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Ag

ric
ult
ura
lBi
ot
ech
nol
ogy2
017,6(3):5
5-5
8 En
vir
onme
nta
lMi
cro
bio
log
y55

Is
ola
tiona
nd Chara
cter
iza
tio
no fVib
riosina
loe
nsi
sfr
om
Penae
usvanna
meiBooneinHa i
nanPro
vinc

nWU ,Yi
YuZHOU,Li fanZHOU,De
ngqi
ngSI
,Ri
wenLI
N,Ti
anl
eTANG
Te
ach
inga
ndRe
sea
rchOf
fi
ceo
fEn
vir
onme
nta
lBi
ol
ogya
ndEc
olo
gy,Ha
ina
nMe
dic
alUn
ive
rsi
ty,Ha
iko
u57
119
9,Ch
ina

Abstrac
t Toe f
fe
c t
ivepreve
ntandc on
trolthedis
easeofPenaeusvanname iBooneinHa i
nanPr ov
ince
,t hemorpho
log
y,c ul
turalcha
ract
eri
sti
cs,p hys
iol
ogic
aland
bio
chemicalr
eact
ionso fthed o
minantstrai
ni s
olat
edf r
omt heh e
p at
opancreastis
sueo fdis
ea s
edP.v annameiwereidentif
ie
db yc onve
nti
o nalmetho
d s
,a nd
16SrDNAs e
quencewa samplif
ie
db ypolymeras
echainreact
io
n(PCR)f o
rs eque
n c
inganal
ysis
.Fu rt
hermore,ar
ti
fi
cia
lexperiment
alinf
ect
ionwa sconduc
tedtod e

e c
tthep a
thog
enicit
yo ft
heisol
ate
.Fi nal
ly,thelyt
icabil
it
yofp ha
g e28TM aga
instt
hei s
ola
tewa sdet
ect
edb yin
vit
rolys
istest.Theresu
ltsshowedthattheisol
at

(HYRJ 2013)wasi denti
fi
edasVibri
osinaloe
nsi
s.Th er
ewe r
es i
milarsymp t
omswi t
hthen at
uralinc
ide
nc eofP.vannameiwh entheis
ola
teHYRJ 201
3wa su s
edto

n f
ectP.v ann
ame ibymu scul
arinj
ect
ion.Th emedianlet
haldose(LD50)wa sme a
suredas6.68×1 07 cf
u/ml.Phage28TM coul
dlyseth
ei s
olateandbeu sedfor
bio
logi
calcont
rol
.Th er es
ult
sindic
atethatV.s i
nalo
ensi
sisano pport
unisti
cp at
hogentoP.v annamei.
Keywo rds Pe naeusvannameiBoone;Vibri
os i
naloe
nsi
s;Pa t
hogen;16Sr DNA;L D50;Phage

Pena eusvann ame iBo o


n e
,a l
sok nowna sL i
topen
aeu sv an t
hel
yt
icabil
it
yo fp
hagei
nv
it
roagai
nsti
twer
einve
sti
gat
ed,soas
namei,h a sthea dvantageso flar
g es i
ze,fastg rowth,l own utr
i t
opr
ovi
decert
a i
nguid
anc
etoe f
fe
cti
vecont
rolo
fdise
ase
sc a
use


ionalre quire
me nta nds tro
ngd iseaseresist
anc e.I th a
ss tronger b
yth
ep at
hogen.
adaptabil
itytoc han geso fwa t
ere nv i
ronmentf actorsandl o wer
requi
reme nttop ro
teinc ontentinfeed,a nditsd ressi
ngp ercentage Ma
ter
ial
sandMe
tho
ds
couldrea ch65%.P.v anname icoulds ur
vivef oral ongtimeo utEx perime nta lanima l

ofwa t
er,a ndi sah ighquali
tyvarietyforint
ens i
v eh i
ghyi
eldc ul V.p enaeic
idawi t
hab odylen gt
ho f1 0-1 5cma ndawe i
ght

u r
e.Wi tht heexpansiono fthec ult
ures ca
leo fP.v anname ia n
d of(9. 5±2. 5)gwe reselect
ed.Di seaseds amp l
eswerec ol
lec
ted

h edeve l
o pmentofh ighdensi
tycu l
turemo de,d ama gebyd iseasesfr
omaV.p e
n aeic
idaf ar
mi nDa n zhouCi t
y,Ha inanProvince
,a nd
hasb ec
o meama j
o rp robl
em,s omep ondss howe dat otallosso fhealt
h ys amp leswe rep ur
chasedf romama rketinHa ikou.
P.v anna mei,a nda sar es
ult,theb reedingofPe naeuson cef e
llMa i
nr ea gents
of
fac l
iff.Int heb r
ee dingprocessofPe naeus
,b acteri
alandv i
ral TCBSme dium;2 216El i
qu i
dme diuma ndb act
eri
almi c
rob
i

n f
ecti
ou sd is
easesb ecomet hemo stprominenta n dh av
ec au sedaochemic alide nt
ific
ationtubes,p urchasedfromGu a
ngdongHu an
hugee con o
mi closst oPe nae
usf arming,a ndt h ep ath
ogen sa rekaiMi crob ia
lS ci
.& Te ch.Co .,Lt d.;DNA Ma r
kera ndTa q
[1]
somes pec i
esinVi bri
os pp.o rAeromo nasspp. . DNAp ol
y me ras
ea n ddNTPmi xt
u re,p urc
h asedfromS angonBio
Ba cteri
ao fVibri
os p
p.a reco mmo np a
thogen soropport
u nist
icte
c h(S hangh ai
)Co . ,Ltd. ;p hage2 8TM (p hagemi c
roecol
ogi
cal
pathoge
n s,wh ichc ouldc aus
ed ecli
neo fPenaeu simmu nit
y ,e venpreparati
o n),p urchasedfromNa njingJ u
b aoBi oengi
neeri
ngCo .,

h edeathr atecouldr each100%[2-4].Th ec o
n tr
o lofVibr
iod iseaLtd.
sesbeco meama jorprob l
emi nPe naeuscult
ure.Th ereareav a r
ieIs
o lat
io na ndpur ifi
c at
ion

ys pecieso fVibri
o s
pp .,mo ret h
a n8 0s peci
esh aveb ee
nk nown Un ders te
rilec on
diti
on, h epat
o pancreast is
sueo fd y
ing
[5]
al
ready ,a nda mo ngt hem,mo ret ha
n1 0s p eciesc o
uldc auseP.v ann ame iwa ss t
reakin
o cul
atedi ntoTCBSa g
arp l
ates
,wh ich
dis
easest oa quat
ica nima l
s,s ucha sV.a ngui
lla r
um,V.a lginowerec ultureda t37℃ f or24h .Do minantsinglec ol
oni
eswi t
hu ni


yticus
,V. p arahaemo ly
tic
us, V. h ar
veyi, V. p enae
ici
daa n
d f
orms hape swe r
ep icked,p uri
fi
eda ndisolat
e dforsever
altimesun
V.c amp bel
lii
.I n2 012,i n
fecti
ousd iseas
eo cc urre
di nac ertai
n t
ilp urec ulturewa sob t
aine
d ,andc ol
on yshap echaract
eri
sti
cswere
cult
uref armi nDa nzhouCi ty,Ha inanPr ovi
nceo nP.v anname i,re
c or
d ed.S i
nglec ol
on ywasp ic
k eda nds ubjecte
dt ograms t
aini
ng,
whichs howe danorexia,e mptyintesti
nea ndstoma ch,s ever
ea t
roandb ac t
erialmo rphologywa sobservedu nde rami cros
cope.
phyo fh epatopancreasa ndwh itef l
esh.I nth iss t
u d
y,p atho ge
n Phy si
olog icala ndbi ochemi caltestso fisolate
wasi so
lateda ndi den t
ifi
edf ro
mh epatopancr
ea so fP.v anna mei Identifi
ca t
ion wa sp erfor
me da c co
rd i
ngt oi nst
ructi
ono f
showingo bvioussy mp t
oms ,a ndp athogeni
cit
yo fth eisol
atea n
d VITEK3 2a utoma t
ici dent
ifi
cati
o ns y
stem.Wi t
hr ef
erencetobac
te
rialid entif
icati
onme thodsin"I de nt
ifi
cationMa nua
lo fCommo n
[6]
Rece
ived:Decemb e
r2 2,20
16 Accept
ed:Ma r
ch18,2 0
17 Bac t
eri
a lS yst
e ms" , var
iousp hysio
logical and bi
ochemicali
tems
Supp
ortedbytheCo ll
egesandUn i
ver
si
ti
esS c
ient
if
icResea
rchProje
ctsofthe ofthei solatewe retes t
edc ompleme ntari
ly.
Educa
tionDepart
mento fNat
ion(2013118
10046);He a
lt
hDe part
mentProje
ct PCRa mpl ifi
c ati
ono fbacteri
a l16Sr DNAs equence
ofHain
a nPro
vince(QW2 013ZU053);t
heCo l
le
gesandUn i
ver
sit
ie
sS ci
ent
if
ic
Ba ct
e r
ia lg enomi cDNA Th ei sol
atewa sc u l
ture
di nliqui
dme 
Rese
archProj
ectsofth eEduca
tio
nDe par
tmentofHainanMe d
icalUniver
si
ty
(HYCX2 0121
3). diuma t3 7℃ f or24h,a ndg en omicDNAwa se xt
rac
tedb yphe
Co r
respo
ndingauthor.E
ma i
l:woshi
wuli
n728@163.co
m. nolchl
o r
o for
me xtr
a cti
onme thod.
56 Ag
ric
ult
ura
lBi
ot
ech
nol
ogy 2
017

PCRa mplific
a ti
ona nds equenc ingo fBa cterial16Sr DNAs e
 Theisol
a t
ewa sc u
lturedonTCBSs ol
idme diumat3 7℃ f or2 4h,
que nce Un iv
ersalp r
ime rsf orb acter
ial16Sr DNAwe reu sed:2 7F androundb ulgedy ell
owc o l
onieswiths moot
hwe tsurfacesa n

5
AGAGTTTGATCCATGGCTCA 3 and1492R5 GGTTACCTTGT neatedgesc ouldb eob s
erved.Af t
erg r
ams tai
ning,g r
am nega
tive
TACGACTT 3
.Th ea n t
icipateds izeo fthea mp l
ifi
edf r
a g
me n twas bact
eri
umc ouldb eo b s
ervedu ndermi cr
oscope,int hes hapeo f
15 00b p,a ndth eprime rswe res ynthes
izedb yInvitr
ogen.PCRa m comma .
pli
ficat
ionwa sp erf
o r
me dwi tht heg enomicDNAo fthei solateas Physiol
ogicala ndbi o c
hemi calreacti
onso fiso
late
temp l
ate.Th ePCRs tarteda t9 5℃ f or10mi n ,foll
owedb y3 0c y
 The p hysiol
ogicala nd b iochemicalc haract
e r
ist
ics o f
clesof9 4℃ f or40s ,5 5℃ f o r4 0sa nde x
tensiona t72℃ f or HYRJ 2013we reh ighlysimila
rt oV.s inalo
ensi
ss tandards t
rai

1mi n,a ndc omp le
tedb yr e e
x t
en si
ona t72℃f or1 0min .Th ea m LMG2 5238T,a ndd if
ferenc
e swe r
eo nlyfoundin2i denti
fic
atio

pli
ficat
ionp roductwa st ransportedt ot heb ranchc ompa nyo fBGI it
ems,i. e
. ,g rowtha t40℃ a ndme li
bio
sef e
rmentati
on.Th er e
Sequ e
n ci
ngi nGu angzh ouf orp urif
icat
iona nds equencin
g . sul
tsweres howni nTa bl
e1.
Seque ncea nalysiso fba c teri
al1 6Sr DNAs eque nce Phylogenetica nalysi
s1 6Sr DNAs equence
Thes equenc e
d1 6Sr DNAwa ssubjectedtoBLAS Ta na l
ysi
s The1 6Sr DNAo fstr
ainHYRJ 2
0 13wa ssequenced,a nde x
onNCBI ,a nd1 0s equen ceswi tht hehighestsimilari
tywithi twe r
e cl
ud i
ngthep ri
me rb i
n di
ngr egi
ons,t heamp l
if
ied1 6Sr DNAs e
down l
o a
d eda nda l
igne dwi tht hes equence.Th es equenceo fc or
 quenceh a
dal e ngt
ho f14 49b p,ass howni nFig.1.Th eo b
respondingg eneso fd iff
erentv ariet
iesint hes ameg en uswe r
e ta
ineds equencewa ss ubject
edt oh omologya l
ignmen tinGe n
down l
o a
d eda sreferences equen ces,a nds imilari
tywa sa na ly
zed Bank,a ndt her es
ultsshowe dthatthes e
q u
enceh adh ighersimi
withs of
twarep ackageMe gAligni nDNAS tar7. 1.Ap hylogeneti
c la
rit
ytoVi bri
o .Th ese q
uenceo fstr
ainHYRJ 2013s howeds i
milar
tr
e ewa scon st
ructeda ccord i
ngt oNe i
gh b
orJoini
n gme t
h o
di ns oft
 it
yu pto9 8% t oth esequenceo fV.s inal
oens
is.Differentst
rains
wa r
ep ackageMEGA6. 0,a ndc onfi
den c
ed etecti
onwa sp erf
o rmed ofthegenu swe reselect
edtogetherwiththeali
gnme ntresult
s,a n

byb ootstr
app i
nga nalysisa ccordin gtob oot
strappingseto f10 00. ap hyl
ogenetictreewa sc on
struct
edwi t
hMEGA6. 0,a ss howni n
Ar ti
fici
alinf e
ctiont est Fig.2.
TheP.v an name iwa sc ulturedi nano xyge nat
eda qua r
iu ma t Itcouldb es eenf r
omFi g
.2t hat
,HYRJ 2013wa sn atur
a l
ly
28℃ f or1we ek,a n dth ei ndivi
d ualsdeter
mi nedt obef r
e eo fdis
 gat
heredwi thV.s i
naloensi
ss t
andards t
rai
nLMG2 5238T,i ndica
easesa ccor
d i
ngt oa pp earancewe r
ei nfec
tedwi tht heb acterium. ti
ngthatstrainHYRJ 2 013wa sthec l
osesttoV.s inaloe
n s
is,a n

Wi threfere
n cetol i
teratures[7-8]
,t heme dianl ethaldose(L D50) coul
db eid enti
fiedtob eV.s i
naloe
nsis.
wa sd et
e r
min e
d.A c ulturedb acteri
als ol
u t
ionwa sc entri
fuged,
andt hes upernatantwa sd i
scarde d.Th er esiduewa sd il
u t
edwi t

st
ro kephys
iologica
ls alines oluti
o ni nt
o1 ×1 06,1 ×1 07,5 ×
107,1 ×1 08 and1 ×1 09 c f
u/ ml ,a n
de achc oncentr
ationwa s
designedwi th3r eplicati
o ns.Th eP.v annameiwa si nt
ramu scular

lyi nj
ectedwi tht heb a ct
e r
iumf romt hea bdome na ccordin gto
25μl /i
ndividual,a ndt hei nd i
v i
d ual
si nt heCK we rei nject
ed
withe quala mou nto fn orma ls ali
n e.Th ewa tert emperaturewa s
control
leda t(28±3)℃,a ndt hes a
lini
tywa s3 5°.Th eP.v a n

M.DL
2000Ma
rke
r;1.Negat
iv
ec o
ntr
ol;2.
PCRpr
odu
ct.
name iwa sfedn orma l
lye veryd ay ,andt hewa t
erwa sa eratedf or
Fi
g.1 PCR ampli
fi
cat
ionresul
to f16S r
DNA geneo
fst
rai

24h.Th eo bs
e r
vationwa sp erforme df o
r1 0dc ont
inuously,a nd HYRJ
2013
thei nf
ecti
onr esult
sa n dn u mb ero fdeadP.v an nameiwe rer ecor

ded.Th eLD50 wa sc alcu l
a t
edwi thS PSSa ccordingtowe ighed
probitme t
h od.
De terminationo fly ti
ca bili
tyo fpha ge2 8TM aga i
nstisolate
Thei sol
atewa sc u lt
ure dwi th2 216El iquidme di
umt oe arl

logari
thmicp hase,a nda ddedwi thp hage2 8TM (ti
ter≥ 4.7 ×
1010pfu/ml)a tthemu ltipli
cityo fi nf
ecti
on(MOI )o f0.1.Th e
nega t
ivecon t
rolwa sn ota ddedwi thp hage.Th eOD600 valueswe r

determined.

Re
sul
tsa
ndAna
lys
is

sol
ati
on,puri
fic
ati
onandmo r
phologi
cobservat
ionofbacte
 Fi
g.2 Cons
truc
tio
nr es
ultofphy
log
ene
tict
reeba
sedo
nba
cte
ria


ium f
rom di
seas
edP.v annamei 16SrDNAs eque
nces

Onedomina
ntbact
eri
umwa sis
olat
edfro
mt hehepa
top
ancre
 Ar
tif
ici
ali
nfe
cti
ontes


sti
ssueofdi
sea
sedP.vannamei,anddesi
gnat
edasHYRJ 20
13. There
sul
tsofi
nfe
cti
ngh
eal
thyP.v
ann
ame
iwi
thHYRJ
201

YuZHOUe
tal
.Is
ola
ti
ona
ndCh
ara
cte
riz
ati
ono
fVi
bri
osi
nal
oen
sisf
romPe
nae
usv
ann
ame
iBo
onei
nHa
ina
nPr
ovi
nce 57


howedt h a
tP.v annameiinjec
tedwi t
h1 ×1 06,1 ×1 07,5 ×
7 8 9

0,1 ×1 0,a nd1×1 0c fu/ mlV.s inaloe
nsi
s,respect
ivel
y,

xh i
bit
edt hed eat
hr at
eso f0,3 6.7%,4 4.5%,6 4.4% a nd

00%,e achofwhichwasa ver
ageof3r epl
icati
ons
.Th eCKg r
oup

howedn od e
ath,asshowni nFig.3.Th es y
mp t
omso nP.v an
nameicau s
edb yi nf
ect
ionwi t
hV.s i
naloensi
swe r
es ubst
anti
all


hes ame,i ncl
udingemptyintes
ti
n eande mp t
ysto
ma c
ha ndwh it


ndt urbidmu scl
e.As tr
ainwi t
htotal
lythes amemo r
p ho
logyand
phys
icochemicalprope
rti
esa stheteste
ds trai
nwa sisola
tedfrom

heh epatopancr
easti
ssueofd i
seasedP.v annameiagain,indic
a

ingthats t
rai
nHYRJ 20
13wa sthep atho
geno fP.v annamei,and Fi
g.3 Re
sul
tso
far
tif
ici
ali
nfe
cti
ont
est

heL D50 wasdeter
minedtob e6.68 ×1 07 cf
u/ml.

Ta
ble1 De
ter
mina
tio
nre
sul
tso
fba
cte
ria
lphy
sio
log
ica
lbi
oche
mic
alr
eac
tio
ns

tem HYRJ
201
3 LMG2
523
8T I
tem HYRJ
201
3 LMG2
523
8T
Gr
ams
tai
nin
g - - Ci
tr
ateu
til
iz
ati
on - -
Gr
owt
honTCBS + + V-P - -
Gr
owthon40℃ - + Ur
eas
e - -
0% NaCl - - Gl
uco
se + +
1% Na
Cl + + Ma
nni
to
l - -
6% Na
Cl + + I
nos
it
ol - -
8% Na
Cl + + S
orb
ito
l - -

0% Na
Cl - - Rh
amn
ose - -
0/
129(1
0μg
) S S S
ucr
ose - -
0/
129(1
50μg
) S S Me
li
bio
se - +
Ph
eny
lal
ani
ned
eca
rbo
xyl
ase + + Amy
gda
li
n - -
Ly
sin
ede
car
box
yla
se + + Ar
abi
nos
e - -
Ar
gin
ine
dih
ydr
ola
se + + Ga
spr
odu
cti
onwi
thg
luc
ose - -

ndo
ler
eac
ti
on - - Ga
lac
tos
e + +

ydr
oge
nsu
lfi
dep
rod
uct
io
n - - Ma
lt
ose - -
Ox
ida
se + +
+i
ndi
cat
esp
osi
ti
ve,-i
ndi
cat
esn
ega
ti
ve,a
ndSi
ndi
cat
ess
ens
it
iv
e;a
nd v
ari
antb
iol
ogi
cali
den
tif
ic
ati
oni
te
ms.

Ly t
ica bi
lit
yo fphage2 8TM invi
troagai
nstisola
te Di
scus
sio


tc oul
db eseenfr
omFi g.4t hataft
erthea ddi
ti
ono fphage Theme t
hodofreduci
ngt heti
me so fc hangingwa t
erinal arge
28TM i
ntothec u
ltu
re,theOD600 v al
ueofthecultu
resh o
we danin quanti
tyc o
uldma i
ntai
nthes tabi
li
tyo fP.v ann ame ic u l
turee co
creasi
ngtrenda tf
irs
t,an d4 0mi nlat
er,wit
ht helysi
so fbac
ter
i syst
em,b u
tthea q
uacult
uresewaged i
sch argedt os u rr
o undinge n
um,t heOD600 val
ueo fthec u l
tur
eb ega
nt odrops t
eadily
.Af t
er vi
ronme ntcoul
dc aus
ewa terde g
radation[9],wh ichwo ul
df urt
her
TM [10-1
1]

h ea d
diti
ono fphage28 ,t hea bs
orba
n c
ewa sfarlowerthanthe inducelarge
scal
eo ut
breako fmicrob ia
ld is
ease s .Fo rin
CK,i nd i
cat
ingthatphage2 8TM h
asa n
tiba
cter
ialeff
ect. st
ance,v ibr
iosi
sh assprea
dt oa l
lc u l
tu r
eda qu atics,a n dn ew
pathog
eno fVibr
iospp.ared is
covere
dc on t
inuou sly,c a us
in glarge
[1
2-13]
economicloss
esforbreedi
ngi ndus
try .
Int hi
ss t
udy,1d ominants t
rai
nwa si so
late df romd iseased
P.v annamei,andidenti
fi
edt ob eV.s ina l
oensisb ymo r
ph ology,
physi
ologi
cala n
db i
oche
micalr eact
ion sa ndmo lec ularb iology.
Theb io
ch e
micalreac
ti
onc haract
eri
st
i cso fs t
rainHYRJ 2013we re
thesamea sthed e
scri
pti
ono fsta
ndar ds t
ra i
nLMG2 523 8T[14](ex
ceptgrowthat40℃ a n
dme l
ibios
er ea c
tion).Th er esu l
tsa ccord
withtheresul
tsobtai
nedbyB. Gomez 
Gi le tal.[15]thatt hee ffe
cts
ofV.s i
n al
oens
isong r
owtha t40℃ a ndme li
b i
o sed ecomp osit
ion
arevari
ab l
e.Itwasindi
catedthattheseb ioc h
emi c alrea ct
ionc har
act
e r
is
ticsmightu nder
gog eneti
cv a riat
ioni ns t
a bleh e
r edit
ary
proces
s.
Fi
g.4 De
ter
mina
tio
nre
sul
tso
fly
tica
bil
it
yofpha
g 8TM a
e2 gai
nst Ther esul
tsofa r
ti
fi
cialinfect
iont e stsho we dt hatwi tht he

sol
atei
nvi
tr
o concent
rati
ono fbact
eri
alc ult
ureinc reasing,t h ed eathr a t
ei n
58 Ag
ric
ult
ura
lBi
ot
ech
nol
ogy 2
017

creased,a nda ft
era rt
ifi
cialinfection,t heP.v an name is ho we d dis
easeo fPe naeusmon odon[J ].Di sAq uatOr g an,2 0 03,5 7(1/ 2):
similarsymp t
omst ot hosea ft
eri nf
e cti
onu nd ern at
u ralc ondition s. 147-1 50.
[5]JI
NCY.I sol
ationa ndi den t
ifi
cationo fVibrios i
na l
o ensi
so fPe n aeusv an

Fu r
th er
mo r
e,s tr
ainHYRJ 2 013wa si s olat
e df rom P.v a nna me i nameiBo one[J]. Ma rine En vironme ntalS c
ienc e,2 013, 3 2(5):
diedf orthed i
sease.Bo thr es
u lt
ss h o
we dt h a tV.s inal
o e
n siswa s 647-6 51.
ther ealp a
thogenc au s
ingt hed iseaseo fP.v anname i
.Th eLD50 [6]DONGXZ.I d enti
fi
c a
tionma nua lo fc ommo nb acterialsystems [M].Be i

wa sd et
erminedtob e6. 68×1 07 cfu/ml ,wh ichc losetot heLD50 ji
ng:S cienc
ePr es
s,2 001.
7 [7]MAFM,ANTONI OLG,CRDOVAAI C,e ta l.I solati
ona ndc h ar
acter

value(3.3 × 1 0c fu/ml )o fV.s inalo e
n sisisolatedb yMae t iz
ati
ono finfecti
ousVi bri
os ina
loe nsiss t
rainsfromt hePa c i
fi
c shrimpL i
tp

al.[7]agai
nstP.v ann amei,b utf arh ighe rt hant hat(8.1 ×1 03 penaeusvanname i(De cap oda:Pe nae i
d ae
)[J ].Re vBi olTro p,2 012,6 0
[1
6]
cfu/ml )o fV.h ar
v eyiagainstP.v anna me i andt hat(1.4 × (2):5 67-5 7 6.
6 [8] [8]ZHANGBC,LI UF,BI ANHH,e ta l.Is
olati
on,i d entif
icat
ion,a ndp at
h
10c fu/ml)o fV.p a r
a hae
mo l
ytic
u sa g
ai nstL.v anna me i .I twa s
ogeni
c i
tyanalysisofaVi b ri
op ara haemo lyti
cuss t
rainf romL i
to pe
n aeus
v an

indicatedthatV.s inaloe
n si
sh a slo werp atho geni
cityt oP.v a n
namei[J].Ma rineFisherie sRe search,2 012,3 3(2):5 6-6 2.
name ithano the
rVi briospp.Fl oresMir andae tal.[17]deeme dt ha t [9]SAMOCHATM,LAWRENCEAL,COLLI NSCA,e ta l
.Pr odu ct
iono f
V.s inaloens
iscou l
ds erv
ea sani mmu n itys timu l
an tforr ed uc ing thePa ci
fi
cwh it
es hrmp,L
i i
to
p enae usv anname i,i nh i
gh densityg r
e e
n
thed eathrateofP.v anname i;a ndi twa sf ou ndla tertha tu nd e r houseencl
osedr acewaysu singl ows ali
n i
tyg r
ou ndwa t
e r[J].JAp pl
poorwa t
erqualit
ywh enPe naeush asl owi mmu ni
ty,e xcess
iv emu l Aquac ul
t,200 4,1 5(3):1-1 9.
[7] [10]ESIOBUN,AMENTAL, IKEJ .An ti
bioti
cr esist
a ncei ns oila ndwa ter
ti
p l
icati
ono fthisba cter
iumc ouldc aused ea t
ho fPe naeus .Th e enviro
nme nt
s[J ].I ntJEn vironHe a l
thRe s,2 0 02,1 2(2):1 3 3-1 44.
patho ge
n e
sisofthisb acter
iu misn otcle ar,b u tith ass i
mi larp h e [11]AKI NBOWALEOL,PENGH,BARTONMD.An ti
mi crobialre si
st
a n
ce
[18]
notyp i
cc har
acter
ist
ic stoV.v ulnif
icu s .Tos umu p,t hisb a cte inb act
eri
aiso l
atedfroma quacu l
tu res our
cesi nAu str
a l
ia[J].JAp pMi 
ri
ai n f
ectsP.v anna meia sa no pportunisticp a th
o g
e n. cro
b io
l,2 0
06,1 00(5):1 103-1 11 3.
[12]WUHB,PANJ P.Th ec h ar
acteristicso fthee xotoxinVM PM p r
odu c
ed
Th eresul
tsofi nvit
rol yt
ict estsho we dt hatp ha ge28TM ise f
byVi br
iomi micus[J].Ac taHy d robiolo
g i
caS i
nica ,2 004,2 8(4):4 09
fecti
v etoisol
ateHYRJ 2013.Th ereh aveb eenma nys tudi
e su sing -4 12.
pha getol y
seb acte
r i
a ,b utp hageh ast hed i
s advantageo fn arr ow [13]KUS HMAROA,BANI NE,LOYAY,e tal.Vi br i
os hilo
is pn ov.,the
lysi
ss pectr
u m whichr estr
ic t
si tsa ppli
c ation.Ph ag e2 8TM mi cro causat
iveagen tofb l
each ingo fthec o r
alOc ulinap at
a gonica[J ].I n t
lJ
ecologicsarea l
soad dedwi thBd e
llovibr
i oa ndi mmu no s
up presso rs, SystEv o
lMi crobiol
,2 00 1,5 1(4):1 383-1 3 88.
[14]CHI METTOLA,CLEENWERCKI ,MOREI RAAP,e ta l.Vi briovari
a
andt hush avedou bl
ee f
fectsofi nhibit
in gp ath og
e nsa ndi mp rov ing
bil
issp.n ov.a ndVibrioma ri
timu ss p.n ov. ,isolatedf r
o mPa l
ythoac ar

immu ni
tyinPe naeu s.Th ou ght heire f
f ecto fl ysi
n gb a
c t
e r
iumi s iba
e or
um[J ].I ntJS y stEv olMi c robiol
,2 01 1,6 1(1 2): 3009-3 015.
[19]
lowe rthanp hageGH1 5a n dKa gainstS tap hylococcusa ureu s , [15]GOMEZ GILB,FAJ ER AVI LAE,PAS CUALJ ,e ta l.Vi bri
os inaloe
n
theya lsocouldbeu sedfori mprovingwa terq u ali
tya n dcon trolling sss
i p.n ov.,i so
late
df romt hes pottedr oses napp er,L utj
a nusg ut
tatus
growtho fp a
thogen. Ste
in da
chner,1 869[J ].I ntJS ystEv olMi cr,2 008,5 8(7):1 621-
1624.
[16]SUH.I sol
ationa ndid entif
icati
o no fVi b
rioh arveyiinc ulturedL itope
n ae

Re
fer
enc
es usva nnameia ndi t
sp re venti
ona n dt re
a t
me nt[D].Ch eng du:S i
chuan
[1]SHENWJ ,YANGHJ ,YINJ X.Di sease
sa ndcontr
olresear
chs t
atusof Agricul
tura
lUn iv
ersi
ty,2 010.
Penaeu
sv an
n ameiBoone[J].Re ser
voirFishe
rie
s,20 0
4,2 4(1):5 8- [17]FLORES MI RANDAMDC,LUNA GONZALEZA,CAMPA CORDOVA
60. AI,e ta l
. Mi cr
obiali mmu no st
imu l
antsr edu ce mo rtali
ty i
n wh i
tel
eg
[2]SOTORODRI GUEZSA,GOMEZ GILB,LOZANOR,e ta l.Virul
ence shr
imp(L i
topenaeusv ann amei)c hallengedwi thVi briosinalo e
n s
iss t
rai
ns
ofVib
rioharve
yirespo
nsib
leforthe"Br ig
htr
ed"S yndro
me i
nt hePacif
ic [J].Aq uacult
u r
e,2 011, 320(1/ 2):5 1-5 5.
whit
esh r
impL it
ope
naeusvannamei[J].JI nver
teb
rPa tho
l,2 012,1 09 [18]STALEYC,CHAS EE,HARWOODVJ .De tecti
ona ndd if
fe r
entiat
iono f
(3):3 07-3 1
7. Vibr
iov ulni
fi
c usan dV.s inaloensisi nwa t
era n do y s
terso faGu lfo f
[3]PHUOCLH,CORTEELM,NAUWYNCKHJ ,etal.Incre
asedsu s
cept
i Mex i
coe st
uary[J].En v i
ronMi crob i
o l
,2 013,1 5(2):6 23-6 33.
bil
it
yo fwhi
tes p
otsyndr
omevirus
inf
ectedL i
to
penaeusvann
a meitoVibr
io [19]LIUXH,GUJ M,HANWY, eta l
.Co mb i
n ati
onu t
ilit
yo fp hage sGH1 5
campbe
lli
i[J].En v
iro
nMi cr
obiol
,2 008,1 0(10):2 718-2 727. andK a ga
instS taphylococcusa ureu s[J].Ac tami crobiol
o gi
caS inic
a,
[4]SELVINJ ,LPI TONAP.Vi b
rioalginol
yti
cusassoc
iate
dwi t
hwh i
tespot 2013,5 3(5):4 9 8-5 0 6.

Edi

or:Yi
ngzhiGUANG Pr
oof
reader
:Xi
aoyanWU
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