
Opening of body cavity
Weight
Internal sampling
Morphometrics and meristics
Size
Gape height and gape width Body height Size of pectoral fin Spines and rays counting
Gonad extraction Stomach removal Otolith extraction Tissue sampling Fillet removal
Presence of cirri
10
Considerations for sampling lionfish All pro edures des ri ed here refer to ish pro essi g u der la orator o diio s see Appe di for a list of aterials . It is e essar to ha e a proper ork spa e ith la orator ta les i order to allo ate the aterials properl . The ish, pre iousl kept froze , ust e tha ed i tap ater. Pre auio s ust e take efore pro essi g the ish. The i spi es dorsal, pel i , a d a al are e o ous, a d the e o reai s a i e e e if the ish is death for a ouple of da s u der froze o diio s. Therefore, spe ial pre auio s ust e take he ha dli g the ish for a oidi g a ide tal pu tures. While eari g thi k glo es is suggested he ha dli g ish at the egi i g he easuri g a d eighi g , it is e essar to ear late glo es for detailed pro edures gut re o al, otolith re o al . I ase of a a ide tal pu ture, it is eessar to ai tai the afe ted area i hot ater u der tolera le te perature , appl a algesi s if e essar , a d lo al a i ioi for pre e i g i fe io .
are take , a d the ish has ee photographed, the it is re o e ded to re o e the dorsal spi es if e essar for pre auio . Ater pro essi g the ish, it is suggested to dis ard the ar asses i se ure o taiers a d get the i i erated.
O e the orpho etri easures a d erisi s eighi g, easure e ts, ou i g
11
Measure e ts size, eight, pe toral le gth, od height, gape idth a d ou i g of e ter al stru tures irri, spi es, a d ra s are re orded i spe ial for ats see Appe di . Ea h for at ust e illed up ith the follo i g data: la el u er, perso re ordi g i for aio , olle io date, pro essi g date, lo aio of olle io , a d ethod of olle io .
Morphometrics and meristics
Size Lio ish´s od size a e easured usi g graduated i e i eters woode ta les. These ta les i hth o eters , uilt i wood workshops, are asi all two pie es of wood o e of 7 of le gth a d the other o e of 5 ; oth with .5 thi k asse led. The larger pie e has a rule. For easuri g the ish, this later is pla ed o top of the ta le with ish´s s out losed tou hi g the s all ta le whi h egi s at the zero ark of the rule. It is suggested to re ord at least to easure e ts of od size: total le gth TL a d sta dard le gth SL . The TL is take fro the s out losed to the e tre e side of the audal i , a d the SL is re orded fro the s out losed to the ase of the audal pedu le. Fish spe i e s ust e le i le e ough a d free of a defor it i order to do the easuree ts properl .
14
Standard length Total lenght
Weight I o de to eight the ish, it is e essa y to use a ele t o i s ale. Ho e e , efo e eighi g the ish it is e ui ed to ta e the disse i g t ay he e the ish is to e pla ed. The s ale ust e lo ated o top of a lat su fa e, su h as the la ta le. The ish tail ust e i side the t ay ithout tou hi g a ythi g outside. O e the ish is p ope ly pla ed o the t ay, it a e eighed i total TW o guted GW , oth easu e e ts i g a s.
15
Gape height and width Gape a e easu ed y ope i g ide the s out of the ish a d i t odu i g a alipe . Re o ds of height top a d oto o de s of the s out a d idth side to side of the s out a e take . Both easu e e ts a e take to the losest illi ete .
16
Body height It is the dista e et ee the ish´s hest , lose to the pel i i s, to the ish´s a k. For re ordi g this easure e t a etri tape a e used a d re ord it i illi eters.
17
Pectoral fin length Usi g a aliper, the pe toral i is easured fro the ase of the i to the e tre e distal side. The easure e t is re orded i illi eters a d oth pe toral i s are o sidered.
18
Fin´s spines and rays counting As part of the ta o o i ide ii aio of the ish, the u er of spi es a d ra s i the dorsal i are ou ted. Also, the u er of ra s i the audal a d the pe toral i s are ou ted.
Caudal in
Dorsal in
19
Pectoral in
Pelvic in
Also, the u er of spi es a d rays i the pelvi a d the a al i s are ou ted.
Anal in
20
Presence of cirri in head and rostrum The irri are i tegu e tar e res e es i arious parts of the head a d rostru of the ish. It is preferred to ide if the a d ou t the . The suprao ular irri are lo ated o top of the e es hile the oper ular irri are lo ated o the rostru . This later irri are lo ated i the i fra or ital a d oper ular regio .
21
Internal sampling
Opening of the abdominal cavity A re e tly tha ed ish ust e pla ed o the disse i g tray. For ope i g the a doi al a ity, s issors a e used irst a d the a surgeo s alpel to further ope i g. The pro edure ust e do e arefully for a oidi g a y da age i the i ter al orga s duri g the a do i al i isio . This pro edure a e i iiated fro the a al regio .
The ope i g se io a e do e for ard fro the a al regio to the pel i i regio . O e this later area is approahed, it is e essary to use s issors to ut the pel i o e. Also, a alter ai e pro edure ay e follo ed y aki g a i isio o the side of the ish to get the ody a ity ope ed. Before pro eedi g to re o e a y orga , it is e essary to ide ify ea h of the a ordi g to their lo aio i the ody a ity. This ill pre e t a y i ide tal daage o a y orga duri g the i isio to re o e the sto a h or the go ads.
23
Gonad extraction O e the od e ide iied.
a it is ope , the go ads eed to
These later are lo ated o the e tral area of the s i ladder a d lose to the a al regio . Whe the ish is se uall a i e, the go ads are eas to e ide iied si e these are s olle . The go ads ust ot to e o fused ith ese teri fat. The go ads are paired orga s. If the ish is ot se uall a i e, the go ads are dii ult to ide if e ause these are ot s olle . I order to ake the a ess to the go ads eas , for eps a d disse i g eedles ha e to e used. Go ads a e hold ith the for eps a d the ase ut ith s issors.
24
O e the go ads are re o ed, the a e stored i a i ed soluio eutrall ufered for ali % i plasi la eled o tai ers. Ma ros opi all , if the go ad is se uall a i e the it is relai el eas to re og ize ish´s ge der. Fe ale go ads are ello ish a d ha e a gra ulate o diio , hile ale go ads are hiish a d ha e o gra ulaio . For details o this, a d a other pro edures, see Gree et al. .
Stomach removal It is e essary to use for eps a d s issors. Before re ovi g the sto a h, ish´s u al area ust e he ked verifyi g a y aterial ite regurgitated a d logged i the esophagus.
The , the sto a h a e ut usi g s issors fro the esophagus a d arefully re ovi g the sto a h. At this poi t, the sto a h a e stored i the ooler i la elled ags a d eve tually the o te ts a e re oved. If the o te ts eed to e re oved, the sto a h a e plaed i a Petri dish.
25
The , the sto a h e ra e a e ope ed i isio a oidi g a da age i the stoa h o te ts. Pour ater i the Petri dish to pro ide a et o diio to the o te ts. Usi g for eps, the o te ts a e separated arefull a d he ked through the stereos ope. For pro eedi g to ide if the o te ts, proper ta o o i ke s a e used. The o te ts a e stored i al ohol 95% .
26
Otolith extraction The otoliths a e al iu a o ate sto es loated i the utriculus of ish´s i e ea . These sto es ake pa t of the sta ilizaio s ste of the ish ith espe t to the ha ges i ho izo tal posiio . Ea h ish has th ee pai s of otoliths: lapilli, asterisccii and sagitae. Of these, sagitae a e elai el la ge a d t adiio all used fo eadi g g o th i gs i o de to i fe ish´s age. The e a e a ious e o e ded te h i ues to e t a t otolith sagitae f o ish hi h i ol e e o i g the head to e pose the oi apsule. Ho e e , i this a ual, the sele ted te h i ue does ot e ui e e o i g the ish´s head. Fo this p o edu e, the ish ust e pla ed side a d o e the disse i g t a . Usi g fo eps a d s alpel, the ope le is lited a d the issue o e i g the oi apsule is eo ed. P og essi el , the o e of the oi apsule is e o ed u il ope i g a hat h i the a ea u il e posi g the otolith sagitae hi h a e e o ed usi g fo eps. This p o edu e ill eed to e epli ated o the othe side of the ish to e o e the othe sagitae.
27
O e the pair of sagitae are removed, it is lea sed ith tap ater. Ater this, the sagitae a e stored dry i ags or i ials laeled. The ethod for readi g gro th ri gs i ol es ea h sagitae to e pro essed reog izi g ea h ri g. This ethod o siders polishi g a d ur i g the otolith a d ui g the usi g a spe ial sa .
28
Tissue sampling Tissue re o al i ol es re o i g either a i lip or us le pie es or e e gill issue. For re o i g these parts, s issors a e used a d ut a out 1 3 of issue. I ase of i lips, the pel i i is re o e ded. I ase of us le, the a do i al area is suggested. For a oidi g possi le o ta i aio a o g sa ples, s issors ust e ashed out ith al ohol e ery i e a porio of issue has ee take . Tissue a e stored i ials o tai i g a preser ai e al ohol 95% or ufered DMSO . Tissue sa pli g is for studies i ol i g ole ular iology.
29
Fillet removal The illet a e re o ed usi g a sharp k ife, a d aki g the i iial i isio fro the a k of the ish to the rear part.
The illet is re o ed as the k ife is ui g the us le – ithout rea hi g the o eu il o plei g the ut to the ase of the tail. At this part, the illet is ei g peeled of a d the ski has to re ai at the ase of the tail.
O e the illet is re o ed, this is stored i a ylo ag la elled a d kept froze . Fillet re o al is for io he i al a alysis of the us le protei , ater, ar ohydrate, faty a ids .
30
What can be done with the recorded data? to k o the fat a ids proile a d the utrie tal alue of the eat.
Data o tai ed fro the easure e ts, ou i g, a d e tra io s a e used to u dersta d the gro th, reprodu io , ole ular iolog , a d he i al hara terizaio of the eat. Data of size a d eight i e i eters or illi eters a e useful for al ulai g le gth- eight relaio ships li eal regressio a d to deter i e the t pe of gro th of the ish. Go ad sa ples a e used to k o the ge der of ish si e the ish does ot ha e se ual di orphis , deter i e the reprodu i e phases, a d ide if the reprodu i e seaso i the ear. Sto a h o te ts a al sis a re eal the diet of the ish a d deter i e hi h pre s the ish is feedi g o . Otolith sagitae a e used to i fer ish´s age a d gro th. Tissue sa pli g a dis lose populaio ge ei s a d o e i it et ee populaio s i the regio . Che i al a al sis of eat a pro ide useful i for aio
31
References Aguila Pe e a, A., Tuz-Sulu , A. . No - ai e, i asi e Red lio ish (Pterois volitans [Li aeus, ]: S o pae idae , is i st e o ded i the southe Gulf of Me i o, of the o the Yu ata Pe i sula, Me i o. A uai I asio s : S -S . Aki s, J.L. . Edu aio a d out ea h: Buildi g suppo t a d e pe ise. Pages - i : J.A. Mo is J . ed. I asi e Lio ish: A Guide to Co t ol a d Ma age e t. Gulf a d Ca i ea Fishe ies I situte Spe ial Pu li aio Se ies Nu e , Ma atho , Flo ida, USA. pp. Bal oa, C.M. . The o su pio of a i e o a e tal ish i the U ited States: a des ipio f o the U.S. i po t data. I : Cato, J.C., B o , C.L. Eds. , Ma i e O a e tal Spe ies. Colle io , Cultu e a d Co se aio . Io a State P ess, A es, Io a, pp. – . Cohe , A.S., Olek, A.J. . A e t a t of lio ish (Pterois volitans spi e issue o tai s a et l holi e a d a to i that afe ts eu o us ula t a s issio . To i o : – .
C té, I.M., G ee , S.J., Hi o , M.A. . P edato ish i ade s: I sights f o I do-Pa ii lio ish i the este Atla i a d Ca i ea . Biologi al Co se aio : – . G ee , S.J., Aki s, J.L., Mo is, J.A. J . . Lio ish disse io : te h i ues a d appli aio s. NOAA Te h i al Me oa du NOSS NCCOS , pp. Ha
e , R.M., F esh ate , D.W, Whifield P.E. . Mito ho d ial to h o e a al sis e eals t o i asi e lio ish spe ies ith st o g fou de efe ts i the este Atla i . Jou al of Fish Biolog : - .
Mo is, J.A., J . Ed. . . I asi e Lio ish: A Guide to Co t ol a d Ma age e t. Gulf a d Ca i ea Fishe ies I situte Spe ial Pu li aio Se ies Nu e , Ma atho , Flo ida, USA. pp. L pez-G ez, M.J., Aguila -Pe e a, A., Pe ea-Cha , L. . Ma a di e -ishe s as iize s ie ists: dete io a d oito i g of the i asi e ed lio ish i the Pa ue Na io al A e ife Ala aes, southe Gulf of Me i o. Biologial I asio s : -
32
Se
e s, B.X., Buhle, E.R., Salo o , A.K., Pate gill-Se e s, C.V. . A hotspot of o - ai e a i e ishes: e ide e fo the a ua iu t ade as a i asio path a . Ma i e E olog P og ess Seies : –
S hoield, P.J. . Update o geog aphi sp ead of i asi e lio ishes Pterois volitans [Li aeus ] a d P. miles [Benet ] i the Weste No th Atla i O ea , Ca i ea Sea a d Gulf of Me i o. A uai I asio s : S -S . S hultz, E.T. . Pterois volitans and Pterois miles: T o alid spe ies. Copeia : - .
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Appendix General Data Sheet FACULTAD DE MEDICINA VETERINARIA Y ZOOTECNIA CAMPUS DE CIENCIAS BIOLÓGICAS Y AGROPECUARIAS UNIVERSIDAD AUTÓNOMA DE YUCATÁN DEPARTAMENTO DE BIOLOGÍA MARINA
LIONFISH PROJECT LABEL______________
COLLECTION DATA LOCATION:
DATE :
COORDINATES:
COLECTOR:
FISHING GEAR:
DEPTH :
BOTTOM TYPE:
OBSERVATIONS:
TOTAL LENGTH: STANDARD LENGTH:
FINS
MORPHOLOGIC AND MERISTIC DATA
TOTAL WEIGHT: GUTTED WEIGHT:
SPINES
DORSAL CAUDAL ANAL PELVIC PECTORAL
STOMACH CONTENTS:
INTERNAL SAMPLING WEIGHT:
OBSERVATIONS: GONADAL PHASE: OBSERVATIONS:
WEIGHT:
VERIFIED: CHECK UP:
DATA OF PROCESSING:
34
RAYS
OBSERVATIONS
List of materials for dissection and measurements I hthyo eters Electronic scale Disse i g trays S alpel For eps Scissors K ife Al ohol 95% Petri dishes Nylo ags Markers Vials Calipers
35