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PORCINESTRESSSYNDROMEAND ITSEFFECTTOSWINE POPULATION Presenter:JessicaG.Manalaysay AnimalHealthUnit,PhilippineCarabao Center NationalHeadquartersandGenepool PORCINESTRESSSYNDROME(PSS) | Ageneticdisorderinpigs | Halothanegeneorryanodine receptor1gene isresponsibleforporcinestresssyndrome(PSS) thatiselicitedwhentheanimalsare experiencingstressorwhentheyhavebeen exposedtohalothane(Rosenvold and Andersen,2003) ~ 17 ~ RYANODINERECEPTOR1GENE PORCINESTRESSSYNDROME (PSS) | PigsunderPSS conditionmaysuffer thefollowing symptoms: y ǣȀȀ Ǥ ǤȀ ȀʹͲͲͺȀ ͺȀ ̴Ǥ ͳǤ heatstress, laboredbreathing, musclerigidity andinworstcases, death(Bandetal. 2005) ǣȀȀǤǤȀ ȀͲͲ͵ȀͲͷȀͲͷͲͷǤ Figure 2. Rapiddeathof positivepigscanoccurwhen exposedtoseverestress PORCINESTRESSSYNDROME(PSS) NORMALANDMUTANTRYR1 GENE | Generalanestheticssuch asthehalothanegascan alsotriggerthe symptomsofPSSthus, thename“halothane gene” ǣȀȀǤǤ ȀʹͲͳʹȀͲͺȀʹȀǦǦǦȀ Figure3.Anesthetic Induction ǣȀȀǤ ǤǤȀ Ǥ ǫαͳͳͺͳͲͻʹ Figure4.Differencebetweentheaffinitiesof cytosolic Ca2+ andcytosolic Mg2+ innormaland mutantRYR1 ~ 18 ~ PORCINESTRESSSYNDROME(PSS) | PSSiscausedbyadefectintheryanodine receptor1(RYR1)gene ǦThedefectiscausedby Cytosine/Thyminemutationatnucleotide 1843intheRYR1gene. Asaresult,the aminoacidCysteine isreplacedby arginine (Jovanovic etal.2005). PORCINESTRESSSYNDROME(PSS) | Favero (2000)addedthattheincreased anaerobicrespirationinmuscles generatesrapiddecreaseinpHafterdeath Ǧaffectsthemeatquality characteristicssuchascolorandwaterǦ holdingcapacitywhicharenotsuitablefor meatpreparationduetoexcessivewater loss PORCINESTRESSSYNDROME(PSS) | TheHALgeneorRYR1genebringsa conditiondescribingthequalityofporkas pale,softandexudative (PSE)whichisa negativeeffect HTTP://WWW.FAO.ORG/DOCREP/003/T0756E/T0756E05.HTM http://www.globalswine.com/assets/images/qualitystandards.jpg Figure5.PorkQualityStandards.NationalPork ProducersCouncil ~ 19 ~ SCREENINGFORPORCINESTRESS SYNDROME | Thescreeningwillenablebreedersto effectivelydeterminewhichpigswillbe suitableforbreeding y HTTP://WWW.FAO.ORG/DOCREP/003/T0756E/T0756E05.HTM Figure6.Dark,firmanddrypork (right);pale,softandexudative pork (left);thenormalporkisinthemiddle. Thiswillthenallowonlybetterqualityof porkthatwillincreasetheeconomicvalueof themeat SCREENINGFORPORCINESTRESS SYNDROME SCREENINGFORPORCINESTRESS SYNDROME | ApplicationofMSǦPCRinScreeningfor PorcineStressSyndrome y MSPCRisaPCRǦbasedtechniquedevelopedby Rustetal.in1993,wherebybothnormaland mutantallelescanbeamplifiedinthesame reactiontube,usingdifferentlengthalleleǦ specificprimers Figure7fromJinet.al.(2005).Ǧͳ Ǥ ͳǡ ͳͺȀHaeǢ ʹ ͵ǡ ǢͶͷǡǢ ǡ Ǣͺǡ ͳͺǡͶͷͳͺǡͻͷǤ ~ 20 ~ PORCINESTRESSSYNDROME(PSS) 1 PORCINESTRESSSYNDROME(PSS) 2 3 4 5 6 7 8 9 10 11 12 | Thegenecanoccurinthreepossiblegenotypes: normal(NN),carrier(Nn)andpositive(nn) | UsingMSǦPCRtheDNAoftheanimalshadthe sizeof: 114bpǦ Normal 114and134bpǦ Carrier 134bpǦ Mutant 125 bp Figure8.Agarose gelelectrophoresisoftheHalgene amplifiedbyMSǦPCR.Lane1Ǧ Molecularweight markerladder50bp;lane2isNC;lane3isPC;lanes 4Ǧ12areDNAsamples PORCINESTRESSSYNDROME(PSS) 1 L 2 3 4 5 MOTHER FATHER 6 ǣȀȀ Ǥ ȀǤȀǦͻ ǣȀȀ Ǥ ȀǤȀǦͻ R r R r 150bp 100bp 50bp Figure9.Agarose gelelectrophoresisoftheHalgene amplifiedbyMSǦPCR.Lane1Ǧ Molecularweight markerladder50bp;lanes2Ǧ12wereclassifiedas heterozygotes,Nn (134and114bp) ǣȀȀ Ǥ ȀǤȀǦͻ ǣȀȀ Ǥ ȀǤȀǦͻ R R NONCARRIER 1OUTOF4CHANCE (25%) ǣȀȀ Ǥ ȀǤȀǦͻ R r GENETICCARRIERS 2OUTOF4CHANCE(50%) ǣȀȀ Ǥ ȀǤȀǦͻ r r AFFECTED 1OUTOF4CHANCE 25% Figure10.Autosomal RecessiveInheritance ~ 21 ~ SCREENINGFORPORCINESTRESS SYNDROME | Manipulationofthebreedingsystemcan bedoneviamolecularanalysissuchas geneticscreeningusinglivepigswhichisa fastandnoninvasivetypeoftesting (Bastos etal.2000) SCREENINGFORPORCINESTRESS SYNDROME | Correlationwithinand amongthebreed shouldbedonesothat onemaybeableto determinethe relationshipbetween PSSandbreedinthe swinepopulation. Æ Thismayhelpinthe formationofanew breedingprogramthat couldpossibly eliminatethePSSin thebreedingstock. REFERENCES | | | | | | ǡ ǡǡ ǡ ǡ ǡ ǡ ǡ ǤʹͲͲͷǤ ʹ ǤGeneticsandMolecularBiology ʹͺǣͻʹǦͻǤ ǡ ǡ ǡ ǡǡǤȋʹͲͲͲȌǤ ǤGeneticsandMolecularBiology ʹ͵ȋͶȌǡͺͳͷǦͺͳǤ ǤȋʹͲͲͺȌǤǤEǦCollectionCenterCanada ͺǣͳǦǤ ǡǡǡǡǡ ǡǡ ǡǡǡ ǡǡǡǤʹͲͲͷǤ ǤAsianǦAustralasianJournalofAnimal Sciences ͳͻǣʹͺͲǦʹͺͷǤ ǡ ǡǡ ǤʹͲͲͷǤ ȋȌ ͳȋͳȌ ȋSus scrofa ferusȌǤActa Veterinaria (Beograd) ͷͷǣʹͷͳǦʹͷͷǤ ǡǤʹͲͲ͵Ǥ Ǧ ǤMeatScienceͶǣʹͳͻǦʹ͵Ǥ ǨǨ ~ 22 ~