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S62/20h
M ^ ^ ^ ^ J ^ " ^ ^ ^ ^ ^ CmETXUATBANG NlTdC VOI Sp- HO TRO Situ AM BEN BOAT TTOH CHONG OXY HOA
CUAD|CHCHIETTfrHAIMl£N/rc»M««te„5
TOM TAT
ABSTRACT
uwivio' ^ extractsJttm manne sponge Ircbiia mutans throu^k nppu 6B-, ™ J - J
appniniale cmuMonfr mtUaldmt eanaia, A™, * , . . . ^ ^ ^ * OOVKB. The emaionWHe. ISmn. 'On-a.o, lemperatm!: SO-Q mracUo,i^paMon:3eme.
I.B«TVAND£
H a mien & met ngu6n t f e i nSng cho vl$c nghiSn a>u khai thSc c i c host ch^t sinh hoc t v nhlSn OS Idij n i n g ijng dgng Irong y hoc do s v da dang v4 c^u Irtic h<ia hpc cOa cac chSt chuySn h6a aS trong chiing. Trong nh&ng nSm g i n dSy, m « s6 nghiSn ciiu d5 phat hl^n ra nhOng hprp chit cd host Unh sinh hoc tCf
h a miSn nhu ctilt chdng oxy h6a, chSt Pch^ng viSm, i d i ^ Idiuan, chong lao, ch6ng ung Ihi/, khang n i m , chSng s6t rSt kh^ng vinjs va khang HiV (Mehbub va cpng sv, 2014). H i i mi6n diiprc xSp vac nh6m cfi chira host chat ch6ng oxy hoa cao Irong s6 cac ngu6n heat chit ch&ig oxy h<Sa trong nr nhiSn. Met s i chat chuyin l»Sa Irang hai miSn ciS hogt tInh
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TOUONGDAI HOC NHATRANG • 3
Tup ihiKhoa hoc - Cow^ nghc Thiiy ^un So 2/2016 chong oxy hoa cao nhu polypeptide, saponin,
sterol, fiavonoid, glycoside va cac hop chSt phenol (Halliwelt, 1994, Chatrman va Singh.
2012) Ngoai ra, Sato va cong su' (2006) da tim thSy cac hap chat carotenoid, polyphenol, glutathione trong mot so loai hai mien, 6ay cOng la nhu'ng hop ch^t c6 hoat tinh chong oxy hoa cao
Viet Nam c6 i5ieu kien rat thuan lai cho cac loai hai mien cung voi cac sinh vat ky sinh tren chung phat tnen Nhu'ng nghien ciiu tru'oc day da cong bo c6 khoang 201 loai hai mien dugc Iim thay 6 vung bi4n Viet Nam (Thai Minh Quang, 2000) Trong do, cac loai hai mien Ircinia spp. da duoc cac nha khoa hoc quan lam nghien ci>u khai !hac hoat ch^t sinh hoc (Kumar va Pal, 2012) Nghien cuu cua Orhan va cong su' (2012) cho thay dich chi4t tu cac loai hai mien Ircinia spinulosa, I fasaculala.
and / variabilis co hoat tinh khang khuan, khang nam, khu' g6c tu- do 2,2 diphenyl- Ipicrylhydrazine (DPPH) va LCC che acetylcholinesterase Hoat linh khang khuan cua dich chi^t tu' hai mien /, mutans da du'p'c Nazemi va cong su' (2014) cong b6 va cac protein co hoat tinh khang khuan da dicoc phan lap tu' hai mien boi Salehi va cong su' (2014) Nhin chung, nhu'ng cong bo v4 nghien cuu hoat tinh ch6ng oxy hoa cua hai mien / mutans con rk han chd Oe khai thac hieu qua nguon tai nguyen quy gia nay, viec nghien cu'u tach chigt cac hoat chat sinh hoc tu' hai mien / mulans 6 vung bien Viet Nam la r§t can Ihiet Huynh Nguyin Duy Bao va Nguyin Khac Bat 12015) da nghien cuu anh hu'ang cua toai dung moi chiet va sieu am den hoat tinh ch6ng oxy hda va ham lu'ong protein cua dich chilt hai mien / mutans, kit qua nghien cu'u cho thay nuoc cat la dung moi Ihich hap de chiet xuat hoat chit ch6ng oxy hoa tu' hai mien va protein la hoat chSl chong oxy hoa chu y l u co
4 . muONG OAI HOC NHA TRANG
trong djch chilt De chiet xuat ho^t chat chong oxy hoa tCf hai mien dat hieu qua cao, nghien cii'u nay da tien hanh xac dinh anh hu'dng cua dilu kien chiet xuat bSng nuac cSt vai sy h6 tra sieu Sm den hoat tinh chong oxy hoa ciJa dich chiet ti> hai mien / mutans nham tim ra dieu kien chiet xuat thich hap de thu du-ac dich chilt CO hoat tinh ching oxy hoa cao.
II D O I TLCgNG, VAT LIEU VA PHl/ONG PHAP NGHIEN CIJ'U
1. Nguyen vat lieu va hoa chat 1 1 Nguyen vat lieu
Hai mien / mutans su dung trong nghiSn cuu nay du'oc lay mSu a vung bten Phu Quoc, tinh Kien Giang
l l i i i h I. H:ii mien /. muian\ lav mau iV vijng bien Pliij Quot Ngay sau khi lay m i u , hai mi§n du'pc u'ap lanh va van chuyen ve phong thf nghiem Tru'ang Dai hpc Nha Trang Tai phong thi nghiem, hai mien du-p'c bao quan dong o' nhi^t do - 20'*C de si> dung cho nghien ci>u nay 1 2, Hoa chat
2.2 diphenyl-lpicrylhydrazme (DPPH), Bovine serum albumin (BSA), Folin-Ciocalteu reagent du'gc mua ti> Cong ty Sigma-Aldnch, Hoa Ky. Cac hoa chat con l^i Id (ogi dat ti§u
lm<MgliMkoc-t»^^ny^
So 2/2016 c h i i o diins Gho pMBacH M a hoc. * « ! c » i u a
* c a i « « Lot>ac»ieMe,,Aii^ vieai^ty
Wako;f«iJtBaii.
2. NiiiongpMp ngMen dm
•nii nghiem xac dinh arifi hirtng eBa aiSu Weii cWh xu4t ( S le nguySh liluftlung moi. thm
^an cW*, nhW g^ chA. s i lin chiit) b&ig nirdc eft MM sgi hS tfigr siSu am d i n hMt trnh s t i i hoc c h ^ 6i(y cfla djdi d A tCr hai mien /. iMlans^ d u j c t i n faift lifnj sau:
2.*. Thf ngUfm xac efnh anh huang cOa If 10
M3i itJM W n g W m . ^ 100 a hai miw dong lanh * -20°C aira di c4t rtio dJn Wch Ihufc 1 - 2 mm i4i dSng hOa wM niiftc cit o c i c t ; l« nguyin Wu/ra»»ceit fg/mt) B: 1/2,1/4,1/6, 1/8 v4 1/10. H5ti h(rp # n g h i ^ ftisrc dtia a c h i t xuil MM a? h&trv cfla stag s«u em iSn s6 a)KH2 trong thdfi gian 10 phat ft 30>C. Sau d6airadHyiamft4°oirong20phiJtviMvent6c 3000 vdng/ptia. Sau khi ly tarn, ach 1 % dWi vaioeqUagllyloeWhMmansdI.BSIecaiiOT tiSn hanh chlSt W 2 Ite nOa viW cac Iheng sa va each ohA nhl/ Ife l^u. d b h t a : Bru Hutfc to cac l4n chilt nhep chung W i& tiln hanh ly tarn ft 4'C ftong 10 ph« vftl vjn l i o 15000 veng/phOt. TSch Sydjchiytativaiocquagily loc Whatman s 4 1 , h i sungttem dong mai vio dich loc cho de 3000 ml HSi chiit Dich chlit hai mlSn thu duoc gi0ft4"C dOng d l phan tfch hoat tinh ch6ng oxy hea trong ngay.
2.2. TtilnghmiSsainhinhhiangctatM gian chiM
MS mlu thI nghiem Hy loc^ hai miSn Mng lanh ft-20« dira dl d t nh« dSn kich Ihuftc 1 - 2 mm i& ding h6a vfti nuftc c ^ ft i ; le nguyen lieuftiuftccfclhWilKip«dUw;xac dinh Irong thI nghiem ft mgo 2.1. H5n h w OSng hda du^re dlra di cNSt x u * vffl sv h8 Sv cfla song
sieu am l a n s 6 2 0 K H z f t 3 0 « trong cac khoang thfti gian khac nhau: 5, 10, 15, 20 va 25 phat Sau M dua di ^ tarn ft 4*0 trang 20 p h « vfti van I f a 3000 vang/phiil Sau khi ly lam. Bed l i y (Jch va loc qua gl4y loc Whahran s6 1 . Ba loc tago Ben hanh chiet
¥ 2 a n nOa vfti cac th6ng so va each chiet ntaf Ian d i u . rach loclhu duwc ttf cac ftn cMet nhap Chung lai rSi liln hanh ly lam ft 4°C trong 10 phut TO! van t i c 15000 vong/phiit.
Tach iSy d i * ly tarn V4 loc qua giSy loc Whatman s6 1 , b6 sung them dung m6i vao
* h loc a i cBm bao the tfch dich chilt thu d u ^ aJa cSc miu bang nhau. D|ch chilt hSi mien Biu duoc gio ft 4<C dDng d l phan tich host tfnh ching oxy hoa trong ngay.
Z3. ThI nghi$m xac Oinh inh hudng da nhi$t tKcHk
^46l m i u Ihl nghiem lly 100 g hai mien
« n g Isnh ft-20<; dira di cat nh6 r4i ding hda d&lg hoa vfti nuftc d t ft t^ le nguyen lieu/
nBfto pit fWch hsni da auprc xac dinh Irong thI nghiem 6 nure Z.1. Hln horp ding hda duvc aira di chUl xuA bing sieu am 6 cac nhiet de khie nhatt 10, 20. 30, 40 va 50°C trong thfti gian Ihlch hgp I E dugc xac dinh trang thI nghiem ft mgc 2.2. Sau d6 dua dl ly tarn ft 4"C trong 20 phiit vfti van toe 3000 vdng/phllt Sau khi ly lam, tach ily dich va Ipc qua glly loc Whatman s 6 1 . Ba ipc duvc tiln hanh chilt ISi 2 iin nOa vfti cac thdng s6 va each chilt nhi/ lin ^ u . Dich loc thu dwjrc ti> cac lln chill nhSp Chung Isi i4i Sen hanh ly tam ft 4°C trong 10 phijt vfti vSn tic 15000 v6ng/phut. Tach lly dich ly tarn va loc qua gily loc Whatman s i 1, b l sung them dung mOI vao dich loc d l aam bao t h I tIch dich chUl thu dufic cQa cac miu bIng nhau. Dich chilt hai mien thu d u w giO ft 4«Cd(lng^phan tfch host tinhching oxy hda trong ngay.
TRUONG D * I HOC NHA TRANG . 5
Tap chi Khoa hoc - Con^ nolle Tluiy san So 2/2016
2 4 Tt)i nghiem xac dinh anh huung cua so Ian chiet
M i l m i u thi nghiem l l y 100 g hai mien dong lanh a -20°C du'a di c i t nho r l i ding hoa ding hoa vai nu'oc cIt a ty le nguyen lieu/
nu'ac cat thich hp'p da du-gc xac dinh trong thi nghiem a muc 2 1. Hln hap ding hoa du'p'c due di chilt xuat bIng sieu am vai thai gian va nhiet dp thich hop da dirac xac dinh trong thi nghiem a muc 2 2 va 2.3. Sau do dua di ly tarn a 4°C trong 20 phut vai van toe 3000 vong/phut. Sau khi ly tam. tach (ay dich va loc qua giay loc Whatman so 1 D l i vai cac m i u chilt ti> 2 - 5 Ian, ba Ipc tu- Ian chilt 1 du'gc tien hanh chilt lai tu'ang irng tu' 1 - 4 Ian niia vai vcn cac thong so va each chiet nhu' )ln dau. MSU chiet 1 Ian khong lap lai qua trinh chiet Dich loc thu du'oc ti> cac Ian chiet nhap Chung Igi roi tien hanh ly tarn o 4''C trong 10 phut vai van toe 15000 vong/phut. Tach lay d|ch ly tarn va loc qua giay loc Whatman so 1, bo sung them dung moi vao dich loc de dam bao the tich dich chiet thu du'gc cua cac mau bIng nhau. Dich chilt hai mien thu du-ac gii>
6 4=0 dung de phan tich hogt tinh chong oxy hoa trong ngay
3. Phipcng phap phan tich
Hoat finh chong oxy hoa cua djch chiet tCr hai mien du'gc danh gia dya vao kha nang khi>
g l c t i / do DPPH du'gc phan tich theo phu'ong phap cua Fu va cong sii (2002) va dya vao tong nang lye khO du'gc phan tfch theo phirong phap cua Oyaizu (1986).
4. Phu'O'ng phap xi> ly so li^u
So lieu trinh bay trong bai bao nay \k gid tri trung binh cua 3 l§n thi nghiem. Tinh gia tn tmng binh va ve 66 thi SLK dyng phin mem Microsoft Excel 2003. Sy khac bi§t co ^ nghla ve mat thing ke (p < 0.05) cua cac gi^ tri trung binh du'ge phan tich tren phan mim thong ke R phien ban 2 13,1,
III. K^T QUA NGHIEN CLfU VA T H A O LUAN I. Anh hu-ffng cua ty fe nguyen lieu/niwyc cat den hoat tinh chong oxy hoa cua djch chiet tij' hai mien
Hoat tinh khir goo t y do DPPH v^ tong nang lye khCp cua dich chiet hai mien du'gc chiet xuat bang nuac c I t vai eac ty I? nguy§n lieu/nugc cat khac nhau co sy h i trg cua s6ng sieu am t^n so 20KHz trong thai gian 10 phut g 30°C duge the hien tren hinh 2.
1
0
| 0 . f i
= 0.4
01 -^112
-i-iis -*-vs
- ^ 1 / 1 0
^ ^
(b)
^ ^
^ s ^ ^
Th^ tich dich chiSt (ml) •* 0.1 02 0,3 0.4 0
Th^ tich dich chitt (ml) Hinh 2. Hoai tmh khir eoc tu tin H P P H l o i , A . '
trong Ih(ri gian 10 phiit ff 30°C
• TRUONG OAI HOC NHA TRANG
S6 2/2016 Kit q i « j * a n ; ; « * phuong ^ i ; « h o « t t #
* lenguyen Heutouiic d l c e i p h huos^ i ^ l i o * t(nh khii g & tiP do DPPH va ting n * ^ IWC i*i> cOa tjch ohilt hai ifiien (P < a o o C i Htnh 2 cho tfily, hoat tinh Bift glc V do DPPH-
^ dfch rtiill hSimienlang ( P < 0,06) khi Siay
« ^ le nguyen lleu/nufc d t tCp i e ain 1 « j , nhmig hoattlnh khOrglctM'do DPPH cila d i * i , chill hai mien khOng ting..Mm (P > 0,05) Wn*"
t j le nguyen lieu/nudc d t thay a l l to 1/4 d|n 1/10. Traijd khi do, tong nang lire khu cua dich chilt hai mien, tang fp < OfiSj khi thay dil t}
le nguyin. Iieu/nu6c d t ttr i e din 1/6, ting nahg-lirc k h * c t o : & i n ^ I aiehdriS hiimien
^ feu
iF
BOd^t
|oa 01
••s
-•-«
-*-«
•»-»
•*« ' ^^
f^
* ^ le. nguyin neu/nute d t 1 / 6 . 1 ^ va 1/10 MlBog c6 svr khac biet (P > 0,05). Tir kit qua n a y ^ Chen ^ if nguyen lieu/nuAc d t thich Sl^rp ctw chiet xuat chat ching o ] ^ h6a t)> hai
m B n » 1 / B .
2. A»H hudiig cda thoi gian ohtSl d i n K ^ tfnti chong o ] ^ h6a cea dfch chilf tip I d t m t l n
Moat tirih khft glc \sf do DPPH va ting hSng 1 ^ kh» cda dich chiit hfi mien duffc chiet xuat hang nwSrc d t vol ty le nguyen lieW ninSc d t la 1/6, co su hS trgr dja s6ng sieu am t i n s i 20KHZ trong d c khoSng th&l gian khac nhau * 30»C duoc M hiln tten hinh 3.
(b)
iMMlldiclilllliiill "• TrtMUdMMSlmll
<><liie«<<l<liidllg UiU gian kliic ghaii » 3 I K Kit qua phan tloh phuang sal cho thiy thM
gian chilled anh hudngdln host tinh khSgIc ttf do DPPH ve ting nSng Itfo khi> oOa d|eh chilt hai mien (P < o,00f), Hinh 3 cho thIy, hoat tinh k h * gio tu"do DPPH cOa dich chilt hai mien tang ( P < #,65) khi tsnglhei gian ohllt ti> 5 phiMfen i a j * a . n h u n g host tinh khftgic tv do DPPH ata'dibh: chilt hai mien khOng tang them (P > 0,0^ khi tang th&l gijn chilt I * 10 phlM din 25 phlit Trong khi d l , khi IgngthM gian chilt ti> 5 phiit din ISphiit llm tang ting nang lireWl^qSa^bttchilttiaimleh (P<©,(B) va ting ningi«c'kh&oOadich chifeljaiinien
giam khi tang th«r| gian chilt KF 15 phM len 25 phlit (P > 0,05). TO kit qua nay da chon thM gian chilt thfch hop cho chiit xuit chit ching oxyhdatirhaimieniaiSphUt.
3. Anh hvdng cua nhift do chilt d i n host tfnh ching oxy h6a cOa dich chiet ti>
hai miln
Host llnh khi> goc tv do DPPH va ting nang iu« khi> cua dich chUl hai mien duoc chiit xult bing nucSc d t vfti 1} le nguySn lilu/
nuftc d t la 1/6, c6 sv h i iro cua sing silu am t i n s i 20KH2 trong 15 phOt ft cac nhiet ae chilt khac nhau d u w thi hien tren hinh 4.
IRl^iNSDAI HOC NHA TRANG . 7
lap chi Khoa h<u - Coiii^ ii^hc Thiiy \iiii
Hinh 4. Hoi liAni; niro-i- ta
Th^ Bch dich chiil (ml) tinh khir goc lir do DPPH (a) \ M H cac t> 1^ nguyin li^u/nirdc c
long It: 1/6 c nhici dg khac nhau
Th* tfch dich chiit (ml) lire khir (b) ciia djch ehict hai mien du^c chiet xu^t ho t r y cua song sieu am tAn so 20KHz trong 15 phiit
Ket qua phan tieh phyang sai cho thay nhiet dp chiet cd anh hu-ang den hoat tinh khLf goc tu' do DPPH (P < 0.05) va tong nang lye khLf (P < 0,001) cua dich chiet hai mien Hinh 4 cho thay, hoat tinh khu' goc ty do DPPH cua cac m i u dich chiet hai mien khong cd sy khae biet (P > 0,05) ngoai tru' m i u chiet xuat a nhi^t do 30°C eo hoat tinh khu' goc tu' do DPPH cao han m i u ehfet xult 6 nhiet do I C C (P < 0.01). Tong nang lyc khLr cua cac m i u chiet xuat a 10, 20, 40 va 50«C khong co sy khac biet dang ke (P > 0,05) va tbip han tong
nang lyc khCi' eua mau chiet xuat d nhi^t dp 30°C. Ti> ket qua nay da chgn nhiet dO chiet thich hyp cho chiet xult chat ch6ng oxy hda ti>
hai mien la 30°C.
4. Anh hu-o-ng cua so Ian chiet den ho^t t(nh chong oxy hoa cua djch chiet tiy hai mifiti
Hoat tinh khu- goc ty do DPPH v^ tong nang lu-e khCK cua dich chiet hai mien du'pc chiet xuat bang nu'ac cat vai ty le nguyen li$u/
nu'O'c eat 1^ 1/6. co su- h6 tra cua song sieu Sm tan so 20KHZ trong 15 phut a 30=0 vai s chiet khac nhau du'p'c the hi#n tren h
I dicti chiit (ml)
Hinh 5. Ho^t tinh k h i i gfic tir do DPPH (a) vi, tflng nang lire khir (b) cua d bang n w c cat v.n eac t j le nguyen lieu/niroc c^t: J/6 e6 su- ho t r y cua soi
ff 30''C viii so Ian chi^t khac nhau
8 • TRL/ONG DAI HOC NHA TRANG
T»cUaoBki/e-CSK!aa£niysa,i
Kit i M pMn M l phuong sai cho thay s6 l b cMlt c6 i n h huSng din hoat I M i kM> glc ttr do DPPH va ting nang Ivc kh» dja <|di chUt hai mien (P < ftlMJ). t M i 5 cho Ihly, host tinh k f * glc Itf do DPPH va £ n g nang Krc khii cua cjch c N i t hai n«i» tang kM tang s i lln cWlt ICrl a i n 3 Bn (P •= 0.01). Hoat tinh k h i j g i c t v do DPPH ra ling nang Urc khft cea cac m i l chilt 3, 4 va 5 a n khong c6 sv khac bi» dang k l ( P > 0 , ( B ) . T O k ^ qua n a y ^ chon s i a n cwlt thfch hop cho cWlt x u i chit ching oxy hfta Kr hH miln a 3 a n .
So 2/2016 IV. K^T LUAN VA K I ^ NGHj
Trong Sesi kien chilt xuat bang nuftc cat vfti su he tr? aia siiu am. t ; le nguyen lieu/
nude cat. that gian chilt nhiet do chill va so a n chilt CO anh huftng dang k l a l n hoat tinh chong oxy hoa cua dich chUt tu hai mien. Cac thong so IWch hop Cho chilt xult chll ching oxy hda tCr hli mien /. mutans liang nuftc cat vfti s(f ho t i ^ oja song sieu am tan so 20KHz nhu sau:
- Ty ie nyjyen iieu/nuftc d t 1/6 - Thai gian chilt IS phlit;
- (Met dp chilt 3 ( K : : - S i a n chilt 3 a n .
T A l U f U T H A M K H A O lUrgnft:
lUiigAmk:
2. ' : ' » ' « ° ^ ' ^ S i , * A . J . A . I L . A l . g ™ « l , „ . G . ( 2 0 I 2 ) . C i a « o x i c . , « l . « i ™ d ™ « i v j ^ s p < n 8 = . A i m H « S ! c J t a ™ / ^ 7 > , y i < ^ D i , e ^ 2 ( 3 ) - 2 3 4 - 2 3 8 " " " l y ot saeded m a ™
,M10:Troi*«d«Fpi«Dn«iesfcdmxmq.rfbioKliv«.ilto™,Dn«,. 12(8): 4539-1577.
I W C N G DAI HOC NHA TRANG . 9
Tap ihi Khoa hot - Ccmon'^hO Thin san Sd 2/2016
N a / c m i M \foiallehi. \ 1 A A . .I.imili. J . Ma-ihincliian, ' \ . G h a \ a m . ivi P. ( 2 0 1 4 ) , Compari.son of a n i i b j c i e n a l acti\ ilici <)i In inui nmains eMracls in iw o diftereni bejsi^ns from Kish Island. Persian Gulf. Iran Iniiiuiii .huniah'f FnliL-iiL's SLWIUC^ 13(4) S23-833
( J r h a n . l t . O z L c h k B . Konuklugil, B Pui7. A . K.it>an. U O . Prokscli, P ( 2 0 1 2 ) B]oacH\ i n screening of the M."k'i:(L-d mrki-h marine spon^c> and three compciund* from .igclc/.^ oroide.s R e c o r d of Natural Products. 6(4|'
0>aiz:u \ 1 (19S6) Studis on p r o d u t l s of brow ning ri-Mlions autioxidativc activities o f products of browTiing reaction prepared from gluco-iainine Japanese Join iwl of \iiiiiiioii. 4 4 , 3 0 7 - 3 1 5
ii; Saiehi. a.. Paionss. R . Ahmad N (2014) Isolation .And Characieni^alion O f S o m e K i n d Bioactive Proieins Sponge A>. .Anli bacteria I .A^ent International Journal of Scientific & T e c h n o l o g y Researcti, 3 ( 3 ) ' 233-236.
J J Sato. S , Kuramolo, \ l . O n o . N Ircinamine, B (20061. Bioactive alkaloid from m a r i n e s p o n g e Dacnlia sp Ti^ualwlnm tellers, Al 7S71 - S T i
12 l a h c n . .\ . Jalaline/liad. S (20151 .Antio\idati\e and C M O I O M C Effect of M a r i n e S p o n g e {Geoilia perarmaUi) L Mracis aijainsi Breasi and Colorectal Cancer Cells 2""' International Conference on A d v a n c e s in Environment.
A!!rtculture& Medical .Sciences ( I C \ E A \ 1 - 1 5 ) . June 11-12, 2 0 1 5 , Antalya ( T u r k e y )
!,•> I hai \linli Ouani! (2(1! 3) \ re\ leu of ihe d n e i s i t y of s p o n g e s (ponfera) in Vietnam T h e 2"'' intemalional u o i k s h o p o n marine bioresourees of \ i e l n a m . Hanoi 5-(> C 2 0 1 3 . 109-J15
11) • TPUONG DA[ HOC NHA TRANG
