ISPshL£h£^ hoc - Cong nghe Thuy sdn So 1/2014
THONG BAO KHOA HOC
ANH H i r d N G CUA TY LE PHA LOANG, NHIET DO, pH VA AP SUAT THAM THAU LEN HOAT LlTC TINH TRUNG CA MU COP
(Epinephelus fuscoguttatus Forsskal, 1775)
EFFECTS OF DILUTION RATIO, TEMPERATURE, pH AND OSMOLALITY ON SPERM MOTILITY IN TIGER GROUPER (Epmephelus fuscoguttatus Forsskal, 1775)
Le Minh Hodng', Hodng Thi Hiin\ Pham Phmmg Linh^, Pham Quoc Hitng*
NgSy nh|n b^: 30/7/2013; Ngay phin bien ihong qua 18/9/2013. Ngay duy?t dang: 10/3/2014
T O M T A T
Muc tieu cua nghien cieu Id tim ra ty le pha loang, nhiet do, pH va dp sudt thdm thau toi uu cho hoat luc tinh trung cd mii CQp (Epinephelus fiiscoguttatus Forsskal, 1775) Tinh triing cd mu cop dugc pha loang & cac ty 14 1:25, I 50. 1:100 vd 1:200 (tinh dich. nirac bien nhan tao). Tien hdnh xac dinh ty lepha lodng toi uu cho hoql life tinh triing cd mu, sau do ty le ndy dugc su dung cho cdc thi nghidm ve anh hudng cua nhiet do (10. 20, 30, 4ff'C), pH (6. 7. 8. 9) vd dp sudt thdm ihdu (200. 300. 400, 500 mOsm/kg) Moi thi nghiem duac lgp lai 3 ldn. Ket qud cho thdy tinh triing cd mu egp hogt lifc tdt nhdt khi duac kich hogt trongmoi truang vai ty lepha loang 1:100, nhiet dd 2(/'C. pH^7vd dp sudt thdm thdu Id 500 mOsm/kg.
Tic khod • Cd mu egp. ty le pha loang, dp sudt thdm thdu. hogt lifc tinh tritng A B S T R A C T
The aims of present study were to find the optimal dilution rate, temperature, pHand osmolality for sperm motility of tiger grouper (Epinephelus fiiscogullalusforsska.\, 1775). Tiger grouper semen was diluted at ratios of 1 25,1:50; I. IOO or 1:200 (semen: artificial seawater). Detennining optimal dilution ratio for sperm motility of tiger grouper, then it was used for further experiments lhat effect of temperature (10. 20. 30 or 40°C), pH (6, 7, 8 and 9) and osmolality (200, 200, 400 and
500 mOsm.kg'). All experiments were replicated three times. Tlie results indicated that tiger grouper sperm was the highest when activating in solution with dilution rate 1.100, 2(fC. pH=7 and osmolality 500 mOsm.kg^'.
Keywords • Tiger grouper, dilution rate, osmolality, sperm motility
I. DAT V A N D ^
Tinh tning cua hau h i t cdc lodi d bat hoat trong buing sg vd dich ti/ong. Hoat dgng cua chung xay ra ngay sau khi phdng thich ra ngodi mdi tnjdng nu'dc (sinh sdn ty nhien) hogc trong moi tru'dng thieh hdp (sinh sdn nhdn tao) Mdt trong nhdng thdng so quan trong d l danh gid chat lu'gng tinh djeh va kha nang thy ttnh eua tinh trdng d Id hoat lyc eua chiing [13], [15], [18]. Tuy nhien, hogt lyc cug tinh tning cd bj dnh hu'dng cua mpt vdi thdng s l trong mdi tnfdng hogt dpng ciia ehung nhu- nong dp cdc cation (K*, Na*. Ca'*, Mg=*), dp suit tham thiu.
nhiet dp, pH va tn# pha lodng [4], [5], [16], [18], [19].
Hilu bilt eac thdng s l ndy cd t h i giup tgo ra du'ge mdi tn/dng hoat lyc toi LFU cho tinh triing eua cd, giiJp qud trinh sinh sdn nhdn tgo du'O'c tot hon [13].
Cd nhilu nghien cdu da dupe thu'c hien trSn tinh tning ca dii vdng [16], tinh tning cd chem [3], cd bd da [17], ea hoi Dgi Tdy Du-ong [9]...
Cd mij cop {Epinephelus fuscoguttatus Forsskal, 1775) la ddi bftyng nudi bien d gid tn kinh t l cao.
Hien nay chiing dang du'oe nuoi nhieu d khu vyc cua chdu A nhu" Ddi Loan, Singapore, Philippines d Viet Nam, lodi d nay dang du-pc nudi d mpt so tinh
' TS. L6 lUlinh Hoang, ^ ThS. Phgm Phuung Unh, ''TS. Ph^m Quoc Hung Vi^ Nuoi trong thiiy san - Trudng D?i hQC Nha Trang ' HoSng Thj Hi^n: Cao hpe Nuoi trorg thuy sdn 2011 - Tnfdng Dai hpe Nha Trang
TRl/ONG OAI HOC NHA TRANG • 19
Tgp chi Khoa hoc - Cong nghe Thdy sdn So 1/2014 nhw Qudng Ninh. Phii Yen, Khdnh Hda vd Bd Rja -
Vung Tdu [1, 2] Mae dii trong nhdng ndm gan ddy, vige sinh san nhan tao cd mu CQp da dippc thye hien nhu'ng hi§u qua kinh t l mang Igi du'a eao do thilu thdng tin ve toi u'u hda d c dieu kign d n thiet cho sinh sdn nhdn tao nhu- dgc dilm sinh san cua loai, mdi tnj'dng toi u'u cho tinh tning hogt ddng vd khd ndng thy tinh vin tnJng... Chinh vi vdy, nghidn cdu ndy nhdm kilm tra anh hudng ciia d c yeu to ti le pha loang, nhiet do, pH va dp suit tham thiu Idn hoat lyc tinh tning ea mu epp Idm tiln de eho cdc nghien cdu tilp theo ve bao qudn tinh tning ea mu epp.
II. VAT LIEU VA PHl/ONG PHAP NGHIEN CtTU 1. Qudn l y c a dye vd vuot tinh
Cd dye du'ge chdm sde vd nudi du-ong tai long nudi (3x3x5 m) Vung Ngdn - Nha Trang eho din khi ed thanh Wiye sinh dye tiL Thde an dugc sd dyng Id d tap vdi khiu phan an la 5% khoi lugng eo thi. Cd mu epp dye trudng thanh cd khoi lugng 4,4 ± 0,53 kg vd chilu ddi 46,7 ± 2,81 cm, mau sde tuoi sdng, hogtddngtit, khdng xay sdtdj tdt vd khdng bj b0nh dugc sd dyng de tien hdnh vuot lly tinh dich. Tnj'dc khi tiln hdnh vuot tinh, d dye diroc gdy me bing Methylene Glycol (Merck, Ddc) 200 ppm Sau dd diing khan lau sgeh xung quanh Id sinh dye giiip trdnh viec Iln tgp nhdm thu dupe mlu dgt ehit lugng. Diing tay vuot nhe byng d td td cho tinh d|eh chdy ra vdo eppendof tube 1,5 ml dd dugc vd trung vd khd. C i n thdn khi vuot tinh khdng d l lan mdu, nude tiiu d l thu duc^e tinh cd chit lugng tot.
Tinh thu xong dugc giu trdn da bdo vd tiln hanh nghidn edu tgi phdng thi nghiem.
2. Ddnh gia chat l u ^ n g tinh
Tinh dieh dupe pha loang trong nude bien nhan tgo vdi ty le 1:100 (lpi tinh dich va 99pl nude bien nhdn tgo), sau dd dung micropipette hiit 1pl hin hgp tren dgt Idn lam kinh va quan sat dudi kinh hiln vi cd k i t noi vdi camera Nhung mau d trdn 85% tinh tning hogt dpng dugc dua vdo nghidn edu.
3. Xae djnh cac y l u t l dnh hirdng den hoat lyc tinh trCing ca mO epp
Kilm tra dnh hudng cua edc y l u to: ti If pha loang, nhidt dp, pH vd dp suit tham thiu len hogt lye tinh trung cd mu epp. Mau dug'c thu tren 3 eon ed dye, mil thi nghifm duac Idp lai 3 lan
D i u tien, hogt lye ciia tinh triing dupe xdc dinh sau khi pha lodng tinh djch vdi nude bien nhan tgo (gim 27 g NaCI; 0,5g KCI; 1,2g CaCi^; 4,6g MgCI,;
0,5g NaHCOj trong mpt lit nude d t vd pH 7,8) d edc ty lg 1:25; 1:50; 1:100; 1:200 (tinh d|Ch: nude biln
nhan tgo). Quan sat hogt lyc tinh tning vd xde djnh dugc ti lg pha loang toi uu cho hoat lyc ciia tinh trung Id 1:100. Ti lg nay dupe sd dung eho d e t h i nghiem tilp theo.
Kilm tra dnh hudng cCia nhiet dO Idn hogt lyc tinh triing bdng d e h pha loang edc mau tinh djch vdi nt/dc biln nhdn tgo d ti 1$ 1:100 vd duvc dilu ehinh d d e nhiet dp khdc nhau: 10; 20; 30; 40 f ^ ) . Anh hudng ciia pH ldn hogt lye tinh tning dirpc kiem tra bdng edeh pha bang cdc mlu tinh djch vol nude bien nhdn tgo d cac mdc pH khdc nhau: 6; 7, 8. 9 vdi ti If 1:100. De dgt dupe cdc mdc pH theo mong muon tiln hdnh bang each su dyng dung djch NaOH 0.01N va HCI O.OIN d l dieu ehinh.
Muli NaCI dupe su dyng d l pha cde dung djcli cd nong dO dp suat tham thiu khdc nhau 200; 300;
400. 500 mOsm/kg. Kilm tra dnh hu'dng cua dp suit tham thiu d l i vdi vgn dOng cua tmh tning bdng each cho tinh d|ch vao cac dung djch ed edc mdc do dp suat thim thiu khac nhau vdi ti le 1:100
Hogt lyc cCia tinh triing duge quan sdt dudi kinh hien vi d dd phdng dgi 400X. Kinh hien vi ndy duoc k i t nil vdi mdy tinh thdng qua camera. Hogt lyc eua tinh tning (phin tram hogt lyc. vgn tic ciig tinh triing vd thdi gian hogt lyc) dugc phdn tich bdng phan mim CASA.
Cdc s l ligu thu thdp dupc xu ly trdn phin mem SPSS 16.0 vd Excel 2003. Anh hudng ciia ti If pha loang, nhift dp, pH vd dp suit tham thiu Idn hogt lyc cua tinh trung dugc so sdnh theo phuong phdp phdn tich phuong sai mpt y l u to (one-way ANOVA), So sanh sy khac biet cd y nghta thong ke giua cdc dai luong trfn phan tieh phuong sai (Post Hoc Test) bang phuong phdp kilm djnh Duncan's vdi mdc ^ nghTa P < 0,05. Moi quan h f giua d e dgi lupng duge phdn tich theo phuang phdp hli quy Cdc gid trj duge trinh bdy dudi dgng gid tn trung binh ± sai so chuin (Mean ± SE).
III. K^T QUA NGHIEN CCTU V A T H A O L U ^ N 1. Anh hirdng cua t i 1^ pha loang len hogt lyc tinh trung
Ti If pha lodng Id y l u t l quan trgng d l kich thich sy hogt dpng va duy tri khd nang thy tinh ciia tinh triing. Ti If pha loang t l i uu cd the giup ehiing ta kiem soat sy ding nhat cua tinh triing ve khd nang vdn dOng [5], [8]. Ket qud nghidn cdu anh hudng cOa ti If pha lodng len hoat lyc tinh triing cd mu epp cho thay, ti If pha lodng 1200 d phin tram tinh trOng hoat dpng Id cao nhat (97,1 ± 1,05%), tilp sau 66 vdi 95.3 ± 0,71% finh tning hogt dfng d ti If 1:100;
tuy nhien, thdi gian hogt lye ddi nhit d ti If 1100
20 • TRUONG OAI HOC NHA TRANG
ISESM ^hoa hoc - Cong nghe Thuy sdn So 1/2014 (538,2 ± 15,48s). Tinh tning vgn ddng vdi vgn toe
tuong duong nhau d ede ti If pha lodng. Phdn tieh so lieu thong ke eho thay, ti If pha loang la 1:100 vd 1:50 khdng cd sai khde giua phan tram vd thdi gian
hogt lyc (P>0,05). Tuy nhien, hai ti If ndy d sy sai khdc (P < 0,05) so vdi hai ti If edn lai. Xet ehung d e thdng so hoat lye thi thay d ti If 1:100 cho k i t qua t i t nhit trong d e ti If phan tieh (hinh 1).
i le pks loing (ttob t r t ^ : n '
Hinh 1. Anh hudng cua ti I f pha Hogt lye tinh triing cd sau khi phdng thieh ra ngoai mfii trudng t y nhien hogc mfii trudng thy finh nhan tgo la mgt trong nhung y l u to quan trpng quylt djnh den k i t qud thy tinh [6, 8. 18]. Ti If pha lodng ndm trong khoang t l i uu se kich hogt dong bd tat ea tinh triing hogt dpng vd kdo ddi thdi gian hogt lyc, trong khi ti If pha lodng thap hodc eao se anh hudng den kha ndng boi cua tinh tning do nd phai canh tranh cao trong mpt khdng gian hgp hodc phai hogt dfing nhilu trong mft khodng khdng gian qua rpng se khiln eho tinh triing tieu hao ndng lupng ldn va nhanh chet han. Do dd, nlu tinh triing dugc pha loang vdi mft ti If thieh hgp se cho kit qud hogt lyc Id toi uu nhit [8], [18].
Bfin cgnh do. ti If pha loang d l i vdi edc lodi ca khdc nhau thi khae nhau. K i t qua trong nghien cdu nay tuong ty vdi nghien cuu cua Le vd etv [16]
tren ca du vdng, Morisawa vd etv [19] trfn ed vdng, Tan-Fennin vd etv [21] trfn d da tran chdu A. Trong khi dd d cd t i m Iran vd ed rd chdu Au fi If 1 '50 [5], [8], ea mil den vd d doi myc Igi Id t i i f 1:10 eho hogt lye eao nhit [10], [12].
loang Iln hoat luc tinh trting cA mii C9p
2. Anh hudng cCia nhift d f Ifn hoat lire tinh trimg Ket qua quan sdt hoat lyc tinh tning cd mil epp cho thiy cd sy sai khdc y nghTa giifa phan tram, vgn toe vd thdi gian hogt lyc d cdc nhift dp thi nghifm (P<0,05) 0 10°C tinh triing hogt lyc tuang doi cao (75.8± 1,48%, 102,2±8,39pm/svd532,8± 11,83s), hogt lyc cao nhat d 20"C (93,8 ± 1,48%;
130,6 ± 2,35 pm/s vd 561,0 ± 8,82s), sau dd hogt lyc finh triing gidm mgnh d 30''C vd thip nhat d 40''C (9.7 ± 3,12%; 120,6 ± 1,81pm/s vd 71,4 ± 6,84s) (hinh 2). Qua ddy cd t h i khang djnh, tinh triing cd miJ epp cd t h i hogt dpng tot d 10 - 20''C, hogt dpng tot nhat d nhift dp 20"C.
Tuang ty nhu fi If pha lodng. nhift dp thieh hpp eho moi lodi cd khde nhau thi khae nhau Ket qua fi-ong nghien cuu nay tuong ding vdi kit qud nghifin d u tren d t i m Iran [4], cd chep [20]. Tuy nhien, thdng d ndy cao hon d d hli ndu [14], cd chem chau Au [3] va cd dii vdng [16]. K i t qua nghien cdu cho thiy, nhift dp mdi trudng d dnh hudng den eac thdng so hogt lyc cua tinh triing ea mil epp dk ddm bdo tinh chinh xdc.
II.
Hinh 2. Anh huthig ciia nhiet dg len boat lgc linh triing ca mu ci^p
TRUONG OAI HOC NHA TRANG - 21
Tap chi Khoa hoc - Cong nghe Thuy s So i/im
3. Anh h u ^ n g cOa pH Ifn hogt lyc tinh triing K i t qua nghien cdu dnh huong cua pH len hogt lye finh trung ea mu egp cho thiy, tinh tning hogt lye t i t nhit d pH = 7 (93,0 ± 1,22%; 129,3 ± 1,22 pm/s;
539,7 ± 15,52 s). (3 cdc dd pH d n lai hogt lye ciia tinh filing la tuong duong nhau. Phdn tich thing ke
cho t h i y d pH 7 vd 8 khdng cd sy sai khdc v l phin tram hogt lyc vd vfn t i c tinh triing (P>0,05), tuy nhien Igi d sy sai khac y nghTa ve thdi gian hogt lyc Clia tinh tning (P<0,05) (hinh 3). Dilu ndy eho thay, tinh tning hoat dfing tot nhat trong mfii tnjdng ed pH = 7.
^ ^ n u d i ii^u " t^u Mil- OUU 1
liTiP l l l l l
Hinh 3. Anh hvomg ciia d$ pH Ifn ho^t l^c tinb triing c i mii egp
Kit qua nghifn cuu ndy cho thay, pH toi uu ciia moi tardng pha loang cho hogt lye ed mii epp thip hon so vdi d dCi vdng [16], d tim Iran [5] vd d chem chdu Au [3]. Tuy nhif n, kit qua nay eung phii hpp vdi cdc nghidn edu tiud-c ddy rang, khoang pH thieh hpp cho hogt lye tinh triing eiia ede lodi cd Id tu5-10[4],[5].
4. Anh hudng cua dp s u i t tham t h i u len hoat lyc tinh triing
Kit qud nghifn edu dnh hudng ciia dp suit thim thiu vdi hogt lyc tinh tning d mii epp eho thay. khi kich hogt hogt lye eiia tinh tning d cdc mdc dp suit tham thiu 200, 300, 400 vd 500 mOsm/kg thi tinh thing khdng hogt lyc ngay ma ed thdi gian
d l tinh Irdng Idm quen vdi mdi trudng (Uil hifn d thdl gian tri hoan). Quan sat khi finh filing hogt lyc cho thiy, mfii trudng pha lodng cd dp suit thSm thau 500 mOsm/kg eho k i t qud hogt lyc eao nhit (93,3 ± 2,29%; 138,4 ± 4,16pm/s; 543,8 ± 10,66s), dong thdi d nong dp ndy, thdi gian tri hodn cho hogt lyc cua tinh triing cung thip nhit (26,3 ± 1,05s);
hogt lyc eiia finh triing ed mu epp cd xu hudng gldm khi eho vdo edc mdi trudng ed ap suit thim thiu thap han (hinh 4). Ket qua phan tieh thong kf cho thay ed sy sai khdc d ^ nghTa giira d c ning df dp suit thim thiu thi nghifm vdi hogt lyc tinh tning (P<0,05). Qua dd, ed the khang djnh finh triing hogt lyc t i t nhat d dp suit tham thau 500 mOsm/kg.
•aPhJnllblbaMluc -*-\^iac • Then gianhoat luc —Ihin gian tli hoin
—-HTl ""! f'" ^ ii I I
• • - "n S 'Si 3a» B H H H
-J I IF dIJJJ
ASTr(BOim/l(e) Hinh 4. Anh huung ciia ap su^t t h a m thau lin hoat 1 ^ tinh triing ca mu egpKit qua nghifn cuu nay da t h i hifn rang, Hnh triing ea mil epp hogt dgng trong dung djch cd dp suit thim thau cao han mft so d nude ngot nhu:
cd chep [20], d hli van [13], d rd chdu Au [8] va thip han mpt so logi d biln nhu: ca mii den [12], cd dil Afiantic [11] v d d chem chdu Au [3]. Ket qua nay
eung phu hgp vdi d c nghien cdu tnjde ddy rdng, d cde lodi d biln, sy vdn dfng ciia tinh tning dupc kich hogt trong mfii trudng uu truang. nghTa Id dp suit thim thiu ngoai mdi tn/dng hoat dfng eiia tinh triing eao hon so vdi dp suit thim thiu ciia djch tlFOng [6], 17], [8], [6],
22 • TRLTONG OAI HOC NHATRANG
IgSShi f^hoa hoc - Cong nghe Thuy sdn So 1/2014 IV. K^T LUAN VA KI^N NGH] ti If 1:100 cho phan tram, van t i c va thdi gian hogt I . K l t i u d n lyc t i t nhit
Tinh tning cd mu cop cho k i t qud phan tram, 2. K i l n nghj
v f n toe vd thdi gian hogt lyc t i t nhit khi dupe Thanh phin djeh tuang trong tinh dieh cd Id pha lodng vdi ti If 1:100 trong nude bien nhan tao khdc nhau doi vdi mdi loai, chiing chju anh hudng
^^^^ bdi thanh phan ede ion eau tgo nen. Do dd, d eac thi Trong dilu kifn pH = 7 cung vdi ti If pha bang nghifm sau d n tiln hdnh nghifn ctFu anh hudng 1:100 hoat lyc finh tning cd mu cop Id eao nhaL cua nong dp cdc ion len hoat lyc tinh triing ea mu Tinh triing cd mii epp dupe pha lodng trong egp nham gdp phin tgo d c dilu kifn toi uu d l qua dung djch ed dp suat thim thiu 500 mOsm/kg vdi trinh bdo qudn tmh triing thanh cfing
TAI LIEU THAM KHAO Tieng Vi^t
1. D o ^ Khdc BQ, 2008. Ky thuat nuoi ca mii NXB D^ Ning.
2. Le Anh Tuan, 2004. Tinh hinh nuoi ca mu 6 Vi^t Nam- hien trang va cac tra ngai ve m|it ky thuSt T?p chi Khoa hpe - Cong ngh$ Thiiy san. Truong Djii hpe Thuy san: 174- 179.
TiingAnh
3 Abascai, F.J., Cosson, J. and Fauvel, C , 2007 Characterization of sperm motility in sea bass: the effect of heavy metals and physicochemical vanables on sperm motility Joumal Fish Biology, 70: 509-522.
4 Alavi, S.M H. and Cosson, J., 2005. Sperm motility in fishes: (I) effects of temperature and pR Cell Biology International, 29 101-110.
5 Alavi, S.M.H and Cosson, J., 2005. Spermatozoa motility m the Persian siui^eon, Acipenserpersicus: effects of pH, dilution rate, ions and osmolality. Aquaculture Research, 36 841-850.
6. Alavi, S.M.H. and Cosson, J., 2006. Sperm motility in fishes: (II) Effects of ions and osmolality. Cell Biology Intemational, 30: 1-14.
7 Alavi, S.M.H,, Cosson, J., Karami, M , Abdolhay, H. and Amiri, B M , 2004 Chemical composition and osmolality of seminal fluid of .Acipenser persiciis; their physiological relationship with spenn motility, Aquaculture Research, 35; 1238-1243.
8 Alavi, S.M.H., Rodma, M, Pohcar, T, Kozak, P., Psenicka, M. and Linhart, O, 2007. Semen ot Perca ftuviatilis L.:
Spenn volume and density, seminal plasma indices and effects of dilution ratio, ions and osmolality on sperm motility Theriogenology, 68: 276-283,
9 Bozkurt, Y., Selcuk, S. and Suleyman, B., 2006, Relationship Between Spermatozoa Motility, Egg Size, Fecundity and Fertilization Success in Salmo trutta abanticus. Tanm Bilmlen Dergisi, 12(4): 345-348,
10. Chang, Y.J., Choi, Y.H., Lim, H K. and Kho, K.H., 1999. Mdt property and sperm motility of grey mullet A/ugiVcep/iu/iii JKor Fish Soc, 32: 238-241.
11. Charles, D. and Peter, T., 1998. Role of Ions and Ion Channels in the Regulation of Atlantic Croaker Spemi Motility The Journal of Experimental Zoology, 281: 139-148
12. Gwo, JC , 1993. Cryopreservation ofblack grouper £p(ne/)/ie/HJ7na/aAflnci« spermatozoa Theriogenology, 39: 1331-1342.
13. Lahnsteiner, F., Beiger, B., Weismann, T. and Patzner, R A , 1996 Physiological and biochemical determination of rainbow trout Oncorhynchus mykiss semen quality for cryopreservatioa J Appl Aquacult, 6 47-73.
14. Lahnsteiner, F., Berger, B,, Weismann, T and Patzner, R A., 1997. Sperm motility and seminal fluid composition in the burbot Lota lota. J Appl Ichthyol, 13: 113-119.
15. Le, M H , 2010, Reproductive cycle and semen preservation of yellow cioakei Larimichthys polyactis, Ph.D thesis. Pukyong National University, Korea: 168.
16 Le, M.H., Lim, H K., Mm, B.H., Park, M S., Son, M.H., Lee, J.U. and Chang, Y.J., 2011. Effects of varying dilutions, pH, temperamre and cations on sperm motility in fish Larimichthys polyactis. Journal of Envu-onmental Biology, 32: 271-276.
17 Le, M.H., Lim, H.K., Min, B.H., Sung, Y.K and Chang, Y.J., 2007. Mill properties and spermatozoa structure of filefish (Thamnaconus modestus). Dev Reprod,, 11(3): 227-233
18. Linhart, O., Cosson, J., Mims, S D , Rodina, M., Gela, D. and Shelton, W.L, 2003. Effects of ions on the motility of fi-esh and demembranate spermatozoa of common carp Ctprinus carpio and paddlefish Polyodon spathula Fish Physiol Biochemical, 28: 203-205.
19. Monsawa, M., Suzuki, K , Shimizu, H., Monsawa, S. and Yasuda, K , 1983. Effects of osmolality and potassium on motility ofspermatozoa firom fi-eshwater Cypnmd fishes J.exp.Biol, 107: 95-103.
20. Suquet, M., Billard, R , Cosson, J., Dorange. C , Chauvaud, L., Mugnier, C. and Fauvel, C, 1994 Sperm features in turbot Scophthalmus maximus: a comparison with other freshwater and marine flsh species Aquat Liv Resour, 7: 283-294.
21. Tan-Fermin, J,D„ Miura, T., Adachi, S. and Yamauchi, K, 1999. Seminal plasma composition, sperm motility and milt dilution in the Asian catfish C/flrinsmacrocepAa/uj Aquaculture, 171:323-338.
TRL/ONG DAI HOC NHA TRANG • 23
