Tgp chi Khoa hoc - Cong nghe ThUy sdn SS 3/2014
THONG BAO KHOA HOC
ANH Hl/OfNG CtlA TY LE PHA LOANG, pH VA NONG DO THAM THAU LEN HOAT L i r e TINH TRUNG CA DIA {Siganusguttatus Bloch, 1787)
EFFECTS OF DILUTION RATIO, pH AND OSMOLALITY ON SPERM MOTILITY IN RABBIT FISH (Siganus guttatus Bloch, 1787>
Vd Thi Ngpc Giau', UMinh Hoan^, Phan Van il^, Ph^m Qudc Hiing' Ngiy nhjn b^: 25/2/2014; Ngay phin bi?n thong qua: 16/4/2014; Ngay duy^t dSng. 13/8/2014
T O M T A T
Nghien cuu ndy da dupc tien hdnh di tim ra ty If pha loang, pH vd nong do thdm thau tdt tm cho hogl luc tinh trung cd dia (Siganus guttatus Bloch, 1787). Ttnh dfch cua cd dia dupc pha ioangv&inu&c bten nhdn tgo dcdc ty le 1:50.1:100.
1.150 vd 1:200 di xdc dinh tyle pha loang ldi uu cho hogt lyre tinh triing cd dia Tiip theo, tyl^pha loang ndy dugc su dung quan sdt apH (6. 7, 8. 9) vd ndng dg thdm thdu (200. 300. 400. 500 mOsm/kg) dexdc djnh gid tripHvd ndng dg Ihdm thdu tdt nhdt cho hogl luc linh triing. Mdi quan sdt ditqc lgp lgi 3 ldn. Kel qud cho thdy, ttnh trung cd dia hogt l^c lot nhai a ty lipha loang 1.50, pH = 8vd ndng dp thdm thdu 400 mOsm/kg Irong nu&c bien nhdn Igo.
Tir khda: cd dia. tyl4pha loang. nong do thdm lhau, hogt lifc ttnh trung, pH ABSTRACT
The objectives of the present study were reviewed lo assess the optimal dilution rale, pH and osmolality for sperm motihty in rabbit fish (Siganus guttatus Bloch, 1787). Semen were diluted wilh artificial seawater at ratios of 1:50,1:100.
1:150 and 1:200 to find the best dilution ratio for sperm motility And then, this bestdilution rate was used for observing onpH(6. 7. 8. 9) and osmolality (200, 300. 400, 500 mosmoi kg-1) to find the best pH and osmolality for sperm motiiity.
All observations were replicated three times. The results indicated that the best performance of sperm were shown in the dilution rale 1:50, pH - 8 and osmolality 400 mOsm.kg-1 artificial seawater.
Keywords, rabbit fish, dilution rale, osmolality, sperm motility, pH
I.DATVAND^
Ca dia Id lodi phdn bo rdng, cd gid trj ^ u o n g pham cao, sdn lupng khd Idn vd dupe nudi d nhieu nudc chdu A [1] {dupc tham khao trong [16]), Mdt vai loai thude giong ca dia dupc nudi theo hinh thuc truyin thdng d Phi-lip-pin nhung vide nudi thuang mgi vln chua dupc hinh thanh [12]. & Vipt Nam, trong dy dn IMOLA, Trung tdm Khuyin ngu tinh Thira Thien Hul dd thye hien md hinh nuoi ed dia, tdm sii vd rong cdu ehi vang [4]. Hipn nay ngudi ddn d tinh Khdnh Hda thudng nuoi thuang phim ed dia vdi nguIn giong thu ti> ty nhidn Id ehii yiu. IVldc dii cd dia Id mdt doi tupng nudi ngdy cdng pho bien vd c6 gid tri kinh te eao d khu vyc Ddng Nam A nhung van d l san xult gilng van chua duoc gidi quyet tdt [13].
Chat lupng sdn pham sinh dye cua cd bd m^
la ca sd quan trpng trong vide ndng eao chit lupng thy tinh [7]. Tinh triing cua hau hit cdc lodi ca bien khdng hogt lyc trong buong se vd djeh tuong [5].
Tinh triing b i t dau hogt lyc khi dupc phdng thich ra ngodi mdi trudng nudc, tinh triing ggp trung tien hdnh qud trinh thy tinh. Vi vdy, hogt lyc cua tinti tnjng Id yeu t6 quan trpng quylt dinh kha nang thg tmh cua tinh triing [11]. Khi dupc phdng thidi ra mdi tardng thu tinh, hogt lyc cda tinh trting phy thuOc rit nhieu vdo dieu kidn mdi trudng nhu ty Id pha loang, dp suat thim thau, pH, nhipt dp, ning dO cdc ion (Ca^*, Mg=*, Na*...)[5,11] The gidi ndl chung va Vidt Nam ndl rieng da cd nhilu nghien cuu ve chit lupng tinh triing nhu cd chem mdm nhpn
1 V6 Th| Ngpc GiSu, n s . LS Mmh Hodng, =ThS Phan V§n Ot, 'TS. Phgm Quoc Hung: Vi$n Nu6t trong thiJy san - Tnjdno Dai hpc Nha Trang
26 • TRUONG DAI HOC NHA TRANG
Tgp chi Khoa hoc - Cong nghe Thuy sdn SS 3/2014
Psammoperca waigiensis [3], ca mu cpp Epineptielus Aiscoguttatus [2], cd bon Hippoglossus hippogk3ssu [9];
cac y i u td mdi trudng ngodi dnh hudng tdi hogt lyc tinh triing cOa nhieu lodi cd nhu cd hoi Salmo gairdnen[^4], ca tam Ba Tu Acipenserpersicus [5],..
Tuynhien.diuacdnghidncuundotilnhdnhfrdncddia Siganus guttatus. Do dd, nghien ciru nay da dupc thyc hidn nhdm tim ra ti Ip pha lodng, pH vd nong dd tham Oiau tdi uu cho hogt lyc tinh triing cd dia nhdm nang cao hieu qud thy tmh trong qud trinh sdn xuat nhan tgo, giam strc dp len nguon Ipi cd dia ty nhidn ddng thdi gop phin da dang doi tupng nghidn cuu hoat lyc tinh trung cd.
H. VAT L l f U VA PHU'CfNG P H A P NGHIEN CLTU 1. Chuan bj nguon tinh djch
Cd dia dye (Siganus guttatus Bloch, 1787) da dup'c thu thdp tu t y nhien, dua ve nudi vd trong long tgi Vung Ngan - Nha Trang - Khdnh Hda mdt thdi gian. Ddn cd ndy dupc djnh ky kiem tra hdng tuin, khi cd dat didt lupng tmh tdt thi dupc vuot tinh de tien hanh cac quan sat. Cdc quan sat nay dupc thyc hien tgi Phdng Thi nghiem Sinh hpe nghd cd - Vipn Nudi trdng thuy san, Trudng Dgi hpc Nha Trang.
2. PhiFvng phdp thu mau tinh djch o l u tien, chting ta dDng khan bing tham sgch nudc d xung quanh Id sinh due cua ed due. Tiep theo, ehiing ta vult dpc ben hdng vd bung ea, khi tmh dich ed chay ra thl hiit tinh dich bang dng xilanh.
Tinh djch trong ong xilanh dupc cho vao dng eppendorf 1,5 ml, ddy kT n i p dng roi dua vdo thiing xdp dyng dd bdo, vdn chuyin ngay ve phdng thi nghidm d l tiln hdnh cdc quan sat. Trong khi vuot, Id sinh due cua cd phai duac tranh khdng de nudc, phan hay nudc t i i u l l n vao tinh dich de thu dupc tinh ed chat luang tot.
3. Anh hudng ciia ty Id pha loang len hoat lyc tinh triing
o l t i l n hdnh quan sat ndy thi tinh djch vdi nudc biln nhdn tgo d cde 0 Ip pha loang khde nhau. Thanh phan nudc biln nhdn tgo bao g i m : 27g NaCI; 0,5g KCI; 1,2g CaCI^; 4,6g MgCI^;
0,5g NaHCOg dupc pha trong 1 lit nudc cat vd pH = 7,8, ndng dp tham t h l u 356 ± 0,5 mOsm/kg.
Tinh djch va nudc bien nhdn tao dupc pha theo ede ti Id 1:50; 1:100; 1:150; 1:200 trong Ing eppendorf.
Sau dd, 1 pl tinh djch eiia ca trong Ing eppendorf
pha lodng vdi nudc biln nhdn tgo dupe hut ra bdng micropipet. Tiep theo, 1 pl tinh dich da pha loang dupc hilt dua Idn lam kinh quan sdt dual kinh hiln vi d dp phdng dgi 400 l l n . Hogt lyc cua tinh triing bao gom cac thing s l : phin trdm tinh triing hogt lyc (PTHL) vd thdi gian hogt lyc (TGHL). PTHL dupc xdc dinh bang so tinh triing hoat lyc so vdi t i n g sd tinh triing quan sat. TGHL dupe tinh tif liic pha loang cho d i n 100% tinh triing bat hogt (dun vj tinh' gidy).
Sau khi xac djnh duac ti Id pha loang tot nhat thi ti' Id pha lodng do duac diing de t i l n hanh ede quan sdt tilp theo.
4. Anh hiramg cua pH ien hogt lyv cua tinh trung Anh hudng eiia pH duae ddnh gid bdng each sir dyng nude bien nhdn tao d cdc gid trj pH: 6, 7, 8, 9 vdi ty Ip pha lodng tdi uu ed nong dp thim thau tuang duang vdi nudc bien nhan tgo. Hoat lyc tinh triing eung dupe kiem tra tucmg tu nhu trinh bay d tren.
5. Anh hu-dng cua ndng dp t h i m thau len hoat l i ^ tinh triing
o l xdc djnh anh hudng cua ndng dp tham thau Idn hogt luc tinh tning, dung djch NaCI d cdc muc ning dO thim thIu' 200, 300, 400 vd 500 mOsm/kg d ti Id pha loang tdi uu cd ndng do pH tuang duang nhu nudc bien dupc su dung. Hoat lyc cOa tmh triing d quan sat ndy bao gdm cdc thdng so: phin tram tinh triing hogt lye, thdi gian hogt lyc va thdi gian tri hoan (TGTH). TGTH Id khodng thai gian khi kich hoat hoat lyc cua tinh triing, tinh triing khdng hogt lyc ngay md ed thdi gian Idm quen vdi mdi trudng sau dd hoat luc. Dung djeh d cdc mirc thim thlu trdn dupc pha lodng vdi ti Id tdi uu d l quan sat dnh hudng cua nong dp tham thau len hoat lye tmh tnJng. Hoat lyc tinh tnjng eung dupc kiem tra tuang ty nhu trinh bdy d tren.
6. Phuvng phdp x u ly so ligu
Cdc so lieu thu thdp duac xi> ly tren phin m l m SPSS 16 vd Excel 2003. D l tim ra ty le pha loang, pH va nong do tham thau tdt nhat eho hogt lyc eiia tmh triing cd dia, chung tdi diing phdn tieh phuang sai mdt y i u td (one - way ANOVA) Sy sai khde giira cdc nghidm thirc dupc so sanh theo kiem djnh Duncan, sai khde cd y nghTa dupe xem xet khi (P < 0,05). Cdc gid tn dupc trinh bdy dudi dgng gid tri trung binh ± sai s l chuan (futean ± SE)
Tgp chi Khoa hoc - Cong nghe Thdy sdn Sd 3/2014
III. K^T QUA NGHIEN CLTU V A T H A O L U A N
1. Anh hudng ciia t^ 1$ pha loang (TLPL) Idn hoat lyc tinh triing
Cdckyhifu cha cai a, b, c
Pha loang tinh trung Id yeu to kich hogt tinh trCing hogt ddng vd duy tri kha ndng thy tinh cua tinh trdng trong mdi trudng thy tinh [5]. K i t qua nghien cuu dnh hudng cua ti le pha loang Idn hoat luc tinh tnJng cd dia d hinh I A cho thiy, thdi gian tinh trimg hogt lyc cao nhit d ti Ip pha loang 1:50 (161.3 ± 15,51 s) vd thip nhat d ty le 1:200 (49,7 ± 3,18 s). Tinh tnJng pha lodng vdi ty le 1:50 cd sal khde ve mdt y nghTa thong ke vdi cde ty le pha loang cdn lai (P < 0,05).
Theo hinh I B , phin tram tinh tnjng hoat lyc gidm din theo thdi gian d cde mde quan sdt. d 10 s dau tlen, tmh triing hoat lye d tat cd ede ty le pha lodng 1:50, 1:100, 1:150; 1:200 nhung phin tram tinh triing hoat lyc cao nhat (86,7 ± 8,82%) d ty le pha loang 1:50, thap nhat d ty le pha lodng 1100 (66,7 ± 8,82%). Tuy nhien. phdn tram tinh tning hogt lyc giu'a cac ty Ip pha loang d thdi diim nay khdng cd sy sai khae ve mat thong ke (P > 0,5). 60 s tiep theo, phin tram tinh triing hoat lye cao nhat (86,7 ± 8,82%) van d tJ le pha loang 1:50 va da cd
TGHL (•) TLPL ISD boat lire tinh trikng c i dia
khac vi m^l ihdng ke (P < 0,05) cho (A) TGHL va (B) PTHL Iheo limg cm thai gian (lOs. 60i. 120s hogc tSOs) sy sai khac v l mgt thdng ke so vdi ede ti Id pha loang con lai (P < 0,5). Oieu ndy dung d 120 s vd 180 s tiep theo, phin tram tinh trting hogt lyc chi cdn d ty le pha loang 1:50 (3 ±3 .3%). cdc ty Ip pha lodng 1:100.1:150; 1:200. tinh triing da ngirng hogt lyc d 180 s. Vgy ty le pha loang tot nhat cho tinh triing ea dia hogt lyc Id 1:50 vdi TGHL trung binh la (161,3 ± 15,51 s)vd PTHL dat (86,7 ±8,82%)d 10s.
Ket qud ndy tuang t y vdi t^ Id pha loang d ed tdm Ba Tu Acipenser persicus [4], ed Perca fluviatilis [5] nhung cung khde so vdi mpt sd lodi nhi/.
ed ehem mom nhon Psammoperca waigiensis [4], cd mu cpp Epinephelus fuscoguttatus [2] ty le pha loang tot nhat 1:200, cd dii vdng Larimichthys polyactis [11] ti Ip pha loang tot nhat la 1:100, ca Mugil cephalus [15] ti Id pha loang tot nhit Id 1:10.
TT le pha lodng 1:50 dupc sir dyng d l kilm tra sy dnh hudng cOa ndng dp tham thau. pH len hoat lyc tinh triing ed dia.
2. Anh huvng cua pH len hogt \ijK tinh triing
I
P*- TGHL (s) Hinh 2. Anb huong cua pH len hoat lire tinb trung ci diaCac ky hi?u chu cdi a. b. c. d chl sir sal khac ve mar thdng ki(P< 0,05) cho (A) TGHL va (B) PTHL Iheo limg c^m thai gian (lOs, 60s. 120s hoac ISOs) Nghien CLPU anh hudng cua pH doi vdi hoat lyc cdc gid tn pH khde nhau thi hogt lyc tinh tmng ca dia tinh trting cd dia dupe tiln hdnh d cdc nong dp pH cd sy khde nhau. Tinh trting cd dia hogt lue tdt nhit (6, 7, 8, 9) vdi tt Id pha loang 1:50. Ket qud eho thay, d pH = 8 vdi thdi gian hoat lyc Id 158,3 ± 12,83 s.
28 • TRUONG OAI HOC NHA TRANG
Tgp chi Khoa hgc - Cong nghe Thiiy sdn So 3/2014
Khdng ed sy khac nhau cd y nghTa thing kd v l thdigian hogt lyc ciia tinh tnJng giua cac gid tn pH. Tuy nhien, phin tram hogt lyc d cac gid trj pH gidm d i n theo thdi gian. 10 s dau tien, phan trdm hogt lyc d pH = 8 idn han phan trdm hogt lyc pH = 6, pH = 7, pH = 9 vd ed sy sai khac cd y nghTa thing kd vdi phan trdm hogt lyc pH = 6, pH = 7 nhung khdng cd sy sai khac pH = 9. 60 s tilp theo, phin trdm hogt lyc d pH = 8 cung Idn han phan tram hogt lyc pH = 6, pH = 7, pH = 9 nhung da cd sai khde ed y nghTa thong kd vdi phan tram hogt lyc d ca 3 gid tn pH = 6, pH = 7, pH = 9. Oilu ndy dung vdi 120 s va 180 s. sau 180 s thi phan tram hogt lyc pH = 8 la 13,3 ± 4.41 % cdn d cdc gid trj pH cdn lgi tmh triing da ngirng hogt lyc.
v l m$t smh thai, viing sinh sdng ciia ed dia trudng thdnh Id d ven bien [10]. pH cua nudc biln dao dpng trong khodng 8.1 - 8.3. Theo kit qud quan sdt thi pH = 8 cOa dung djch pha loang Id phii hpp nhit d l kich hogt sy vdn dpng cCia tinh trtmg ed dia. Ket qud ndy tuang ty nhu nghien ciru tren cd tam Ba Tu Acipenser persicus [3] ed ehem mdm nhpn Psammoperca waigiensis [A], cd dii vdng Larimichthys polyactis [11], cd h l i vdn Salmo gairdneri [14] nhung khac so vdi ed da tran chdu A Clanas macrocephalus [8] thi ty Id pha lodng t i t nhit khi pH = 7.
3. Anh hudng cua ndng do tham thau (NDTT) len hoat lyc tinh triing
300 400 500 NDTT (mOsm/kg)
Hinh 3. Anh hirdng ciia NDTT len ho^t l^v tinb triing ci dia
khde vi mgl ihong ki(P< 0,05) cho (A) TGTH. a.b.c.dcho (B) PTHL theo lung cu I Ihdi gian (lOs. 60s. 120s h
K i t qud nghidn ciru anh hudng eila nong dd tham t h l u tdn hogt tyc tinh frting cd dia cho thay, khi kich hogt d cac mirc ndng dp thim thlu 300, 400 vd 500 mOsm/kg thi tmh triing khdng hogt lyc ngay ma tinh trting can thdi gian Idm quen vdi m i l trudng (the hien d thdi gian tri hoan). 6' mdc nong dd tham thau 200 mOsm/kg tmh triing khong hogt lyc. Quan sdt khi tmh trting hogt lyc cho thay, mdi trudng ed ndng dp thim thlu 400 mOsm/kg cho k i t qua hoat lyc cao nhat (201,7 ± 13,34 s) vdi thdi gian tri hoan (225 ± 9,2 s). thdi gian hogt lyc vd thdi gian tri hoan d ndng dd thim thau 300 mOsm/kg va 500 mOsm/kg lan lugt Id (114,7 ± 17,75 s vd 248,3 ± 18,03 s);
(148 ± 19,14 s vd 188 ± 30,02 s). Thdi gian hogt lyc ciia tinh trtmg c6 sy sai khac co y nghTa thong ke giiVa ning dp thim thlu 400 mOsm/kg vdi ede nong dp tham t h l u cdn Igi (P < 0,05) nhung giCra nong dd tham t h l u 300, 400 mOsm/kg thi khing c6 sy sai khac v l mdt thong ke trong thdi gian hogt lyc tinh fnjng (P > 0,05).
Phan tram tinh trting hoat lyc dgt tot nhit tpl 400 mOsm/kg (51,7 ± 4.41%) d 10 s vd ed sy sai khac
vdi cdc ning dp 200, 300, 500 mOsm/kg. Sau 180s, tinh trtmg d cdc gid trj 200 mOsm/kg, 300 mOsm/kg va 500 mOsm/kg khong edn hoat lye, trong khi do 400 mOsm/kg tinh tnJng van cdn hogt lyc 10,7 ±5,02%.
Qua dd, cd t h l khang dinh tinh triing ed dia hoat lye tot nhdt d dung djch co nong dp thim thlu 400 mOsm/kg, eao han cd hoi van Salmo gairdnen [14], cd Perca fluviatilis [5], gid trj NDTT cdn dao dpng Idn va rat cao nhu theo nghien ciru ctia Harald vd ctv (2001), NOTT t i t nhit cho tinh trung cd Hippoglossus hippoglossus tu 400 - 1000 mOsm/kg [9].
IV. K^T LUAN VA KI^N NGHj i . K l t l u I n
Trong nghidn ciru nay, tmh djch ed dia pha lodng vdi nudc bien nhdn tgo d ti lg 1:50 cho k i t qud tdt nhat ve PTHL {86,7 ± 8,82%) d 10 s dau tien vaTGHL(161,3±15,51 s).
d mdi trudng nude biln nhdn tgo ed pH = 8, tinh trung hogt lyc va PTHL toi uu trong nghien euu ndy l l n lupl Id 158,3 ± 12,83 s vd 63,3 ± 8,82%.
Tgp chi Khoa hoc - Cong ngh^ Thuy sdn So 3/2014
Tinh djch ca dia pha loang trong dung djch c6nong dp tham thdu 400 mOsm/kg vdi ti Ip 1:50 thl thdi gian va PTHL tinh trimg tdt nhat ciJa nghidn ctru nay lln lupl Id 201,7 ± 13,34 s vd 51,7 ± 4,41%
d 10s.
2. K i l n nghj
Ngodi ty Id pha loang, nong dp tham thlu. pH thi nhipt dp, nong dp eac ion (Ca^*, Mg^*, K*, Na*..)
cQng anh hudng Idn hogt lyc cua tmh tnJng trong mdi trudng thu Unh. Do dd, nghien edu dnh hudng cua ndng dd cdc ion len hogt lye tinh triing cd dia c^n dupc nghien euu thdm nham gdp phin tao dilu kidn mdi tnrdng toi uu trong qud trinh sdn xult thdnh eflng giong cd dia.
Ngodi ra, nghidn ciFu ddc tinh 1;^ hda hpc eua tinh djch cd dia cOng can thilt gdp phan vdo qud trinh bao qudn tinh trting lodi cd nay trong tuang lai.
TAI LIEU THAM KHAO ning Vift
Le Van Dan, LS Van Ngoan, 2006. Ngliien cuu suphat triSn tuyen sinh due ci dia (Siganus guttatus Bloch, 1787) a vimg dim pha Thira Thien Hu6 Tap clii Nong nghidp vji Ph^ tnen nong thon, 2: 61-64
Hoang Thl Hiin, 2012. Nghien cihi mpt s6 d ^ tinh ly, hoa hgc ciia tinh trimg ca mu cgp (Epinephelus fiiscoguttatus Forsskal, 1775). Lu3n van Th^c Sl. Traong Dp hgc Nha Trang
Nguyin Thl Hdng Nhung, 2012. Nghien ciru danh gii chdt lirgng tinh tning va anh huong cda mgt so yeu to len boat luc tinh triing ca chem mom nhgn (Psammoperca waigiensis Cuvier &ValencieQes, 1828). Luan van Thac si. Tnrdng Dai hpc Nha Trang.
Trung tam KhuySn ngu Thiia Thien Hue, 2007 Bio cio ket qua thuc hign m6 lunh nuoi ca dia giong sinh san nhan tao (Siganus guttatus) ket hgp voi rong cau chi ving (Graciiaria verrucosa) va torn sii (Penaeus monodon).
Tieng Anh
Alavi. S. M. H. Cosson. J, Karami. R, Amiri. B. M and Akhoundzadeh M A, 2004, Spermatozoa motility in the Persian sturgeon (Acipenserpersicus): effects of pH, dilution rate, ions and osmolality. Reproduction 128:819-828.
Alavi. S. M. H, M. Rodina M, Policar. T, Kozak. P, Psenicka. M and Linhart 0, 2007. Semen of Perca fluviatilis L:
Sperm volume and density, seminal plasma indices and effects of dilution ratio, ions and osmolality on sperm motility.
Theriogenology, 68: 267-283.
Cabrita E, Robles. V and Herrasez P, 2009 Methods in reproductive aquaculture marine and freshwater species. CRC Press Taylor & Francis Group
Fermin J. D. Miura T, Adachi. S and Yamauchi K, 1999. Seminal plasma composition, sperm motility and milt dilution in Harald. B. T, Tillmann. T, Benfey. J, Deborah. J, Robichaud. M and Power, J. 2001, The relationship between spenn density, spennatocnt, spenn motility and fertilization success in Atlantic halibut, Hippoglossus hippoglossus. Aquaculture.
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10. Lam, T. J, 1974, Siganids: Theu biology and mariculture potential. Aquaculture: 325-354.
11. Le M H,Lim. H.K,Min, B. H, Lee. J UandChang. Y. J,2011, Effectsofvaryingdilutions,pH,teraperacureandcationson spermatozoa motility in fish Lanmichthys polyactis, Joumal of Environmental biology 32: 271-276
12. Pillai. T. G, 1962. Fish farming methods in the Philippines, Indonesia and Hong Kong. FAO Fish: 51-52.
13. Rachmansyah, Usman, Lante. S and Ahmad T, 2007. Rabbitfish Siganus guttaWs breeding and larval reanng trial Aquaculture Asia Magazine, July - September 2007: 39-41.
14. Van der Horst. G, Dott. H. M and Foster G C, 1980. Studies on the motility and cryopreservation of rainbow trout (Sahno gairdneri) spemiatozoa, SouthAfncan Joumal Zoology, 15- 275-279.
15. Yeganeh S, Mojazi A BandAlavi S M. H, 2008. Motility of Mugil cephalus L. spermatozoa in coelomic fluid, seminal fluidandsalinemedia, Joumal of Applied Ichthyology 24: 517-518
16. http://wwwfishbase org. Access in December 2013,
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