Tap chiKhoa hoc va Cong nghe Ndng nghiep Viet Nam So 06(127)/2021 ANH Hl/ONG CUA VIEC BO SUNG OLIGO 0 GLUCAN TAO RA BANG PHUONG PHAP CHIEU XA TIA y VAO THUC AN LEN TANG TRUONG, TY LE SONG VA CAC CHI T[.]
Tap chiKhoa hoc va Cong nghe Ndng nghiep Viet Nam - So 06(127)/2021 ANH Hl/ONG CUA VIEC BO SUNG OLIGO-0-GLUCAN TAO RA BANG PHUONG PHAP CHIEU XA TIA y VAO THUC AN LEN TANG TRUONG, TY LE SONG VA CAC CHI TIEU MIEN DICH TOM HUM DA (.Palinuridae hom arus) Le Quang Luanu , Nguyen Trong Nghia1, Le Thi Thu Thao1, Nguyen Thanh Vu1 TOM TAT 01igo-/3-glucan co khoi ldpng phan td (Mw) khoang 15 kDa tao bang phdong phap chieu xa tia y ddpc sd dung de danh gia hieu dng tang trddng va kich thich mien dich d tom hum da (Palinuridae homarus) Ket qua cho thay tom hum dddc cho an bo sung che pham oligo-/3-glucan nong 1.000 - 3.000 ppm dat toe tang trddng va sinh khoi thu hoach cao hdn co y nghia so vdi doi chdng Khi cho an bo sung oligo-/3-glucan d nong noi tren cung lam tang 13,6 - 16,0% ty le tom song va giam 0,9 - 1,0 he so tieu ton thde an Viec cho an bo sung che pham oligo-/3-glucan lam gia tang dang ke cac chi so mien dich nhil tong so lddng te bao mau, hoat thde bao, hoat enzyme phenoloxidase va superoxide dismutase tom hum Ket qua nghien cdu cung cho thay nong oligo-jS-glucan cho an bo sung d mile 1.000 ppm la thich hdp Che pham oligo-j3glucan che tao bang phiidng phap chieu xa rat co trien vong de lam chat thuc day tang triidng va mien dich td nhien nuoi tom hum da Td khoa: Tom hum da (Palinuridae homarus), ktch thtch miin dich, oligo-/3-glucan, chieu xa tia gamma ld At v a n d e Nghe nuoi hum dang cang phat trien d midc ta va da dong gop quan vao tang triidng kinh te d khu vdc mien Trung, tom hum da (Palinuridae homarus) diidc nuoi bien Theo thong ke cua Tong cue Thuy san, nghe nuoi tom hum long bat dau phat trien tii nam 2000 va tap trung chu yeu d cac tinh Binh Dinh, Phu Yen, Khanh Hoa, Ninh Thuan va Binh Thuan, den cac tinh co khoang 10 nghin ho nuoi vdi khoang 53 nghin long (Nguyen Phu Hoa, 2019) Tuy nhien, nhdng nam gan day, djeh benh tren tom hum xay tran lan va dien bien rat phde tap gay thiet had rat nghiem cho ngddi nuoi tom Td van de dich benh thddng xuyen xay ra, ngddi nuoi tom hum thddng dung cac loai khang sinh nhd Doxycycline base, Strepto-Tetramycine, v.v va do gay nhieu van de ve an toan cho ngddi tieu dung Trddc quy dinh cam sd dung mot so chat doc hai va khang sinh nham dao bao chat lddng va an toan ve sinh thde pham doi vdi tom thddng pham, ngddi nuoi tom da va dang doi mat vdi khong it kho khan nhflng san pham thay the, nhat la che pham thay cho khang sinh chda da dang va chda that sd co hieu qua nhd mong dqi (Vo Van Nha, 2017; Nguyen Phu Hoa, 2019) Chinh vi vay, viec nghien cdu tao san pham co nguon goc td nhien giup tang ty le song, tang nang suat va chat ldqng nhdng dam bao an toan ve sinh thde pham la het sdc cap thiet hien doi vdi nganh tom Viet Nam /?-glucan la hop chat polysaccharide ddOc tao nen td cac don phan td D-glucose bang lien ket /3-glycoside, la phan cau tao nen te bao thde vat, cac loai nam va nhieu nhat la nam men (Sacchromyces serevisiae) /3-glucan da ddOc biet den nhd la mot chat kich thich mien dich bao, tang eddng mien dich, khang khoi u, khang viem, khang khuan, virus, v.v (Chan et a l, 2009; Bacha et al., 2017) /3-glucan cang ddOc sd dung nhieu nuoi thuy san nham tang eddng phan dng mien dich td nhien (thde bao, san xuat anion superoxide va hoat dong cua lysozyme), kha nang chong lai cac tac nhan gay benh (vi khuan va virus) va cac ap ldc td moi trddng, va thuc day tang trddng (Suphantharika et al., 2003; Ajadi et al., 2016) Trong nuoi tom, /3-glucan ddOc sd dung nhd la mot chat kich thich mien dich chong lai cac mam benh vi sinh vat (Chang et al., 2013; Li et al., 2008; Tran Viet Tien va Dang Thi Hoang Oanh, 2020) Bai va cong tac vien (2014) da bien tinh /3-glucan bang each carboxymethyl hoa va sulfoetyl hoa de tao cac san pham /3-glucan tan nddc co hoat tinh kich thich mien dich cao d tom the chan trang Litopenaeus vannamei /3-glucan co nguon goc td cac loai nam men bien cung ddpc sd dung de bao ve tom the chan trang chong lai virus gay benh 1Tmng tam Cong nghe Sinh hoc TP H6 Chi Minh : Trucmg Dai hoc Tai nguyen va Moi truemg TP H6 Chi Minh 102 Tap chi Khoa hoc va Cong nghe Nong nghiep Viet Nam - So 06(127)/2021 dom trang WSSV (Ochoa-Alvarez et al., 2021) Tuy nhien, kha nang hoa tan cua /Tglucan la rat thap co khoi lilpng phan td (Mw) cao dan tcfi kho hap thu, hoat tinh sinh hoc thap va kho ap dung rong rai quy mo cong nghiep Gan day, mot so nghien cdu da cho thay /Tglucan co Mw thap va tan nil he co hoat tinh sinh hoc cao hdn so vdi /3-glucan co Mw cao (Byun et al., 2008) De che tao oligo-jS-glucan, ngiidi ta thpdng sd dung cac phxldng phap hoa hoc, sinh hoc va bde xa Trong phildng phap cat mach bde xa ddpc cho la phdOng phap rat hdu hieu va co nhieu du diem nhd tiet kiem nang lilpng, nguyen lieu, tin cay cao de dang dieu chinh khoi ltipng nhii mong muon, san pham co chat ldpng cao (co the sd dung ma khong can phai tinh che), dap dng nhu cau bao ve moi trddng, gia thap, hieu qua kinh te cao va de san xuat d quy mo ldn (Luan et al., 2012; Long et al., 2019) Muc tieu cua nghien cdu la xac dinh diipc nong oligo-/?-glucan che tao bang phddng phap chieu xa tia y bo sung vao thde an thich hpp co kha nang kich thich tang trilbng va tang eddng mien dich tren tom hum da, nham tien tdi dng dung che pham nhd mot chat thuc day tang triidng va khang benh td nhien nuoi tom II VAT LIEU VA PHUONG PHAP NGHIEN CUU 2.1 Vat lieu nghien cdu Tom hum da (Palinuridae homarus) ddpc khai thac td vung bien ven bd Nha Trang, Khanh Hoa Cong ty TNHH TM DV Anh Hong Res cung cap Tom hum giong ddpc dong den dat khoi ldpng trung binh 2,6 g de tien hanh bo tri thi nghiem Mau oligo-/i-glucan dang dung dich co Mw~15 kDa che tao bang philong phap chieu xa ddpc cung cap bPi Trung tarn Cong nghe Sinh hoc TR Ho Chi Minh 2.2 Phddng phap nghien cdu 2.2.1 B o tri th i nghiem, cham soc va quan ly Tom hum da sau ddng, dong deu ve khoi ldpng (~ 2,6 g/con), kich thdPc ddpc tha vao long nuoi (co kich thildc m x m x 1,5 m - dai x rang x cao) vdi mat 100 con/long ddpc chia nghiem thde (lap lai lan) gom: 16 cho an thde an thong thddng (doi chdng) va 16 cho an thde an co bo sung oligo-j8-glucan vdi nong lan ldpt 1.000,2.000 va 3.000 ppm Long nuoi ddpc dac each vung trieu khoang 300 m sau m co nhiet trung binh khoang 25 ± 0,6°C va man trung binh khoang 34 ± 0,5%o Tom ddoc cho an (chu yeu la ca tap co bo sung ghe va so tddi) moi mot lan vao buoi sang, oligo-/3-glucan ddpc tarn trpe tiep vao ca cho an bo sung vao cac thd ,4 va hang tuan Oligo/1-glucan ddpc tarn trdc tiep vao ca nhii sau: Ca ddpc lam rao nddc, sau ddpc tarn vcfi toan bo 25, 50 va 75 mL dung dich oligo-/?-glucan (4%) giP de dat nong oligo-jS-glucan ca lan ldpt la 1.000, 2.000 va 3.000 ppm Ca sau tarn ddpc tiep tuc phu vPi dau mdc va de rao 30 phut trilPc cho tom an Trong qua trinh nuoi, tom ddpc kiem tra hang va ve sinh long nuoi hang tuan 2.2.2 X ac dinh cac chi tieu tang tnidng, ty le song vd h e so tieu ton thvtc an Cac chi so ddpc xac dinh bao gom: - Khoi ldpng tom ddpc xac dinh hang thang bang each bat ngau nhien moi long con, sau can tom ddpc dda trd lai vao long de nuoi tiep - Ty le song cua tom ddpc xac dinh ket thuc thi nghiem, theo cong thde: N Ty le song (%) = -— x 100 No Trong do: N : So tom luc bat dau thii nghiem, Nt: So liidng tom tai thdi diem t - Mdc tang dilpc xac dinh nhd sau: KL - KL Mdc tang (%) = — -—— x 100 KL - Tang trddng tddng doi (SGR) ddpc xac dinh theo cong thde: SGR In (khoi liidng cuoi)-ln (khoi liidng dau) (%/ngay) So nuoi x 100 He so tieu ton thde an (FCR): FCR Tong so kg thde an (tiidi) da cho an Tong so kg thu hoach - Sinh khoi long (kg/long) ddpc xac dinh bang each can toan bo tom moi long thu hoach 2.2.3 X ac dinh ede chi so m ien dich Chi so mien dich cua tom hum da ddpc danh gia sau thang nuoi gom: Tong te bao mau, hoat thde bao (PA), hoat enzyme (PO), hoat enzyme (SOD) 103 Tap chi Khoa hoc va Cdng nghe Nong nghiep Viet Nam - 56 06(127)/2021 Tom ddpc thu mau (5 con/long) de xet nghiem cac chi so mien dich theo quy trinh cua Yudiati va cong tac vien (2016) Cu the: Dich huyet tddng ddpc thu tii xoang bung tom bang kiem tiem mL ddpc chong dong Tong te bao mau (HTC) diiqc xac dinh bang buong dem hong cau dilcfi kinh hien vi sif dung vat kinh 40X Hoat thqc bao (PA) diiqc xac dinh bang each hoa 20 pL dich huyet tiidng va 20 pL PBS Hon hdp dildc bo sung them 20 pL dich vi khuan Bacillus subtilis d nong 108CFU/mL Trai pL h6n hdp len lam kinh, co dinh bang ethanol 95% va nhuom vdi giemsa 10% 20 phut Cac lam kinh dildc rda lai bang nddc, phdi kho va dem dddi kinh hien vi Hoat enzyme phenoloxidase (PO) diiqc xac dinh bang each tron 100 pL dich huyet tiidng vdi 100 pL PBS, ly tarn d 400 g, 4°C 20 phut de thu nhan tua Rda tua vdi 100 pL dem cacodylate va tiep tuc ly tarn de thu nhan tua Bo sung lan lddt 100 pL dem cacodylate va 100 pL trypsin, tien hanh u 10 phut Sau do, them 50 pL L-DOPA vao hon hdp va tiep tuc u 10 phut trddc doc ket qua tren may ELISA (Molecular Devices, My) d bddc song 490 nm Hoat enzyme superoxide dismutase (SOD) dildc xac dinh bang each hoa 40 pL dich huyet tiidng vao 360 pL dem phosphate, tien hanh ly tarn 6.000 g, 4°C phut de thu dich noi U dich thu dildc 65°C phut, sau bo sung them 50 pL NBT va tien hanh doc ket qua tren may ELISA (Molecular Devices, My) d bildc song 630 nm 2.3 Phildng phap sii ly thong ke Tat ca thi nghiem dildc lap lai lan Ket qua diiqc sd li thong ke bang phildng phap ANOVA sd dung phan mem SPSS 27 Cac ket qua trung binh diiqc so sanh dpa vao mdc khac biet toi thieu co y nghia (LSD) vdi sai so 5% 2.4 Thdi gian va dia diem nghien cdu Thi nghiem dildc tien hanh tai Trung tarn Cong nghe Sinh hoc TP Ho Chi Minh va Cam Ranh, Khanh Hoa (toa 11°55’43.5”N; 109°11T7.0”E) td thang 9/2020 - 4/2021 III KET QUA VA THAO LUAN 3.1 Anh hilo'ng cua che pham oligo-jS-glucan co Mw~15 kDa che tao bang phildng phap chieu xa len sd tang trildng cua tom hum da Bang Tang tnidng, ty le song, sinh khoi tom va he so tieu ton thde an sau thang nuoi Nghiem thde Nong oligo-/l-glucan bo sung vao thde an CV (%) ppm (Doi doing) 1.000ppm 2.000ppm 3.000ppm Khoi lddng dau (g) 2,607 2,65* 2,55* 2,60* 1,57 Khoi ldpng cuoi (g) 267,85* 313,73b 313,43b 316,78b 7,74 2^1* 2,27* 2^9* ± 1,63 2^9* 1,73 10.201,9* 11.738,6b 12.191,2C 12.083,7C 7,98 Sinh khoi tom (kg/long) 21,45* 29,1 lb 28,61b 29,39b 14,03 FCR 5,82* 4,92b 4,90b 4,83b 9,21 Ty le song (%) 80,3* 93,0b 91,4b 93,0b 6,84 SGR (%/ngay) Mdc tang (%) Ghi chu: Cdc gid tri trung binh cimg mot hang co ky tu khac thi khde biet co y nghia thong ke (p