• Nenhum resultado encontrado

RESULTS Wound surface colonization by Penicillium expansum

N/A
N/A
Protected

Academic year: 2019

Share "RESULTS Wound surface colonization by Penicillium expansum"

Copied!
11
0
0

Texto

(1)

6RPHFHOOXODUFRUUHODWHVRIKDUSLQLQGXFHGUHVLVWDQFHWREOXH

PROGRIDSSOHV

Guy de Capdeville

1

, Steven V. Beer

2

, Charles L. Wilson

3

& James R. Aist

2

(PEUDSD5HFXUVRV*HQpWLFRVH%LRWHFQRORJLD%UDVtOLD')%UD]LO'HSDUWPHQWRI3ODQW3DWKRORJ\&RUQHOO 8QLYHUVLW\,WKDFD1<86$8QLWHG6WDWHV'HSDUWPHQWRI$JULFXWXUH$56$SSDODFKLDQ)UXLW5HVHDUFK6WDWLRQ .HDUQH\VYLOOH:986$

&RUUHVSRQGLQJDXWKRU*X\GH&DSGHYLOOHHPDLOJX\#FHQDUJHQHPEUDSDEU

RESEARCH ARTICLE / ARTIGO

ABSTRACT

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

Keywords3HQLFLOOLXPH[SDQVXP0DOXVGRPHVWLFD5HG'HOLFLRXVDOWHUQDWLYHFRQWURO RESUMO

$OWHUDo}HVFHOXODUHVFRUUHODFLRQDGDVFRPDUHVLVWrQFLDLQGX]LGDSHODSURWHtQD+DUSLQHPPDomFRQWUDRPRIRD]XO

$SURWHtQDKDUSLQHVWiHQYROYLGDFRPDSDWRJrQHVHGH(UZLQLDDP\ORYRUDHPPDom8WLOL]RXVHPLFURVFRSLDSDUDHVWXGDUDOWHUDo}HV FHOXODUHVLQGX]LGDVSHODKDUSLQTXHVHMDPFRUUHODFLRQDGDVFRPDUHVLVWrQFLDGHPDomFRQWUD3HQLFLOOLXPH[SDQVXP0LFURVFRSLDGHYDUUHGXUD GDVXSHUItFLHGHIHULPHQWRVLQRFXODGRVPRVWURXTXHRIXQJRLQLFLRXFRORQL]DomRGRVIHULPHQWRVKDSyVDLQRFXODomRHTXHSURIXVD FRORQL]DomRGRVWHFLGRVRFRUUHXGHQWURGHKQRVFRQWUROHV1RVIUXWRVWUDWDGRVFRPKDUSLQJHUPLQDomRGHHVSRURVHFRORQL]DomRGRV IHULPHQWRVIRUDPHYLGHQWHVHPK6RPHQWHDSyVHVWHWHPSRIRLSRVVtYHOREVHUYDULQYDVmRGRVWHFLGRVSHODVKLIDVTXHIRUDPEORTXHDGDV SHODGHSRVLomRGHPDWHULDLVDPRUIRVQRVtWLRGHSHQHWUDomR$PLFURVFRSLDGHOX]PRVWURXTXHQRVFRQWUROHVFUHVFLPHQWRPLFHOLDORFRUUHX GHQWURGDVSDUHGHVFHOXODUHVHPKHTXHLQWHQVDFRORQL]DomRGRVWHFLGRVRFRUUHXHPK1RVIUXWRVWUDWDGRVDJHUPLQDomRGHHVSRURV HDFRORQL]DomRGHIHULPHQWRVRFRUUHXKDSyVDLQRFXODomR1XPHURVRVYDF~RORVGHWDQLQRHGHSRVLo}HVGHPDWHULDOGHSDUHGHIRUDP REVHUYDGRVQDVFpOXODVGDHSLGHUPHHGDKLSRGHUPHGHDPRVWUDVWUDWDGDVPDVSRXFRVIRUDPHQFRQWUDGRVQDVDPRVWUDVGRVIUXWRVFRQWUROH ([DPHVIHLWRVFRPPLFURVFRSLDGHWUDQVPLVVmRPRVWUDUDPTXHRFUHVFLPHQWRGRIXQJRGHQWURGDVSDUHGHVRFRUUHXIUHTHQWHPHQWHQRV FRQWUROHVPDVHPIUXWRVWUDWDGRVDVGHSRVLo}HVQDSDUHGHRFRUUHUDPPXLWRIUHTHQWHPHQWHHHPDOJXQVFDVRVEORTXHDUDPRDYDQoRGR SDWyJHQR2VUHVXOWDGRVVXJHUHPTXHHVWDSURWHtQDSRGHDWLYDURXLQWHQVL¿FDUUHVSRVWDVFHOXODUHVGHUHVLVWrQFLDHPPDomV

Palavras-chave:3HQLFLOOLXPH[SDQVXP0DOXVGRPHVWLFD5HG'HOLFLRXVFRQWUROHDOWHUQDWLYR

INTRODUCTION

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

LQFUHDVLQJSUREOHPVUHODWHGWRIXQJLFLGHWR[LFLW\GHYHORSPHQW RI UHVLVWDQFH E\ SDWKRJHQV DQG SRWHQWLDO KDUPIXO HIIHFWV RI WKHVHFKHPLFDOVRQWKHHQYLURQPHQWDQGKXPDQKHDOWKDVZHOO DVWKHQHFHVVLW\RIUHGXFLQJORVVHVZKLOHUHGXFLQJWKHLUXVH QHZVWUDWHJLHVIRUFRQWUROKDYHEHHQSURSRVHG:LOVRQHWDO $OWHUQDWLYHFRQWUROPHDVXUHVXVLQJQDWXUDOFRPSRXQGV VXFKDVKDUSLQKDYHEHHQHYDOXDWHGDVSURPLVLQJDOWHUQDWLYHVLQ FRQWUROOLQJSRVWKDUYHVWGLVHDVHVE\LQGXFLQJUHVLVWDQFHLQIUXLW &DSGHYLOOHHWDO<DQJDHWDO

+DUSLQLVDQDFLGLFKHDWVWDEOHJO\FLQHULFKN'D SURWHLQHQFRGHGE\WKHKUS1JHQHRIWKHEDFWHULXP(UZLQLD

(2)

HOLFLWWKHK\SHUVHQVLWLYHUHVSRQVH+5LQSODQWV%DNHUet

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

,W KDV EHHQ VKRZQ WKDW KDUSLQ WULJJHUV D YDULHW\ RI FHOOXODUUHVSRQVHVVXFKDVDFWLYDWLRQRIDFWLYHR[\JHQVSHFLHV DQGFHOOPHPEUDQHGHSRODUL]DWLRQZKLFKDUHNQRZQWREH LQYROYHG LQ UHVLVWDQFH UHVSRQVH PHFKDQLVPV RI V\VWHPLF UHVLVWDQFH 'HODQH\ 'RQJHW DO +DUSLQ KDV EHHQ SURGXFHG FRPPHUFLDOO\ XQGHU WZR WUDGHPDUNV ³0HVVHQJHU70´ DQG ([WHQG® DQG LW LV FXUUHQWO\ EHLQJ

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

MATERIAL AND METHODS

7RH[SORUHFHOOXODUFRUUHODWHVRIUHVLVWDQFHLQGXFHG E\ WUHDWPHQW ZLWK KDUSLQ ULSH DSSOHV FY 5HG 'HOLFLRXV ZHUH KDUYHVWHG DW WKH &RUQHOO 8QLYHUVLW\ 2UFKDUG ,WKDFD 1<EURXJKWWRDFRQWUROOHGHQYLURQPHQWFKDPEHUDWR& ZDVKHGWKRURXJKO\ZLWKGLVWLOOHGZDWHUDQGDOORZHGWRGU\ ZKLOHDFFOLPDWL]LQJIRUK7KHIUXLWZHUHWKHQVSUD\HGZLWK ZDWHUFRQWURORUDQDTXHRXVVROXWLRQRI0HVVHQJHU£WKH

FRPPHUFLDOIRUPXODWLRQRIKDUSLQDWDKDUSLQFRQFHQWUDWLRQ RI PJ/ 1LQHW\VL[ KRXUV DIWHU WUHDWPHQW D ZRXQG PP GHHS DQG PP LQ GLDPHWHU ZDV PDGH LQ HDFK IUXLW (DFKZRXQGZDVWKHQÀRRGHGZLWKDP/GURSRIDVSRUH

VXVSHQVLRQ RI3 H[SDQVXP DW [ VSRUHVP/ FD VSRUHV SHU ZRXQG &RQWURO IUXLW ZHUH WKRVH WUHDWHG ZLWK KDUSLQZRXQGHGEXWQRWLQRFXODWHGZLWKWKHSDWKRJHQDQG XQWUHDWHGDQGQRQLQRFXODWHGIUXLW

Inoculum preparation

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¿OWHUSDSHUDQGSODWHGRQ3'$PHGLXPDPHQGHGZLWK VWUHSWRP\FLQ7KHSODWHVZHUHSODFHGLQDJURZWKFKDPEHU DWR&DQGDIWHUFRORQLHVZHUHIRUPHGGLVNVRIP\FHOLXP ZHUHUHPRYHGIURPWKHHGJHRIWKHFRORQ\DQGWUDQVIHUUHGWR DVVD\WXEHVFRQWDLQLQJ3'$SOXVVWUHSWRP\FLQDOORZHG WRJURZIRUGD\VDWR&DQGWKHQVWRUHGLQWKHUHIULJHUDWRU DWR&7KHVHVWRFNFXOWXUHVZHUHWHVWHGIRUSDWKRJHQLFLW\ HYHU\ IRXU PRQWKV 7KH VWRFNV ZHUH XVHG IRU JHQHUDWLQJ QHZGD\ROGFRORQLHVRQ3'$SOXVVWUHSWRP\FLQ IURPZKLFKVSRUHVZHUHFROOHFWHGLQVWHULOHZDWHUDPHQGHG ZLWK7ZHHQ7KH¿QDOFRQFHQWUDWLRQRIWKHVSRUH VXVSHQVLRQWREHXVHGZDVDGMXVWHGXVLQJDKHPDF\WRPHWHU

Scanning electron microscopy studies

)RU VFDQQLQJ HOHFWURQ PLFURVFRS\ ZKROH ZRXQGV ZHUHGLVVHFWHGRXWXVLQJDVLQJOHHGJHUD]RUEODGH(OHFWURQ 0LFURVFRS\ 6FLHQFHV )W :DVKLQJWRQ 3$ IURP VL[ LQRFXODWHG IUXLW SHU WUHDWPHQW DQG K DIWHU LQRFXODWLRQ 7KH ZRXQGV ZHUH LPPHUVHG LQ JOXWDUDOGHK\GHLQ0VRGLXPFDFRG\ODWHEXIIHUDWS+ IRUPLQDWURRPWHPSHUDWXUHGXULQJZKLFKHDFKZRXQG ZDVFXWLQWRWZRKDOYHV7KHVDPSOHVZHUHWKHQWUDQVIHUUHG WRDIUHVKVROXWLRQRIWKHVDPH¿[DWLYHIRUPLQPRUHDW URRPWHPSHUDWXUHDQGWKHQIRUDQDGGLWLRQDOPLQDWR& 6DPSOHVZHUHULQVHGZLWKWKHVDPHEXIIHUDQGSRVW¿[HGIRU KDWURRPWHPSHUDWXUHLQYYRVPLXPWHWUR[LGHLQ 0VRGLXPFDFRG\ODWHEXIIHUS+7KHVDPSOHVZHUH WKHQGHK\GUDWHGLQDJUDGHGHWKDQROVHULHVDQGGULHGXVLQJ D%$/7(&&3'&ULWLFDO3RLQW'U\HU%$/7(&$* %DO]HUV=ULFK'ULHGVDPSOHVZHUHPRXQWHGRQDOXPLQXP VWXEVDQGFRDWHGZLWKJROGSDOODGLXPXVLQJD%$/7(& VSXWWHUFRDWHU%$/7(&$*%DO]HUV=ULFK7KHVDPSOHV ZHUHH[DPLQHGXVLQJD=HLVV'60VFDQQLQJHOHFWURQ PLFURVFRSH /(2 (OHFWURQ 0LFURVFRS\ /WG &DPEULGJH 8.DWN9

Transmission electron microscopy studies

(3)

VXUIDFHDQGDOVRIURPDERXWPPRIXQZRXQGHGWLVVXHDW WKHHGJHVRIWKHZRXQGLQFOXGLQJHSLGHUPDOFHOOVZKHUH LQ VRPH FDVHV WKH IXQJXV WULHG WR SHQHWUDWH 7KH VDPSOHV ZHUH FROOHFWHG RU K DIWHU LQRFXODWLRQ DQG LPPHGLDWHO\ LPPHUVHG LQ JOXWDUDOGHK\GH LQ 0VRGLXPFDFRG\ODWHEXIIHUDWS+IRUPLQDWURRP WHPSHUDWXUH 7KH VDPSOHV ZHUH WKHQ WUDQVIHUUHG WR IUHVK SUHSDUDWLRQVRIWKHVDPH¿[DWLYHDQGKHOGIRUPLQPRUH DWURRPWHPSHUDWXUHDQGWKHQIRUDQDGGLWLRQDOPLQDW R& 6DPSOHV ZHUH ULQVHG ZLWK WKH VDPH EXIIHU DQG SRVW ¿[HGIRUKDWURRPWHPSHUDWXUHLQYYRVPLXPWHWUR[LGH LQ0VRGLXPFDFRG\ODWHS+7KHVDPSOHVZHUHZDVKHG LQWKHVDPHEXIIHUGHK\GUDWHGLQDJUDGHGHWKDQROVHULHVDQG WKHQHPEHGGHGLQ6SXUU¶VHSR[\UHVLQ(OHFWURQ0LFURVFRS\ 6FLHQFHV)W:DVKLQJWRQ3$8OWUDWKLQVHFWLRQVZHUHFXWZLWK DGLDPRQGNQLIH0LFUR6WDU+XQWVYLOOH7;XVLQJD5HLFKHUW 20XOWUDPLFURWRPH/HLFD0LFURVFRS\6\VWHPV+HHUEUXJJ 6ZLW]HUODQGFROOHFWHGRQIRUPYDUFRDWHGFRSSHUJULGV[ PHVKDQGVWDLQHGZLWKXUDQ\ODFHWDWHDQGOHDGFLWUDWH7KH VHFWLRQVZHUHH[DPLQHGZLWKD3KLOLSV(0WUDQVPLVVLRQ HOHFWURQPLFURVFRSH)(,+LOOVERUR25DWN9

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¿HOG SKDVH FRQWUDVW RU 1RPDUVNL GLIIHUHQWLDO LQWHUIHUHQFH FRQWUDVW RSWLFV $ERXW WKLUW\ VHFWLRQV SHU WUHDWPHQW ZHUH H[DPLQHG DQG IURP HDFK WUHDWPHQW WKUHH VHFWLRQV ZHUH UDQGRPO\ VHOHFWHG DQG H[DPLQHG IRU WKH SUHVHQFHRISXWDWLYHFRQGHQVHGWDQQLQYDFXROHV7RFKHFN LI WKH LGLREODVWV DFWXDOO\ FRQWDLQHG YDFXRODWHG WDQQLQ ZH VWDLQHG VRPH IUHVK VHFWLRQV ZLWK IHUURXV VXOSKDWH WR ORRN IRUEURZQLVKRUDQJHVWDLQHGSKHQROLFFRPSRXQGV %XVVRWL HWDO7KHGDWDREWDLQHGZHUHDQDO\]HGVWDWLVWLFDOO\ XVLQJDQDO\VLVRIYDULDQFHSURFHGXUHV

RESULTS

Wound surface colonization by Penicillium expansum

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

,QZRXQGVRIKDUSLQWUHDWHGIUXLWVSRUHJHUPLQDWLRQ DQG ZRXQG FRORQL]DWLRQ ZHUH QRW IRXQG XQWLO K DIWHU LQRFXODWLRQ ,Q WKHVH ZRXQGV D ÀRFFXOHQW PDWHULDO FRPPRQO\FRYHUHGVSRUHVRI3H[SDQVXPDQGVWDUFKJUDLQV IRXQGIUHTXHQWO\LQWKHZRXQGV)LJXUH$7KHÀRFFXOHQW PDWHULDORFFXUUHGDVHDUO\DVKDIWHULQRFXODWLRQ)LJXUH % DQG EHFDPH PRUH DSSDUHQW DQG ZLGHO\ VSUHDG RQ WKH ZRXQG VXUIDFH DV WLPH EHWZHHQ WUHDWPHQW DQG LQRFXODWLRQ LQFUHDVHG)LJXUHV&'6SRUHJHUPLQDWLRQDQGP\FHOLDO JURZWK FRXOG EH VHHQ RQO\ LQ DUHDV RI WKH VDPSOHV ZKHUH PLQLPDODPRXQWVRUQRÀRFFXOHQWPDWHULDOFRXOGEHIRXQG )LJXUH ($ SHQHWUDWLRQ DWWHPSW K DIWHU LQRFXODWLRQ PD\ KDYH EHHQ KDOWHG E\ VRPH NLQG RI GHSRVLWLRQ DW WKH SHQHWUDWLRQSRLQWZKHUHGDPDJHWRWKHK\SKDFDQEHVHHQ )LJXUH ) 1R DOWHUDWLRQV ZHUH REVHUYHG RQ FHOOV LQ WKH ZRXQGVRIFRQWUROIUXLW

Tissue colonization by Penicillium expansum

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¿FDQWO\VPDOOHUQXPEHUVWKDQLQWKHKDUSLQ WUHDWHG IUXLW EXW QR GLIIHUHQFHV EHWZHHQ VDPSOLQJ WLPHV ZHUHREVHUYHG7DEOH7KHFRPSRVLWLRQRIWKHVHYDFXROHV FRXOGQRWEHSUHFLVHO\GHWHUPLQHGZLWKRXWDPRUHGHWDLOHG ELRFKHPLFDODQDO\VLVEXWVWDLQLQJIUHVKVHFWLRQVZLWKIHUURXV VXOSKDWHGLGUHVXOWLQWKHVWDLQLQJRIWKHVHYDFXROHVZLWKD EURZQLVKRUDQJH FRORXU FKDUDFWHULVWLF RI VWDLQHG WDQQLQV GDWDQRWVKRZQ6LQFHWKHSUHVHQFHRIWDQQLQLQDSSOHWLVVXH KDVEHHQVKRZQDOUHDG\/HHVHWDOZHLQIHUUHGWKDW WKHVHYDFXROHVZHUHLQGHHGIXOORIWDQQLQ

/DUJH ZDOO DSSRVLWLRQV ZHUH VHHQ IUHTXHQWO\ LQ WKH KDUSLQWUHDWHG VDPSOHV DQG LQ VRPH LQVWDQFHV WKHVH DSSRVLWLRQVPD\KDYHWUDSSHGWKHIXQJXVZKLFKDSSHDUHGDV DEODFNHQHGDUHDLQWKHFHQWHURIWKHDSSRVLWLRQ)LJXUHV' DQG)2YHUDOOKDUSLQWUHDWHGVDPSOHVUHPRYHGIURPWKH DUHDVXUURXQGLQJWKHLQRFXODWHGZRXQGGLVSOD\HGEHWWHUFHOO LQWHJULW\LQWKHHSLGHUPLVDQGK\SRGHUPLV)LJXUH'WKDQ GLGWKHFRQWUROV)LJXUH$

(4)
(5)

FIG. 2 - 6FDQQLQJHOHFWURQPLFURJUDSKVRIZRXQGVXUIDFHVRIKDUSLQWUHDWHGDSSOHIUXLWLQRFXODWHGZLWK3HQLFLOOLXPH[SDQVXPKRXUV DIWHUWUHDWPHQWA.6SRUHVRI3H[SDQVXP6DQGVWDUFKJUDLQV6*FDQEHVHHQFRYHUHGE\VRPHNLQGRIÀRFFXOHQWPDWHULDODUURZKHDGV KRXUVDIWHULQRFXODWLRQ6FDOHEDU PPB.&KDLQVRIFRQLGLDFRYHUHGE\ÀRFFXOHQWPDWHULDODUURZKHDGVKRXUVDIWHULQRFXODWLRQ +&: KRVWFHOOZDOO6FDOHEDU PPC.)ORFFXOHQWPDWHULDO)0FRYHULQJVSRUHVRI3H[SDQVXPDQGVWDUFKJUDLQVKRXUVDIWHU LQRFXODWLRQ6FDOHEDU PPD.$EXQGDQWÀRFFXOHQWPDWHULDO)0RQWKHVXUIDFHRIWKHZRXQGKRXUVDIWHULQRFXODWLRQ6FDOHEDU

(6)
(7)

TABLE 13HUFHQWDJHRISXWDWLYHFRQGHQVHGWDQQLQFHOOVLGLREODVWVLQWKH¿UVWWZROD\HUV RIFHOOVRIKDUSLQWUHDWHGDQGQRQKDUSLQWUHDWHGIUXLWSHHOVDPSOHV D6DPSOHVHFWLRQVZHUHFROOHFWHGIURPDOOVDPSOLQJWLPHVXVHGIRUWKHOLJKWPLFURVFRS\DQG7(0 ZRUN E6DPSOHVZHUHFROOHFWHGIURPQRQKDUSLQWUHDWHG&WUODQGIURPKDUSLQWUHDWHG+DUSIUXLW F2QHPLFURPHWHUWKLFNVHFWLRQVZHUHUDQGRPO\VHOHFWHGIURPWKHWRWDOVDPSOHXVHGLQWKHOLJKW PLFURVFRS\ZRUNDQGWKHQXPEHURILGLREODVWVZDVFDOFXODWHGXVLQJDOLJKWPLFURVFRSHDWWKH PDJQL¿FDWLRQRI; d7KHSHUFHQWDJHVRILGLREODVWVZHUHGHWHUPLQHGE\FDOFXODWLQJWKHQXPEHURIFHOOVFRQWDLQLQJ FRQGHQVHGWDQQLQYDFXROHVLQWKHHSLGHUPLVDQGLQWKH¿UVWOD\HURIFHOOVRIWKHK\SRGHUPLV 6LJQL¿FDQWGLIIHUHQFHVEHWZHHQWKHFRQWURODQGKDUSLQWUHDWHGVDPSOHVZHUHGHWHFWHGP EXWQRVLJQL¿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ÀRFFXOHQWPDWHULDOEHFDXVHLQERWKWKHOLJKWPLFURVFRS\ DQGWKH7(0ZRUNZHGHFLGHGWRIRFXVRQFKDQJHVEHORZ WKHZRXQGVXUIDFH DISCUSSION ,QWKLVVWXG\ZHKDYHWULHGWRUHYHDOVRPHRIWKHSRVVLEOH PHFKDQLVPV LQYROYHG LQ WKH DELOLW\ RI KDUSLQ WR LQGXFH UHVLVWDQFHLQDSSOHIUXLWWREOXHPROGFDXVHGE\3H[SDQVXP E\ ORRNLQJ IRU VWUXFWXUDO FRUUHODWHV RI UHVLVWDQFH 8VLQJ PLFURVFRS\ WHFKQLTXHV ZH VKRZHG WKDW KDUSLQ WUHDWPHQW HLWKHUSUHYHQWHGRUGHOD\HGWKHFRORQL]DWLRQRIWKHIUXLWE\ WKH SDWKRJHQ ,Q KDUSLQWUHDWHG IUXLW FRORQL]DWLRQ RI WKH

ZRXQG E\ WKH IXQJXV GLG QRW RFFXU XQWLO K IROORZLQJ LQRFXODWLRQZKHUHDVLQWKHFRQWUROIUXLWFRORQL]DWLRQZDV GHWHFWHGZLWKLQKIROORZLQJLQRFXODWLRQ

'XULQJ H[DPLQDWLRQ RI ZRXQG IURP KDUSLQWUHDWHG IUXLWVDÀRFFXOHQWPDWHULDOZDVIUHTXHQWO\VHHQSDUWLDOO\RU WRWDOO\FRYHULQJWKHVSRUHVRI3H[SDQVXP,QWKHVHZRXQGV FRORQL]DWLRQ ZDV OLPLWHG WR DUHDV ZKHUH WKH ÀRFFXOHQW PDWHULDO ZDV QRW SUHVHQW ,Q ZRXQGV RI FRQWURO IUXLWV WKHÀRFFXOHQWPDWHULDOZDVQRWVHHQ7RWKHH[WHQWRIRXU NQRZOHGJH QR VXFK ÀRFFXOHQW PDWHULDO KDV EHHQ UHSRUWHG LQ DQRWKHU SODQWSDWKRJHQ DV D UHVLVWDQFH UHVSRQVH %DVHG VROHO\RQWKHZRUNZHKDYHGRQHLWZRXOGEHGLI¿FXOWWR H[SODLQ ZKDW HIIHFW WKLV ÀRFFXOHQW PDWHULDO PD\ KDYH KDG RQWKHIXQJXVEXWLWVFRQVWDQWSUHVHQFHLQWKHWUHDWHGIUXLW PD\KDYHLQVRPHZD\LQKLELWHGWKHDELOLW\RIWKHIXQJXV WRJHUPLQDWHDQGFRORQL]HWKHZRXQG2QHFRXOGVSHFXODWH WKDWWKLVÀRFFXOHQWPDWHULDOFRXOGEHVRPHW\SHRIGHIHQVLQ SURGXFHGE\WKHIUXLWFHOOV

(8)

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

PPD.0XOWLSOHZDOOGHSRVLWLRQVDUURZVRFFXUULQJLQWKHLQWHUFHOOXODUVSDFHV,6LQGHSHQGHQWO\ RIWKHSUHVHQFHRIWKHIXQJXV&\ F\WRSODVP+: KRVWFHOOZDOO6FDOHEDU PP

SKHQROLFFRPSRXQGVRIKLJKPROHFXODUZHLJKWZKLFKKDYH DQWLR[LGDQWSURSHUWLHVDQGPD\IXQFWLRQDVUHVHUYHPDWHULDOV IRU FDUERK\GUDWH PHWDEROLVP DFFRUGLQJ WR (VDX )XUWKHUPRUH/HGEHWWHUDQG3RUWHUSRLQWHGRXWWKDW WDQQLQPLJKWDOVRDFFXPXODWHLQSODQWWLVVXHLQUHVSRQVHWR DWWDFNE\SDWKRJHQV,QGXFWLRQRIDFFXPXODWLRQRIWDQQLQLQ UHVSRQVH WR SDWKRJHQ DWWDFN ZRXQGLQJ DQG RWKHU IDFWRUV

(9)

FIG. 5 -7UDQVPLVVLRQ HOHFWURQ PLFURJUDSKV RI QRQ KDUSLQWUHDWHG DSSOH IUXLW VDPSOHV LQRFXODWHG ZLWKP.

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

,Q ZRXQGV RI FRQWURO IUXLW LW ZDV YHU\ FRPPRQ WR

(10)

REFERENCES

$LVW -5 *ROG 5( %D\OHV &- 0RUULVRQ *+ &KDQGUD 6 ,VUDHO +: (YLGHQFH WKDW PROHFXODU FRPSRQHQWV RI SDSLOODH PD\EHLQYROYHGLQPO±RUHVLVWDQFHWREDUOH\SRZGHU\PLOGHZ 3K\VLRORJLFDODQG0ROHFXODU3ODQW3DWKRORJ\

%DNHU&-2UODQGL(:0RFN10+DUSLQDQHOLFLWRURIWKH K\SHUVHQVLWLYHUHVSRQVHLQWREDFFRFDXVHGE\(UZLQLDDP\ORYRUD HOLFLWV DFWLYH R[\JHQ SURGXFWLRQ LQ VXVSHQVLRQ FHOOV 3ODQW 3K\VLRORJ\

%HQKDPRX1(OLFLWRULQGXFHGUHVLVWDQFHLQWRPDWRSODQWV DJDLQVWIXQJDOSDWKRJHQV8OWUDVWUXFWXUHDQGF\WRFKHPLVWU\RIWKH LQGXFHGUHVSRQVH6FDQQLQJ0LFURVFRS\

%XVVRWWL ) *UDYDQR ( *URVVRQL 3 7DQL & 2FFXUUHQFH RI WDQQLQV LQ OHDYHV RI EHHFK WUHHV )DJXV V\OYDWLFD DORQJ DQ HFRORJLFDO JUDGLHQW GHWHFWHG E\ KLVWRFKHPLFDO DQG XOWUDVWUXFXUDO DQDO\VHV1HZ3K\WRORJLVW

&DSGHYLOOH * %HHU 69 :LOVRQ &/$LVW -5 $OWHUQDWLYH GLVHDVHFRQWURODJHQWVLQGXFHGUHVLVWDQFHWREOXHPROGLQKDUYHVWHG µ5HG'HOLFLRXV¶DSSOHIUXLW3K\WRSDWKRORJ\

&DSGHYLOOH*%HHU69:DWNLQV&%:LOVRQ&/$LVW-5 3UHDQGSRVWKDUYHVWKDUSLQWUHDWPHQWVRIDSSOHVLQGXFHUHVLVWDQFH WREOXHPROG3ODQW'LVHDVH

'HODQH\ 73 *HQHWLF GLVVHFWLRQ RI DFTXLUHG UHVLVWDQFH WR GLVHDVH3ODQW3K\VLRORJ\

'RQJ+'HODQH\73%DXHU':%HHU69+DUSLQLQGXFHV GLVHDVH UHVLVWDQFH LQ$UDELGRSVLV WKURXJK WKH V\VWHPLF DFTXLUHG UHVLVWDQFHSDWKZD\PHGLDWHGE\VDOLF\OLFDFLGDQGWKH1,0JHQH 3ODQW-RXUQDO

(O *KDRXWK $ 0DQLSXODWLRQ RI GHIHQVH V\VWHPV ZLWK HOLFLWRUVWRFRQWUROSRVWKDUYHVWGLVHDVHV,Q:LOVRQ&/:LVQLHZVNL 0((GV%LRORJLFDOFRQWURORISRVWKDUYHVWGLVHDVHV7KHRU\DQG SUDFWLFH%RFD5DWRQ)/&5&3UHVV,QFSS

(VDX.3ODQWDQDWRP\QGHG1HZ<RUN1<-RKQ:LOH\

6RQV,QF

)RQVHFD-0.OLQH:/:\HQDQGW&$+RTXH0$MZD+ (YDOXDWLRQRIDQHZKDUSLQSURGXFWRQPLFURELDOTXDOLW\DQGVKHOI OLIH RI PLQLPDOO\ SURFHVVHG OHWWXFH KWWSFDOVDUL]RQDHGXSXEV FURSVD]D]BGSGI$FFHVVLQVHSW

/HGEHWWHU0&3RUWHU.5,QWURGXFWLRQWRWKH¿QHVWUXFWXUH RISODQWFHOOV%HUOLQ6SULQJHU9HUODJ

/HHV*/:DOO.0%HYHULGJH7+6XWWLOO1+/RFDOL]DWLRQ EHWZHHQ FHOO ZDOOV ZLWKRXW DQ\ YLVXDO UHVLVWDQFH E\ WKH

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¿QGRXWZKDWVWUXFWXUDOKRVWUHVSRQVHVRFFXUUHGZKHQKDUSLQ LQGXFHGUHVLVWDQFHZDVEHLQJH[SUHVVHG%HFDXVHRIWKLVZH KDYH H[DPLQHG WKH LQIHFWLRQ SURFHVV RQO\ LQ KDUSLQWUHDWHG YHUVXVZDWHUWUHDWHGDSSOHV7KHLPSRUWDQWSRLQWLQWKHSUHVHQW ZRUNLVWKDWZLWKWUHDWHGDQGLQRFXODWHGDSSOHVZHZHUHDEOHWR ¿QGGLIIHUHQFHVUHODWLYHWRWKHXQWUHDWHGDQGLQRFXODWHGDSSOHV

7KH UHVXOWV RI WKLV VWXG\ VXSSRUW WKH QRWLRQ WKDW EHVLGHV RWKHU HIIHFWV WKDW KDUSLQ PD\ KDYH RQ DSSOH IUXLW LQGXFHG UHVLVWDQFH LV RQH RI WKH PRVW LPSRUWDQW HIIHFWV DQG PD\FXOPLQDWHLQGLVHDVHFRQWURO7KLVLQLWLDOJURXQGEUHDNLQJ VWXG\ LV WKH ¿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

ACKNOWLEDGMENTS

7KLVUHVHDUFKZDVVXSSRUWHGPRVWO\E\WKH&RUQHOO $JULFXOWXUDO ([SHULPHQW 6WDWLRQ )HGHUDO )RUPXOD )XQGV

(11)

RIFRQGHQVHGWDQQLQLQDSSOHIUXLWSHHOSXOSDQGVHHGV&DQDGLDQ -RXUQDORI%RWDQ\

0DUVKDOO 05 6PDUW 0* $LVW -5 ,VUDHO +: &KORUWHWUDF\FOLQH DQG EDUOH\ SDSLOOD IRUPDWLRQ ORFDOL]DWLRQ RI FDOFLXPDQGDOWHUDWLRQRIUHVSRQVHLQGXFHGE\(U\VLSKHJUDPLQLV &DQDGLDQ-RXUQDORI%RWDQ\

0XOOLQ3:DQJ-64LX':HL=0'LVHDVHFRQWURODQG JURZWKHQKDQFHPHQWHIIHFWVRIKDUSLQRQWREDFFR3K\WRSDWKRORJ\ 6XSSO6

3HQJ -/ 'RQJ +6 'RQJ +3 'HODQH\ 73 %RQDVHUD -0 %HHU 69+DUSLQHOLFLWHGK\SHUVHQVLWLYHFHOOGHDWKDQGSDWKRJHQ UHVLVWDQFHUHTXLUHWKH1'5DQG('6JHQHV3K\VLRORJLFDODQG 0ROHFXODU3ODQW3DWKRORJ\±

3UXVN\'%D]DN0%HQ$ULH5'HYHORSPHQWSHUVLVWHQFH VXUYLYDO DQG VWUDWHJLHV IRU FRQWURO RI WKLDEHQGD]ROHUHVLVWDQW VWUDLQV RI3HQLFLOOLXP H[SDQVXP RQ SRPH IUXLW 3K\WRSDWKRORJ\

5LRX['%LJJV$5&HOOZDOOFKDQJHVLQKRVWDQGQRQKRVW V\VWHPVPLFURVFRSLFDVSHFWV,Q3HWULQL22XHOOHWWH*%(GV +RVWZDOODOWHUDWLRQVE\SDUDVLWLFIXQJL6DLQW3DXO01$363UHVV SS

6QRZGRQ $/ &RORU DWODV RI SRVWKDUYHVW GLVHDVHV DQG

5HFHLYHG0DUFK$FFHSWHG0DUFK733 $VVRFLDWH(GLWRUV:ROIJDQJ2VVZDOG(OLRW:.LWDMLPD GLVRUGHUV RI IUXLW DQG YHJHWDEOHV 9RO %RFD 5DWRQ )/ &5&%RFD 5DWRQ )/ &5& 3UHVV,QF

6SLHUV$*%UHZVWHU'7%XV9*+RSFURIW'+6HDVRQDO YDULDWLRQ LQ VXVFHSWLELOLW\ RI [\OHP VDPSOHV RI0DOXV3\UXV 3UXQXVDQG6DOL[VSHFLHVWR&KRQGURVWHUHXPSXUSXUHXPLQ1HZ =HDODQG0\FRORJLFDO5HVHDUFK

7HUU\ /$ -R\FH '& (OLFLWRUV RI LQGXFHG UHVLVWDQFH LQ SRVWKDUYHVWKRUWLFXOWXUDOFURSVDEULHIUHYLHZ3RVWKDUYHVW%LRORJ\ DQG7HFKQRORJ\

:HL =0 4LX ' .URSS 0- 6FKDGLQJ 5/ +DUSLQ DQ +5HOLFLWRUDFWLYDWHVERWKGHIHQVHDQGJURZWKV\VWHPVLQPDQ\ FRPPHUFLDOO\LPSRUWDQWFURSV3K\WRSDWKRORJ\6XSSO6 :LOVRQ&/(O*DRXWK$&KDOXW]('URE\66WHYHQV&/X-<.KDQ 9$UXO-3RWHQWLDORILQGXFHGUHVLVWDQFHWRFRQWUROSRVWKDUYHVW GLVHDVHVRIIUXLWDQGYHJHWDEOHV3ODQW'LVHDVH

<DQJD%6KLSLQJ7-LHF=<RQJKRQJ*+DUSLQLQGXFHV ORFDO DQG V\VWHPLF UHVLVWDQFH DJDLQVW7ULFKRWKHFLXP URVHXP LQ KDUYHVWHG +DPL PHORQV 3RVWKDUYHVW %LRORJ\ DQG 7HFKQRORJ\

Imagem

FIG. 1 -6FDQQLQJHOHFWURQPLFURJUDSKVRIZRXQGVDPSOHVRIFRQWURODSSOHIUXLWWUHDWHGZLWKZDWHUDQGLQRFXODWHGZLWK3H[SDQVXP KRXUVDIWHUWUHDWPHQW A
FIG. 2 - 6FDQQLQJHOHFWURQPLFURJUDSKVRIZRXQGVXUIDFHVRIKDUSLQWUHDWHGDSSOHIUXLWLQRFXODWHGZLWK3HQLFLOOLXPH[SDQVXPKRXUV DIWHUWUHDWPHQWA.6SRUHVRI3H[SDQVXP6DQGVWDUFKJUDLQV6*FDQEHVHHQFRYHUHGE\VRPHNLQGRIÀRFFXOHQWPDWHULDODUURZKHDGV KRXUVDIWHULQRFXODWLRQ6FDOHEDU PP B
FIG. 3 -  /LJKWPLFURJUDSKVRIFRQWURO$%&amp;DQGKDUSLQWUHDWHG'()DSSOHIUXLWVDPSOHVIURPWLVVXHFROOHFWHGDWWKHZRXQGVXUIDFH DVZHOODVDWWKHHGJHRIWKHZRXQGLQRFXODWHGZLWK3HQLFLOOLXPH[SDQVXPKRXUVDIWHUWUHDWPHQWA.,QWUDPXUDOJURZWKRI3H[SDQVXP FRXOGEHVHHQKRXUVDIWHULQRFXODWLRQ
FIG. 4 -  7UDQVPLVVLRQ HOHFWURQ PLFURJUDSKV RI KDUSLQWUHDWHG DSSOH IUXLW VDPSOHV LQRFXODWHG ZLWK3H[SDQVXPKRXUVDIWHUWUHDWPHQW A
+2

Referências

Documentos relacionados

Posteriormente, no Capítulo IV, é abordado o que constitui o objeto central do trabalho, infere-se da importância do SGI recorrendo, para o efeito, à análise de

7KHDQLPDOVFROOHFWHGLQ$QJUDGRV5HLVED\5- ZHUH VWRUHG IUR]HQ XQWLO H[WUDFWLRQ $IWHU IUHH]HGU\LQJ JRUJRQLDQV J ZHUH H[WUDFWHG ZLWK 0H2+&amp;+ &amp;O [DQGSXUH&amp;+ &amp;O

PDMRUFRQVWLWXHQWVLGHQWLILHG7DEOH2WKHUFRPSRQHQWV IRXQG ZHUH KH[DQDO SKHQ\OHWK\O DOFRKRO EXW\ULF DFLG DQG ( HLFRVHQH

(2009) Influence of storage temperature and apple variety on patulin production by Penicillium expansum. Subacute toxicity study of patulin in the rat: effects on the kidney

VHHG PLFURELROL]DWLRQ ZLWK UKL]REDFWHULD LQFUHDVHG 3$/ DFWLYLW\ DQG WKDW QR DOWHUDWLRQV ZHUH REVHUYHG LQ HQ]\PH DFWLYLW\ LQ UHODWLRQ WR FRQWURO DIWHU SDWKRJHQ LQRFXODWLRQ +RZHYHU

7KLVUHVHDUFKH[DPLQHVWKHWKHRUHWLFDODQGHPSLULFDOFRQVHTXHQFHVRIWKHJORFDO SKHQRPHQRQ±DK\EULGLVPRIWKHORFDODQGJOREDOWKDWHQFRPSDVVHVWKHFRQWHPSRUDU\ FRQGLWLRQ RI GHWHUULWRULDOL]DWLRQ

DIWHU KDUYHVW DQG FRPSOHWHV LWV ULSHQLQJ ZLWK FKDQJHV LQ TXDOLW\DWWULEXWHV(WK\OHQHZDVQHFHVVDU\IRUWKHVNLQFRORXU GHYHORSPHQW DQG ¿UPQHVV ORVV GXULQJ ULSHQLQJ 7KHVH FKDUDFWHULVWLFV

To sum up, in this study we had two main aims: (1) to develop a Portuguese version of the FMM and test the construct validity of this instrument in a sample of Portuguese young adults