Tap chi Khoa hoc va Cong nghe biin TIO (2010). S6 1. Tr 51-65 MOT SO KET QUA NGHIEN CUtJ VE VI SINH VAT
TAI VUNG VEN BIEN HAI PHONG
DO MANH HAO, PHAM THE THU
Vien Tai nguyen va Moi trirtmg bien
Tom tdt: Tic 50 mdu nude vd trdm tich viing ven bien Hdi Phong da phdn lap duac 65 chung vi khuan dien hinh thuoc 31 lodi, 16 chi. 8 ho vd 2 bo. So luang nhom vi khudn hieu khi rieng ngdp man Bdng La, Trdng Cdt khd cao, bien ddng 10 -10 tb/ml,g; xq khudn tir 0 - l&tb/ml,g; nam men tir 0 - 2.10^tb/ml,g vd ndm soi cd mat do dat tai l&tb/ml,g. Sd luang vi khudn hiiu khi vd xa khudn trong cdc mdu trdm tich thuang cao hem mdu nuac bi mat tic 10 - 100 Idn, nhung ndm sai vd ndm men lai cd xu huang nguac lai, dac biet vdo miia mua. Nhieu chiing vi khudn cd hoat tinh nitrat hod, phdn nitrat, phdn gidi protein, tinh bdt vd khd nang doi khdng vai vi khudn gdy binh Vibrio parahaemolyticus vd Vibrio fuoniss cao. Chi pham dang long vd bdt duac tao ra tic td hap mot sd chung vi khudn trin khi thir nghiem trin nude thdi dam nuoi trdng thuy san cho thdy cdc chi tieu mdi truong duac phdn tich Id DO, COD, BOD} vd NH4* cd su cai thiin ddng ke so voi mdu doi chieng sau 10 ngdy thd chephdm.
Tie khoa: vi khudn hiiu khi; xq khudn, ndm men; ndm sai: rieng ngdp man Bdng La vd Trdng Cat: chi pham dang long vd dang bdt.
I. MO DAU
Vung ven bien Hai Phong nam trong pham vi he toa do 20''35'00"-2l''00'00"N va 106''35'00"-107°10'40"E, phia Dong va Dong Bac giap voi ven bien Quang Ninh, phia Tay va Nam giap voi viing ven bien Thai Binh. Trong do, Viing nay bao gom doi ngap trieu rgng 242 km^ va dai bo ngam voi do sau 6 - 10 m rgng tren 500 km^ [8] da tao ra nhieu kieu he sinh thai khac nhau nhu la rung ngap man (RNM), cira song, bai trieu cat, khu vuc nuoi trong thuy san va cang bien. Vung ven bien nay nam trong khu vuc nhiet doi gio mua voi mua mua nong am mua nhieu va mua kho lanh kho it mua, cac thong so tram tich lo limg, do muoi, pH, nhiet do va cac hgp chat huru co boa tan ciing co sir bien doi theo mua rat ro ret.
Cho den nay, da c6 nhieu cong trinh nghien ciiu ve da dang ciia he thong dong thuc vat, danh gia nguon Igi thuy san va cac dieu kien thuy ly, thuy boa, nhung vin con thiiu nhiing nghien ciiu ve vi sinh vat (VSV), nhom sinh vat dong vai tro rit quan trgng trong
qua trinh chuyen boa vat chat, dong thai vi sinh vat ciing la nguon gen gia tri cho cac nganh cong nghiep thuc pham, y dugc, boa my pham va xir ly moi truong. Chinh vi vay, chung toi da tien hanh nghien ciiu sir da dang vi sinh vat, dong thai danh gia va tuyen chgn mot so chimg vi sinh vat hiiu ich tai khu vuc nay.
II. PHl/ONG PHAP NGHIEN CLTU
1. Nguyen lieu
Bao gom 50 miu nuoc va trim tich dugc thu thap tai cac dam nuoi tring thuy san, rimg ngap man, cira song trong viing ven bien Hai phong, thuoc de tai co so Vien Tai nguyen va Moi truong bien nhiing nam 2003, 2004, 2005 va 2007.
2. Phuorng phap
2.1. Thu mau ngoai hien trircmg
- Mau nuoc dugc lay bang bathometer sau do chuyen vao Ig thuy tinh dung tich 100 ml da khir triing va bao quan lanh trong hop xop co chiia da truoc khi mang ve phong thi nghiem phan tich.
- Mau tram tich dugc lay bang thiet bi lay bun chuyen dung hoac bang thia inox da khir trung, khoang 500 g tram tich dugc chuyen vao tiii nylon va bao quan lanh trong hop xop CO chiia da truac khi mang ve phong thi nghiem phan tich.
2.2. Phuong phap trong phong thi nghiem
- Dem so lugng cac vi sinh vat va phan lap vi sinh vat hiiu ich tren moi truong chgn Igc: Vi khuan hieu khi (HK) tren moi tnrong hieu khi tong so; Nam men (NMn) tren moi truong Hansen; Nam moc (NM) tren moi truong Czapek; Xa khuan (XK) tren moi truong xa khuan bien (1).
- Xac dinh hinh dang te bao duai kinh hien vi thuang (Ba Lan) va kinh hiin vi dien tu quet JSM-5410 LV, JEOL (Nhat Ban) (1).
- Xac dinh cac dac diem sinh ly va sinh hoa cua vi khuan theo huang din ciia Smibert&Krieg, (1994)(10).
- Doan gen 16S rRNA sau khi dugc nhan len va thuan khiet se dugc dgc true tiip tren may dgc gen ABI 3100.
- Phan loai vi khuin theo khoa phan loai ciia Bergey, (1957) (9).
- Xac djnh ham lugng amoni (NH3) va nitrit (NO2") trong nuac biin dugc xac dinh bing phuang phap so mau phenat tren may do quang pho ke DR/2000 (HACH, My) voi dai do: tir vit (0,0 |igN) tra len, do phan giai: 0,lpigN va sai so: ± 10% gia tri do.
m . KET QUA NGHIEN CUU 1. Thanh phSn loai
Bang 1: Thanh phin loai cac chiing vi khuan phan lap tir ven bien Hai Phong Stt
1 2 3
4 5
6 7 8 9 10
11
12
Thanh phan loai Class: Schizomycetes Order I: Pseudomonadales Sub-order: Rhodobacteriineae Family I: Nitrobacteriaceae Genus 1: Nitrosomonas Nitrosomonas spp.
Genus 2: Nitrobacter Nitrobacter spp.
Genus 3: Nitrospina Nitrospina spp.
Family II: Athiorhodaceae Genus 4: Rhodopseudomonas Rhodopseudomonas capsulate BLV-01
Rhodopseudomonas capsulate BLV-02
Sub-order: Pseudomonadineae Family III: Pseudomonas Genus 5: Pseudomonas Pseudomonas aeruginosa Pseudomonas pseudomallei Pseudomonas caviae Pseudomonas fluorescens Pseudomonas putida Order II: Enbacteriales Family IV: Rhizobaiceae Genus 6: Rhizobium Rhizobium spp. 2V2 Family V: Brevibacteraceae Genus 7: Brevibacterium Brevibacterium healii
Stt
13 14 15
16 17 18
19
20 21 22 23 24 25 26 27 28 29 30
31
Thanh phan loai Family VI: Bacteroidaceae Genus 8: Bacteroides Bacteroides capillosus Genus 9: Streptobacillus
Streptobacillus moniliformis 2VK2 Streptobacillus moniliformis 1VK3 Family VII: Achromobacteraceae Genus 10: Alcaligenes
Alcaligenes faecalis Genus 11: Flavobacterium Flavobacterium marinum Flavobacterium marinotypicum Genus 12: Achromobacter Achromobacter liquefaciens
Genus 13: Lactobacillus Lactobacillus spp.
Genus 14: Escherichia Escherichia sp.
Genus 15: Bacillus Bacillus subtilus STCK99 Bacillus subtilus VK07 Bacillus lichenniformis Bacillus cereus
Bacillus brevis Bacillus megaterium Bacillus sphaericus Bacillus spp. 2VK5 Bacillus spp. 2V8 Family VIII: Bacillaceae Genus 16: Clostridium Clostridium spp. 2V1
Dua tren cac dac diem hinh thai khuan lac, te bao, dac diim sinh ly, sinh hoa, ket qua giai trinh tu 16S rRNA va theo khoa phan loai ciia Bergey's 1954. Chiing toi xep 65 chiing vi khuan phan lap dugc tir vung ven bien Hai Phong vao 31 loai, 16 chi, 8 hg, 2 bg (bang 1). Tat ca cac chiing giong nay da dugc luu vao bg suu tap giong vi sinh vat cua Vien.
Hg Achromobacteraceae c6 s i chi nhieu nhat - 6 chi, trong khi do cac hg khac chi CO tur 1 - 2 chi. Chi Bacillus c6 toi 8 loai vi khuan khac nhau, tiep den la chi Pseudomonas voi 5 loai vi khuan, cac chi con lai chi co tir 1 - 2 loai.
2. Phan b i cua VSV trong RNM Bang La va Trang Cat
Kit qua cho thiy ca 4 nhom vi sinh vat nay deu co mat o trong vung nghien ciiu voi mat do khac nhau (bang 2):
s i lugng t i bao vi khuin hiiu khi biin dong tir lO"* - lO^tb/ml; g, mat do te bao vi khuin hiiu khi it nhat trong mau nuoc Bang La vao miia kho - 10 va cao nhat la mau dat Trang Cat vao miia mua - 10^ (hinh 1).
s i lugng xa khuin dat mat do cue dai la 10^tb/ml;g trong mau dat Trang Cat vao miia mua. S i lugng t i bao nim men dat mat do cao nhat trong mau nuoc Trang cat vao mua mua - 2.10^tb/ml;g, nim men khong thay xuat hien trong tat ca cac mau vao mua kho.
Trong miu nuoc Trang Cat vao miia mua mat do te bao nam sgi dat toi 10^tb/ml;g, nhieu mau khong thay co mat nam sgi (hinh 2).
Bang 2: S i lugng mot so nhom vi sinh vat trong RNM Bang La - Trang Cat
STC WTC SBL WBL
Mua mua (thang 8/2007) HK
10^
10*
10^
10^
XK 10^
0 10' 0
NM 15 2x10^
0 10
NS
50 10^
0 2X10^
Miia liho (3/2008) HK
10*
5x10"
10*
10"
XK 10^
0 10 0
NM 0 0 0 0
NS
0 50 0 0 Ghi chu: STC: Dat RNM Trdng Cdt, WTC: Nuac RNM Trdng Cdt, SBL: Dat RNM Bdng
La, WBL: Nuac RNM Bdng La, HK: Vi khudn hieu khi, XK: Xa khudn, NM: ndm men, NS: ndm soi
Nhin chung, t i bao vi khuan hieu khi, xa khuan, nam men hay nim sgi diu co mat do cao vao mua mua va co mat do thap horn nhieu vao miia kho. Dac biet la nim men va
nim sgi, chiing hiu nhu khong co mat trong cac miu vao miia kho. Co the vao miia mua, nuoc tir cac luu vuc song mang theo mot lugng Ion nguon dinh duang va ca cac vi sinh vat luc dia phan tan vao viing RNM nay lam cho khu he VSV 6 day phong phii them.
RNM Trang Cat co si lugng VSV ca 4 nhom vi khuan hieu khi, xa khuan, nam men, nam sgi diu cao ban VSV a RNM Bang La. Day c6 the la ket qua ciia su khac nhau ve thanh phan moi truong tu 2 RNM nay.
Cac mau nuoc co so lugng nam men va nam sgi cao hon cac mau tram tich, dieu nay CO the ly giai la do 2 nhom VSV nay co the di nhap tam thai tir cac viing luc dia ra.
12000000 -
10000000
8000000
6000000 -
4000000 -
2000000 -
^
^-W#^-w^ VA
1100 -1 1000
900 800 700 600 500 400 300 200 100 0
^
j\
— • — X K — i — N M — • — N S/ A \ •
O X.O X.O x ^ rtV * ^ rtV rtV
Hinh 1: Phan bo vi khuan HK trong RNM Trang Cat va Bang La mua mua(R) va kho
3. Dac diem hinh thai
Hinh 2: Phan bi XK, NM va NS trong RNM Trang Cat va Bang La miia mua (R)
Tir khoang 50 mau tram tich va nuoc viing ven bien Hai phong, chiing toi tuyen chgn dugc 65 chung vi khuan khac nhau (bang 3). Cac khuan lac co mau sac rat khac nhau, tir mau trang sua, mau dat den mau vang, luc va mau do. Hinh dang va kich thuoc khuan lac ciing kha da dang, co nhung khuan lac c6 kich thuoc tu nho den to va lan toa len khap be mat thach, mep khuan lac co the tron deu den rang cua, be mat loi hoac 16m.
Da so cac chiing phan lap dugc c6 te bao dang hinh que ngan den dai, ciing co mat cac te bao co dang hinh cau, xoan va hinh ovan, mot so chiing co dang da hinh va bien doi theo chu ki phat trien hay dieu kien moi truong. Cac chiing nay co thi phan ung am hay duong khi nhugm Gram.
A. Chiing vi khuan PLTa B. Chiing vi khuan PLTb'
C. Chiing vi khuin PLTE D. Chiing vi khuan KT8e Hinh 3: Anh ti bao mot so chiing vi khuin duai kinh hien vi dien tir quet
4. Dac diem sinh ly
Cac chung vi khuin nghien ciiu dugc phan tich kha nang phan ling vai oxy, vai nhiet do tai 4, 30, 45 va 55°C, phan ling voi nong do muii tai 10 nong do khac nhau tir 0% din 10%, va ba pH khac nhau la 5,5; 7,0 va 9,5.
Hiu hit cac chiing vi khuin dugc phan lap la vi khuan vi hiiu khi din hiiu khi.
Chiing deu la nhiing vi khuin chiu muii va sinh truong tot nhat tai nong do muii tir 2 - 3%. Chiing CO kha nang sinh truong tit nhat tai 30 C, mot so chiing c6 kha nang sinh truong tai 45°C, nhung nhung chiing vi khuin nay khong mgc dugc tai 4 V va 55°C.
Gioi han pH ciia cac chung vi khuin nay kha rgng, tir 5,5 den 9,5, tuy nhien chiing c6 kha nang sinh truong tit hon tai cac pH kiem.
Bang 3: Dac diim hinh thai mot si chiing vi khuan diim hinh
Mau
Dam nuoi Kien Thuy Dam nuoi Kien Thuy Dam nuoi Do Son Dam nuoi Do Son RNM Gia Minh Cua song Gia Minh
RNM Phu Long RNM Phu Long
RNM Vinh Quang
RNM Trang Cat
RNM Trang Cat RNM Trang Cat
RNM Trang Cat
RNM Trang Cat
RNM Bang La
RNM Trang Cat
RNM Bang La
D§t RNM Trang Cat
Chung
KTla KT8a' DSla*
DSlc GMTa GMDa PLTa PLTb'
VQT
STCK6.22
STCV3.13 STCV3.10
STCM3.29
WTCVL25
SBLV4.15
BLV-02
WBLV2.15
STCK99
Dac diem khuan lac
Tron nho li ty, mau sua Khuan lac tron mau luc, be mat nhan
Khuan lac tron, mau vang be mat nhan
Khuan lac nho li ty, mau vang Khuan lac tron, mau sua Khuan lac tron, mau do, te bao de
tach
Khuan lac tron, mau do Khuan lac tron, mau vang nhat, be
mat nhan
Khuan lac tron, d = 2 - 3 cm, mau da cam, be mat nhan
Mau nau lan toa, mep khong deu, de tach, khong thay doi mau ducri cac moi truong khac nhau va khong hoa
tan vao moi truong thach Khuan lac mau nau dat den vang nhat
Khuan mau lan dat co sac to hoa tan tiet vao moi truong thach Khuan lac to, mep tron khong deu, be
mat hoi loi, mau vang nhat Mau da cam to lan toa mep khong deu, be mat phang, khong thay doi mau dudd moi truong khac nhau va khong hoa tan vao moi truong thach Khuan lac mau nau dat, co sac to hoa
tan tiet vao moi truong thach Khuan lac mau do, nho, mep tron khong deu ducri dieu kien ki khi co
chieu sang
Mau nau dat, co sac to hoa tan tiet vao moi truong thach Mau trang sua to, khong thay doi mau duoi moi truong khac nhau va khong hoa tan vao moi truong thach
Dac diem te bao H. thai
Cau Que Que Que Cau Que Que Que
c i u
Que
Que Que
Que
cau
Que
Que
Que
Que
Gram - - - - - - - - -
-1-
_ -
-1-
-
-
-
-
-1-
Bang 4: Dac diem sinh hoa mot si chiing vi khuin diin hinh
Ten chiing
STCM3.27 SBLK3.6 STCK6.22 STCV3.13 STCV3.10 STCM3.29 SBLV4.2 WTCV1.25 SBLV4.15 STCV3.7 BLV-01 BLV-02 SBLV2-12 STCK99 WBLV2.15
1VK3 1VK5 2VK2 2VK5 2VK8 4VK1 4VK2 VK07 1VK8 2V2 2V1 2V8
Djch hoa siia Trung hoa
Axit hoa Axit boa Trung hoa
Kiem hoa Axit hoa Axit hoa Axit hoa Kiem boa
- Axit hoa Axit hoa Axit hoa Axit hoa Trung boa
0 0 0 0 0 0 0 0 0 0 0 0
Catalaza
+ + + + +
-1-
+ + + + + +
-1-
+ + + + + +
+ + + + + +
-1-
LM glucose
- +
-1-
- - +
- - - - + + +
-f
- + + + + - -
+ + -t- + + +
LM lactose
-1-
+
-1- -1-
+ + + + + - + + + + + + + - - - - - +
4-
+
-1-
+
Mannose
+
-1-
+ +
-1-
+ - + + - - +
-1-
0 0 0 0 0 0 0 0 0 0 0 0
Axit axetic
- -
-1-
- - +
- - - + + - + 0 0 0 0 0 0 0 0 0 0 0 0
Sucrose
-1-
+ + + + + + + + + +
-1-
+ - - - - -
-1-
+ + + + + +
Etanol
- - - - - - - - -
- + + + + + + + - -
-1- -1-
-
-1-
Starch
- - +
- -
-1-
- +
-
- +
- - - - - - + + + + + +
Gelatine
+ -t- + - + + + + - +
- +
- - - - - - - + + + + +
Indole
- + 0 - - 0 - - 0 -
-1-
0 0 0 0 0 0 0 0 0 0 0 0 0 0
s
- 0 0 - 0 0 0 - 0 0
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
*Ghi chii: (+): duang tich, (-): dm tinh, (0): chua xac dinh
5. Dac diem sinh hoa
Cac chiing vi khuin nay ciing dugc tiin hanh phan tich cac dac tinh sinh hoa nhu la kha nang dich hoa siia, phan ling catalaza, len men glucose, len men lactose, kha nang dong boa cac loai duong dan (mannose, sucrose, etanol, axit axetic .v.v.), kha nang phan buy cac dai phan tir (tinh bgt, gelatine) va mot so dac tinh sinh boa dung de dinh loai khac nhu la nitrat hoa, phan nitrat, sinh indole, khir H2S.V.V. ciing dugc sir dung (bang 4).
Mot so ket qua dugc trinh bay a bang 3 cho thay, tat ca cac chung deu co phan ling catalaza duang tinh, da so cac chiing co kha nang kiem hoa dung dich siia, nhieu chiing CO kha nang len men glucose va lactose, nhieu chung co kha nang phan buy gelatine hon la tinh bgt, co nhieu chiing co the phat trien dugc trong moi truong chi c6 etanol lam nguon cacbon duy nhat.
6. Phan tich trinh ty 16S rRNA chung vi khuin VK 07 va STC99
Trinh tu 16S rRNA ciia chiing vi khuan VK07 va STC99 dugc nhan len bang cap moi: 5' AGAGTTTGACCTGGCTCAC 3' va 5' CGGCTACCTTGTTACGACTT 3' tai nhiet do gan moi la Ta 54''C, sau khi tinh sach san pham nay dugc phan tich trinh tu tren may phan tich DNA.
58 80 99
B.subtilise.4 B.subtilise.2
B a c i l l u s . s p . e P I
100 77
B.subtilise.07 B.megateriumC.99
B.megateriumG.3 B.megateriumC.2 68 I B.megateriumC.1
Bacillus.aeolius
• Bacillus.caldo\«lox Bacillus.sp.Na101-1
• Bacillus.hwajinpoensis
Bacillus.spJA[VI-FM2901 I 1
0.005
Hinh 4: Cay phat sinh chiing loai cua chung
Cay phat sinh chiing loai ciia 2 chung vi khuin nay voi cac loai vi khuin co mii quan he di truyen gan giii dugc xay dung bing phin mim MEGA 4.0 (Kumar et al.,
2006). Ket qua trong hinh 4 cho thiy chiing VK07 thuoc loai Bacillus subtilis va co moi lien he gan giii voi loai Bacillus subtilis C.2, Bacillus subtilis C.4 va Bacillus sp. CPl, chiing STC99 thuoc loai Bacillus megaterium va co moi quan he gin giii voi loai Bacillus megaterium C.3.
7. Hoat tinh mot so chiing vi khuan ven bien Hai Phong 7.1. Hoat tinh nitrat hoa
Sau thai gian 42 ngay nuoi vi khuan oxy hoa amon trong be thuy tinh thii nhat dung tich 20 1, nong do amon tir 45,86 mg/l giam xuong con 0,12 mg/l, tiic la chi bang 0,26% so voi nong do ban dau (bang 5). Tir hinh 5 cho thay nong do amon giam manh nhat trong 22 ngay dau, tir 45,86 mg/l xuong con 4,01 mg/l, chi bang 8,7% so vai nong do ban dau.
Nong do nitrit tir 0,052 mg/l sau 42 ngay nuoi giam xuong con 0,013 mg/l, bang 25%
nong do ban dau.
50 -1 45 * 40 - 35 - _ 30 -
|> 25 -
= 20 - 15 - 10 - 5 - 0
c
0,16 - 0,14 - 0,12 - 0,1 -
•3)0,08 -
^0,06 ^ 0,04 - 0,02
0 -
(
NH4+
1 1 1 T ^
10 20 30 40 ngay
N02-
D 20 40 ' '
ngay
50
60 '
Hinh 5: Hoat tinh nitrat boa trong cac bi thuy tinh thi nghiem
Bang 5: Kit qua do boat tinh nitrat hoa trong cac be thuy tinh thi nghiem STT
B i O l B i 0 2
Hoa chat (mg/l)
NH4^
NO2"
Thoi gian do hoat tinh (ngay) 0
45,86 0,052
14 23,98 0,138
22 4.01 0,104
29 3,88 0,021
42 0,12 0,013
7.2. Hoat tinh phan giai protein, tinh bdt va phan nitrat hoa
Tam chiing vi khuan dugc phan lap tir dam nuoi trong thuy san Kien Thuy va Do Son dugc tien hanh phan tich kha nang phan giai protein, tinh bgt va phan nitrat. Ket qua trong bang 6 cho thay, ba chiing KTOOl, KT003 va DS003 c6 ca ba boat tinh phan giai protein, tinh bgt va phan nitrat, trong do KTOO1 va KT003 c6 boat tinh phan giai protein va tinh bgt manh.
7.3. Kha nang doi Idiang doi voi Vibrio
13 chung vi khuan phan lap viing ven bien Hai phong dugc tien hanh phan tich kha nang de khang hai chiing vi khuan gay benh Vibrio parahaemolyticus va Vibrio fuonissi.
Ket qua bang 7 cho thay, 9 chiing co kha nang doi khang vai Vibrio parahaemolyticus va chin chiing co kha nang doi khang voi V. fuonissi. Dang chu y la cac chung B1H6, L1H3, B1H5 va DS002 co kha nang lie che ca 2 loai Vibrio cao.
7.4. Ket qua thir nghiem che pham HIO dang long va bot vao viec xir ly niro'c thai nuoi tdm in quy mo phong thi nghiem
Hai loai nuoc thai la nuoc thai dam nuoi trong thuy san thuoc dia ban buyen Kien Thuy (NTKT) va dia ban thi xa Do Son (NTDS) sau khi dugc dua vao be thuy tinh voi lugng the tich nhu nhau 10 lit. Cac be kiem tra se dugc bo sung 30 g che pham/10 m^
nuoc doi voi che pham dang bgt va 01 lit che pham/1000 m^ nuac doi voi che pham dang long.
Sau 10 ngay tha che pham, ket qua phan tich mau dugc trinh bay o bang 8 cho thay ca hai che pham c6 tac dung cai thien moi truong nuoc thai dam nuoi trong thuy san Kien thuy va Do son kha tit, chi phim HIO dang bgt to ta co tac dung cai thien moi truong nuac tot ban.
Bang 6: Hoat tinh phan giai protein, tinh bgt va phan nitrat hoa mot so chiing vi kh STT
1 2 3 4 5 6
Chung VK KTOOl KT002 KT003 DSOOl DS002 DS003
Phan giai protein
+++
+ -I-++
++
-I-+
+
P h a n nitrat
+
- +
- - +
Phan giai Tinh bot +++
++
4-H-
++
+ +
Ten loai V K
Bacillus lichenliformis Lactobacillus sp.
Bacillus cereus Bacillus subtilis Lactobacillus sp.
Bacillus sp.
*Ghi chu: (+++): hoat tinh manh, (++): ht trung binh, (+): htyeu, (-): khong hoat tinh
Bang 7: Kha nang lie chi Vibrio cua mot so chiing vi khuan ven bien Hai Phong Chiing
B5H5 B1H4 B5H10
B1H6 B7H3 L1H3 B1H5 KTOOl KT002 KT003 DSOOl DS002 DS003
Doi k h a n g V.p
-1- + - ++
- ++
++
+ + - + ++
-
V.f - + + -I-+
- -I-+
++
+ -1- + - ++
-
Ten loai vi khuan
Bacillus cereus B. alvei
B. Polymyxa B. licheniformis B. subtillis Lactobacillus sp.
B. licheniformis Bacillus lichenliformis Lactobacillus sp.
Bacillus cereus Bacillus subtilis Lactobacillus sp.
Bacillus sp.
Ghi chi: V.p: Vibrio parahaemolyticus, V.f Vibrio fuonissi
Bang 8: Thong si boa ly trong nuoc thai nuoi trong thuy san sau 10 ngay tha che pham
Thong so T°C
pH D0(mg/1) B0D5(mg/l)
COD(mg/l) NH4^(mg/l)
Nu-o-c thai NTTS Kiin Thuy Bot
30 7,6 5,5 2,2 15,5 0,13
Long 30 7,5 5,0 3,0 15,9 0,14
DC 30 7,32
4,5 4,8 16,7 0,20
Nuo-c thai NTTS D6 Son Bot
30 7,65
4,6 3,9 14,5 0,14
Long 30 7,60
4,5 4,0 14,7 0,15
DC 30 7,50
4,3 4,3 15,0 0,17
IV. KET LUAN
Chiing toi da xac dinh dugc 31 loai vi khuan thuoc 16 chi, 8 hg va 2 bg cho vimg ven bien Hai Phong. Vi khuan kieu khi, xa khuan, nam men va nam sgi thuang c6 mat do cao vao miia mua va thap vao mua kho, so lugng vi khuan hieu khi va xa khuan trong cac mau tram tich thuang cao ban mau nuac be mat tir 10 - 100 lan, nhung nam sgi va nam men lai co xu huong ngugc lai.
Nhieu chiing vi khuan co boat tinh nitrat boa, phan nitrat, phan giai protein, tinh bgt va kha nang doi khang voi vi khuan gay benh cao. Che pham dang long va bgt dugc tao ra tir to hgp mot so chung vi khuan c6 cac boat tinh cao nay khi kiem tra tren nuoc thai dam nuoi trong thuy san cho thay mot so chi tieu moi truong nhu DO, COD, BOD5 va NH4^ c6 sir cai thien dang ke so voi mau doi chiing sau 10 ngay thir nghiem.
TAI LIEU THAM KHAO
1. Nguyen Lan Dung, Doan Xuan Mu-oii, Nguyen Phung Tien, Dang Due Trach, Pham Van Ty, 1972-1978. Mot so phuong phap nghien cuu vi sinh vat tap 1,11, III. NXB. KH
& KT Ha Noi, 471Tr., 430Tr., 441Tr.
2. Lai Thuy Hien, Dang Phu-ong Nga, D6 Thu Phirong, Pham Thi Hing, Kilu Quynh Hoa, Vuoiig Thi Nga, Nguyen Thi Yen, Nguyen Ba Tii, 2008. Mot si kit qua nghien ciiu da dang vi sinh vat tai biin dao Cat Ba. Hoi nghi khoa hgc toan Quic vi sinh thai va tai nguyen sinh vat lin thii hai: 284-293.
3. Do Manh Hao, Dam Due Tien, 2003. Buoc diu nghien ciiu nhom vi khuan
hoa trong dam nuoi trong thuy san Hai Phong. De tai co so Vien Tai nguyen v. loi truong bien, 57tr.
4. Do Manh Hao, Dam Due Tien, 2004. Buoc dau nghien ciiu nhom vi khuan phan giai protein, tinh bgt va phan nitrat boa viing ven bien Hai Phong. Bao cao de tai co so 2004, 40tr.
5. Do Manh Hao, 2005. Nghien ciiu thir nghiem san xuat chi pham xir ly nuoc thai dim nuoi tring thuy san. Bao cao de tai co so 2005, 50tr.
6. Do Manh Hao, Pham The Thir, 2007. LTng dung phuong phap giai trinh tu gen 16S rRNA vao viec phan loai mot so chimg vi khuan rimg ngap man Bang La, Trang Cat.
Bao cao de tai co so 2007, 49tr.
7. Hao Do Manh, Matsuo Yoshihide, Atsuko Katsuta, Satoru Matsuda, Yoshikazu. Shizuri and Hiroaki Kasai, 2008. Robiginitalea myxolifaciens sp. nov., a novel myxol-producing bacterium isolated from marine sediment and emended description of the gentis Robiginitalea. Intemational Journal of Systematic and Evolutionary Microbiology 58: 1660-1664.
8. Tran Due Thanh, Nguyin Dire Cu, Nguyen Chu Hoi, 1993. Bao cao moi truong dia chat ven ba Hai phong. Thu vien Phan vien Hai duong hgc Hai phong. 151 Tr.
9. Breed Robert, S.; Murray, E.G.D.; Smith Nathan, R., 1957. Bergey's manual determinative bacteriology, seventh edition. London, Bailiere. Tindall & Cox, Ltd.
USA. 1094p.
10. Smibert, R. M., & Krieg, N. R., 1994. Phenotypic characterization. In Methods for General and Molecular Bacteriology, pp. 607-654. Edited by P. Gerhardt, R. G. E.
Murray, W. A. Wood & N. R. Krieg. Washington, DC: American Society for Microbiology.
SOME RESULTS OF STUDY ON MICROBE IN HAI PHONG COASTAL ZONE
DO MANH HAO, PHAM THE THU
Summary: From 50 water and sediment samples collected in coastal zone of Haiphong, we have isolated 65 bacterial strains and identification belong to 31 species, 16 gena, 8 families and 2 orders, the genus Bacillus, there are the highest specie^ number - 8 species, Pseudomonas being 5 species and the other genus has only 1 - 2 spect. ^rrobic bacterial amounts in Bang la, Trang cat mangrove are quite high, range of 10 - h I'ii.g;
Actinomycetes is 0 - l&tb/ml,g: yeast is 0 - 2.10^tb/ml,g and fungi density can reach up
lO^tb/ml.g. The number of aerobic bacteria and actinomycetes in sediment samples were higher 10 - 100 times than those in water samples, but the numbers of yeast and fungi were opposite orientation, especially in raining season. Many bacterial strains were tested the ability of nitrifying, denitrifying, degradation of protein, starch and resistibility to pathogens of Vibrio parahaemolyticus and Vibrio fuonissi indicated that had high activities. Liquid and powder probiotics made from a the bacterial combination were evaluated in the wastewater of shrimp aquaculture pond indicating that analysed chemical parameters of DO, COD, BOD;
and NH4 significantly approved in comparation with control samples in 10 days.
Ngdy nhan bdi: 05 - 7 - 2009 Ngudfi nhan xet: TS. Dam Diic Tiin
