Tp ch Khoa hc Trng i hc Cn Th

S chuyn : Nng nghip (2014)(2): 111-115

TNH HNH NHIM VI KHUN VIBRIO SPP.

TRN TM BC (PENAEUS MERGUIENSIS), TM S (PENAEUS MONODON),
TM RO T (METAPENAEUS ENSIS) TI MT S CH
THUC QUN NINH KIU THNH PH CN TH
Nguyn Thu Tm1, Nguyn Phc Khnh1 v Phan Th Hng Nhung2
1
2

Khoa Nng nghip & Sinh hc ng dng, Trng i hc Cn Th

Sinh vin Th Y K35, Trng i hc Cn Th

Thng tin chung:

Ngy nhn: 26/9/2014
Ngy chp nhn: 07/11/2014

Title:
Contamination of Vibrio
spp. on Penaeus merguisis,
Penaeus monodon, Penaeus
ensis in some markets in
Ninh Kieu District, Cantho
City
T kha:
Tm, Vibrio
Keywords:
Vibrio, shrimp

ABSTRACT
From December 2012 to May 2013, 120 samples (43 Su shrimp, 43 Bac
shrimp and 34 Rao Dat shrimp) were collected from some markets in Can
Tho city to isolate Vibrio spp. bacteria by using ISO/TS 21872-2:2007
standard. The results showed high propotion positive with Vibrio spp.
16,66% (20/120). The rate of Vibrio spp. from Su shrimp, Rao Dat shrimp
and Bac shrimp was 20.93%, 17.65% and 11.63% respectively. By
chemical tests, Vibrio cholerae (11.11%) and Vibrio fluvialis (22.22%)
were found in Su shrimp. However Vibrio cholerae and Vibrio vulnificus
were found in Bac shrimp with 40% and 20%, respectively. Vibrio fluvialis,
Vibrio cholerae, Vibrio vulnificus were found in Rao Dat shrimp with the
same rate (16.66%). Vibrio spp. were sensitive with Doxycyline,
Gentamicin, Bactrim, Amoxicillin and Norfloxacinin in antibiotic test.
Vibrio spp. were multiresistance with (Ax+Bt), (Nr+Bt) (10%) and (Bt, Ax,
Ge)(5%).
TM TT
T thng 12/2012 n thng 5/2013, 120 mu (bao gm 43 mu tm S, 43
mu tm Bc v 34 mu tm Ro t) c thu thp ti mt s ch thuc
qun Ninh Kiu Thnh ph Cn Th nhm xc nh s hin din ca vi
khun Vibrio spp. theo tiu chun ISO/TS 21872-2:2007. Kt qu ghi nhn
c nh sau: t l nhim vi khun Vibrio spp. cc mu kim tra kh cao
16,66% (20/120), trong mu tm S nhim 20,93%, tm Ro t
17,65% v tm Bc nhim 11,63%. Bng cc th nghim sinh ha c bit,
xc nh c s hin din ca 2 chng vi khun Vibrio cholerae v Vibrio
fluvialis tm S vi t l ln lt l 11,11% v 22,22%. cc mu tm
Bc, t l nhim Vibrio cholerae v Vibrio vulnificus ln lt l 40% v
20%. S hin din ca 3 chng Vibrio fluvialis, Vibrio cholerae v Vibrio
vulnificus trn tm Ro t l nh nhau (16,66%). Kt qu kim tra khng
sinh , cho thy vi khun Vibrio spp. phn lp c nhy cm hon ton
vi 2 loi khng sinh l Doxycyline, Gentamicin, nhy cm cao vi
Bactrim, Amoxicillin v Norfloxacin vi t l ln lt l 80%, 80% v 95%.
C hin tng a khng vi 2 loi khng sinh (Ax+Bt), (Nr+Bt) chim t l
10% v khng vi 3 loi khng sinh (Bt, Ax, Ge) chim t l 5%.
111

Tp ch Khoa hc Trng i hc Cn Th

S chuyn : Nng nghip (2014)(2): 111-115

qu trnh th nghim l 120, gm: 43 mu tm S,

34 mu tm Ro t v 43 mu tm Bc.
2.3 Phng php nui cy phn lp
2.3.1 Tng sinh

1 GII THIU
Vi khun Vibrio spp. l mt trong nhng vi
khun nguy him v n khng nhng l tc gy
bnh quan trng m cn c th gy ng c thc
phm ngi. Hin nay, bnh truyn nhim do
nhm vi khun Vibrio spp. trn thy sn t bit l
trn tm c xem l tc nhn gy bnh ng c
quan tm, lm nh hng rt ln n sn lng
tm nui hng nm (Nguyn Vn Ho, 2000). Bn
cnh , c quan ch ca vi khun Vibrio spp.
thng l khi gan ty ca tm (Chan, 1989). y
cng chnh l nguyn nhn lm ly truyn nhiu
bnh ng tiu ha, trong c bnh dch t, cho
ngi n cc mn n ti hoc sng t tm.

Mu c cn v lm nhuyn, sau cho vo

mi trng tng sinh. Mu sau khi cho vo mi
trng ASPW c cha nng mui NaCl l 2%,
c 370C trong 24 gi.
2.3.2 Phng php xc nh khun lc
Cy chuyn mu t mi trng tng sinh sang
mi trng chn lc TCBS, 370C trong 24 gi.
Hnh thi v mu sc khun lc xc nh. V.
parahaemolyticus, V. vulnificus, V. mimicus c
thc hin theo phng php ca tiu chun ISO/TS
21872-2: 2007.
2.3.3 Kim ra cc c tnh sinh ha

Nghin cu ny nhm xc nh t l nhim

Vibrio spp. trn tm, nh danh mt s loi vi
khun Vibrio spp. v xc nh tnh nhy cm i
vi mt s loi khng sinh ca vi khun Vibrio spp.
phn lp c trn tm ti qun Ninh Kiu - Thnh
ph Cn Th.

Cc chng vi khun phn lp c tin hnh

kim tra c tnh sinh ha da theo tiu chun
ISO/TS 21872-2: 2007.
2.3.4 Khng sinh

2 PHNG PHP NGHIN CU

2.1 i tng nghin cu
Tm S, tm Bc v tm Ro t c thu
thp t mt s ch thuc qun Ninh Kiu, Thnh
ph Cn Th.
2.2 Phng php ly mu
Tng s 120 mu, bao gm 43 mu tm S, 43
mu tm Bc v 34 mu tm Ro t c ly ti
cc sp mt s ch nh ch An Ha, ch An
Nghip, ch Ci Kh, ch Hng Li, ch Tn An,
ch Xun Khnh, mt ca hng hi sn ti sng
v mt siu th thuc qun Ninh KiuThnh ph
Cn Th. Mu c ly 3 v tr khc nhau, mi v
tr ly 1 con tm. i vi tm sng ca hng hi
sn ti sng v siu th mu c ly ngu nhin
3 con. Cc mu c bo qun v em v phng
th nghim phn tch. Tng s mu thu thp trong
Bng 1: Tiu chun ng knh vng v khun ca
for Antimicrobial Susceptibility Testing)
Tn Khng

K hiu

Hm lng

Norfloxacin
Nr
10
Doxycyline
Dx
30
Amoxicilin
Ax
10
Bactrim
Bt
1,25/23,75
Gentamicin
Ge
10
Phng php x l s liu: S liu c x
l bng chng trnh Excel 2007 v phn mm

112

Kim tra nhy ca cc khng sinh

Amoxicillin (Ax), Doxycycline (Dx), Gentamicin
(Ge), Norfoxacin (Nr), Bactrim (Bt) i vi vi
khun phn lp c bng cch tri u canh
khun (108 CFU/ml- c tng ng vi ng
Mc Farland 0.5 ln mt thch MHA. t cc a
khng sinh sao cho hai a cch nhau 2,5 cm 3,5
cm v cch mp hp thch 2 cm 2,5 cm, 370C
trong 24 gi.
Kt qu khng sinh c xc nh bng cch
o ng knh vng v khun sau so snh vi
bng tiu chun ng knh vng v khun ca
Vin tiu chun lm sng v xt nghim (CLSI Clinical and Laboratory Standards Institute) (CLSI,
2011).
khng sinh (CLSI, 2011. Performance Standards
ng knh vng v khun (mm)
Nhy
Trung gian
Khng
17
13-16
12
14
11-13
10
17
14-16
13
16
11-15
10
15
13-14
12
thng k Minitab 16. S liu >5 dng Chi square .
<2 dng Fisher exactly test.

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Bng 3: T l nhim Vibrio spp. trn cc loi

tm theo a im ly mu

3 KT QU V THO LUN
3.1 Kt qu nui cy phn lp vi khun
Vibrio spp. trn tm S, tm Bc, tm Ro t

Ch

Kt qu Bng 2 cho thy t l nhim vi khun

Vibrio spp. trn tm S, tm Bc, tm Ro t
chim 16,66%. T l nhim ny kh cao, iu ny
cng c th gii thch l do tm S, tm Bc, tm
Ro t sng trong mi trng nc c s bin
ng v nng mui m vi khun Vibrio spp. li
l loi a mui, cn mui pht trin, chng phn
b rng cc khu vc nc mn, vng nc ven
bin, ca sng v cc khu vc nui trng thy hi
sn (Nguyn Th Hin v ctv., 2009). Mt khc,
theo Oliver and Kaper (1997), vi khun Vibrio spp.
thng sng cc vng n i v nhit i m
nc ta li c kh hu nng m, iu ny rt thun
li cho vi khun Vibrio spp. pht trin.

A
B
C
D
E
F
G

S mu phn
lp
Tm S
43
Tm Bc
43
Tm Ro t
34
Tng

120

T l
(%)
13,33
5,88
5,55
20
12,5
27,77
28,57
NS

3.3 Kt qu nh danh mt s loi Vibrio

spp. phn lp c trn tm S, tm Bc, tm
Ro t bng phn ng sinh ha
Kt qu Bng 4 cho thy t l nhim vi khun
Vibrio spp. trn tm S kh cao chim 33,33%,
ng thi c 2 loi Vibrio spp. c tm thy l V.
fluvialis (22,22%) v V. cholerae (11,11%). S vy
nhim ca 2 loi ny trn tm S c th l do nhm
vi khun Vibrio spp. tn ti mi trng nc ao
nui tm S. Tuy nhin, s khc bit ny khng c
ngha thng k (NS). Theo Chan, (1989) cho rng
thi gian nui tm cng di th thnh phn v s
lng cc loi vi khun cng cao, mt vi khun
Vibrio nhiu nht giai on 90120 ngy nui.
ng thi cng c th b vy nhim vi khun
Vibrio t mt s loi thy hi sn khc. Bi loi vi
khun Vibrio spp. thng sng ven b sng, gn
ca bin ni c nc mn vo v thng gy
bnh trn cc loi thy hi sn khc nhau, nn kh
nng tm S b ly nhim l rt cao. Theo Bi
Quang T, (2006) th vi khun Vibrio spp. thng
gy bnh ng vt thu sn nc mn v nc
ngt: c, gip xc, nhuyn th...

Bng 2: T l nhim vi khun Vibrio spp. trn

tm S, tm Bc, tm Ro t
Mu

S mu phn S mu Vibrio spp.

lp (mu) dng tnh (mu)
15
2
17
1
18
1
15
3
16
2
18
5
21
6

S mu
dng tnh
9
5
6

T l
(%)
20,93
11,63
17,65
P> 0,05
20 16,66

T l nhim vi khun Vibrio spp. trn tm S l

20,93% cao hn tm Ro t l 17,65% v thp
nht l tm Bc l 11,63%. Tuy nhin, s khc bit
ny l khng c ngha thng k (p>0.05). iu
ny c th l do cc tm c ly cng mt thi
im, cng ngun cung cp tm cho cc sp bn
cc ch (v qua vic iu tra khi thu thp mu c
bit tm bn cc ch hu ht c ly t mt ch
u mi).

Bng 4: T l nhim mt s loi Vibrio spp. trn

tm S

3.2 Kho st t l nhim Vibrio spp. trn tm

S, tm Bc, tm Ro t theo a im ly mu

Chng
Vibrio
V. cholerae
V. fluvialis

Kt qu Bng 3 cho thy t l nhim vi khun

Vibrio spp. trn tm ch G cao nht chim
28,57% v thp nht ch C c 5,55%. S khc
bit ny khng c ngha thng k (NS> 0.05).
Nguyn nhn c th l do cc sp thu mua tm
t mt ngun cung cp ng thi s mu nghin
cu cha ln th hin s khc bit v t l
nhim Vibrio spp trn tm cc ch.

Tng

113

S mu
phn lp
9
9

S mu
dng tnh
1
2

T l
(%)
11,11
22,22
NS
33,33

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l do tm c ly cc ngun ging nhau. Loi

vi khun V. fluvialis c tm thy trn tm Ro
t c th l do vy nhim t mi trng nc ao
nui tm. Theo Lee v ctv. (1981) th vi khun
V. fluvialis phn b rng ri khp th gii, cc
vng nc ngt v nc l ca sng.

Bng 5: T l nhim mt s loi Vibrio spp. trn

tm Bc
Chng Vibrio
V. cholera
V. vulnificus

S mu
phn lp
5
5

Tng

S mu T l
dng tnh (%)
2
40
1
20
NS
3
60

Bng 6: Kt qu nh danh cc chng Vibrio

trn tm Ro t

Bng 5 cho thy t l nhim vi khun Vibrio

spp. trn tm Bc cao (60%). Trong V. cholerae
chim 40%, v V. vulnificus l 20%. Tuy nhin, t
l nhim 2 chng ny l khc nhau khng c
ngha thng k. V. cholerae chim t l kh cao,
y l mi nguy c i vi sc khe ngi tiu
dng c bit l ngi hay n hi sn chn ti v
y l loi gy bnh dch t ngi. V. cholerae
c tm thy trong tm Bc c th do b nh
hng t ngun nc b nhim khun.

Chng Vibrio
V. cholera
V. vulnificus
V. fluvialis

S mu
phn lp
6
6
6

Tng

S mu
dng tnh
1
1
1

T l
(%)
16,66
16,66
16,66
NS
3 49,98

3.4 Kt qu kim tra s nhy cm ca vi

khun Vibrio spp. phn lp c i vi khng sinh

Qua kt qu bng trn cho thy cc chng

vi khun Vibrio phn lp c u nhy cm
cao vi 5 loi khng sinh s dng Gentamicin
(100%), Doxycyline (100%), Norfloxacin (95%),
Amoxicillin (80%) v Bactrim (80%).

Cc chng Vibrio c tm thy trn tm Ro

t l V. cholerae, V. vulnificus, V. fluvialis u
chim t l ging l 16.66%. Nguyn nhn c th

Bng 7: Kt qu kim tra tnh nhy cm ca vi khun i vi khng sinh

Nhy
Trung gian
Khng
Tn khng
Tng s
K iu
sinh
mu S mu T l (%) S mu T l (%)
S mu T l (%)
Norfloxacin Nr
20
19
95
0
0
1
5
Doxycyline Dx
20
20
100
0
0
0
0
Amoxicillin Ax
20
16
80
1
5
3
15
Bactrim
Bt
20
16
80
3
15
1
5
Gentamicin Ge
20
20
100
0
0
0
0
sinh chim t l 15%. Trong , khng vi 2 loi
3.5 Kt qu kim tra tnh a khng ca vi
khng sinh (Norlfoxacin + Bactrim, Amoxicillin +
khun Vibrio phn lp c trn tm S, tm
Bactrim) c 2 mu v khng vi 3 loi khng sinh
Bc, tm Ro t i vi cc loi khng sinh
(Amoxicyllin+Doxycycline+ Bactrim) c 1 mu.
Kt qu Bng 8 cho thy, vi khun Vibrio
phn lp c khng ng thi nhiu loi khng
Bng 8: Kt qu kim tra tnh a khng ca vi khun Vibrio phn lp c i vi cc loi khng sinh
(n = 20)
S loi khng sinh b vi khun Vibrio spp.
khng (loi)
2
2
3
Tng

S lng mu
(mu)

Nr+Bt
Ax+Bt
(Ax+Bt+Ge)

S lng mu
T l (%)
(mu)

20
1
5
20
1
5
20
1
5
20
3
15
(11,63%) l khng khc nhau. Trong 20 mu
dng tnh nh danh c mt s loi Vibrio spp.
hin din trn tm S, tm Bc, tm Ro t l
V. vulnificus, V. fluvialis, V. cholerae. c bit loi
V. cholerae chim t l cao nht (40%) tm Bc,
20% tm Ro t v 11,11% tm S.

4 KT LUN
C s hin din ca vi khun Vibrio spp. trn
tm S, tm Bc v tm Ro t vi t l kh cao
(16.66%), v t l nhim Vibrio spp. trn tm S
(20,93%), tm Ro t (17,65%), tm Bc
114

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S chuyn : Nng nghip (2014)(2): 111-115

6. Nguyn Vn Ho, 2000. Mt s vn k

thut nui tm S cng nghip. NXB Nng
nghip, TP H Ch Minh, trang 7-14.
7. Nguyn Th Hin v ctv, 2009. Vi sinh vt
tp nhim trong lng thcthc phm. i
hc Bch Khoa H Ni.
8. Nguyn Vn Duy v Nguyn Th Cm Ly,
2011. Phn lp v xc nh gen c t ca
Vibrio parahaemolyticus trong hi sn ti
sng Nha Trang. Tp ch Khoa hc
Cng ngh thy sn, trng i hc Nha
Trang, s 2.
9. Chan, K.U., 1989. V. parahaemolyticus and
other halophilic Vibrios associated with
seafood in Hong Kong. J. Appl. Microbiol.
66: pp. 57-64.
10. Lee, J.V.; Shread, P.; Furniss, A.L.; Bryant,
T.N, 1981. Taxonomy and description of
Vibrio fluvialis sp nov (Synonym Group F
vibrios, Group EF-6. J. Appl. Bacteriol: pp.
73-94.
11. Microbiology of food and animal feeding
stuffs-Horizontal method for the detection
of potentially enteropathogenic Vibrio spp.
Part 2: Detection of species other than
Vibrio parahaemolyticus and Vibrio
Cholerae. ISO/TS 21872-2, Firt edition
15/4/2007.
12. Oliver JD, Kaper JB, 1997. Vibrio species.
In: Doyle MP, ed. Food microbiology,
fundamentals and frontiers. Washington,
DC, ASM Press: pp. 228-264.

Vi khun Vibrio u nhy cm cao

Doxycycline (100%), Gentamicin (100%),
Norfloxacin (95%), Amoxicillin (80%) v Bactrim
(80%). Tuy vi khun Vibrio phn lp c c t l
khng thp nhng c biu hin a khng vi 2
loi khng sinh (Amoxicillin + Bactrim, Bactrim +
Norfloxacin) 10%, khng vi 3 loi khng sinh
(Amoxicyllin + Doxycycline + Bactrim) 5%.
TI LIU THAM KHO
1. 1. Bi Quang T, 1996. Bnh tm c v gii
php phng tr. Tp ch thu sn s 4/1996.
2. ng Th Hong Oanh v on Nht
Phng, 2006. Xc nh v tr phn loi v
kh nng khng thuc khng sinh ca vi
khun Vibrio pht sng phn lp t hu u
trng tm s (Penaeus monodon). Tp ch
Nghin cu Khoa hc, i hc Cn Th,
trang 42 52.
3. Hunh Kim Diu, 2010. Th Y C s
dc l trong iu tr. NXB Nng nghip,
TP HCM.
4. Nguyn Th Xun Trang v Nguyn Ngc
Tun, 2012. Tn s xut hin Vibrio
cholerae trn tm v nhuyn th, xc nh
serogroup O1, O139 v biotype ca V.
cholerae bng k thut multiplex- PCR. Tp
ch Khoa hc K thut th y, Khoa Chn
nui th y, i hc Nng Lm TP. HCM,
9(3), trang 51-59.
5. Nguyn Cng T v Nguyn Duy Phong,
2006. Nhng bnh min nhit i thng
gp. NXB Y hc H Ni.

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