how to attract any woman in 6 days or less

7/30/2019 How to attract any woman in 6 days or less

1/9

Veterinarni Medicina, 55, 2010 (1): 19 Original Paper

1

IS900 RFLP types ofMycobacterium avium subsp.

paratuberculosis in faeces and environmental sampleson four dairy cattle farms

I. Pavlik, A. Horvathova, L. Bartosova, V. Babak, M. Moravkova

Veeny Reec Inue, Bn, Czec Repuc

ABSTRACT: Mycobacterium avium up. paratuberculosis (MAP) e cue gen puecu,

n ecnmcy eu, cnc nd pgee nen dee umnn. MAP ed n e enn-

men pcuy n e ece umnn. e pupe e peen udy w mn MAPccuence

n nm ece nd deen cmpnen e ennmen n u ed neced ce nd nege

e dun deen ecn gmen eng pympm (RFLP) ype. Fec nd ennmen

mpe wee eed y cun nd dened ung Myccn J dependence e nd IS 900 pymee

cn ecn. Rndmy eeced MAP e wee yped ung ndded IS900 RFLP med. A

2 410 mpe wee exmned y cuue. MAP e wee deeced n 111 (6.4%) ec mpe, 25 (3.9%)

ennmen mpe nd w (5.1%) m ee mmm: cmmn e (Microtus arvalis) nd ee we-

ed ew (Crocidura suaveolens). On m A1 nd A2 (nm wee ed eween m A1 nd A2), e

me u IS900 RFLP ype (A-C10, B-C10, B-C1 nd B-C9) ec e wee deeced. ee RFLP ype

(excep RFLP ype B-C10) wee deeced n e e ennmen n m. On m B1 nd B2

(nm wee ed eween m B1 nd B2), w denc IS900 RFLP ype (B-C1 nd E-C1) wee deeced

n ece nd n e e ennmen. e MAP nmn ug we w n cnmed n u

udy. Anm cud e neced w deen RFLP ype MAP e ennmen w gy cnmnedw e cue gen. On me ce m,MAPucen eennmen cud ep expn e eey

ng-em peence MAP n u nm ujec.

Keywords:MAP; cw; cun; epdemgy; nmn; peence; Jne dee

Supped y e Mny Agcuue e Czec Repuc (Gn N. MZE 0002716202 nd N. QH81065),

e Mny Educn, Yu nd Sp e Czec Repuc (Gn N. CZ 1.05/2.1.00/01.0006) nd Bu-e, EC (PB FP6-2004-FOOD-3B-023106).

Puecu (Jne dee) n ecnm-

cy eu, cnc nd pgee nen

dee umnn cued yMycobacterium avi-

um up. paratuberculosis (MAP; Hn nd

P, 2006). MAP nmed pnneuy

m du juene mny y e ec-

ue (Whittington et al., 2004). MAP necednm (m en ce) edMAPn ny n

ece (P e ., 2000) u n m (Ayee e

., 2005) nd emen (Ayee e ., 2004).MAPde

n ppge n e ennmen (Whittington etal., 2003). mn cmpened y

y ue ude e gnm

cndee me ped: n e we up

270 dy, n ece nd c 11 mn nd

n dung-we 5C up 252 dy (Cdn

e ., 1984).

MAPcn e ped n e ennmen y -

u cnmned me (e.g. mnue, ec.)

nd numeu ec (P e ., 2009). In e

ennmen w neced nm (e.g. ce

wd umnn),MAP een deeced n m

ee mmm (Geg e ., 1999; Kpecn

e ., 2008), n dpeu nec deen


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Original Paper Veterinarni Medicina, 55, 2010 (1): 19

2

ge deepmen (Fce e ., 2001) nd

n ewm (Fce e ., 2003). Sme ee

ng nm cn e cndeed pen-

ec nd py uce MAPn

pue nd n e wd. ee e wd

(Oryctolagus cuniculus) n Scnd (Geg e .,

1999) nd ey ee ng umnn pece

n Cnnen Eupe: ed dee (Cervus elaphus),

w dee (Dama dama), e dee (Capreolus

capreolus), mun (Ovis musimon), wd

(Sus scrofa) nd wn e (Lepus europaeus)

pued y P e . (2000), Mcc e

. (2003, 2004) nd Kpecn e . (2008).

Fwng deecn e pecc IS900 eemen

(Geen e ., 1989) peen n e MAPgenme

n 15 20 cpe (Bu e ., 2000) e ny

n y e RFLP med w cmned wydn IS900. med w gduy

pped epdemgc ude puecu-

n Eupe, USA nd New Zend (Wppe

e ., 1989; DeLe e ., 1992). e med w

nddzed n 1999 (P e ., 1999)

e eu ned m deen e

cud e cmped. Suequeny, med

een ued n e ude e nmn

pgen eween deen neced .

I een empyed e cmpn

MAPe gnng m nm ece, eennmen nd ewm m deen c-

e (Fce e ., 2003) e negn

MAPpge ug e nen c n-

eee uc nymp en ccce

ewm (Fce e ., 2003,).

Due e c ed umnn (mny

ce) wee neced w ne RFLP ype MAPn

m ee ude, we neged e equency

ccuence deen RFLP ype n ece nd n

e ennmen n deen neced ce m.

Accdngy, e pupe e peen udy w

mnMAPccuence n nm ecend deen cmpnen e ennmen n

u ed neced ce nd nege e

dun deen RFLP ype w e ue

nddzed IS900 RFLP med.

MATERIAL AND METHODS

Studied cattle farms

e udy w pemed n w ce ed nu m e e cue ne ye. Hed A w

ep n m A1 nd A2 n Een S nd

ed B n m B1 nd B2 n Ween S.

On ee u m, u yem mng,

eedng nd c eng wee pped.

Herd A farm A1. On m A1 (140 cw),

nm gzed n djcen pue. Dung m

nun ( w ee mn), ce wee

ep n pen w e en nm n ec pen.

Ae e cnge n de m pn nun

(eween w nd ee mn e e ),

e eedng u cnnued un e ge 18

20 mn wen ey wee d e .

Hee wee med e n m A2.

Herd A farm A2. On m A2, e nm

wee ep n ee pen w 50 nm n ec

(150 cw n ). Dung m nun, yung

u nd ee wee ep n pen w e enn ec pen. Bee e cnge m m pn

nun, u wee med ne e n

m A1. Yung ee wee med nd ed

e me e (ued n A2 m) w ee

m m A1. I necey, e e-pegnn e-

e (epece e m wee ey wee n)

wee med ndm e e n m A1

nd A2.

Herd B farm B1. On m B1 (170 cw), e

nm wee ep n u xe pe 40 50 n-

m. Cw wee ed up ug w dne e ye und. Dung m nun, ec c

w ep n epe pen (ne cnc pe).

Ae e wcng m m pn nun,

e u nd ee wee med n w epe

e n m B2.

Herd B farm B2. On m B2 (400 cw), e

nm wee ep n e w 50 60 nm

n ec e ye. Ec c w ep n ep-

e pen (ne cnc pe) w ee

mn dung m nun. On m,

eed (ened) u eceng pn nun nd

ee wee ep n w epe e. Beepun, cw n dnced ge pegnncy

wee ndmy (epece e m wee ey

wee n) med m B1 B2.

Examined samples

Fece wee ceced ndduy m e ec-

um nm de n 18 mn ung d-

pe ge. Smpng w pemed n

m wce ye. A 1 735 ec mpewee ceced n dpe pe-eed 20-m


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Veterinarni Medicina, 55, 2010 (1): 19 Original Paper

3

pc . Ae cecn, ec mpe wee

ced 6C nd deeed e y wn

24 u.

Ang w e cecn nm ece n e

m, 636 mpe m e m enn-

men wee en ec me: 346 cpng m e

nen ennmen e e, 260 m-

pe m pddc nd 30 mpe dnng w-

e m e dne. y-nne m ee

mmm, .e. 35 cmmn e (Microtus arvalis)

nd u ee we-ed ew (Crocidura

suaveolens) wee pped und u nd w

c n pue nd n e m uundng.

Culture examination

Fec nd ennmen mpe wee ed n

e y 20C up x wee ee

cuue exmnn. Appxmey 1 g mpe

w need 50 m cnnng 30 m

ee ded we nd ged n zn

e 30 mn. Suequeny, e e w

e undued ue 30 mn e

ge pce mg ee; 5 m e upenn

w need 50 m e cnnng 25 m

0.9% HPC (Hexdecy Pydnum Cde:

N-ceypydnum cde mnyde, Mec,USA) un (Wppe e ., 1991; P e .,

2000). Ae 72 u e edmen (200 ) ec

decnmned mpe w cuued n u pe

deen Hed egg y med (HEYM) w

Myccn J (2 mg/ medum) nd ncued

37C. e cuue wee eued e ped

24 mn.

Soil from the pastures/pens and scrapings

from the stables. Appxmey 10 g nd

cpng m e e (cpng m e ,

cd, w, d nd e) wee n-

eed 50 m cnnng 30 m eeded we nd ged n zn e

30 mn. Fue pceng een deced

e.

Water samples . weny-e m ec we

mpe wee e ee 24 u m em-

peue. Ae decnn upenn, 2 m

4% NOH w dded e pee. e mpe w

e undued 15 mn m empeue

nd cenuged 15 mn 3 000 pm. Ae e

decnn e upenn, 2 m 5% xc

cd w dded e pee 15 mn mempeue nd cenuged gn 15 mn

3 000 pm. e pee w ncued n ee pe

HEYM w, nd ne wu, Myccn J.

Tissue samples from small terrestrial mam-

mals.Ae g exmnn, emed gn

wee ed 20C up w mn ee

cuue exmnn. ue mpe wee exm-

ned mccpcy e nng e me c-

cdng Ze-Neeen (Z-N) cd- d

(AFR) deecn. w mpe m ec nm

wee exmned y cuue: pencymu gn

(e, peen nd dney gee) nd e ne-

n c (ge nd m nene gee).

Ec ue mpe (u 1 g) w mgened

n mce (Kened Lecn GmH,

Gemny) nd decnmned n 0.75% HPC

72 u (P e ., 2000); 200 e ed-

men w ncued deced e.

Isolate examination

Species identification.A 138 mycce

e wee dened y e Myccn J depen-

dence e: ucuue e wee ed n ee

HEYM w Myccn J nd ne HEYM wu

Myccn J ccdng P e . (2000) nd

uequeny dened y e IS900 pymee

cn ecn (PCR; B e ., 2006).IS900 RFLP analysis. A 127 ndmy

eecedMAPe (104 ec nd 23 enn-

men e) wee yped y e ndded

IS900 RFLP med (P e ., 1999). Dgen

DNA w pemed y e ecn endnu-

ceePstI nd BstEII.

Statistical evaluation

w-ded Fe e (InS 3.06, GpPd

we, Inc., USA) w pped e ceun eu.

RESULTS

MAPw ed m 138 (5.7%)

2 410 neged mpe gc me

(e 1). e deence eween m nMAP

deecn n ed wee gncn (P< 0.01):

In ed A,MAPdeecn n m A1 w ge

n n m A2; n ed B,MAPdeecn n mB1 w ge n n m B2.


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Original Paper Veterinarni Medicina, 55, 2010 (1): 19

4

By cuue,MAPw deeced n 6.9% cpng

m e, 0.4% pue/pen mpe nd n

e gn 5.1% m ee mmm: ne

(2.9%) cmmn e (Microtus arvalis) nd ne

(25.0%) ee we-ed ew (Crocidura

suaveolens). MAPw n deeced y cuue n

ny e dnng we mpe en m e

m (e 1).

On m A1, gncny me equen (P

how to attract any woman in 6 days or less

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