The salmon louse problem and salmon louse research … 1979: Diklorvos (Nuvan, ... 1000 1500 2000...
Transcript of The salmon louse problem and salmon louse research … 1979: Diklorvos (Nuvan, ... 1000 1500 2000...
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Lakselus:Behandling og resistens
Tor E. Horsberg
Norges veterinærhøgskole
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Lakselus ? Friis ca. 1600
“Lakseluus”
Strøm 1770
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Kjemisk avlusning
1974: Formaldehyd
1976: Metrifonat (Neguvon) oralt!
1978: Metrifonat (bad)
1979: Diklorvos (Nuvan, Skottland)
1987: --- " --- (Norge)
1987: Ivermektin (Ireland)
1993: Hydrogenperoxid
1993: Pyrethrum top coating (Norge)
bad (Canada)
1994: Azametifos (Salmosan)
1996: Cypermetrin (Excis)
1996: Diflubenzuron (Lepsidon, Releeze), teflubenzuron (Ektobann)
1998: Deltametrin (AlphaMax)
1999: High-cis cypermetrin (Betamax)
1999: Emamektin benzoat (Slice)
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Pyretrioder
Deltametrin (AlphaMax, Pharmaq)
High-cis cypermetrin (Betamax, Novartis)
Low-cis cypermetrin (Excis, Novartis)
Mål: Nervesystemet (Natrium-kanaler)
Badebehandlinger
Kan brukes hele året (forsiktig ved temp. < 6 °C)
95 - 99 % effekt på mobile stadier
~85 % effekt på larvestadier (chalimus)
Dreper parasitten
Bra sikkerhetsmargin
Sterk binding til partikler
Hydrolyseres i vann
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Avermektiner Ivermektin (Palmer & al. 1987)
Dårlig sikkerhetsmargin
Emamektin benzoat (Slice, Schering-Plough Animal Health)
Mål: Nervesystemet (klorid-kanaler)
Behandling via fôr
Kan brukes gjennom hele året forutsatttilstrekkelig fôropptak
95 - 99 % effekt på alle stadier
Varighet av effekt: Opp til 10 uker
God sikkerhetsmargin
Langsomt opptak, langsom utskillelse
Bindes sterkt til partikler
14 days
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Organofosfater Azametifos (Salmosan, Fish Vet
Group)
Mål: Nervesystemet (AChE)
Badebehandling
Utmerket for voksne Lepeophtheirus hunner
OK for preadulte
Ingen effekt på larvestadier ...? nauplier og chalimusstadier har
AChE-activity > 50 % av det preadulte og adulte lus har Walday&Fonnum 1989
45 % effekt på chalimus Jøssund 2009, pers. comm.
Brukes med forsiktighet ved vanntemperaturer > 10 °C, og bør unngås > 15 °C
Begrenset sikkerhetsmargin
Liten binding til partikler
Hydrolyseres raskt i vann
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Kitinsyntesehemmere Diflubenzuron (Releeze, Ewos)
Teflubenzuron (Ektobann, Skretting)
Mål: Kitinsyntesen
Behandling via fôr
~ 90 % effekt på chalimus- ogpreadulte stadier
Ingen persisterende effekt
Ingen effekt på voksne lus
Minst 12 uker mellombehandlinger
Bør ikke bruker i månedene juni -august
Svært god sikkerhetsmargin
Langsom nedbrytning i vann ogsediment
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Hydrogenperoxid
Badebehandling – fortrinnsvis i brønnbåt
Begrenset sikkerhetsmargin
50 % løsning – ekstremt etsende!
Effektivt (> 90 %) mot preadulte og adulte
Noe effekt mot chalimus
Uklart hvor god drapseffekten er
Bør ikke brukes ved vanntemperaturer > 13 °C
Hydrolyseres raskt i vann
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Kombibehandlinger
Azametifos etterfulgt av kitinhemmer
H2O2 etterfulgt av kitinhemmer
Azametifos og pyretroider samtidig
Full dose av begge (fra Skottland)
Lav dose azametifos etterfulgt av full dose pyretroider (patentsøknad fra Nettforsk – Baard Johannessen)
+
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Statistikk, lakselusmidler
Folkehelseinstituttet
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Behandlingsintensitet
Data fra Folkehelseinstituttet og Statistisk sentralbyrå
0
500
1000
1500
2000
2500
3000
3500
1000 t
on
Salmonids treated for salmon lice Production of salmonids
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SEARCH – EU prosjekt
2001 - 2004 Partnere
1. NVH, Oslo
2. National University of Ireland - Galway
Irish Marine Institute, Galway
3. Rothamsted Research, UK
4. Marine Harvest Norway
5. Marine Harvest Scotland
6. University of Prince Edward Island, Canada
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Resistens:
“... nedarvet egenskap hos
en stamme av en
organisme som gjør denne
i stand til å overleve doser
av et giftstoff som ville
drept majoriteten av
individer i en normal
populasjon av samme
arten.”
(WHO, 1957)
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Resistens:
“... nedarvet egenskap hos
en stamme av en
organisme som gjør denne
i stand til å overleve doser
av et giftstoff som ville
drept majoriteten av
individer i en normal
populasjon av samme
arten.”
(WHO, 1957)
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Diagnose Behandlingseffekt
Lusetelling før og 14
dager etter behandling
> 90 % effekt: vellykket
(?)
Sensitivitetstester
Bioassays
Deteksjon av
resistensfaktorer
Biokjemiske assay
Molekylærbiologiske
assays
Treatment efficacy, deltamethrin
0
0.5
1
1.5
2
2.5
3
J F M A M J J A S O N D J F M A
2006 2007
ad
ult
fem
ale
s
avg. before treatment
avg. after treatment
98 %88 % 93 % 96 %
94 %
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Bioassays
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Bioassays er
velegnet for å
sammenlikne sensitivitet
mellom stammer
robuste og reproduserbare
(lab. assays)
uavhengige av
resistensmekanismen Bioassays er ikke
velegnet til å oppdage
resistens på et tidlig
stadium (EC50)
direkte overførbare til
behandlingsdoser
tilstrekkelig validerte for
bruk under feltforhold
EC50 EC50
sensitive resistant
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Grenseverdier
Sensitiv Nedsatt følsomhet Resistent
Azametifos < 8 8 – 40 > 40
Deltametrin < 0.3 0.3 – 1 > 1
Cypermetrin < 1.5 1.5 - 6 > 6
Emamektin < 100 100-400 >400
EC50 (µg/l; ppb)
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deltamethrin (N=304)
0
0.5
1
1.5
2
2.5
3
3.5
4
2001 2002 2003 2004 2005 2006 2007 2008 2009 2010
EC
50
Deltamethrin 2009
0
1
2
3
4
5
6
7
8
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49
EC
50
(p
pb
)
Sensitivitetstester fra ulike norske laboratorier
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cis-cypermethrin (N=103)
0
0.5
1
1.5
2
2.5
3
3.5
4
2001 2002 2003 2004 2005 2006 2007 2008 2009 2010
EC
50
Cypermethrin 2009
0.00
2.00
4.00
6.00
8.00
10.00
12.00
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49
EC
50 (p
pb
)
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Emamectin benzoate (N=195)
0
50
100
150
200
250
300
350
400
2003 2004 2005 2006 2007 2008 2009 2010
EC
50
Emamectin 2009
0
100
200
300
400
500
600
700
800
900
1 8 15 22 29 36 43 50 57 64 71 78 85 92 99 106 113 120 127
EC
50 (p
pb
)
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Azamethiphos (N=84)
0
5
10
15
20
25
30
35
40
45
2001 2002 2003 2004 2006 2006 2007 2008 2009 2010
EC
50
azamethiphos 2009
0
10
20
30
40
50
60
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43
EC
50 (p
pb
)
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Ingen resistens oppdaget mot
Hydrogenperoksid
Kitinsyntesehemmere
(enda …)
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Deltamethrin
0
0.5
1
1.5
2
2.5
3
3.5
Measured Sensitive
Azamethiphos
0
10
20
30
40
50
60
70
Measured Sensitive
Emamectin
0
50
100
150
200
250
300
350
Measured Sensitive
Multiresistens
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Mål for nye feltassays
Enkle (èn protokoll uansett middel)
Arbeidsbesparende (20 - 30 parasitter pr. assay)
Raske (24 timer)
Robuste (samme svar for samme stamme hver
gang)
Fleksible (uavhengig av utviklingsstadium)
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Tentativ protokoll
em
am
ektin
cyperm
etr
in
deltam
etr
in
azam
etifo
s
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Resistensutvikling
Behandling ReproduksjonBehandling Reproduksjon
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Behandling ReproduksjonBehandling Reproduksjon
Best case scenario
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Behandling ReproduksjonBehandling Reproduksjon
Worst case scenario
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Area with sensitive lice
Area with reduced sensitivity (1 or more)
Area with resistance to one drug
Area with multi-resistance
Not tested
Nov 2008
Marine Harvest
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Area with sensitive lice
Area with reduced sensitivity (1 or more)
Area with resistance to one drug
Area with multi-resistance
Not tested
Nov 2009
Marine Harvest
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Andre diagnostiske tilnærminger
In vitro diagnostikk
Avhengig av kunnskap
om virkningsmekanisme
Mekanismer
Økt detoksifisering
Mutasjoner som gir
nedsatt binding av midlet
til målproteinet
Nedsatt akkumulering
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Lakselusresistens i Europa
Organofosfater
Massive resistensproblemer i Norge, skottland og Irland på 1990-tallet.
Mekanisme:
OrganofosfatresistentAChE
Pyretroider
Sporadiske tilfeller i Norge, Skottland og Irland siden1998
Alvorlige resistenstilfellersiden 2007
Mekanismer:
Økt detoksifiseringsevne
Pyretroidresistent Na+-kanal
0
0.5
1
1.5
2
1 2 3 4
sealice strain
EC
50 (2
4h),
cype
rmet
hrin
EC50 EC50, prebeh. PBO
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Emamektin benzoat 1999-2006
Sporadiske tilfeller med behandlingssvikt i Irland, Skottland og Norge
Sannsynligvisbehandlingsfeil
2006
Resistens verifisert i Irland(og Skottland?)
Emamektin tatt ut av bruk i to år i Irland
2007
Sterke holdepunkter for
resistens i ett distrikt i Norge
2008
Resistens påvist i to distrikt i
Norge
Resistensproblemene ser ut til
å spre seg
2009
Resistens verifisert i en rekke
områder opp til Lofoten
Ukjent mekanisme (foreløpig …)
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Hva nå?
Kort sikt Reintroduksjon av “gamle”
medikamenter organofosfater
kitinsyntesehemmere
hydrogenperoksid
kombinasjonsbehandlinger
Optimalisering av behandlingsmetoder
Fôrtilsetninger som øker slimproduksjonen
Ikke-kjemiske metoder f.eks. leppefisk
Systematisk effektmonitorering
Medium lang sikt
Synkronisering av
produksjonen i større
områder
Nye produksjonsmetoder
Nye behandlinger
Lang sikt
Vaksine
Lakselusresistent laks
Øverlie, unpublished
√
Synkronisering av behandlings-
tidspunkt
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Resistensforebygging i en ideell
verden
Minimal bruk av kjemiske midler Leppefisk
Synkronisert produksjon og brakklegging
Branngater
Oppvelling av vann
Funksjonelt fôr
Rotasjon av behandlinger Ulike virkningsmekanismer
Korrekt avlusning Riktig dose
Tilstrekkelig eksponeringstid
Optimal fordeling av medisinert fôr til fisken
Oppfølging
Regionale strategier Vinteraksjoner
Koordinering sommerstid
MonitoreringPhoto: John Burka
APR Kitinsyntese hemmere
MAI
JUN Emamektin
JUL
AUG
SEP Pyretroider
OKT
NOV Azametifos
DES
JAN H2O2
FEBMAR
APR KitinsyntesehemmereMAI
JUN PyretroiderJUL
AUG
SEP AzametifosOKT
NOV
DES UTSLAKTING
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Unngå å drepe den
siste følsomme
lakselusa...x
Den bør i stedet
fredes!
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