KHOA HQC KY THUAT T H O Y TAP XX S 6 6 - 2013
Sir PHAT TRIEN M O HQC TUYEN tfC CUA CA CHEM dates calcarUer, Bloch 1700) NUOl TAI VIET NAM
Nguyin Huu Ddng, Trdn Vy Hlch Tmng tdm Nghiin cmi Gidng vd Dfch binh thuy sdn -Tnrang Dgi hgc Nha Trang
T6M TAT
S\f phdt triln tuyen ftc cua ca chSm trong dieu ki?n uong nudi nhan tgo tai Vigt Nam dd dupe theo ddi bdng phuang phdp md hgc. Mau cd chem dugc thu lien tyc tu khi cd mdi nd din khi ddn cd dat 40 ngdy tu6i (khodng cdch gifta cdc lan thu mau Id 12 gid) vd dupe c6 djnh trong dung djch formalin 10% trung tinh. Cdc Idt cdt md cd d cdc ngdy tudi dupe nhiigm haematoxylin & eosin vd quan sdt dudi kinh hiin vi quang hpc. Kit qua quan sat cho thay, mdm tuyin lic cua ca chdm bdt dau hinh thdnh til ngdy thii 10 sau khi nd. Din ngdy thu 14, tuyin lic dd khd hoan chinh ve mdt hinh thdi. Tuy nhien, tuyin lic cd chem chi hodn thi?n vl mdt tl bdo hpc d ngdy 30 sau khi nd vdi sy phdn hda rd r?t vimg vd vd viing tuy. Kit qud nghien ciiu ndy cho thdy rdng, vi?c dp dyng li?u phdp vacxin de phdng b?nh cho cd chem trong dilu ki?n uong nudi tai Vi?t Nam chi cd thi dat hieu qud cao tu khi cd dgt 1 thdng mdi, khi co quan lympho trung uong cua cd Id tuyen Ore dd phat triln hoan thi?n vl cdu tnic vd chiic ndng.
Tukhoa: Cd chem, Tuyin dc, Phdt trien md hpc.
Histogenesis of tliymus of tlie barramundi (Lates calcarifer, bloch 1790) in Vietnam
Nguyen Huu Dung, Tran Vy Hlch
SUMMARY
Development of the thymus of hatchery-reared barramundi larva was examined using light- microscopic histoioglcal method. The larva samples were collected continuously from hatching to the day 40'^ thereafter at 12 hrs. interval The fish samples were fixed in 10% phosphate- buffered formalin solution, serially sectioned and stained with haematoxylin and eosin. The stained sections were observed under light microscope to detect the changes of thymus organ.
The time-serial observation showed that thymus germination appeared In fish at the day 10*", and the thymus seems to be morphologically completed at the day 14"* after hatching. However, the thymus was only developed completely on cytology feature, and therefore may be functionally matured at the day 30'" with the clear differentiation of the cortex and medulla regions with matured T-lymphocytes. The studied results indicated that vaccination of hatchery produced barramundi may only result in good protection if the application is conducted to fish when fish is older than 30 days of age, when the thymus is structurally and functionally matured.
Key words: Barramundi, Thymus, Histogenesis.
I. MCJ D A U phd ven biln nhdm khdc phyc tinh trang djch
^ . , . ,, . , -^ t., u ,nnns 1- 1^?"^ dang lan rdng chua khdc phyc dugc cua Ca chgm (Lates calcanfer, Bloch 1970) la , i .. „ , .f . , , . . , ^, ^T. u - - , \..-, • t. ' . * * - . •; nghe nuoi tom bien a Viet Nam. Tuy nhien, loai ca CO gia tri kmh te va dang duac phat tnen f . . . ,- - .. . - • • ' „ „ ,., . , , i . -• I . . - -• . *i cung vai viec ma rong dien tich nuoi va nang thanh doi tuong nuoi chu luc a cac vung dam , * , , . ', ' . - i . - - l u t i
cao muc dp tham canh mang lai hieu qua kinn te 82
KHOA HQC KY THUAT T H O Y TAP XX S 6 6 - 2013
cao, dich b§nh da bdt ddu xuat hi?n tr6n cd chem nuoi, tgo nguy co cho s\r phdt triln ben vihig cua ddi tugng nuoi ndy. Cdc b?nh phd bien tren cd chSm bao gdm b?nh do ky sinh trimg ngogi ky sinh, b^nh hogi tu thdn kinh do virus vd cdc b$nh do vi khudn, d$c bi^t la do lien cau khudn Streptococcus iniae gdy ra (Dd Thj Hda vd ctv, 2004). Trong do, cdc b?nh do vi khudn, virus thucmg ddn den thi?t hgi nghiem trpng vd cdn phdi cd cdc bi^n phdp phdng trj hihi hi$u.
Trong sd cdc bi?n phdp phdng b?nh truyln nhiem, v i ^ sii dyng vacxin Id bi^n phdp tdi uu gdp phdn phdt triln nghi nudi cd bin vftng. Dl nang cao hi^u qud phdng b$nh ciia vacxin, cd nudi cdn dupe gdy miin djch sdm dl tao khd ndng khang b?nh ngay tir giai dogn con non. Tuy nhien vacxin chi cd thi tgo ra khd ndng dl khdng cho cd khi cdc ca quan thupc h? miin dich ddc hi^u cua cd dd phdt triln (Gravningen, 2002).
C) cd, tuyin lie Id mpt co quan lympho tmng uang ddng vai trd quan trpng trong viec biet hda cdc te bao gdc thdnh cdc te bao lympho T trudng thdnh, nhan td quan trpng trong co chi ddp ling mien dich dac hieu cua ddng vgt. Khi tuyen lic xudt hien va thuc hiSn chiic ndng cua no c6 nghia la cd da hinh thanh miln dich dac hi?u (Bowden vd ctv, 2005). Cac nghien ciiu ve phdt sinh tuyen lic d mdt sd lodi cd cho thay rdng thdi gian phat trien hodn thien tuyen lic d cd thay ddi theo loai vd mdi trudng sdng, ddc bi6t la nhi?t dp do cd la loai dpng vat bien nhiet (Tatner, 1996). Do vdy, nghien ciru ve su phdt
trien tuyen lic ciia cd chgm Id bet siic can thiet nham tgo ca sd khoa hpc cho vi?c xdc dinh thdi dilm thich hgp dl su dyng vacxin phdng b?nh ddi vdi lodi ca ndy.
IL PHUONG PHAP NGHIEN ClTU
Cd chgm mdi nd dugc thu m§u tu cdc bl uang nudi cd bdt tai Trung tam nghi8n ciiu Gidng vd Dich benh thiiy sdn, Trudng Dai hpc Nha Trang theo dinh ky 12h mot Idn (10-30 cd/ldn thu) cho din khi cd dat 40 ngdy tudi. Mdu cd dupe cd dinh trong dung dich Formahn (10%) trung tinh trong h? dem phosphate, vd diic paraffin de cdt Idt (6nm) lien tuc todn bg thdn cd. Cac lat cdt dugc nhupm Haematoxylin & Eosin va gdn tieu ban bdng Canada balsam.
Tat cd cdc lat cat mo cua timg cd the dugc quan sat dudi kinh hien vi quang hpc de nhdn dinh ve su hinh thdnh vd phdt triln cua tuyin tie tuang ling vdi tudi cd.
IIL K E T Q U A N G H I E N CUTJ V A T H A O L U A N
Kit qua kiem tra tieu ban md ciia tdt ca ca chem dudi 10 ngdy tudi khdng cho thay su xudt hien ciia tuyen lic. Mdm tuyen lic ciia cd chgm bdt ddu hinh thdnh tu ngay thii 10 sau khi nd.
Ldt cdt dgc theo hudng lung bung cho thay su hien dien cua tuyen lie d bilu md viing hau, sat vdi xoang thinh giac vd gdn vdi tien than ciia cd (Hinh I)
%w
J'
Hinh 1. M^m tuyen tic cd chem 10 ngay sau khi nd T: tuyen uc; B: nao; M: ca ho hdp; HK- tiin than; Ot- xoang thinh gidc;
Oc: Xoang ndp mang; E: mdt; G. mang
KHOA HQC KY THUJiT THtJ Y TAP XX S 6 6 - 2013
0 cd tir 12 ngdy tu6i trd di, tuyen lic phdt triln mgnh mg ve kich thudc. Liic ndy tuyen ire dd nhd ra khdi Idp bieu md vdng h^u vd cd dgng hinh kh6i dgng elip (Hinh 2a; 2b). Tuy nhiSn, tuyin lie vIn cdn Id mdt kh6i dong nhat, chua cd sy phdn chia thdnh cdc viing riSng bi$t. Din ngdy 14, cd thi nh^n thiy khd rd sy phdn viing d
tuyen dc cua cd chgm. Phan vd vdi su tgp tnmg ddy ddc ciia cdc tl bdo lympho bdt mdu tim sdm ciia hematoxylin trong khi d phdn tuy mgt dp te bdo lympho gidm rd r?t d l 10 nhilu khodng khdng vd t l bdo bilu md (Hinh 2c, 2d). Trong giai dogn ndy kich thudc hiyin dc van tilp t^c phdt triin nhung vln ton tai d dang hinh elip.
- • -*
Hinh 2. Tuyen Ctc cd chem 12 (a, b) vd 14 (c, d) ngdy sau khi nd : M: ca ho hda: HK: tien thdn; Ot: xoans thinh gidc; Oc: xoang ndp mang;
T: tuyen dc; M: ca ho hdp; HK: tiin than; Ot: xoang KT: dng thgn; IZ: vimg tuy; OZ vitng vo; G • mang Tir ngay thii 18 trd di, tuyin lie cd xu hudng phat triln keo dai ra va den ngdy 30 sau khi nd, tuyin lie cua cd chem cd dang ddc tnmg cua lodi, do la ca quan lympho gdm hai thiiy hinh qua dau ndm gdn xoang mang vd keo ddi li8n tyc din bieu md hdu. Tuyin lic cd chem nam rat sdt be mdt tilp xuc vdi ddng nude ra vdo xoang mang. Cdu tnic cua tuyen lic dugc phan chia thanh 3 vung ro ret vdi sy xuat hipn them cua viing dinh chiia cac tl bdo bieu md bdt mdu nhgt (Hinh 3).
Trong hdu hit cdc loai cd xuang, tuyen lic la mpt CO quan gdm 2 thuy ndm gdn xoang mang.
Tuy nhien d mdt sd loai ca, tuyen lie cd the tdn tai nhilu han hai thuy. 6 ca dp (Sicyases sanguineus), mdi khoang mang chua 2 thiiy cua tuyen lic vdi mpt thiiy nam d vi tri be mat, con
thuy kia nam sat vdi bieu md mang (Bowden et al. 2005). Nhihig nghien ciiu vl su phat sinh tuyen lie d cd hdi Dai Tay Duong cho thdy trong qud trinh phdt tri8n, tuyin iic ciia cd hdi co xu hudng nhd hdn vdo trong xoang ndp mang trong khi nhung loai cd khac nhu cd chem chdu Au, ca chep {Cyprinus capio L.) sy phdt triSn cua tuyen lie diln ra ben dudi lop bieu md trong co thi cd (Botham and Manning 1981; Bowden et al. 2005). Kit qua quan sdt su phdt triln hiyln lie cua cd chem cho thdy gidng nhu phdn 16n cdc lodi cd xuong khdc, tuyin lic cd chem la mot CO quan gdm hai thiiy ndm gan xoang mang va trong qud trinh phdt trien, tuydn lic md rdng vao ben trong ca the nhung vdn cd mpt phdn nh6 ra ngoai hudng vao xoang nap mang.
KHOA HQC KY THUAT THtJ Y TAP XX SO 6 - 2013
Hinh 3 TuySn C(c cdi chem 30 ngay sau khi nd T: tuyen uc; M: ca ho hdp; HK: tiin than; B: ndo; KT: ong than;
AZ: viing dinh, IZ. viing tuy; OZ: vimg vo
Cd rat nhieu tieu ban ve tuyen tie ciia ca chem d ngay thu 14 sau khi nd cho thay vi tri ciia tuyin lie vd tiln thdn cua cd rat gan nhau, giua chiing xudt hign "cdu ndi" tl bao (Hinh 2d).
Sir tdn tgi cua cdu ndi lien ket gifta tien than va tuyen lie da dugc bdo cdo d nhilu lodi cd khdc nhau nhu cd cam. Spams aurata, cd rd phi Tilapia mossambica, cd bdy mau Lebistes reticulatus hay ca ban sao (Pleuronectes platessa) (Sailendri 1973; Josefsson and Tatner 1993; Bowden et al.
2005) nhung chua cd tdc gid nao chiing minh dugc CO su di chuyen tl bdo tir tiln than din tuyin uc hodc ngugc Iai hay khdng.
Viec xac dinh cdc tl bdo trong tuyin lic va cau tnic cua tuyen dc phdn Idn dugc thyc hien dua tren co sd kiem tra md hpc. Tuy nhien cd the do d giai doan ca con, viec quan sdt tuyen lic bang phuang phdp md hoc khd thuc hien hodc do quan sat khdng ddy du cac lat cat md hpc nen khong danh gid day du ve tuyen uc nen da cd nhftng sai khac trong bdo cao ve hinh thdi tuygn lie (Bowden et al. 2005). Kit qud nghien cihi ciia Liu va cdc cpng su nam 2004 tren mpt lugng Idn
mdu cd ban {Paralichthys olivaceus) cho thdy tuyen uc cua loai cd nay tu khi nd cho den 7 thang tudi khdng xuat hien phan tiiy (Liu et al.
2004), trong khi bao cao cua Chantanachookin va ctv (1991) lai ghi nhan su khdc biet gifta cdc vung ciia tuyen lie ciing ngay chfnh loai cd nay.
Dieu tuang tu ciing da xay ra d cd zebrafish (Lara et al. 2002). Dii vay cdc nhd nghien ciiu dd thira nhdn cdu tnic tuyen lic ciia cd khac nhau tuy theo loai va ngay trong ciing mpt loai, cfing cd nhung khac biet ve cau tnic ciia tuyen uc theo dg tudi khac nhau. Sy phan viing trong tuyen lie xuat hien d hai lodi ca ban Scophlhalmus maximus va Hippoglossus hippoglossus trong khi tuyen lie cua cd hdi {Salmo salar) khdng thdy xuat hien sy phdn viing nay (Ellis, 1977;
Padros and Crespo 1996; Foumier-Betz va ctv., 2000). 6 ca mandarin (Siniperca chuatsi) tuyen lie dupe phan thanh 3 viing; vimg dinh la viing cd nhiing te bdo bieu md hinh khdi bat mau nhat ndm ben tren cdc tl bdo bilu md hinh thon dai.
Viing vd dupe xdc dinh la vimg cd mat do te bao lympho nhd cao trong khi vung tuy dac trung bdi sy thua thdt cua cdc tl bao lympho va tl
85
KHOA HQC KY THUAT T H C Y TAP XX S 6 6 - 2013
bdo dgng bilu md (Xie vd ctv, 2006). Tuy nhiSn, d cd ban {Scophlhalmus maximus), tuyen lic chi dugc chia 1dm 2 viing: viing tuy vd viing vd (Padros vd Crespo, 1996).
Cac tieu bdn md hpc ciia cd chgm trong nghien cdu ndy cho thay ciu tnic tuyen lie ciia cd chgm tuong ty vdi cau tnic tuyen ire ciia cd mandarin nhung ed hai khdc bi?t dd Id vung dinh tuyin uc cd chgm khdng cd te bdo bieu md thon dai ndm gifta vdng dinh vd viing vd, khdng phdt hi?n the Hassatl d viing tuy tuyen lic.
IV, K E T L U A N
Tuyin uc cua cd chgm (Lates calcarifer) bdt dau hinh thdnh tu ngdy thii 12 sau khi nd vd hodn thi?n vdo ngdy thii 30 sau khi nd. Khi dd hodn tbi^n, tuyin uc ciia cd chgm Id co quan lym- pho gom hai thuy hinh qud ddu ndm gdn xoang mang vd keo ddi lien tyc den bieu md hau. Cdu tnic cua tuyen lie dugc phdn chia thdnh 3 vung:
Viing dinh, vung vd vd vung tuy. Trong tuyin lie khdng thdy tieu the Hassall.
Qua trinh phat trien hoan thien cua tuyen ire, CO quan lympho tmng uong cua cd chgm trong dieu kien nudi dudng tai Vi$t Nam cho thay vi?c su dung vacxin phdng benh cho ca chem chi cd the mang lai hieu qua thyc sy sau khi cd dgt tir 30 ngdy tudi trddi.
T A I L I E U T H A M K H A O I. Do Thj Hoa, Bin Quang Te, Nguyen H&u Dung,
Nguyen Thj Mupi (2004). Benh hpc thuy san..
NXB Nong Nghiep
^. Botham J. W. and Manning M.J (1981) Histogenesis of the lymphoid organs in the carp, Cyprinus carpio L., and the ontogenetic development of allograft reactivity Journal of Fish Biology, 19:403-414
3. Bowden TJ., Cook P and Rombout J.H. (2005) Development and function of the thymus in teleosts.
Fish and Shellfish Immunology. 19:413-427.
4. Chantanachookin C, Seikai T. and Masaru T. (1991) Comparative study of the lymphoid organs in three species of marine fish.
Aquacullure, 99:143-155.
5 Ellis A.E. (1977) Ontogeny of the immune response in Salmo salar Histogenesis of the lymphoid oigans and appearance of membrane immunoglobulin and mixed leucocyte reactivity.
In: Solomon J.B. and Horton J.D., editors.
Developmental Immunobiology: Elsevier/
North-Holland Biomedical Press; p. 225-231.
6. Foumier-Betz V, Quentel C. Lamour F.
and LeVen A. (2000) Immunocytochemical detection of Ig-positive cells in blood, lymphoid organs and the gut associated lymphoid tissue of the turbot (Scophthalmus maximus). Fish &
Shellfish Immunology. 10:187-202.
7. Gravningen, K. (2002). Immunoprophylaxis at eariy life cycle stages of marine fish - with emphasis on IPN virus and nodavirus infection.
Havbruk - en nering 1 vekst, Avsluttet 2002.
8. Josefsson S. and Tatner M,F (1993) Histogenesis of the lymphoid organs in sea bream, Spea-us aurata L Fish and Shellfish Immunology, 3:35-50.
9. Lam S.H., Chua H.L., Gong Z., Wen Z., Lam T.J. and Sin Y.M. (2002) Morphologic transformation of the thymus in developmg zebrafish. DevDyn, 225:87-94.
10. Liu Y., Zhang S., Jiang G., Yang D., Lian J. and Yang Y. (2004) The development of the lymphoid oigans of flounder, Paralichthys olivaceus, from hatching to 13 months. Fish &
Shellfish Immunology. 16:621-632.
11. PadrosFandCrespoS.(i996)Ontogeny of the lymphoid organs in the turbot Scophlhalmus maximus- a light and electron microscope study Aquaculture. 144:1 - 16.
12. Sailendri K.. 1973, Studies on the development of lymphoid organs and immune responses in the teleost, Tilapia mossambicus [Ph.D. thesis].
University of Madurai, India.
13. Tatner, M.F. (1966). Natural changes in the immune system of fish. In: Iwama, G. and Nakanishi, T. (Eds.) "The fish immune system:
Organism, Pathogen, and Environment", pp 255-287. Academic Press.
14. XieH.X., NieP., Zhang Y.A., Sun B.J., Sun J„ Yao W J. and Gao Q. (2006) Histological and cytological studies on the developing thymus of mandarin fish Siniperca chuatsi (Percifonnes:
Teleostei). J Appl Ichthyol, 22:125-131.
Ngay nhdn 15-4-2013 Ngay phan bien 12/6/2013
