Tp ch Khoa hc Trng i hc Cn Th
TC DNG CA XUNG NH SNG TRN Bacillus subtilis
DNG HUYN PH V GY NHIM TRN GIA V
Nguyn Bo Lc1, Nicorescu Irina2, Chevalier Sylvie2 v Orange Nicole2
1
Khoa Nng nghip & Sinh hc ng dng, Trng i hc Cn Th
2
LMSM, University of Rouen, EA 4312, 55, rue Saint-Germain, 27000, Evreux, France
Thng tin chung:
Ngy nhn: 31/07/2014
Ngy chp nhn: 27/04/2015
Title:
Inactivation of Bacillus
subtilis vegetative cells in
suspensions and spices by
pulsed light effects
T kha:
Xung nh sng, kh nhim,
gia v, B. subtilis
Keywords:
Pulsed light,
decontamination, spices, B.
subtilis
Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 37 (2015)(2): 1-10
ABSTRACT
The aim of our study was to assess the impact of a pulsed light (PL) technology
on B. subtilis growth on two model matrixes, through bacterial inactivation
and cell integrity. First, the effect of PL treatment (light flux of 0.6 J.cm-2/flash)
on B. subtilis vegetative cells in liquid state (DO580nm = 0.8) was evaluated. We
found that PL treatment leaded to 8 log reduction of B. subtilis vegetative cells
while bacterial morphology was not affected. When DNA extract from B.
subtilis vegetative cells was analyzed, a significant difference was found
between the control and the PL-treated samples. Second, spices, among which
ground caraway, ground red pepper and ground black pepper were artificially
inoculated with a known concentration of B. subtilis in vegetative state (at
logarithmic phase) and then the spices were exposed to PL (light flux of 1 J.cm-
2
/flash). In this case, PL treatment leaded to a bacterial reduction of about 1
log and serious damage of the microorganisms parietal structure occurred.
To conclude, this work highlights that pulsed light had a potential for
inactivation of B. subtilis in both, liquid and dry state but through different
mechanisms.
TM TT
Mc ch ca th nghim l nhm nh gi tc ng ca xung nh sng
(PL) trn B. subtilis trong hai dng mi trng (dng huyn ph v gy
nhim trn gia v) thng qua hai ch tiu chnh l kh nng tiu dit vi
sinh vt v tc ng ln cu trc t bo vi sinh vt. u tin, th nghim v
tc ng ca phng php x l xung nh sng (vi cng nng lng
0,6 J.cm-2/xung) trn huyn ph ca t bo sinh dng B. subtilis
(DO580nm = 0,8) c thc hin. Kt qu cho thy phng php ny tiu dit
c 8 log t bo trong khi hnh dng ca t bo khng b nh hng. Khi
phn tch s di chuyn ca ADN trn gel agar, th kt qu cho thy c s khc
bit ng k gia mu x l v mu i chng. Tip theo, th nghim nghin
cu tc ng ca xung nh sng (vi cng nng lng 1 J.cm-2/xung) trn
ba loi gia v (tiu hi, t v tiu en) c gy nhim nhn to vi t bo
sinh dng ca B. subtilis (giai on pht trin logarit) c thc hin. Th
nghim ny cho thy phng php x l bng xung nh sng c th tiu dit
c khong 1 log t bo vi sinh vt, quan st hnh nh t bo bng knh hin
vi in t cho thy hnh dng bn ngoi t bo vi sinh vt b ph hu di tc
ng ca phng php x l ny. Tm li, th nghim ny cho thy phng
php x l bng xung nh sng c th tiu dit c t bo sinh dng ca B.
subtilis, nhng c ch bt hot th khc nhau.
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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 37 (2015)(2): 1-10
1 GII THIU
Ngy nay, cc yu cu thng mi v php lut
lin quan n s an ton, cht lng v bo qun
cc sn phm thc phm gp phn vo vic pht
trin v ci thin cc phng php kh nhim. i
vi gia v, phng php kh trng thng xuyn
p dng nht l thanh trng (bng nhit kh hoc
hi nc), ngoi ra cn c phng php chiu x,
ht chn khng Tuy nhin, c nhiu nghin cu
cho thy rng nhng phng php kh trng
thng s dng ny c th gy ra s mt mu sc,
ph hu vitamin, thay i hng v hoc lm mt
mt tinh du trong cc loi tho mc v gia v
(Gould, 1964; Vajdi & Pereira, 1973; Grahl &
Markl, 1996). ng dng ca xung nh sng gy
c s ch c bit i vi cc nh khoa hc l m, rau qu v tri cy) (Sauer & Moraru, 2009;
do mc s dng nng lng thp hn so vi cc
qu trnh x l nhit (Barbosa-Canovas et al.,
1998) v s thn thin vi mi trng ca phng
php ny. K thut xung nh sng l k thut s
dng cc xung nng lng cao ca ph nh sng
rng vi thi gian cc ngn tiu dit vi sinh vt
(Woodling & Moraru, 2006). Tu thuc vo mc
nng lng x dng, cu to ca thit b,
khong cch gia ngun sng v mu, vi sinh vt
mc tiu, tnh cht v thnh phn ca mu x l.
Tc dng dit khun ca xung nh sng gy ra ch
yu bi ph UV (l tc nhn to ra cc lin kt nh
hp trn ADN ca vi khun), v s tng nhit
cc b do tc dng ca ph hng ngoi to nn s
ph v t bo vi sinh vt (Dunn et al., 1995;
Wekhof 2000; Takeshita et al., 2003). Mc d, c
ch chnh xc v tc dng ca xung nh sng trn
t bo vi sinh vt vn cha c lm sng t hon
ton. Tuy nhin, nhiu nghin cu cho thy mt
ng gp quan trng ca phn ph UV ngn (250-
260 nm) ca xung nh sng trong bt hot vi khun
(Uesugi & Moraru, 2009).
Bt gia v c s dng rt rng ri trong
thc phm vi vai tr ng gp mi v v mu sc
cho thc phm. Tuy nhin, nhiu nghin cu cho
thy cc loi gia v ny thng b nhim khun vi
mc rt cao, c th k n cc loi nh Bacillus
spp., Salmonella, Escherichia coli, Clostridium
perfringens v cc c t ca Aspergillus (USFDA,
2002). Nguyn nhn ca s nhim khun ny l do
iu kin trng trt cng nh qu trnh sn xut,
bo qun, vn chuyn v phn phi km v sinh.
Bacillus subtilis l mt trong nhng loi vi khun
thng gp trong cc loi gia v dng bt. Cc loi
gia v ny c mc nhim khun thng xuyn
ln ti khong 8 log CFU/g (McKee, 1995; Keith,
1998). B. subtilis c kh nng sinh bo t v l loi
vi khun gy ng c thc phm, c th gp cc
loi nh i ma, au bng v tiu chy. Hu ht
cc nghin cu cho thy lin quan n xt, hnh
ngm, rau u ng hp v cc loi bt (Gilbert et
al., 1981; Kramer et al., 1982 ; Kramer & Gilbert,
1989). B. subtilis v bo t ca chng c kh nng
chng chu rt tt vi mt s cng on trong qu
trnh ch bin v c th sng st sau qu trnh
thanh trng. Chnh v vy, B. subtilis c th c
coi l mt trong nhng loi vi khun mc tiu
trong qu trnh thanh trng nhm m bo an ton
thc phm.
Xung nh sng cho hiu qu tiu dit vi sinh
vt rt cao trn cc loi b mt, bao gi v gn y
c nghin cu nhiu trn thc phm (tht, bnh
Uesugi & Moraru, 2009) nhng rt t ti liu cp
n tc dng ca phng php ny trn cc loi
thc phm dng ht v dng bt (Fine & Gervais,
2004). Nhng nm gn y, mt vi nghin cu
(Keith et al., 1997; Takeshita et al., 2003; Fine &
Gervais, 2004; Staack et al., 2008) hng n vic
nh gi kh nng kh nhim trn thc phm dng
bt bng cc k thut khng s dng nhit. Keith et
al. (1997) nghin cu hiu qu kh nhim ca
xung in trng (PEF) trn gia v. Cc tc gi
nghin cu nh hng ca cc thng s PEF (v d
nh thi gian, s lng xung, hnh dng xung) v
phng php x l lin tip n mc gim mt
s vi khun trn hng qu, th l v bt hnh ty.
Hiu qu dit khun ca tia hng ngoi trn bt t
cng c nghin cu bi Staack et al. (2008) v
aw 0.8, hiu qu kh khun t c t 1 2 log
CFU/g vi nng lng x l l 11 v 5 kw/m2. Mc
d vy, cc nghin cu lin quan ti kh nng kh
nhim ca xung nh sng trn thc phm dng bt
vn cn rt khim tn. Mc ch ca th nghim
ny l nh gi kh nng kh nhim ca xung nh
sng trn mt s loi gia v c gy nhim vi
B. subtilis.
2 PHNG PHP TH NGHIM
2.1 Chng vi khun v iu kin nui cy
Chng B. subtilis (ATCC 6633, vin Pasteur,
Php) c nui cy trong mi trng tng sinh
M17 (Merck, c) c b xung 0,5% glucose vi
iu kin nui cy khuy o t ng (180
vng/pht) nhit 30C v thi gian 24 gi.
Tip theo, mi trng c ly tm 10.000 g,
20 C v thi gian 20 pht, sau pha long phn
t bo thu c vo dung dch nc mui sinh l
0,9% v iu chnh mt quang hc (OD580nm) v
0,8. Sau , 10 ml dung dch s c x l bng
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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 37 (2015)(2): 1-10

xung nh sng (c cp trong mc 2.3). Mt theo dy thp phn bng dung dch mui sinh l
n v log vi khun l logarit ca mt s vi khun (Biokar Diagnostics, Php). Cui cng, dy pha
(cfu/mL hoc cfu/g). long thp phn c cy trang trn b mt mi
2.2 Phng php gy nhim gia v vi trng PCA (AES, Php) v m s khun lc t
B. subtilis c sau khi nhit 30 C trong thi gian 24
gi. Th nghim c lp li 3 ln.
Cho 3 g gia v (tiu hi, tiu en v t )
2.3 Phng php x l xung nh sng
(Colin Palc, Php) vo 25 mL dung dch huyn ph
ca B. subtilis (Pha cn bng), hn hp c khuy Thit b x l xung nh sng c cung cp bi
o lin tc (180 vng/pht) trong thi gian 4 gi Claranor (Php) gm c 1 b phn tch in v 1
nhit 30C. Sau hn hp c ly tm iu bung x l, trong c 4 n xenon dng hnh tr
kin 10.000 g, 20 pht v 20C. Tip theo, phn (Massier et al., 2011). Thit b th nghim ny to ra
gia v sau ly tm c tri thnh mt lp mng trn mt lot cc xung nh sng c bc sng t 200
a Petri v sy kh trong thi gian 15 gi iu n 1100 nm v thi gian ca mi xung l 300 s.
kin v trng v nghin nh bng my dp mu Trong th nghim ny, cng nng lng
(Smasher AES, Php). Sau , nguyn liu c s dng x l huyn ph vi khun l
x l bng xung nh sng (c m t trong mc 0.6 J.cm-2/xung v gia v l 1 J.cm-2/xung.
2.3). Mu x l v mu i chng c pha long
Bng 1: iu kin x l theo cu hnh ca thit b
S lng Khong cch t Cng nng lng t Cu hnh
xung in p (V) n ti mu (cm) c (J.cm-2) ca thit b
12 0.6 4 n
1 3000
10 1 3 n
12 1.2 4 n
2 3000
10 2 3 n
12 2.4 4 n
4 3000
10 4 3 n
12 3.6 4 n
6 3000
10 6 3 n
12 4.8 4 n
8 3000
10 8 3 n
12 6 4 n
10 3000
10 10 3 n

Huyn ph
vi khun

Mu
H thng o trn

(a) (b)
Hnh 1: S minh ho bung x l xung nh sng (Claranor): (a) cu hnh 4 n s dng cho huyn
ph vi sinh vt v (b) cu hnh 3 n s dng cho bt gia v
Huyn ph vi khun c ng trong mt hp V, 1 Hz vi 1, 2, 4, 6, 8, 10 xung c cu hnh 4 n
hnh ch nht bng thch anh v c x l 3000 v cng nng lng s dng l 0.6 J.cm-2/xung

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 37 (2015)(2): 1-10
(Hnh 1a v Bng 1). Trng hp ca gia v c
gy nhim nhn to vi B. subtilis, th mu c
x l trong iu kin o trn 3000 V, 1 Hz v
10 xung vi cng thit b nh trn nhng c cu
hnh 3 n t c cng nng lng l 1 Th nghim u tin c thc hin nhm mc
J.cm-2/xung (Hnh 1b v Bng 1). Sau khi c x
l, tt c cc mu s c phn tch ngay lp tc.
Th nghim c lp li t nht 3 ln.
2.4 Phng php trch ly ADN
ADN ca t bo B. subtilis c trch l theo
phng php c s dng trong phng th nghim
LMSM (Laboratoire Microbiologique Signaux et
Microenvironnement). T bo thu c sau khi ly
tm huyn ph vi khun iu kin 10.000 g,
20C, 20 min c cho vo ng eppendorf cng
vi 2 x 650 L dung dch m P1 (10 mM tris base
[pH 8], 2 mM EDTA, RNAse I 100 g/mL) v
lysozyme (5 mg/mL), sau mu c 37C.
Sau khi 5 10 pht, mu c thm vo 25 L
proteinase K (20 mg/mL) v 50 L dung dch SDS
10%, tip theo mu c 60C trong thi gian
45 pht. Sau khi , hn hp c thm vo 625 L
phenol bo ho v ly tm 13.000 g, 20C trong
thi gian 10 pht. Phn trong pha trn c thu
gi v cho vo ng eppendorf mi cng vi 625
L of phenol-chloroform, hn hp c trn u
v ly tm 13.000 g, 20C trong thi gian 10 pht.
Phn trong pha trn c thu nhn v cho vo mt
eppendorf mi cng vi 625 L chloroform, hn
hp c trn u v ly tm 13.000 g, 20C
trong thi gian 10 pht, phn trong pha trn c
thu nhn v cho vo mt eppendorf mi. Tip theo,
isopropanol c cho thm vo v trn o u
nh nhng gip cho cc si ADN kt hp li vi
nhau to thnh mt mng mu trng. Dng pipet
thu tinh vt mng ADN cho vo mt eppendorf
mi v sy kh nhit phng, sau ADN
c ho tan trong 50 100 L dung dch m TE
(10 mM tris base, pH 7,5; 1mM EDTA). Sau khi
trch ly, hm lng ADN (g/mL) c xc nh
bng thit b quang ph bc sng 260 nm v
280 nm, phc v cho vic tnh ton lng ADN
cn thit cho phng php in di mt chiu trn
gel agar (0,8%) vi cng dng in 90V, thi
gian 1 gi trong mi trng TAE 1X (0,04 M tris
base, 0,196 mM acid acetic, 1 mM EDTA). Tip
theo, gel c nhum mu trong dung dch BET,
thi gian 15 pht v cui cng hnh nh ca gel
in di c chp bng thit b s dng tia UV v
iu khin bng my tnh.
3 KT QU V THO LUN
3.1 Hiu qu x l huyn ph B. subtilis ca
xung nh sng
ch nh gi hiu qu ca s lng xung nh snh
trn huyn ph ca vi khun B. subtilis giai on
pht trin n nh. Kt qu th nghim (Hnh 2) cho
thy ch cn x l mt xung (0,6 J.cm-2) tiu
dit hon ton 8 log vi khun B. subtilis. Kt qu
th nghim ny hon ton trng khp vi kt qu
nghin cu ca Feuilloley et al. (2006), ch cn x
l mt xung tiu dit hon ton 6 log vi khun
Pseudomonas aeruginosa. Vi 5 giy x l bng
xung nh sng, Krishnamurthy et al. (2004) c th
tiu dit c trn 8 log vi khun Staphylococcus
aureus trong mi trng m pht pht. Mt iu
cn lu l huyn ph vi khun trong phm vi th
nghim ny c pha trong mi trng nc mui
sinh l (0,9%), y l mi trng gn nh trong sut
i vi ph nh sng k c nhng tia nh sng thy
c v cc tia t ngoi, c ngha l s hp th tia UV
ca dung dch nc mui sinh l rt thp, to iu kin
cho cc tia ny c c hi tip xc vi vi khun cao hn.
iu dn n hiu qu kh khun cao ca phng
php x l ny (Oms-Oliu et al., 2010).
b sung cho kt qu trn, th nghim nghin
cu tc ng ca xung nh sng trn hnh dng bn
ngoi ca t bo B. subtilis c thc hin vi
phng php qut nh bng knh hin vi in t
(scanning electron microscopy, SEM). Hnh 3a v
3b cho thy xung nh sng khng c nh hng
n hnh dng bn ngoi ca t bo B. subtilis ngay
c vi mc x l nng lng xung nh sng
6 J.cm-2 (10 xung). Tht s, hnh dng bn ngoi
ca t bo vi khun hon ton khng c s thay i
trc v sau khi x l (6 J.cm-2), iu cho thy
vi cc iu kin x l trong th nghim ny th
mc d xung nh sng tiu dit rt tt vi khun B.
subtilis dng huyn ph, nhng n khng gy ra s
ph v hay thay i hnh dng bn ngoi ca t bo
x l. Kt qu th nghim ny tri ngc vi
mt s kt qu th nghim c cng b trc
y trn t bo nm men. Cho ti nay, cha c mt
bo co no lin quan ti tc ng ca xung nh
sng trn vch t bo B. subtilis dng huyn ph.
Tht vy, Takeshita et al. (2003) nhn thy c s
thay i cu trc ca t bo Saccharomyces
cerevisiae dng huyn ph khi x l vi 2 v 3
xung ( cng 0.7 J.cm-2/xung). S thay i ny
cc t bo sau x l bao gm s gia tng kch
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thc ca khng bo, s bin dng mng t bo v ngha l mng t bo nm men khng cn ton vn
s thay i mt s hnh dng khc. Bng nhiu sau qu trnh x l xung nh sng. Tuy nhin, cn
phng php khc nhau, Farrell et al. (2011) nhn mnh rng kt qu th nghim ny c thc
nghin cu tc ng ca xung nh sng n mng hin trn t bo B. subtilis, c bit n nh mt
t bo nm men. Cc tc gi ny nhn thy tia UV loi vi khun c kh nng chng chu vi xung nh
lm thay i tnh thm thu ca mng t bo v sng tt hn so vi nm men (S. cerevisiae). Tht
lm gim ng k s lng t bo sng st sau khi vy, t bo nm men ch c bo v bi mt lp
x l. Sau khi x l cng 90 v 100 xung mng mng, trong khi lp mng bo v ca t
(tng ng vi mc nng lng x l UV trn bo vi khun rt dy v chc chn. Mt gi thuyt
mi cm2 l 2,4 v 4,1J.) th 90 v 99% t bo nm khc c th c dng gii thch cho kt qu th
men b x l bt mu vi thuc nhum PI nghim ca chng ti l s h hi ADN c th l
(Propidium Iodide), iu cho thy tnh thm nguyn nhn dn n s tiu dit vi khun bi x
thu ca mng t bo b nh hng ng k, c l xung nh sng.

8
Log mt s vi sinh vt, Log(cfu.mL )
-1

0
0.00 2.E-05 0,06 0,12 0,2 0,3 0,5 0,6
2 -2
Nng lng xung nh sng x l trn mi cm (J.cm )

Hnh 2: Tc dng kh khun ca xung nh sng trn huyn ph t bo B. subtilis

a) b)
Hnh 3: nh t bo B. subtilis : a) mu i chng; b) mu x l 3 000V, 1Hz, 10 xung
(0.6 J.cm-2/xung)
Th nghim tip theo c thc hin nh khun. Kt qu th nghim c th hin trong
gi tc ng ca xung nh sng n ADN ca vi Hnh 3, kt qu ny cho thy phng php x l

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Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 37 (2015)(2): 1-10
xung nh sng nh hng n kh nng di
chuyn ca ADN trn gel agar. Tht vy, ADN
c trch ly t t bo x l bng xung nh sng
dch chuyn xa hn so vi ADN c trch ly t t
bo ca mu i chng. iu ny cho thy c s
thay i trong cu trc ADN ca cc t bo c
x l bng xung nh sng. S thay i ny c th
l s gy on ca cc si ADN, gi thuyt ny
ph hp vi kt lun ca (Giese & Darby, 2000;
Takeshita et al., 2003), cc tc gi ny cho rng tia
cc tm l nguyn nhn gy ra cc thay i ho hc
v gy v trong cu to ca ADN. Theo
Krishnamurthy (2006) th ADN hp th nng
lng ch yu bc sng khon 254 nm, y l
khong bc sng chnh ca tia UVC v cng l
tc nhn chnh gy ra nhng thay i ho hc trong
cu trc ca ADN. Tm li, kt qu th nghim ny
cho thy phng php x l xung nh sng gy
ra nhng thay i trong cu trc ADN ca t bo
B. subtilis, y chnh l tc dng chnh ca phng
php x l xung nh sng.
Hnh 4: nh hng ca x l xung nh sng
trn tc di chuyn ca AND t bo B. subtilis
(M-marker; NF - Khng x l; 6F mu x l 6
xung v 10F mu x l 10 xung)
3.2 Hiu qu x l ca xung nh sng trn
gia v c gy nhim bi B. subtilis
Hiu qu dit khun ca xung nh sng trn
gia v c gy nhim nhn to vi B. subtilis
c th hin trong Hnh 5. Mc kh nhim ca
xung nh sng khc nhau ng k gia cc loi gia
v th nghim iu ny c th c gii thch bi
mi loi gia v c cc cht c ch khc nhau,
chnh cc cht ny nh hng n kh nng gy
nhim ca vi khun B. subtilis ln gia v cng
nh kh nng kh khun ca xung nh sng (Arora
& Kaur, 1999; Indu et al., 2006). Kt qu th
nghim (Hnh 5) cho thy vi cng nng
lng 10 J.cm-2 (1 J.cm-2 x 10 xung), th xung nh
sng tiu dit c 0,8 log vi khun trn mu tiu
hi v tiu en v t c trn 1 log vi khun i
vi bt t . Kt qu ny khng ng nht vi
mt s nghin cu trc y (Bialka & Demirci,
2008; Wekhof, 2003). Tuy nhin, iu ny c th
c gii thch bi gii hn nghin cu ny, cng
nng lng s dng ti a ch t 10 J.cm-2, vi
cng nng lng ny th mc kh khun
t c (khong 1 log) l t yu cu. Tht
vy, mt s nghin cu (Keith et al., 1997) nh
gi kh nng kh nhim ca trng in t (PEF)
(100 xung vi nng lng 80 kV/cm) trn bt gia
v (hng qu, th l v hnh ty) v kt qu t
c l di 3 log vi khun. Mc kh khun
thp ca xung nh sng trn bt gia v c th c
gii thch bi kh nng tip xc thp ca vi khun
vi nh sng, c bit l tia cc tm. Tht vy,
nhng vi khun nm lp trn s tip xc vi nh
sng v b tiu dit, v chnh nhng vi khun ny
s cn tr s tip xc ca nh sng vi phn cn li
ca mu. L thuyt ny ph hp vi kt qu nghin
cu ca Gmez-Lpez et al. (2007), cc nh
nghin cu ny cho rng kh nng to che khut
ca vi sinh vt cng cao khi cc vi sinh vt ny hp
th nh sng mnh. iu ny lm cho cc vi sinh
vt nm lp di rt kh b tiu dit v ngc li
vi cc vi sinh vt nm lp trn. Hn na, tiu
dit c vi sinh vt th photon v vi sinh vt phi
tip xc vi nhau trong qu trnh x l. Do , cc
vt th nm gia ngun sng v vi sinh vt s hp
thu cc photon v n s hn ch tc dng kh
khun ca phng php ny. Hn na, hn ch ny
s rt khc nhau gia cc mu thc phm dng rn,
dng lng hay huyn ph vi sinh vt. Tht vy,
t c hiu qu dit khun cao khi x l thc
phm dng bt v d nh gia v, th i hi phi c
mt gii php k thut hp l nh thit k thit b
x l xung nh sng kt hp vi o trn thc
phm trong qu trnh x l (Gardner & Shama,
2000).
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Hnh 5: Hiu qu dit khun ca xung nh sng trn gia v c gy nhim vi B. subtilis

Tip theo, tc ng ca xung nh sng trn t
bo B. subtilis gy nhim trn gia v c nh gi
qua hnh nh chp c bng knh hin vi in t.
Hnh 6a, 6b, 7a v 7b cho thy kt qu tri ngc
vi vic x l t bo trong huyn ph, hnh dng
bn ngoi ca t bo vi khun c x l trng
thi kh b thay i r rng trong qu trnh x l
xung nh sng. S nng ln qu mc ca t bo
(June et al., 2003) gy ra cc vt nt trn mng
t bo (Hnh 6b). Hnh nh t bo vi khun B.
subtilis c th hin trong hnh 6b v 7b hon
ton ph hp vi kt qu nghin cu ca Wekhof
(2001) v Sonenshein (2003). Nhng tc gi ny
thy rng bo t B. subtilis b ph hy hoc bin
dng di tc dng ca phng php x l bng
xung nh sng. Mt s nh nghin cu cho rng
nguyn nhn chnh gy ph hy t bo vi sinh vt
khi x l bng xung nh sng l do hin tng
quang ha gy ra do tc dng ca tia cc tm
(Wekhof, 2000; Wekhof, 2001). Theo ti liu tham
kho, c gi thuyt cho rng xung nh sng cng
gy ra tc dng nhit ln t bo vi sinh vt trong
trng hp s dng ngun nng lng cao v thi
gian di. iu ny gy ra s bc hi nc ca t
bo v to thnh nhng dng hi nc nh, lm
cho mng t bo b ph hy (Takeshita et al.,
2003). ng quan im, Wekhof (2000) cho rng
c ch tiu dit vi sinh vt ca xung nh sng bao
gm c hai tc ng quang ha v quang nhit.

a) b)
Hnh 6: Hnh nh t bo B. subtilis trn bt tiu en
a) mu khng x l; b) mu x l 10 xung (1 J.cm-2/xung)

7
Tp ch Khoa hc Trng i hc Cn Th Phn B: Nng nghip, Thy sn v Cng ngh Sinh hc: 37 (2015)(2): 1-10
a).
Hnh 7: Hnh nh t bo B. subtilis trn tiu hi
a) mu khng x l; b)mu x l 10 xung (1 J.cm-2/xung)
4 KT LUN
Th nghim ny c thc hin nhm mc ch
so snh hiu qu kh khun ca xung nh sng trn
t bo B. subtilis dng huyn ph v gy nhim
trn bt gia v. Kt qu nghin cu cho thy, i
vi huyn ph vi khun, ch cn 1 xung (0,6 J.cm-2)
tiu dit ton b 8 log t bo B. subtilis.
Ngc li, iu kin gy nhim trn bt gia v,
vi cng 10 J.cm-2 (10 xung), phng php
ny ch tiu dit c trn di 1 log t bo vi
khun. Qua quan st trn hnh SEM, hnh dng ca
t bo B. subtilis trong bt gia v b bin dng v
ph v bi x l xung nh sng. Ngc lai, i vi
huyn ph vi khun th khng c hin tng ny
mc d ton b t bo b tiu dit.
Kt qu nghin cu ny cho thy tim nng
phong ph ca cng ngh x l xung nh sng
trong vic tiu dit vi sinh vt trong huyn ph v
c trong thc phm dng bt. iu ny a n
nhiu ha hn cho vic s dng xung nh sng
thay th cho phng php v hot vi sinh vt bng
nhit hin nay. Tuy nhin, i vi bt gia v,
tng hiu qu ca phng php ny, th thit b cn
phi c nghin cu v pht trin hn trong tng
lai nhm tng kh nng tip xc ca gia v vi
ngun sng.
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