530/1

CHAPTER FIVE

5¢0¢ EFFEQT O3 1'57“-AT CH TAB; 3URVIVAL .2 '

LE} VIBRIJ P YTICU3.­who survival of §,;°” m=In}_;g and

g___br_;g §% during hating, Ln relation tothonatu:cott.hnhnntLnqnlnnt.mawan\|ou:'lundby outby I011 authors (Hanna and Manna. 1963: Stand-an 1949:Schmidt 1957; Iflqtli and Batu”, 1853! Bilttlaum 19533

Bran 93 3,5” 1960: Goopiort 9; 91.. 1970: 1972;Bouchat lad H0tbb!..nqt@¢ 1976) . ‘rhino studios flirt

adnly as.-rice out by Inning vmm parulltflrl lwh isInto: ccnccntratiun. soluble cazbohydzatno (mainlymama); aaltl. tutu andproialzlnl in thomediuuvhlchptcmlct flu nictoorqmiau or hnlp than to developthttnll roalltanol. ilhlti £1952) found out t-hitm&M £Q%.Qm¢resist-ant ta hut. in 1:111: 1 _% than in Ringers auiatian.

205

g__t_ ,§_l_.. (19691 sham that aux Ihuy and rup­tlaaao aay brwth(1*.sBJ ptntcctod P app tronhoot. I-uodcakc an Baum (1966) rupenwd that thcamount. at tat in nilk did not alhct thn survival atgi dunnq hating. couceznl and clan-2(19692 rnperud em: tho thltmfl. ritlstancn at §incqqproducta mcnandbytheaddltianoioammalt (Had). rhoo£hcto£nam:nauohuaucroal.n‘W Pfllfilfitfl In cmaldcrablly lavcrinq the main-men.Gocptcrt g_§ 3;” (1970) rtpattd that tho boat. raaiatmcn0£ wu meruueéauuuuauez activity attho

r\

beating mnotzum in rofiuoul. var.-tum wanna:-0 (Buucmt1971: Hlmchat. ad Uorthinqtm 1976: and flooflbum1972; vanderant and lilflkilntll» 1972; vmdoraaat gg 5;”1914) RI! Q Q.’ 1971) Slaunllq 1974: int: and Ingram1962), tlpfirtnd the thermal smaitlvlty cl gggg gggywhln heatcll in etch: labwntwy prlpared-nan arm um»: toudprflductfl. _~;mgqwhun gram in higher euoaatzatim Q! #061!!! Qhl-flrifllG! than: 7-57$ Ind ahflnfl H.950!’ manual stability(£113 and flcrthzaqta 1976) - Hevovcr g_g_t_§_­iggi is omstdomod to bu acuitlva to thermalehanqu wt-an hated in dthnr labontcry nadiun. atlhd-Q nubltrntnn (Bmchat 1973: Covert ad woadbum 1972!vmorunt and uickolnm 1912) . their raceway altar

Ii)CD<33

I

flflflli aalanltla greatly Lntlndactd by thu lubltratllnutrient! Ln rneavory India tvaniurnnt Q 5;,» 1914) .

It 1.0 I011 mun that bactitinlaluuunazorouazaezodzyrnnzenaacunauutmaximum mu: Q 5.. 1910: trance. newand scan. use: eoapmngg 53,-. 1910: -:-mu cg 5.. 1911)dunutratld that tho vcgoutlvo bacterial calla qua morntlnrml stability u?aol.ut-.0 emelattatia at tho huntingmantra: Lnercuco. 'th.ta1nc:0a.acha0hoan'a£ta _tnurprnufl zobothu%<:;\nouo£to@mcadwaeoz:act1­vii] 1!.) Q1-QG‘filntgm... Itvna aumaumzmqm-aal autumn! thosaint: nu more lnflamdnl at tho wan: act:lv1t:y'1on1aabavo 0.1Smtmmatr¢lMunmMmoapun 9,539. 1910). rm anamqn ac 1mmPath: (19101 and Holt: gs Q». (1911) um:-0 tn 19:00­nmt lath the chow manna.

Gavnrt and ildilfiiltll (1972):

oatahlishod tho rolatlmahip with amperamru and I061omomtratim an tho am--1 ml at 11$in czlptlcuc soy bmth mu fish hamwnatoc, Q4 aboundzhatumtlvdthwtlalrmqu (Stall!) protccudthnGills at 40; 1°: . so an Iutvlval ac pazhoquu anfilnhnty pnoduaa 11.1.11 emu cmcom aineuthuc

organism: are the major cause at gaatronntarttit-.10.In taunt. ycara the saatood:

1%” worn trying to praducl divaraiiiefl flshqryprcfiuatlandcatcsboththoinlmd mtkitlaafithu%t markltb R-nltmruatl and Cahhda usuallyprlpure randy ta: sown toad» IlPOG1al1Y instead: whcih. \ ‘ avnaybocamm thonear tuturo.oi chimtcal emltltuuutl at instead: Ihclh intlumuobut taint-men in pnthoqnna lib ;_;8a_;_%L;§ and1&8 1" <l=1=¢ 11-'I1"¢~ Th"!amdialanreenlvtngaorniqart-mcntnladia. withiaoxauiao ecnacm the pruqco at certainpl|bl1¢ much-manna aicrenrqanisu in flan aaé fisheryptqbctl. Thu scufiy cm thl attack at variant tilhttypcoductl 1-: " which cantata rich Ozzganlfl nuttinbmpnbln at lnarnaninq the hunt rtliatmcc at thanarqanilnl worn cntrlofl out.

5-1: EE‘2H®~5 E HAERIkLS¢5-1-1­All the strata: at Q

flpp. and in this stufiy Inn: isalatod firm: filthsad fllhory products. atock cultural 110:0 nalnuindan nutrint agar slants at roan tcwerazuro and zautincly

rwltarod men in three weeks. Warkinq cult-uraa weretranstarnud daily intn nutrient breath at pH 7.0 andincubated at 37°c.

I) __1_]_q_§_§_ ;_l_g§_._i_§_: L-Aminfllmidl used in

this cxperimant: ware E..@4arck ax Co. dd B.D.H. pcptenoTtyptam and Yeast. extract wart obtaiacfl from Difeo.The sugars used in this study warn para chcmicalareceived free B...1.1-I. A11 than nutricnt-s were dissolved

in 0.1 M phosphate bulb: hawinq a pl-I 7,0. All the mediawere atetiliuod: by autaclavinq at 15 lbs» tor 15 minuura.

b) » Fish and Qholliiahhomogenates were prepared frcam Cat fish ( app)°*"=k'1‘°ll \ = .E¢..!.=.i3- 511"-W 1?igdicua) and crab (Scylh aerratn) as par mathefiologygiven in auction 4.2.2.

5.1.2. Dctnsninagg 25 hat resisgg.All up strains mentioned in

tabla 4i wore inaculatad from alaatl ta ‘rripticaac anybmth and mmmim 12¢: 24 noun at s1°c. -an call: anharvests! by eoatrifiuqinq and waahnd twicn in Ii"-Brillaalinc anci kept at zoom te@nwr0 till tho ineculatianstudicl were carriad ant.

The heating tempdrattirea

chosen for this study. flora 50%, 55°c and 68°C»

alnen the restaurant! md tiafintstla in ;htropo11tIncities usually lung: at those tomontuzn. the pmparnd

acaiondl in Qdvallfllé-\0;3‘§I!-’1I§¢ It 18 igooaiblc £9unsure the boat resistance at organs in pro­patid dlahts all lovcls Gt buctarlal ¢®¢Glltrlt1@l¢but calaalatim at tha ros1=:t% based anlidtqd n§@n: at Itfifltilfi is paaliblq.

Tm :1. at the wan gigcnntrituqcd cutturo was addnd to 2u0m1 of the hat

mam amt klpt Ln I water mun and q 0qu1l11­bratd to dlfitetant uapanmra oz so°c. ss°c. and 60°cfor than Qorinanta. ‘Inc rnaung vuaol used inthan clpnrlnanta was La 500:1]. three necked vclnclalfl

was aqitatod fitly an at to have a unifiotn ttwot-‘~aturctn ttntcataudiahyuainqnaanll nlchunlcalstirs‘: pontttcuad through tho ant:-0 acct at the fluk­Aqialathnitorwaamnnmiandflmmaqhaaceuldport

E3

to limiter tho tfifltamm during the eatpctlwta.thild part was and for the mtroductim oi mat org:-vnuns and for saqalinq tho material tar tho ovalunudon at oatvivlnq organism during tha courlo at thoboating poroid.

Ti (7)­'1'ha initial inocula lava]. 0!

3 sezatypno pm-aunt in tbs hating mantra anclmltmoully cvaluatnfl by afidinq int oi anatrltaqadeo11cto200m1a£taat:aud1aand1nhoun%tnsa£tub w shall fishes and determined the all camtafor V~3l.'10\II sax-otypos at and tiny ware Ln thanrags at 2.1110‘ to a.u1o'p¢ =1.

11-uriag hating period ans a1at su@.'l.ol1nro~ni|Kd:mn andaddad to 9 ml at atnrilosaline an n diluent at ream t%:.-ature. Sawla at0.5 al at serially dilutnd aaqalea. were surfaced att­cakd in dmlicitll m prqared pt%rad and autfiacldriud Ttlptlaanc may yeast extract. agar (Yeast. extract0.2510. 1'1» ntruaknd places ware incubated ¢e :rr°c£0: 24 hmatl. In mutt 1:2.-lulu. streak plat!!! Iltlnail!‘

h tho can at haauqnatll.tho three hobo 14.9.8. tnehniquo wan cuployad to oaumiratotho aurviven. m this pro-uuiit proaafiurnc mo:1 nwlnl at npptaprtau dilu-flea unto iacnbatnd inuuuzanc brath to: 6 so a naun at 37°c and aqua annlpeationwutrauatazrnd totuhaacuza1n.i.ng9m1rnttathimata and aclenito brathn. This mticlnd anflia

I D;~..,¢..

;.-=1;

won at :1°c ta: 24 mun. A l<I=P£=Il Q1 Iwhbreth were streaked nithtr in Brilliant Gan I9!-‘I’: W-'Hoktocn must! mu ad mamas at :1°c :0: 24 mun.1‘yp.1.¢al solutes are caaflrnnfl qualitatively as lg;by unllq pelyvalcnt '0' anthota. the I-1.9.8. values werecalmlatafl an an man oz the vnflm Q! Pfiflfl-1"

fillutluu in U10 cartil­­I; va1ual(t-.11: death rat: at n

organism at qlvun temperature. ulna taznnfi as daathcnnntnnta:<i0c1.nal:oductJ.ant1ae)war0calculat0flutallans tan virtual bauoqnnata at £1011 and null£10110.

The number at aurvlvqra are

plottld as ncnilflfl PIP!» an the lagrlthdc lainaqaidlttllninntnutasuatimlincarscalcaadtlwbnttatralqhtlinttlnlfitmll tbtmqhthtl. Thtflvalull 10thn tin Ln dnuus remixed for this arcs to traversemu logritrnte cycle. It 16 aleo the than in minutes atacmltmt teupcrnturimncnlnrytndoatroy 9030163orqanlan present. Trials warn cmductafi for a period attint sufficient. 1.-.0:-eault in 5 log cycle drop in viable:

all-M Mm swratrpu M §.- ma’. ­

{D[F1--:.:_\n

IQ

_§. _;___;IIb_g9a__. 9;. g_hg_n_gg_§_. §-_. woltnvrudea and Q. _I_I_a_¢L§9_§'__6_¢

can diphaaic "cu:-vc. 1.0. in the initia-.1 phase theplutl shard rapid Geathl iellound by 1 piano in Uhchdaathjproeoofid at ch slaw taut. In Bch»typI¢ curveat thn decimal redutlnn (B) va1uns~unrn obtained tramthn_partlan at tho curves chafing slave: ratns. D»vn1uuuaura calcuiatad by thn iollauinq oqnatiun rleammlndndby Ratianal annex! IIi~h8I¢G16C1OD Rnncamnh Laboratories

(Laboratory Annual tor toad cannon and Procouara, p.190.1968) .

L‘ '"""ra';%='raa'w';"

IhtttL u death rate in numbers,U I hating flan in d.nut0l¢0 1- initial numeral oi organismb I auto: at @1110

whldm Iurvivd t1._ lb

g QM 19%;‘Thu extant at repair of injury

and laiciatim at growth attur heat. treated s$dopmdo an 1-J1: iaolatlm and unumratim oi the midi;

.49 I‘}?_;_,\5

cwloyed . Iandolo and Ordal (1966). shard mat §__§ggg­gig wag no 100$ talorant 1» sali tor isolatingtknn after bunt tmatmmt. M00001 and Ratto (1970);

praposnd a aanlnlnctivc Iatiehmlat mdll as ptolisnnryrcnucttatlm. aonally this praaodurn shale alien tbstceovozy at antabolllaally Lnjutcd mun. which azur.difhtmt physical strains. an unable to multiply inI I010¢tl.v01y defiant! India md :m;|.|.1r0 nutrimt I@p1:­unuuen. (seam and 81:01:00 1959). hmnvu Gena5 g_1_.¢ (1913). abound that thé phnaoalnan af mtaholicrinjury depend an the pretreated growth canditiaaa Ihcihalso has influcncc as the mam-ezy man.

‘I110 anumnratim at rupait ofinjury after heatiaq at diticrant tnmperauroa -at 50°,ss° and so°c tar age; minutes and nan» mum Q:various scrocypoa at § Nflanisa which UIIO 100­10126 £:cn'vn:.teua aafooda, warn used in this stuciy.~;I¢-qan1=.:m at ninth were in late namaczt-101Phi-II ID!‘ 91% in Ttipticasn say broth and hazvuntldby coatrilugatian. washed Ln stqrild saline water andin startle sultan water.

A Imam cancmtrauoa cl all!(18108 pa: ml) wort lnaeulabod in Tripticalo soy broth$111-O6 1+-9 ii (A1133 1959) whclh

/llHlaHPO‘ -7.0w: --3.09:» B&Cl --0.53: Bi—1‘C1 -4.09:;lflz-i0‘7B2O - o.zsqm Laxtroln -2.0;» pi]: litre at auc­illod umtar-and flan zéaaaqanata and ahallflshgenus. For the lone: cwramre at so°c. umtak~m&th£el:flwhuat.1nju:ynhadywas Sflmlnutnlmd 2-»: higher wupemem.-a at 55° ad s0°c. 1s mm­mu daratim nun null. rinatad aamlos an atudiodfor their all dnath . injury and rcpu: in Trlpflcaltnay broth as wall as in their rcspocttvo heeuaqwtosOf flab ii» E.e.E£1-.2 $1,191 M4 *é .* )and =h¢11 £181» <mnm3&@2=nfl&r.1i-5&9by ucnptiaally imiqhinq Itiwtliflflin 225 quottho reapocttvo hint“ and 10 ml 1:00 90 ml atroapcctlvc liqtflfl broth. Tho Inrlally filllltlfi dttneve»: noecuary in phospmtc bath: and 0.5 nil. at aqlnswar: atrnulnd In xyluoa lyaian agar. and Xylmc Iayaiannamzyamzaa agar and incubated at a1°c tor 24 mus.

who central was pupa-all by

taking late mmnnttally gran culturn 0! 1___1;§strains in triptic-an aq yaaat mtract broth: mdhenohcaatnn and plated in xylosc Lysine near and in Xylo:-:0

Lysine Imounyeholats agar at mmlar intlrvall. ‘rhop1ntucam\:oua:ctalwaa£tc:1.%um0I24neanat 11° c.

215

T110 strillnd all reaavorystufi&uwnrcaa:r1ed§n ant wt» liflmmatnnofincubatlm at teem twrataaro with an interval at 15llhlilllh In this study» tiara]. injury and 219:1! bygran:-h ptwtlaq coqcmdu mach 8813186 acids. vitaminsand nuclncnidcn llllll and cltbflhydratal IO!‘ Qvaluatnfidiftormt aatatypol 0! . The preparationat than India lchndulc is sham balms. In this auaflyit is stand to find out tin ability at lndilnldualqrowthpmaaotlnq earmaunfil taaldthuelaaatcal

anabolic Lpjury of. @011: in theta: otqanig mdla.In tht 30611106 hthth 0! :4-9,

a@1nmunt0d with the £01 lasing aupalttiana at nucleon

aides at bun (admins. quaninc and aacilfz. 10/9;/:1Oach.) t vttan:-an ibifiil-fl; chalice. £0111»: ma. iaositel

nicdtlnic acid . pan ulna bonsai: acid. pyrlémcinm

paatatralnio acids. thialln ad vitamin B12 a 10/uq/mlat nah) and aatneactdn (alanine. arqiaino. anputlc _14:16 , egstinm qlntmnia acid, qlycinm hiatldinm ice­lcueiao, lcuclnm nathluninm puny]. almium ptullnuscrim, throminm tryptnphant. tyzoslam valtncc 100,119per ml at @) .

£415.: schumlo to: the zoeavory

at boat. injured 0: stressed culls oi Balgolla.1) 14-9 broth mime.

5318

2) e-1--9 bx.-ma with nuclcolidoa bun.

3) broth Iith vitamins 6! anaem­anu-Ean re uq/-u ouch.

4) #4-9 broth with all ‘lino said! oiammtrattm at 100 sq/an .

5) M-9 broth with ycast Imtt-aw! of9.5%­

6) Tziptlaani say bmth (Misc)7) Mun I-hart Intusim bratt­

§__,_g,_1 aasums.

its tabla 41 tho dntallaoi tha Iurvival o£ nmtmnQilflififi buwcraturna. G: a glam oi tho rtt-‘tilts thnLass: aurvival in tn: liquid bretho at rnpueun soybzothndm if-9bt0ththm1nth8rO8p¢ct1\¢G~ltll la ivifidnt. In this Quay. initially 1.-.n¢ hlct-23101load as mm an we tanqn at 2.1 to 1.6110‘. at 50°:hlltlnq ¢ Triptiam may broth abound rnductlon of chant3 log cyclca whilo H-9 btath abound as rnhctim to 4log qclufltiql. 13 um . titan the tcqaontazo waslncruasod ta ss"'c and a0°c. an zoducetcn as e legcyclonta 1~.bo1'$!b:othanfl71<>qey¢J.aLna-9b:ath­of rcductlans were noud in thefliihzflt homoqmatu at fish md atmtlflahal also, but

53 1

tho diffarcucn wan Q the higher survival (Figs 16 ab) .tarthilatullyuzhlbltndthaplanonmmofi rcduut1au%t£o:§. gwhich “manna tho hiqhat maistuzm to boat at so°cthan at 55° and 60%. 21» zelultl indicatnd mere sur­vival ammq thl acrotypm in the variaua humo­gmauatuqa. 15. 16 a 11). 1: has been and that thoIurvival at vcmius strains warn act P1‘.'OPGl't16­aaltethntiananacpttarthaacwotgpcm §.@gg.5- s-u§»=-1. .§­

‘rho cttncta oi variant amino

add: at a amcmtratton lcval of I00 nq pa:n1 unto shun in tabla 42 and 43. Orcator ptotoctlmwas exhibited by the aflno acid!’ alumina. qlycint. lyalnoprelim: and auxin: at a tn@e:amr0a of 5;-3° and 60°C.80 aimi. flcmt protoctim was notnd at 60°42. ‘rho l\‘l1~phat anno acids ays&0 ad should lassprcudtim. arlacuhnzuaufia. Lttlaliwlyinazuuadthi Imlitlvity ‘£6 hilt. 21:10 pmhctivc afloat 91 infil-uv1¢ua1un1nca:::1dsuna%u~ndand£o\md1ttob01uactthat: um; at 'rr1pu.can lay yum.-. attract broth» At eo°c.the pratbctim was loss in all pan mug: acids aoluttonl.

ED;..¢31'".

‘Sh studying the constraint pup­tldol which an ccnnmly used £0: aha bucterialoqicalmidst!» ttuasobanrvedthatpmutoctiaa wunorcthantor pun amine acids. and in this stutiyitlblc 44). M.55°C. ycalt attract (LS lcvol) qava non fpratoctlon thanthorrlpttcun. poptnnoamtriptnnc. B\*tat60°caJ.1than pnptidaa ahand aasttivtty to hut.

‘rtm anal: protcia of fish andshall fishes gave subataaeial ptwocum to hat at ll 55°and ao°c (table 41). rm eemnteinl ptipntatlm oirandy to IOIVO sea £0060 in Cafeteria and Restaurantswhats than pg-qpu-atims but tho pantbility of cuta­mnatlm at than plthcqlnic: organism and and torshorter duratlcn at C oi nun: 55° or60°C.. abound the chines: at survival of the Mstrain! (nqa. 15. 15. a 11). -mu: 18 a vaxy slmiflcmtabsczvattm that am uncle premium aoed protocticnfirm the lethal clients of thus: organisms­

Rwarfiino @111 wrbwrdrawl

(tabla 44) no slmifiianat protnctian was sum Qlthlr atss° or ao°c. any meson and Rhunolo ggvfl analpmucum Q: as°c. but at 6o°c Ltjlnctnaaqlthlamsittvlty ta heat.

219

‘rho tins to 1:111 in manta

oi variant organisms at a lava). at 10' pea: %n toundoteetablc lea). in variant haoqmatfl at £1011 andshell £11110: one prnsmtad in tabla 45. Tho hlqhcrt~knt.moratutnthe1aaaor'l:hnt1nltok111\:l'|oarqa—

alum 0 under study... tbs 43 so vn1un(unut.u at unpas­uro at 60°C: rtrnlitud ta ottoct 100% danttuettau) anll fiollmuz g. 1_ in fish 11001.0; crab muscleand pram meals he|naquaatul505) 80: £0: othc: atrainaof this ‘IPIIIIII ‘-1:106 trap 35 1:0 N. Thoand points of survival-1:111 at all tho teat homema­"I" “I W“ §.- ""1 "WP Iwvaried QQQ

‘thermal death time (avnxaqo

L valunn) al various & aorotypaa m dlfifioreattirataas at aqueous: abound hiqhtt valull tar hono­qmatnn than the liquid broth! oi Ttipticanc any breath.aim :4-9 breath (tabla as md 47). At so°c can amzaqn1» valuu of dlttea:-ant species at in Trll-Pi’-1-¢a8Osay breath, main H-9 truth mqiabnrd valun kc: 61.6to 86.4 ad aorzuspmdiag tumaqanatu sham! high“values af 65.8 to 99.3. Simlarly the B valuna (tabla:46 471 at highs: taperutsutcs shunt! ¢:o:ros@1nqmun: valuu. than result: are cnwrcaaed as nan

3530

Inlunuatt-he thrcourtourcqanrinunta coaduatnd. ‘rhoind-icatd that _‘_‘-j- ‘ was highly £0810­zmeenmunounzapaeia ma:-a.app. t-ht aim D valucl at 55° and 60°C were an or la:the scan ad In sigziiieant. Myra: tesiltmm ma natd.

Tluao éata lndlcatnd that puttin­abloarmaaoatoadl oflthotyposatadiodinthetornof hmqcaahos at 65°cand balding uvcry partials 0!a0afi9@dsatth18t§erat=nt'0£ou.'at.101st15d.a8t0lrI­duend the nmmm no mlflul. Tb! sans daqroo at dustzuctlm was clan seldo­vii slmlarly whit nmqmam wore ape at so°c ms?5to80ni.natan£or_§- a<nd£orathe:tp0­eta o£ £wmmuu.

"hut thfl 5a;gg11-a. orqnniillware suapcadcd in variaus h%a1::u of £1012 and shall­flahon md in Ttlptleaa say utter hunt trnntnlnttbs classical nutshell: injury runaway an abazvd{tabla Q ad 69). sh afirwarlnq the hmqmatu nth:heat troatnmt, at so°c ma bacterial 1-and ‘mm thatthc shall fish (pram ml club mat ) ticaabtt ladl­Qatnll a higher survival than the flan %ina1:II- Ingeneral 11:1»: ebncnndtnatall at

* 0I O§,.....~.

9/

50°C 1101.604 highs:-' recevary of thnrnally stroasfi

_§g_q1_;_§ than in frrtptieaso soy broth ad :4-9

broth. In -at but rcaiatmt organismat . a highs: par.-cmtaqa recovery than thothat 0! tho 8a1mrz01§_;. organism was taund.

I-icatlnq at 55° and ao°c £0:30 nl§a\atn8 and 1.5 nlnuhnn toapoctivcly revealed na almi­

tlcantrflea-*O!y huthinhmoqcnatnumdlnliquldbrothaat rzzpuma lay broth ma M-9 bx.-omiabu so). matn the hlqhnt tnworatum of hunting. tho tetal bact­Otial laad cam: dam to tho sands at 160 at 55°C fill 01m at eo°c. Individual Sirétypfl vntitd away 1»thi initial emclntratims inoculated in thi baatlnqnnaatrua. At the higher tcwaratun alto _§. gshowed highs: x-os1a@cc uh: ether Icrotypu undid.At hlqhcr tmoratuzu. thn rncavnry at gggg acro­typtliltrilaascr Lnthtkwnnoqmatoa, hutmtlfn cm-­'-’-I-'-ll-'Y¢ 5;. in hiqhar awbczt htt-ht flab tmaagcnatd.

whim cultures gran invariant homaqnatni um mum at m°, 55° and ao’cthe thozanlly injured calla actually rueevercd withintwo hours. Tbopaothatdhmaqcnatu during incubati.-ma for 130 mutton at tom

6") Q)/H01 ft’ ‘,(,°__’

tllqoxaulso, shoved the recovery of injured onus alsoplated Q Xylolo lqllao I>cx@cho1ato scar (tabla 48M 40). At 50°C. the death culls worn less whq cul­puaa to thou of the ugh» tozmoraturoo 01 55° andso°c:. All tho mm-. strand 001.10 Into xcpuno inTriptlcuo soy broth and thou: rwpocttvo homoqmatol.

53!: Iii 8S>;zI.

‘tho roonltl oi tho presentst\|d1QthovaI:.lounb%qIllotIIptOlmt:l.n haltiii protect the organism oi 335$ do variousaototypoo fitom hoot. Fish and shall am I-lmoqunaanand Trlptlaaao say brofim qavb more protncrtlmn t-hm H-9

broth which La more chemically defined audio. lbw“,th0p:otoot1=JnwasnoI.'oLntho8Cfl.lo11¢!$d.i.othlnI.ntho liwld 3061.0-. Tho oarliar tqlottl ivldl IQII)Iuqgolt that the total solid ocntato of honoqaatuan higher thm in tho ehaatoally defined nulls ant that

0

may pa.-cue; thoonlll tmntnoheat ottoduoodwatorwuvuy (1,) m cm» (Colhoorn and Franc: was;Hanson and Holman. 1963: 1-loot: Q5 5;“ 19711 Pierson

__I_t_ Q” 1971: hmdoka and Hmsaa 1966). Earlilr Soott

(1957) lhouod that the unto: activity (I ') qroator

53733

*han 0.99 in all midi; gave loos éuriaqlumunq. Gbflpfitrt Q5 _§_1_.. (1910). cmncludcd that umcmdcalaatflndtmampnflnqmuinn \|nIasrc1.Q­crtat thm the? unto: activity at tho tang: 0.75 Q 1.00.&|0vor, tho mluatlaa at Manta Q5 5},» (19112, INQ­qaatnd that tho commands giving the protuctivc affect:tn thi mat amsitin pmtdn nalcculea in the cullsbocawl stablc to hcat. and hrthlur. thug haw! at-atcflthat ml-ynl nultcnllcl an by brat. Stillarcher. Read 5 5.. (use). shunt! um». _§.g_Qw32- EH22: §- £25» §;-1=.Q£¢=\¢.$.- mWlmtvlvad during pastanriutlen when the -0011 enough­rattan and ans" pit ml at tho mu.

0: the than tuporaturnnstudied. tin protcctim ms less at 60°C chm at thoothnr mwanuna at 55“ ma 50%. It is @180 can thata htqh ememuattm at flan unlela hflflllulnntm pm.-aural:8a1z1r\§11a cull! film hilt...

It was tend that tho cubo­txydtatfli alsm have littlt antral Q thll ralatlvn pim­acung ctfloctn of the crqanilas (600p£ert. Q Q,“ 1919) .an tho &t1ys1z QQ amoayamuu 1:. A u-pencnm or ss°c.largo dififiarnneol in the pmutnctlva nthct at boatan observant . Xyloan. flimmane and qluconu protnct

/~,-' r'

"" 5' ""1="='¢'=- 52- @_21£1§.§_g&;- .-‘;-V £315»§. md§.gg5 toaanucutcut.Salmonella use mum atuhrly tha an an notpreafiflud 110:0. Cllhom and 31-3810!» (1966) aboorvd

similar p:0otnul:.lu\ by glucose for _ i and, _£__;_g,o_;_9_I__acn;_;_g_. At a highs: t-warns:-e ateo°c:. no pmtectlm was aband­

Anmq tho amino acid vpannutnra

ltlfll-Cl; Ifllphut wulnn acids lilithlmlni &d egsttaoshoved incmascd amsitivity to an ll 55° ad se°c.Similarly. ccqfqz am! triptmb alno abound leaner pra­uuztan tenant. Thiamyboduntosulprn-hydryl groupswhich might. IPL118 3-Sun bad: in ptotninl Brim: nonamsitivc to heat. floats gg Q” (1971). also thatcud-n, qrntain. qlutatlltunl and sodium citratt lncrlllild1:110: aasiuvwty ta hunt for 34;. gg Thu; tux-thatuplunua mt era Qefiium cltratn any pull ca” and nu“Loan out oi the calls and chubby menace thn sensitiv­ity to hat... strain ad Shun (1959). tepattaull thatan” lass dncrtascd thnrml death nu and em: ¢h0la­tinqfiacmpcmd) aqua: cthylino diamlao htraneutic acidfil10hb¢llfiICa:+al\dEQa* isms smnanduwunmudath ram.

Individual amino acids, alaalam

aapat-.1: acid. qlutantc acid. qlyeinm flydrcty prfllll-III.

225

lylimm proltao ad 002130 shard butts: protectingolfinct than thu ethos mm acids. ac lawn: teqontuaat so"c:, um: .11 Quinn acid! pmu-Qua mu gigezqmasan. mam 5 Q“ (1911). mpnnod um: prawnhad l1ttl¢ otfoct althmqh pnptldd mfi aatnao acidsplug anbsitllmial tall 1.. ptatlictlcn. Earlicr in 1964»Gerhardt and Judqo gave an oxplan-attoa that the bacterialall all act. an hotmparun nalueular cloves to encludnnncrmoloculcs. Enlnon ptotatlll are meta mloaulosand ulnhlc to puma-no thn an heat. aanuvgfiieuin thl all wall; tht bantnrial coll bocmlil inamaltiwtfi hilt.

The atfeet of than ml! h@­amatnrn in nun»; mam altotypol of ;I81!‘ Ifllllid Ln dlfifinrclt honoqiuatcl of fish md shillflshll. Ihl tl%a1 duath stndiil ihullfi that byiaetuuinq up tuqacramrc 2:-an so°c to ss°. tho dnuthrats uaniacrouodovnthaughuntlantalaauulnss.In fact tho um pmu survival-kill at all an antmqozamta £o:ublarqan1amt.est.odw0:0cml.tdor­ably less thm tar the echo: £006.: studied Chllqllatt-1Q55;-w 1961). It mat. bu rnnfiorad hora that thandata thdicatu that hilt porilhablc toads at thl typosshadlndhozbinuufiaraatlnenaqanauinalatataoflat

5353f;

temozatuzc at 65%.‘: tor at least 10 nl_.nut:-us, tocazdndahfiucttm fi&108 P“fl@ unman­nctablo Laval. who lunar tags:-amt; at 60°C to:20 miautot refined lafioly the aim bummrial nficrs tolovel­

It is observes! from tho awaits

Obtlinnd with but assistant _§¢ 9 that alargo ditficrmm in tbs boat ruistanoo was nut. taunt}.$. 77.98 (Salaam Suntan and Calualck.1948: c__3,=_ 5,1,... 1954: Anqalotti. Q 5-. 1959:mqulnm-.1 25 21..-. 1961) is maopumalxy tolerant to heatTtarcaultao£t.h1sstuc1yar9a9tnfi1av1awo£tlnOitlilt work abmnm» ‘EDDY fiainfi qteattt resist-lnet in

tho mustard and mum Z 1; k1nq.1 "moi: cm mm.'l.aa:ata:flmdLach1chIl;lak1aq nu-¢1a.o=ma1.stnspocttnly. Ia this owhnt it is 1%!-mat ta autothan organism ultra ariqinally laolatad Iran eqq pra­ducts: aharnften 11-. is niaaumalu ta aim: no awnqroatlr zeaintlnflc to hat, while or§Lm ta thislflldy warn;-T. Lsollbtl % flsknzy predicti­

‘thin otblr serotypca 0! éilamanella

amqaailm an tho least. ham: matltancc studied and

yfilldcd mu cw valull in uupmq wit-'.ht-‘hose qanerallyrncamiuod ta: £5-‘ialmpy Q3 Q“ 1948: AnollisQ; 55.. 1954: Gnbaumo Q 3;“ 1954) .

i_$

-1-)r'4F?" xv {

In that azporimnntl tho U!!!‘­nnl death tint vaxid Em: tho mm pure culturc. batmanatculuaromnipulatlnn. ttanybnduotothoaunhotQ: variables which cannot ho amualgid such u munnuxfitrs 02 viable calls talnm fiat the Laacnlltlm andthe gravth amdltl.-ans md thnir ditfiaxcat growth phaaa.Sllllar vattatlma at thcrml dtath than at pun cultureVII-‘il rvwrtnd by OJ-an gt 5;,» (1952). ea J31. fig and byMonti Q5 Q,“ (1911). ma _§_. £1. $0 la: an possibletho “pennants an eunductci with var.-lawn mutt:alimnly flth aiaqlc mlturn. float! Q, 5;» (1911):amidndandahu\ndthatthc1:=vaJ.u0o cmhosatlhfactutily eaqntd llutvlval) in diitarint Q81; at aqlvm mpramm at 50°. ss° and eo°c 100:0 “aquahumus: little verb was carried ant m the survivalQ1 vgziuua agzotypaa at Lsalatd iron Ilatinbsalads. run roqultd tnworwmtos at most at tin1-only to an-vs aatoodsan nuntmtnofi Ln ad-vanes tatattnzlnq in vazicaa notrapolitna city hatch andratmnntn. an-owes. tbs aurvinl 6.11:0;-anon aremore at that tun and towetatnrn c@i.nati.<:nI- the

D60 valid (tabla 41) aqhania the relative chartperiod ottiantronthatulthowhulotrcatanmttequlz-odto1d.11!0% o£th0n:l.ctuhl.a1pqn1afi.m and

soso

11-ho valnus urn indwnndant of thfl initial in-ad thnorganisms (Ymlmd ma 19538 P1510; and EIIOQ1954: same» Anqlllntti _c_§ Q,” 1961). Although canwidnnnol a1::'i*\u.zn mu inmrmnn. (Rood 2; 5.. 1951;E1-bisi and man 1956 1. an a gram flexibility 1»lvailabln inc interpreting hunt. rlrsiat-ancu data than bydeath tin! clean. Ea:-liar reportsoi Biqolml md (1980) ronnminod that heat ruistaneoat arqanism was indnpunsiant at the initial. ­Tho C values to: different waponaiml were not 10.mloss tho initial 1-l:6mI1o1’n',i;- lcvnlswnwc idcntical.studying D vain“. it is pmibln tn éctnrdnn tillhuatmsiatancuotan nrgmismaénahadiroctcmrarisen poalibli nausea data in whnin variflua aanunts atinwla at used tor the smdy. Inerdc: to avaid the on inoculun aim. Hints:it Q10: (1951); md Onbntnc Q QL» (1954) iollullfl noizhorlsin flhich thl thnrnnl rInist% ell: ho determined inqivm snbstrnt-cl mdnt various cmditima at zdetnhinlload. This tact is non iwwtmt partiunlarli; in amany at this type pmasantd tan. Item the practinalacme paint. it is iqoaaiblo to manual the but swish­nmo£ a¢ewm8mpupud£mamdMawand-or all luvnil at nicnrehiai lend. Hmnvan tbs calml­amdusmlamcanbousedtoprdiatnhctnnrmlraaisianoo.

532%)

on the umaa mums: ot axporiamta. it ispaniblo ta gin 1: valuoa ta: may bacurial ceaccnt.r­atluaa which an attalnalblc saaiaoda .­

2110 C65 vnlucl 0! ethos:Q strains tepertld hora arc in aqronnmt with~.-was mparua by em qua: wqtluro (magi; 5;»1951: osnoma gggu 1954 ma 811111: 55 5.5.. 1954)adthox-alucaaroofthooréarctl-5. Butthuovalues warn caasldorahly loan than tho vallil at _§._. ‘rho uuztuzdod tin: at 10 nlauta will 1:111tbc arqanism to an wndetnctablc level uni ttnl boccunadvaatagaous also. ‘rho ttlulti 106-lentil hurl thatpcriahablc floods 111:0 marina satoada at tho qpo employ­ad in this study mm. at ss°c tor loos than 12 ntnutnlsafinly teducod g organism ta anlavol.

It was intenttnq to auto thatg. gggg menu hnat reaiatnaoo Lapzavnandcrabkxanqonatoaandtrziswaaisulatcfltluuflslfial and pram shall waatla. This would ht ill!tn apuculato bneannl this ccliataacn ambled 1:210 £­ntraln to autvivn chm proccllinq trontnnnt.Thtl aspect of ca:-rolatica bosom resistance to nutm­nnatal emdltiml and iruqueacy oi iaolatlm has bull:augqaatd Oi‘-nkltl. and AIEOI6 1911) . Outtnril. and

(“'1 4‘)/J <-J?

G1-more raporced aiaulu: behaviour to: §_. gggggnggheld in clthtr Ln Wlpuaa nay broth or Oqq whit:emtainiaq amiifi chloride print tn huatlnq. Laga.»

gg Q“ (1912) nporua em: cmaeulitant q:#th by anMcteflwa Q nllk was Lnfiltlwnd by thn heat roalatanco‘Ii =“>‘.££_¢_.Q.2­

‘rho rwovczy at heat treatndiii}; crqanilfl dapqaded upa: the lsolatim 0! thnamrnnratlm 0! thi mdla @1.0g0é. M08201]. mt! Ratta(1970) ¢ auqyattd it! nan to: chi rcsuseltttlm in am­alecttvo radium or satin!-Illnt broth. This procli­urn ahuuld allaw the rocavory 01 antabollically injuredctlls (sunk: ad stokes. 1959) i which nae: avapicq at physical stratum urn manly to mltlplyon a single doflnofl media or ssoloctlvn media and ruqluro

rich auttlctt to bl nwplttlll £0! tho Input: 0! tht1318806 0011!. Thi Ittabaillc injury Qlill upmthe protroatnnnt growth au:<i$.1:.tcaa(Gann Q_§_ g1_¢,;33­

Thi hint treatcd _Bcalla nun studied for thclr coll rapalr by lnaubatlaq<Il thl rnapecttvo hanoqflatgs ‘md Ttiiiticast any brothand 2+-9 broth. It was ebiorvod that metabolicallyinjures 00110 abound qrallth. it canparinq the liquidbroth at rzlptleasc nay and M-9 broth, the Iona: yloldad

23.5.

highs: of injured calla. This result Ls inaqranmt nth the findings at Gonna 5 g_1_.. (1973) .TM: suggest. that the lava: recovery my be mo totho unavailability at this npoclal. nutrient! nah ascllno acids. malcostdas ad ¢=arb¢!'mirat.ns .

punhd in various hmnqmnuns of fish and ahnllfilshna,tlaeinitlal qroltlahwaa upto 15n1:mtll imflinfiflminutes the repair was rapid and beech lgcffoctiwdeta the salautlvo mmbicau such as doaaycholatt.rm rate oi injury oi 0011.: and death 1:111 dopnad upon

tbs temorazural to liwvhlch it ma, Quad. At. 50°Ctr»: rah: of death at ggggg son.-atypoa audlcd aheadabout 2 log cycles rducttm. At ss°c s0°c. um rmoductims warn very 2:191: am no s lag qaefiapeuwzy.Thy injured surviving @115 started qrmfie up attcrone heat. A o1m:l.1a: abserwratiun was flaund in Trlpttcaao

any m-an . ‘tho raw» tun;-cratnrl oi 60°c shouldcmaldcrablo ptromugin at daath md the majority ofculls could nut memo: 1:: a naaiuhibitory aufiia(Holman 1943: Allmud md Rusanll. 1966: Harriet and

Ruflllll. 1966 ma msllll and Hlttiim 1968} .

‘rho ofiuzt. at thazaal. injury

ropnlrbythagravtzhpromotmqcmpomadasuehasanlno

$0

7i»$

3

ands. vzu-mu. auclmtiéos ma azmnynauqinauaowuvnda alum would not gnu thep:Qc:hooat£o:thnan11:epa1r.A11thnl0Lncan­bhattun in M-9 brath maahad slant the sumo artistml in rripueanc lay broth rapntr.

Thlll £:l.n&qs auhtmtiatlthc afiezcnau oi Allwaufi and imast-11 (ass). I-lattice

and aaaau £1966). and Gama: _g__+,g._.. (1913). who

hadxoperudtintthnaddttim nfiyoastaxtraczv20 amino acids; caamhno acids. vitamins and nualoanidcl

U111 twat: the stihlnthally hnatld mills of Qélilcwmand gig. i-hkarjao and azaucamrja (1910).hadnlaoiandmtncrealliaocmtlinnutricnt agarduring ineubnttan in phoaphatt bath» at huat. trtatnd5,. fig, emu» and sum (mes: 1911) nctnnniwnlyIwdiofi the tnamlbqr at lathally injured 0111:/at§_. gig; by rauiatian. ‘fhty cmclufied that tho injuryphnnclnnnen is duo tn llnlneular hat hitting duringhuatlaq which injures the poxipcral all mantras.Hha than all: (injhtid) warn mbjoctad ta thocowl»: znquizn the synthesis at daug­ribmuclnslc mid: (i»fiN¢ and resulted in m:LnLan1

nncmun r0cc*~'¢t1v. vloadeock aad 9:199 (1913) and 1

Sndjvick m6 urinal! (1972). abound the prunes atsingle and double strand and bxaalm in the maln­

7333

W10 at g. E5. stint hoat troatmmt ad tho 201%grow showed the rentltutlm at than organisms daring.i.n%t.tan in phosphate butter.

Idol: ad 0:601 (1966)­ahauud that boating of £5 ta 55° <2stopped till loakngo at Lnttaccllnlar eunatltuati andqt ; mqmr a-puamro at so°c. tho um eollululeakage and 1000 at viability at nicrourqanlam weremm. '1‘h¢y£utthl:ropor1:dthatthoprosenou0£euc—gun gain 0% protoctlcn against Qht thtnlnlly indualddeath but sum-we tuna to, prawn: tho lcakaqn at eo°c.

ovnnuwaqncnaalz mun:-mnaamqnapp¢a=¢a=9B¢rclhntd . rho groatcr reciucttm oi _;j3g.;_=g;¢_1;§ survi­

val at 609C any bu in the Ltqht ofribowctact! that tntraotllular prdbtns of the @011: caaqulatcand 11511106 with BIA 111:0 materials and tlms

firthnr prcvmtl lcakaqo. as this txeraturo. thanllnaelo protons at fish and ahnllflshns cald allflruducn thn ptatntn aonqulattm ;~-; so that increasedlaxaqo cnuld tnsult. an 50°C. aha partial ptotcctlcmagainst lcalaqu as attozdnd by tho mach protein.

23/1égg:(tho It-rain at 395;; Lag

_;_hag_ng,a_;yg._g_q£¢:thtant.u6yvnattasamnaathlaa0Ln thn nccttau 6.4. m the at treating.Pam alfllbl fibre groin in Tripticaso say broth withof 1% aadium chlorida (pti 1.0) at roam tir­aharotunlltoiihunzs. rhemllswurchnzvaatodmdrabid in statue 1% sodium‘ fihlorida soiutlfi and lus­pqdaé in aturilc 1% sodium chloride salutlm.

‘rho affect. at gggg gag;glgggg on the thermal stability varium encum­rationa 02 scauum in Tripticast soy bratb andin various lumagmates at flan shc1.=. fiahns. variousamino acids mixtzurcl. yeast attract. vita.-ains ad nucleo­sidos in phosphatase buiiicrs at taemoraturea at 50°C and55°C ware carricd art.

kinatiaq alum such as Trip­deaan say broth apri 7.2» omtalninq various salt caacaa-»tzattma at sadism ehlertdc (_D,5%¢ 3%. and 7.5%): Blahad shcllflahna timatos having tn: ab0~o saltemumt:.::at.1.<ma¢ 'r1.t.am1.aa» PODQQQI and

235

olanlhcru with above salt cancmtratima anstnrlliand and us-ed. £01‘ this study.

P;!.§€££IQ.§$.#.Q...21 r.ss§.§.ms­

rarthuligfittraatrannt» the“W111 lama ~11" M ¢=<==<=¢m=ra=1w=used in the boating mnnsttua at the raga of 1.01108parqranarnlmaprdotarminoi. tiaatlnqmialttuawcnhapt in a. water bath at ditficrcnt tcmoraturaa at 50°Cmd 55°C. ifhcn tho rcqulxbd tgoramro was attainflfip5 ll at waehad yibrio _g_\_p_Q ails wereto -1.; 200 all or gram of huatdi must-.rua and £rn%t1yuqltatnfl as as to aakao the tarzporatuxt unltotn. At.regular intervals of 15 tflnutus. an ml Qt arm an thacan any at oamplc an aseoptlcally and anti.­nny dilutnd with 0.114 xzavo‘ (7.2) caatainlnq 3% mm‘ma assay indium asafi in tho ms tripticwnsoy izflb agar 3% aadium chloride. Sortallyduucna 8a@l08 at 0.5 an was stfiabld in 2.5ago: containing 3% in cl. Tia szraiund plata war: tn­cubatod at room terqnraturc (303-_ 2°C) far 18 tn 24 hairs

In the aaac at hmagllnntu atfish and shalltishaa three tube MPH method ms used toronmratim at £~11t‘V1vGtI¢ In this case prc-cnziehmnt

Q36

proccdurm one ml of apprapriate sazmlo wan added toTripticasa any broth and kept for 12 hours at roamtewerature and than transferred one n1 portlm to 9 ml‘1‘r1pt1caso soy bcath. .‘1'h:l.s qnrichmnt was incubatedat room toweratzura. Further, a loopfiul of each breath

\I

was streaked Ln Thiasulphatc citrate bilc salt agar.

5 4 2 Enumaratian of thorn]. death timagi ' J;‘_'?_ij;::.t"'---- ;-:_;:.:»:: ; 1;’ ___::;‘;";fi'-:; ~¢-n-i1--‘=:---- - -*-~ '#&~i

@2159 _L12;v§.lsm> ­

The death rat: at Vibrig ga_:;§_‘ 0 - Olcarqanismn. at t%:at:urea at 56 C and :5was: studied in the abavo liquid broth and in ditfercathamoqnilnates cf fish and ahollfliahco. Thu number of

heat surviving vtablc coll: (109 10) warn plotted insemilaq graph an the logritrunic seals with time inminutos at 50°C and 55°C. réductim tiara of1:50 and D55 (that in minutes roquirod to dcntroy 90%at the viablu calla at are£1-on the graph. Tho values represented IIIJO tho munvalues at than 0: mm indopcndant trials.

5-4-=-??v:§Yl£!e§1;9.1;Ll¢£_&=11a=;¥..-@¢___22:!.£_= *1"mi" ­‘Preliminary cmcrtmnntn ax

the recovezy oi strosaod calls at 1. g ahaflwlltimlfl

Tft 3

nhaund no appreciable ditfiorcnoo banana nutrient aqu­Ilth 3% IIQCI ad. Trlptlcalc any agar cmtalninq 3% lad­lumwlortdn. $o£arthntota1 rlawozy atlnjutndQ0111. ‘IIIQMQAIQ say agar with 3% sodium chloride was

tiled in this study.wlalhnfl calla an rcnuapcndud

~\

£a3i6lIa¢1aad Snliaouulatndinphncphatn bath:(pfl 7.0) containing 3% In cl. 1' s broth. haaognnatoaat flab and shall. {talus an finlcribd in tho coctim5-2.1. oi mthds and tutorials and antaenclds mnéia.an-. the dance auponura ac s0°c ad ss°c em 00110at ccncntratlm :2 1.0110‘ pl! =1/qn wan Lmnmlatodad heated tor J0 arlmltcl. filth lroqunnt aqitattan.M‘. the termination at hcatlnq pried» ample: of 1 :1IIII imudlntnly withdrawn and axially dlluimd adattiahld in ptnpuuzbd Trlptieaai lay aqua‘ Ilth 3% flaC.'Land in Thlosulphnt-0 cltrata M10 salt! agar tar the“Wm ¢¢!1..b£4.2m$.;.I-.21.2.§-.1_2!.- Twnlewk Q1rztpttcaln Buy agar ad 263$ an incubated in chali­car.osat.:oaaca:|pc:at.urotat24l1au:sor4BhuurIuthocan may DI baton Gmllltinq. Whoa Qtlbfilifi I-Dh1b.‘l.tfl'

lilac N85 0006., tho tnjurad culls cuuld act. ha mauv­crod. who Mttozmm oi um raceway batman 2:-an and18$ indie-stun the injury lava].­

4‘J°"§.’"‘@306

1121......Thirclnltl otthostudyottha

auzvival at y_.g._ng9_ attains in vari­ous annstrua ccntainini dififiorunt amcuatratlanwat sedan cl\1%o:1do aim. medium and blah) are givnn in

tablli 49 and 50. A mlativdy i*.1qh lncculua at0011: ¢: (mos pt: =1 nu: gnu)III midi Ill! at m eenpatinq thfi diifiormtbrathl and atca oi fish and ahnlliiahcn (v-1.60supra) the lattnr gave qre ate: prwtnetim thm tholunar. of fish and ahnllflshoc withLncxlashq amcmtraflms at Nam aim}. ticmtly inatt­uod the a\u:v.i.v Lag nflszsbltl at g_;_b_z_1._gThe calla mu rapidly ddcruaainq at ss°c. am to mahmhwr tiwlrazuza. rm data pa-eanud hormdth iam­attains and tnd we typo strains (tabla 49) aboundthnlounzcmmcavinnhnatmqproanoda. rsbmth.chemically dofland sluts. pnptcn and try}:­tnnc mqisturd lessor survival than thl hcaagnnatsnl.A wry shaxp dzq was nfltlfl in thu lawn: cmonntratiauof aodim (0.5%; in broth! and lit“out thus: eaworiamts. tr s broth heated at59°C shard a htghcr survival in 3% and 7-5% sodiumehlorido cmcmtratimn.

533$)

On euwarmq the increasingsalt ummntratlaa at 3% 7.5%. the oi survivaln£_fl3§g waadshaneldlnqrcator pm­pazuma in cm n%am than in ma nqum balm.-am.Ghnnlcally addld slime acids. wit-mflna ad nucloosidoa

Qt bans in bath: at pfl 7.0 did not cahmcatho survival as in thb hcmoqnaatna. A “Mia: patternat result-J was %a1nd um and triptcno. Tintdata at thll prism: 3.n*-matiqatien lndicatd that 3% lafl7.5% hallo raaiatansn to the at:-and eunditimaand lnhmeo the Ifltvivll at culls at 50° and 55°C.

Imzinat rchctian tin: D valucsmm ealculntnd item tho thnzml survivors curvnn. Th!

did not shell: props: smooth 81096 md it wasottaa diplaaic md at 1:-mqular fits. 1:: value calmhtluworn tales: the Lnacttvaticn auzvos which had thogzoatnlt alopll and tranclnciod at laast 5 log roduetlaaaat the viahlo mils <1! g% attain!­At 55% . em 1., values warn toducld and tht all:tdéowded lcssat valun at hunt iurvival Wmthuuqh muttaedtum chloride was added. "2110 was true tor all tun

taut: aarrai whore 1a the pommtaqn of sodium chlerléuwas Lncraasod (7,553. The D 55°C values are givm intabla 53. ‘mo u values an pwmomccd shun callsat ggbrga are heated in hanoqmaza at

ii O

fish and shall ziishal. £11}! dcfincd mifili 02antioacidu L! vitamins, nuallnsiaelol‘ balms, plptvmn

Ind tzyptmn havt lesser valflil 1:-hm thi T53 ad

ti?! h%gIaataI.­The affect <9! inflating the

Lnjurad calls firing hating at 50°C and 55%: aresham in tabla 54. at 56% and 55°¢ £0: tnaim-ntna in wbltratnl. swab an 12; brati».hcaapnatos at fish and shollfitlhno w 1n ehcnlcallycmmumdefi mum acids. vitamins. plptano andbroth: indicator! ditflrcat nus at injury. At 50° 6%

;; in 1'8 brath the percent-.aq0 ofi injury as nazcinlesser ccncentraticna at salt (0.5%). rho can tradwas also can in all the media when tho salttiuas wore lass (0.5%). with increasefi salt aanamtraa­tuna. tho injury has loss with and 1.5% fer the==='@1==== *1 m=._s.e iwlvdino firmattain 0! B613 1902 and 3525‘

Heat injured calla in pliedin Triptlcaao any 1 broth Itth 3% 906-tun chloridnuncovered 811*-.h.Ln up to 1:11:00 hours (tabla 55) . Slaw

recmoty within 30 ntnuta was mawed by dltfiaront altauneqntratlcnn oi lay broth. thly

recovered fully “Q4; 30 minutes to 90 ninfitas. Q:

/‘K

V.-fit; /Z/\'J TL _

Q amazing growth. tho rqaovory in hint» adTripttcaa bx.-nth. an alalflcunt iaanao have canIVIB attnr adding 7.5% sedan chlas.-160.

Q; 6: ;.l1I$(IU8B§Q.

float rnalatmel at both iso­latnd attains typo strain: at j_(;g% ;3% increased uhca salt emwtratlaa nu inexcusa­This obaaxvattcns an in with the findingsof Muahat and ietthiaqtm (1916)-» In qnnnral tho.‘!.&£.1_2 Qrqmiml hm mm Mat "B­s1flwmutm¢humtmgmaamhagi;_ and Clanttgg 1. by thltnalheat. than wq-miss: were in either1abarat.orynnd1aeri.nhema%una£fl.nhanflch:'1Qabaemuu (blmchat 1913: Covert and ueoamm 1912:

Yafltrllnt and Bielnllam 1972) I 38%» thill amissan rezmlcttatn at-be: thruul assault. by the influlaooat an threat. chanted. nabatrataa and rich nutrientspaw 1606. in the raeovary media (Vaiadcrnmt at al »1974: Cmott am 1972: and Harries1967: ibtlchlt mil iintthlnqtm 1976) Bfidlllltv Plttlfianti 1511011008 19763 fifillp Huucbat and Jfifl 1977)»

This is also in agroenent m the m tin

53

influmco at toaporamro m tho growth at _‘iQ,_brl.0 gg,_§_a_­

; and en the nutrient supply in the qtowthIndia an aha survival. oi than orqantaus during mating(Mam g_t__ 5;-¢ 1970! Kite at cl" 19738 mt: adIngram 1962: Ray 35 3;" 1911: atnnaky 1974: Buuchatad Hathtuqtnn 1976)­

In this invatlqatlfln thoisalathd ml tan» "=01-M #1 .t-..h.£!-.2survived la 3% and 7.5% out canantratlcnl in vatiaua

@:I.a. w ospoaially in substrates of flan and ahrlwuThane aubstpatu intluaea tho prctnction at than cull!during hnatinq to so°c ma 55%. Gwioullju ___vLg;_gis 'nabapn:l.J.1.tc and it ro=fli.:oa salt£0: their i tin salt prmatat than duringheating. Christian md mm» (£974) alas aotad thatthe salt conecntrattoru and tmnraturca atfluctadthn:at:c0tqrau~cho¢1I_g;_gspp. Inthozml survivalstudies, the calls of tho lath mcpmlntial qrarth wattuser! as aaspcnliaas. In roam: yuan. dittenat rtaarehrather» umsaoll and tartan 1967: Covort and Bentham1972: muchat and hlorthtngtczn 1916) amt?-ind tho ctfoet

at lunar temperatures an tm survival ot @313 £5:aMwn£mumasmwma&mmsutvhai an these at higher tnmuxnturui at 50° and‘;ss°c 1» \.»~ anon: broth: ma flllbfitl-‘fitll with dlfihrnnt

53-4E3

salt cmccnttattma. thnrmal. rusistmct valfund vmm a mmltlinl amtaiainq highs: putamtaqo atulu (nu cl) was usaa. Thain an manna cm: cmaddition at sodium chlartda to haatlnq amatrunhad 0 protective afloat aqntnut dastructlcn. Calhounand rrallcr (1966) abused similar bchaviuur fa: g. 501;.that was tnportod tar mils haatndin mndta amaze in into: act1v.tty(a‘) was romldafl byQmttalld at aflflium chlctidn (Bnuchat adiwI.'th.‘l.nqt¢| 1976) L. Thu pratniathrc ctfoct at aadium

ahlcrién in hnatinq nuéia against dcattuci-.101 of alani­tla also ape:-bud (Baird-Parka: at :1”1910) .­

iiciiltalml to hlit st’-tillof is dqndnt. m thn chlM.ca1 ah upeithncail asunllutknplmteloqicalnamnofitluhunting Qav1t%t. Scvcral. lnvoltiqatcrn (filuchatmil Hcrthtaqtm 1976» Baird--Ptthlr at &ln19?D¢ cialhmnaad Frasier 1966, Christian and Hahn 1974) abound ao­

mlatlm big-an cellular mm-um ccqaaittm mama: rnzataaeo. who ahuaqoo at tatty acid! eantmtin mo eollular ceqmcntn Mn nnrlnad Lnflucaczden thomt nluwlm #1 ~11‘­(Hduahat and Horthtnqtm 1976} . Covert and Uoodburn

soH2.ilm

£1972) reported the protective nffnct and atabllllltian-ma at uwnmmn 0! 40¢ a.°c with we mm: em.»anntzatlua of {alt at 3% to 6%. A a1m:l.1u' result asmponuu by Matchnl 35 53,. £1910) and range (1966)

11 "ii" #1warn incxascd with Laura wing eaaenntratimat _¢m@=1a¢ a=so° ma ss°c anal» as). Bauchntma Northiaqtm (1916). in that: .=.m.;¢.P@-ua n valuuor than GIQELSK that warn htqlmr than this amdy.an lawn: vain” may ho duo to the mazizolazmcn asthose strain!‘ and tiny my also aepmdant an thechemical nah: up Q1‘ the flflfbflll G2 tn: mating lnvil-t'@—mats. MorO~0~_'&r, Lt was taund that mintalaiaq the

sun mating mv.i2*.'amInt fa the study at ad 1.135was diifiicult sinus tho lubfltxahfl 91 aim-anflab and shell tlahca IIIO caught in dlftnrent mvirm­

wt: and saikfimn wmah cannot bu cantrallod phyaxcally.Hevnv-on the raslatancc oi gggg totasting Qwllcmttatlinl of ladlilln chlflide Ital limi­ticmtly m,qm=: in all substrates at fish and shallfilalillthanlnthehnathfltmdnrstxfly.

Bnmtly nlnswilcr Q5 Q” (1976)tand that tho but injmmdcalla at 41% nun mm £0: so mum in pnapmu

ft, z"

butiez (100 U!) with 3% sodium ehluidd recavond in

higher name: in 'rr1pt'4cu0 any broth. similar result:Into also obtainod by thin in glucoa salt audit! £0814)which as and to remover than cello injured at 45°C.8-=> thn I-=Jvr¢¢.Q.P_£;2 ¢=11= “I blmcovubd olthnr plating than 1:1 rrlptlcaao may brothcontaining 3% sodium chloride or glucose salt undmn(G244) with 3% salt amen tat 3 hears. rho runway nuLit. m than ptntlhl, acid and 606:7­ribmucleic acid aynthais (Emilio: g_§_ _q_1.__., 1976)

‘rho alrbotaelan at anabolic

nuchanian of iniurod calla 10 ralaud ta an increasedlag tin: tnknn vhm the Lnjutod call! -u-uplacnd Lnautriint broth md Triptlcalo soy broth. met: war]:vucarridmtmunsmdyottmotfoouatmblcthal heat stress ca rdcroargmiarns and tho dams­linatian of these calla of bioaynthetic precast involvedto the roeovory at injured culls. ray researchwanker: had taund that the injury in gi­Lamas .!!_1'£.'I_¢. ='>-1%@;e.;.1_=;- ii­§.~ ‘*2-1_1_¢. _1*-Z!..'..‘!d:..%9.!; ii.-.!?;£3.>_2 $92?)

cloaggurg agfriggs 1n':1-znad altoratiana Ln that:attucmrél pormtability and bio synthetic elurnctcra(i-;msI|d.1O: Q_§ g_1__” 1976)

ZZ /14}

G011 nnnbruc dummy! had bnun

xuportnfi to be inc to increased salt. cumnutratlmInnlitivity at these orqwnlsnn ifitllln and wlttur19615; Imdalo méi Qsdal 1966; clash ad 1969:Clark 5 Q...» 1959). '2» loulmqi at the aungaa intra­cellular uvnfifludu inch as prntltal. auIao»nc1da andmtal lma 01:0,, hd hllln dxstratlfl Inridul,Icicntiiic urknro (soqis and Gtfial 1967: Strict uni shun1964: Blight and auxin 1966: hates ad Merit: 1960:Alluuofi n6.uuaol1 1969;. The mqpnlt at thest thor­mmlly injnrcd culls was fauna to occur during rvcewaryin thn r1ch»nut:1t1ana1 audit.

rm flab pmouasinq tllilrnaloqyuse-ci in ma mautaamzo oi fishery praductl awe stressfir injury W @118 ‘-‘**== =1-91*ho present. éiublathal starts: on theme calla cm:aitact the .mata;"201Lc and tne reprofiuctiva flmctlona

which the -qzuwth in the ulocttvo @611 witch: are comumly used to the pr.-saunas at theseorganisms 1h ioodfi-»

53 /1 S)

tabla 40. 30108.“ attach! 0! 3- hilnoido on tbs variousurotnln of 0}}; hp boating at 55% tar30 Iififilo

Boating S .1n1tn- 8 .0nt0r1- 8 .nntt0a- S .n:L11t 8 .011­6 aha $1.01: bar; pail. bumlloaltl-nu cu“ P”, -1.Control200' 2.51105 0.41102 4.71105 3.01105 0.01102011104 0.21105 4.21105 5.01105 1.91102 3.9110220‘ 1111111 2.11102 6.41102 4.41102 5.61102 7.511021.-11106101011‘ 00'“ mt.’11111.1» 0.51105 5.5110‘ 2.41105 1.91105 4.91105111111» 1.21102 0.0110‘ 1.4110‘ 1.51105 7.0110211111111110 5.61102 1.41102 1.0110‘ 4.91102 4.21102up:-010 1010 4.71102 0.91102 1.2110‘ 0.51102 6.311020111114 001 3.11102 1.91105 3.6110‘ 6.01102 5.41102111111101010 1.52105 2.51105 2.41105 4.01105 7.1110501101.16 0.21105 4.61105 0.41105 3.61105 4.71105HJd1'0t1pI'014IlI51h102 3.91105 5.11105 6.11105 1.91105010110111 3.31102 4.71102 2.01105 5.71102 7.211021101410111 4.71102 5.71102 2.11105 0.11102 6.511021101101110 6.m102 0.11102 4.01105 4.51102 5.71102mm 4.0110‘ 3.01105 7.91105 5.91102 4.01102111111161111 1.51105 6.2110‘ 2.71105 4.0110‘ 5.711051.51105 6.2110‘ 3.91105 3.61105 4.91102P1011110 3.41105 4.01105 7.71104 7.31105 0.311058021110 7.4110‘ 6.71105 5.01105 4.91105 6.7110‘:11-101110 2.31105 5.01105 4.71105 6.7110‘ 7.01102111114 2.71105 0.5110‘ 4.91105 6.0110‘ 9.41102

Allan 00110 aonouutatzloau 11004 1: this are 200 11¢/:1.* Erlptioua 001 aprimfi

-to<1:c

tabla 41. Rolntivn affect at In-Amino 00140 on ta variousumtypon of figljllg by heating at 60°6­tor 15 I111.

Bcatlng 8.wo1t0v- 8.nnto:-1- swath B.aai.at- Laban:2060: tzldh Qatar; pqlnfiu“ Bantu per I1­Oaatrolram 40:10” 6.4.110‘ 5.4110” “mo” ?.az1o'Hum: a.6z1o' 9.5:1o' 4.1110‘ 5.9110‘ 9.2110‘K3‘ 39" 5°“ 3°" 5°" 3°“1- mm muin butter!­nnm 44:10‘ 1.9210‘ a.5z1o' 6.9:1o' sane‘lrgtnlnn lc no $0" I6 noAipntglll $0" so" 6.91:1 0' 30" 54:10‘AIpIl'i1¢ acid: 1.1:1o' 5.5110‘ 6.1110‘ 30» 5.9110‘Gyntlat ac). ac no ac ac noezumuo ma sumo‘ mezzo’ 8-9210' sumo‘ 5.9110‘emu 5.5110‘ so" 5.6110‘ 30" so"nyamay palm $0" 4.9:1o' 6.2110‘ so" 30"Hiatflim I0 I8 I0 M2 H0mmman no no ac ac ncI-ouctnc so" we 50" ac IoIq-am no no ac lc acllnthtontnn la ac re ac nuPhony]. alanine are ac ac no nomum 5.9:1o' 1.3z1o' 9.0110‘ 4.1110‘ 5.9z1o'80:1» sumo‘ 9.1110‘ 9.a=1o' a.'mo' 1.u1o'mrmum me no ac no acnu» 50" :0" 4.9:w' we 50"Z"_'**-1' Amino acids conoontmtzoaa 200 ug per 11., ' trlpttcnoany broth», " (Bout Iona than, I8 Io aoullt as nab plaidviflout Minna.

tabla 42. flu stint 01' simple prataan Q carbo­hydrates by boating at 55% tor 15 limita­

Bun" smoltcv Banter!-~ B.untt- 8.na1at- 8.011115aunt:-an I060: ‘C1111: amber; palcoats not :1.Gontlnl983'HostelI33Toast cztnot(15)Payton (175)Iryptonl (1!)

3.01108

20111037.9x105

2.111032.41105

Garbihafiaton in0- 1! pholphnilImtturanma (4!)Sauna (415)Laetou (4!)181000 (4%)Rlzamosdfi)

’o731°,WI’6.21102W.1 .7110:

6.9210”

4.911056.a:103

3.7310’‘01Z‘°$

0.91103WI5061‘ 016.1110‘6.1210‘

5.21108

3.51104153110‘

1 .0110‘2.91104

'!.u10‘$0"3.92106.9110‘5.61110‘

2

5.11108

1.1110’3.91105

3.4111032071‘ Q2

4.e:103sol.6.5310‘3’!4.9210‘

‘$3 triptioaa 001 broth,-1 " Gout in Ion than.

4.11110”

309810,‘q5I' O,

2.9111033053192

4.6110339"5.91110‘W.9.6110’

60:18.” not

252

Heating S.vn1tw- 8.emar1- B.son1'- 8.ad.nt- Bmubcaafill! 8141! tanker; pal..”wu Onmii pf! I1.Hutu; at 60% tor 15 alanin­Goltrol288*Heat“I-‘SB

Yeast extract(Ii)Payton (ii)frnwflii)

l.az1 0°

709:‘ O‘

$.4:1o'4.6zao'WI‘

On-bolmdatoi ll0.11-Ibutts:Glucose (4!)8uu'oaa (#9I-acboao (4!)Xylene Hf‘Bhamoaaifi)

BC

$0!!­iolfHO

BC

sane‘ 6.2:1o° 5.a:1o° 3.9mo°

0.1110‘ 6.72102 5.6110‘ 1.3210‘

‘M681 O‘ ‘O 91‘ 0‘ 5 0,1‘ 0' 60%| 0‘5-3:10‘ 4.'mo' 6.:x1o’ 3.92m‘3°” 39"‘ $0" 5°"

nc ac ac noac gawr no acyoflv 50*» ac .Iac no no ncI0 ac to no“EB Triptiaan any broth" “ Cont in 10:0 till”EC Ho tabla cont an nab pistol wlflllmi dilution.

C13

Cf!Q;

tabla 43w linnxco taknn.t I111 the va:1ouaiflg}|gg}}auroiypoa 1a\d1t!br0n$ substrates a£‘t1dhand uh11!1ahnl­

Ozqpllnlu Pa H. HIE PHI Gil

5°'05§'550'G65°O

50°055'¢60*06590

8. Oliflriiiltl50*¢55‘960'665‘¢S. ‘§0'¢55'¢60°C65'¢

8, aonttunbag50%55'060*665°¢

$3. lllfi50'055'060*065'¢

30$

Q9020°25

50

530210

15Sin

50220

2050

52021 0

25,0

680507010

500210

155

4802i0$950

490200

20sf

500200

2050

59020$950

68049°

6510

51021029

50

49°210355

45°220

15

I"

49°210

2059

510220

255:

5804907010

510200

155

5002105°50

430250

3U

50

C99‘210

2050

51°200

2550

680499

7510

5002iO20

5

c§‘na0 once

Orgnalann Pull .HIB 2'Hl OilS. arisen: app

50'¢55'360'065°C

30 QhQI'§lI'5O'¢55°960*‘6596

so Uufiltl50‘G55'¢6O'065°G

S. hnilllhczg§O'C55°¢5O'G

55‘0Q 0 II]-£0“

50'055‘95O'065°C

Result! oxpruuaed as lsIlII‘IlI§I of thruo oxportnnatl" 100 fill» PI a@ EbaI1.lpot h0I0‘iIl$lI§.H B. uIItII\1honnlpntol; YIB. rrlma.hnIogpnn$0lL1._;|fl1gig): CIRCrab hoaogpnntnl (flgg1]ghgggg|j|).

49°190

255%

500190

255

480190305

490190

25SI

4901803°

5%

$001903°

50

490199$0

5

43°130

$05:

5001903°50

500.180

3Q

5*

500190

25SC

500196

305

49018

$05

50019930

5!’

501803°

5%

£01116 44. Variation ad D IIIIDI during but in-oatluatat at virtual tnpoam-no in£8! bmth mi I!-9 broth.

lamb 1111100 lnabnlausOrganism £128‘! Dawn I-9 bathe5011: 5511: 60'! 50111 5511: Q22

a.1nu1um-am 64.4 12.5 1.5 61.9 10.4 1.56. 6611111161111 62.6 14.1 1.7 61.6 11.2 1.4s. vclhvrodun 69.5 11.5 1.6 64.3 10.1 1.5s. naftaborg 66.4 22.6 1.6 79.6 16.5 5.96. mm 66.5 11.5 1.4 65.4 11.5 1.4$6 Oflfifltlllflifi 1005 165 5169 1063 163s. 661411111»; 62.4 11.5 1.6 62.2 11.6 1.4s. zypm 65.6 12.6 1.6 65.4 12.1 1.6s. arm 66.5 11.4 1.5 62.7 10.5 1.2s. ndiport 66.4 10.5 1.4 65.5 11.4 1.56. 1.6161166. 69.2 11.4 1.6 64.2 10.5 1.6S 0 155‘ 7°05 ‘on, Q 04 ‘1 Q5 902 ' 03

I8!-= Trtptica-00 Soy ‘least extinct broth.

256

Rabin 45- 11:11? or thorns]. ran:-nines of virtualaamotypoa at ,@jm];|,Q in differenthnlomanu.

0:-gunman tong. an 11.8 1'8 GI‘*0 Dvulauflvalubvnlurnvalnn

3 0 50 8°05 3°02 B‘u§55 15:9 14.3 1501 13¢?60 209 2;? 203 Z0‘1'94 1'08‘ 50 720‘55 120‘ H209 1305 11a6C0 200 20$ 206

8 0 Blilltfllul 50 6907 71 05 770‘ 7°0355 1108 11 04 1303 13.35' 2'1 2.3 Z04 2:4

30 UQIQUGQGIII 5° 9°05 9703 99:3 950455 15¢8 16¢? 1104 15.750 3.0 2¢3% 2'6 Z¢6

SI 6Olfi@§8l'Q 5° H004 1$B¢6 "Q05 104:555 44o5 55-7 57¢2 59¢‘­$9 3.] 9-? 8'7 9433 0 558 3'70’ 07 8,0‘ 860555 1 907 21¢‘ 23¢‘ 29¢‘5° 309 405 402 408

D values were of imrrago or than ezparimaim:5-= nackenl hono%tea: 1'a.l.n Pearl. spot homgmaton;EH: r:-a\maQ- 111111 homgonateo; Ga O1-at (1. 3135)h0%§flltQI¢

00l$8......

Organism! !O:g030 5°5560

8. hlldalhorgr 505560

S. typhi 50S560

S. cubnnn 5055.60

S. nswport .50-5560

£3. vqgoroaa 50S560

T257

ii

ii

E

g.

86.311.55.2

55.817.43.6

76.319.24.7

79.511.84.1

89.618.95.9

39.218.92.7

81.210.75.4

73.8780‘2.9

79.218.65.5

59.416.83.7

90.419.74.8

92.319.73.1

78.411.83.9

78.317.64.2

80.419.75.5

91.417.43.5

91.620.25.3

92.820.73.4

77.911.03.5

77.418.53.8

78.‘18.96.1

89.818.93.7

88.919.55.8

88.319.83.7

QEQ3

table 46. Repair at tnjuai 0011: 0! dltteaat etrntfl013.121,]; after thorn! stun at 50%59¢ mi 60°C in fish homogenates of Pearl.'ov¢% UE-.an:llll|llJ­/

Oljlllull nnacanx

31 03

lunar; slate count in IIJJagaa-at 5 '0.30111:. 6031:-.,90n1n. .120I1n­1:104 :10’ :10‘ :10‘

soso50‘U;W.(.1­“IK‘?"O51';NHIffL)‘

~0QJoifweVI

1101401501‘;wlltevrndfibcrllllqcarporttrizzbnfi appvqgaz-umanintpaul.breadnoyanaimaroanannitenbcrgclusteraalxnra

4.15-92.54-92.91 .44.34.84¢52441.1‘5'13.3

5.} 5-5 1 Q: ,1 07608 703 307 .3491,‘ 1.7 3;‘ ,‘06

5.8 607 702 ,7n6306 50? 3061.9 2.5 5.5 H5094.3 §¢5 1.3 1§.6 ‘cg 3.6 _5I,4.9 7.1 5-3 4:93:” 5-7 2:5 459

2.7” 4.6“ 5.1‘7,1 506 ‘O93;‘ 50‘ 40$ 50‘

I

1.1”5.:

an 1:104; b a :10‘; an :10’; 4 O 1105

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