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TECHNICAL REPORT NATICK/TR-78/002
THERMAL INACTIVATION OF VIRUSES
OCTOBER 1977
Approved for public release;
distribution unlimited.
UNITED STATES ARMY NATICK RESEARCH and DEVELOPMENT COMMAND
NATICK, MASSACHUSETTS 01760
Food Sciences Laboratory
FSL
Approved for public releasej distribution unlimited.
Citation of trade names in this report does not constitute an official indorsement or approval of the use of such items.
Destroy this report when no longer needed. Do not return it to the originator.
UNCLASSIFIED SECURITY CLASSIFICATION OF THIS PAGE (TWi-i Dmtm Xntfd)
REPORT DOCUMENTATION PAGE READ INSTRUCTIONS BEFORE COMPLETING FORM
4. TITLE r«mfSUMMI«3 —— —*"7^
THERMAL INACTIVATION OF VIRUSES
3. RECIPIENT'S CATALOG NUMBER
■ yi inn» t-^ayw wuiuntL
* Jan 9 —'Junfj 77^) Final
6. PERFORMING ORG. REPORT NUMBER
7. AUTHORtjjL __..-
Edward P.A,arkin ^ ...~ ' y
S. CONTRACT OR GRANT NUMBER/«)
DRXNM 77-115
r PERFORMING ORGANIZATION NAME ANO ADDRESS / Food and Drug/Administration Cincinnati, Ohio 45226
10. PROGRAM ELEMENT, PROJECT, TASK ~ INIT NUMBERS
11. CONTROLLING OFFIC3 NAME AND ADDRESS
US Army Natick Research and Development Commanc Natick, MA 01760 ATTW; DR.XNM-YMM
*4. MONITORING ACENCY NAME ft ADDRESS/*/ dltUrmnt trom Controlling Olllc») IS. SECURITY
Unclassified
is«, OECLA'SSIFICATION/DOWNGRADING SCHEDULE
1«. DISTRIBUTION STATEMENT (of thta Rmpnrt)
Approved for public release; distribution unlimited.
^- 17. DISTRIBUTION STATEMENT (ol tfi* abatfet totmrmd In Block 20. It dlttotmnt from RMHl\" \W" X\
§^> 18. SUPPLEMENTARY NOTES
It. KEY WORDS (Continue* on rnvrio »idm It r»i:»«»«ry and Identity by bl:nk ntmbmr)
THERMAL INACTIVATION INACTTVATTON TEMPERATURE (S) VIKU3FS
THERMAL RESISTANCE FOODS FLUIDS FOOD PROCESSING
FOOD PRESERVATION CONTAMINATION HEAT VIRAL NUCLEIC ACIDS
ao'.AjUsTNACT (Conilnum an rm**— •**» It nmc+nmy m>d Id+atttr by M«o* fmbm)
A review of the literature pertaining to thermal inactivation of virus in fluid media» fluid foods and solid foods indicated that the majority of viruses are inactivated at 7iC for 1 minute when normally expected levels of contamin- ation occur. A limited number of viruses have been reported to require a higher b.e&t process. Whether low level contamination by such viruses, would be ex- pected in foods, would survive the ?1 iTT-evel is not known because of the limited data available. Most food processes will not inactivate viral nucleic "
*SL 00 ,'
FOMM AM n 1473 EDITION OF i R9V M It &I9ROLETE
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unclassified IKCUWITV CLASSIFICATION OF THIS PAOlfWfc— P— CRIOTO
N Item 20 (Ccntinued)
Qacids. However, the presence of nucleases, pH extremes in foods and long storage times probably reduce their potential for retaining infectivity.
AC" 55" N 'w
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SICUKITY CLASSIFICATION OF THIS F-AQEfWhwt Dim Bnfr*4)
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PREFACE
The U. S. Army's food irradiation program has as its primary
objective the use of high doses (>1 Mrad) of irradiation to make pre-
packaged enzyme inactivated meats stable and safe from a microbiological
health hazard when stored under nonrefrigerated conditions. In this
process (radappertization) foods are formulated, placed in cellulose
casings or metal molds, heated to an internal temperature of 73 to 77 C
to inactivate autolytic enzymes, and chilled to -3 to 5 C. The product
is the vacuum packaged in cans or in flexible pouches, frozen to ca.
-40 C, and irradiated within a temperature range of -40 C to -8 C to
obtain the desired minimal radiation dose (MRD). Inoculated pack studies
with 10 strains of £. botulinum spores provide data for the computation
of the MRD, the dose required to reduce the number of viable spores of
£. botulinum by a factor of 1 x 1012.
Although viruses are more radiation resistant than C_. botulinum
spores,they should not present a health problem in radappertized meats
because; due to their reported heat sensitivity, they should be destroyed
during the preirradiation thermal treatment to inactivate autolytic en-
zymes. This review of literature, performed under project order number
DRXNM 77-115 was deemed essential to confirm the reported heat sensitivity
of viruses.
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Table of Contents
page
Preface 1
Report , 5
Figure 1. Time-Temperature profile for inactivation of virus single stranded nucleic acid or essential protein in a fluid medium 8
Table 1. Some characteristics of animal viruses 10
Bibliography 11
Tabls 2. Virus thermal inactivation . 12
Bibliography 20
Table 3. Thermal inactivation of viruses in foods 25
Bibliography 31
Table 4, Agents modifying virus thermal inactivation 34
Bibliography 36
Table 5. Naturally occurring virus concentrations in Food Animal Products 38
Bibliography 39
Table 6. The effect of temperature on nucleic acids 40
Bibliography 41
Table 7. Viruses producing systemic infections in food animals (North America) 42
Bibliography: Nucleic Acids , 43
Bibliography: Viral Thermal Inactivation Theory 44
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1
THKRHAL INACTIVATION OF VIRUSES
The thermal resistance of VJ rus in foods is t\.t lumvH >v the presence of protective agents that reduce the lethal effect of heat on the viruses at temperatures below 60 C In addition, whtn solid foods are processed heat transfer is by conduction,and the required internal temperatures must be attained to effectively inactivate viral contaminants.
In Table 1 some of the characterlottos of aniiT»! viruses arc depicted. The various f ant lies have been seoarated by their Luovant don sities and are listed alphabetically- The families 11 through 15 h-we been separated and also listed alphabetically. This same system was used to de- pict the information published in the literature pertaining to the familial resistance of viruses in laboratory media and in foods.
Examination of the data shown in Tables 2 and 3 indicate that the vast majority of viruses within the various families are inactivated at temperatures of about 60 C or less This literature review on v'ral in- activation indicates that resistance to thermal denaturation appears to be i restricted to viruses within the last five fanilles depicted in Table 1. i Greater resistance to temperature inactivation has been reported for the parvoviridae, the papoviridae and the nicornaviridae. The adenoviridae and the reoviridae appear to be only slightly more resistant to thermal inactivation than the first ten fanilies. Only the parvoviridae and the picornaviridae contain sufficient numbers of viruses to be of importance in studies on vlr.il t? ermal inactivation. The greatest number of viruses ! of nublic health importance are in the familv of picornaviridae. At the present time the exact position in the classification system of the hepa- titis and ^aatroentertitis viruses is not known but nrobab'v thev will be included in the parvoviridae or the picornaviridae.
The thermal inactivation data published in the literature have been concerned chieflv with inactivation of high titers of viruses in fluid preparations. There is p.reat variation in the techniques and procedures j used to evaluate the thermal resistance of viruses. In the majority of studies the viral suspensions were heated in test tubes that were held in constant temperature wate-: baths. In some cases the vixal suspensions were mixed in an attempt to equalize the temperature in the suspending media. In ] addition, a number of containers were used to process the virus suspensions j such as. capillary tubes, ampules an'' varvin~ tyi-es of flasks and bottles. It has been our experience that in evaluating the thermal resistance of \ viruses that even distribution of the heat occurs only when the viral su, pension is processed in ampules or capillary tubes that are submerged in a constant temperature bath. When other tvpes of non-submerged containers are utilized, there is uneven distribution of the heat and nossibie contamina- \ tion of the heat processed fluids by viruses surviving on the walls and closures of the vessels, buch conM unition may be interpreted as apparent survival of low level virus after the !;cnt treatment process In some cases these viruses are not detected bv cell culture systems but were observed j when animal inoculation was utilized. It is possible that free virus nucl'lc < acid may have been the causative agent in some of the renorted virus per- j sistence papers, especially when the heat treated preparation was injected | into an animal. j
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In Table 4 a nu iber of substances are listed that have a protec- tive effect on viruses when low temperatures (40 to 60 C) are used in thermal inactiveLion studies. In addition, this protective effect has been observed with other substrates such as serum, ice cream mix and other high protein-carbohydrate containing suspensions. It has been shown that with higher teraoeratures, in excess of 60 C, the protective effect on the virus decreases significantly. TThen virus susnen? .ons containing less than 10^ particles/ml are processed, normal inactivation occurs in a short period of time at temperatures of 60 to 70 C ant! appear to be inactivated at relatively the same rate.
Only a limited number of viruses have been reported to have ex- tremely high thermal resistance. These viruses are foot-and-mouth disease virus, hepatitis "A" (infectious hepatitis), hepatitis ,TB" (serum hepatitis) and the limited number of viruses in the napova and parvo virus families. In the reported foot-and-mouth disease studies a nunber of investigations were performed using high titers of viruses and in the malority of investi- gations tubes or bottles were used to process the suspension in the water bath. Apparent survival of low levels of viruses is possible with this pro- cedure. Only limited or no viruses were reported to survive the heat proces- sing of foods naturally contaminated by foot-and-mouth disease virus. Ex- pected virus levels in naturally infected animals are shown in Table 5. He- search in the Food and Drug Administration laboratories in Cincinnati indicates that virus levels of this magnitude were inactivated by temperatures less than those required for pasteurization of ice crean mix (6**.3 C for 30 n!.n or 79.A0 C for 25 sec).
The reported thermal inactivation data on the hepatitis viruses are United. In most cases viruses of unknown titer wer*» heat-treated in blood or blood components in an attempt to inactivate viruses, and infectivity was determined by human or primate feeding or inoculation studies. In one series of studies using hepatitis "A" in marmoset serum a temperature of 60 C for one hour was not sufficient to prevent infectivity in marmosets iniected with the heated product, "hen the sane virus was suspended in water ard heat pro- cessed, a reduction in infectivity was reported for the same time-temperature process. Because of the limitations in the reported data and the scarcity of information, the thermal resistance of hepatitis viruses is presently unknown.
It is possible that viral persistence may occur in heat processes using long time-temperature procedures such as cooking solid foods where periods of hours are required to reach internal temperatures of 60 C. Some viruses that could be present in animal foods are shown in Table 7. If viral protective agents are present in food, the lethality of the heat treatment process at levels below 60 C may have onJy a slight effect on the viruses,and the actual kill will commence only at temperatures above the 60 C. This could cause persistence of viruses in the food unless such considerations are made in determining the total food process, and the
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All the reported thermal inactivation data have been incor- porated into Figure 1. Because of the variation in the inactivation rates of the different viruses, a range of inactivation data is shown. Normal virus concentrations expected to be encountered in food should be inactivated at the time-temperature points shown in the figure. Only limited virus inactivation data have been reported in solid foods and consideration of this deficit was given, as well as the questionable persistence of some viruses, when the data were used to plot the solid food slope. This slope is close to that of the USPHS Pasteurization Standard for ice cream mix.
Thermal inactivation of viruses is directly related to th* chemical composition of the particle. The major organic constituents changed by temperature are lipids, proteins, and nucleic acids. The density of the particle is related to the varying compositions of the three organic constituents. Particles containing lipids or lipid complexes will teru to be more buoyant than particles composed of carbohydrate, protein, and nucleic acids.
Low temperature (20 to 35 C) inactivation of viral infec- tivity is dependent upon disruption of the nucleic acids. At higher temperatures ( > 50 C), inactivation is due to denaturation or dis- ruption of the proteins of the virus particle.
The nucleic acids present in the virus are of two types: ribonucleic acid and deoxyribonucleic acid. In a minority of the cases the nucleic acid may be double stranded. The nucleic acid is highly resistant to thermal inactivation at high temperatures for short periods of time. However, the nucleic acid molecule is dis- rupted gradually at low temperatures over relatively long time spans.
Some of the thermal characteristics of nucleic acids are shown in Table 6. In the case of the RKA containing viruses, nucleic acid inactivation is probably due to breakage of the phosphate bonds, Vhereas, in the case of the DNA containing viruses, inactivation is due to cleavage of the purine or pyrimidine bases of the nucleic acid complex. At low temperatures, DNA is about 30-fold more resistant than the RNA. In Figure 1 inactivation of the RNA occurs in a period of 13-25 hours, whereas the DNA inactivation requires 13-35 days. Complete inactivation is dependent on the viral titer: the higher the. nucleic acid content - the longer the period of time neces- sary for inactivation. At temperatures of approximately 30 - 45 C it is possible to inactivate nucleic acid with little apparent de- naturation of the protein containing units of the virus particle. Little information is available pertaining to resistance of single s.randed nucleic acid as compared to double stranded nucleic acid.
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However, it appears that the comple:: structure of the double stranded DITA is more resistant to disruption than is single stranded DM, and double- stranded RNA is more heat sensitive or equivalent to that of the single- stranded RNA.
Virus particles whose outer surface contain envelope structures are susceptible to mechanical iniury. Such -»articles appear to be very sensitive to thermal inactivation. In many cases the envelope is composed of lipid complexes. Whether loss of infectivity is due to a mechanical injury of the envelope or to a change in protein or lipid structures is at present unknown.
Viruses with protein outer surfaces are thermally inactivated by denaturation of the protein that occurs at higher temperatures. In some cases the coat is ruptured and the nucleic acid is liberated into the medium. A certain percentage of particles entrap the nucleic acid in the core as the protein components on the surface are denatured. This parti- cle has lost its ability to attach to the cell due to disruption of the structural integrity of the attachment site. However, these particles still containing nucleic acid may enter into the cell system by some me- chanical process such as pinocytosis, or some other means of engulfment, and the nucleic acid may be liberated within the cell and infection occurs.
It has been demonstrated that the nucleic acid from a suspension of virus particles is liberated into the medium during a thermal inacti- vation process. If nucleases are present, the nucleic acids are rapidly inactivated. Howevers in the absence of nucleases, the liberated nucleic acid is infectious. Laboratory studies have shown that demonstrated in- fectivity by nucleic acid is 2 to 4 logs less sensitive than that of the intact viroid. Thus, Kp virus particles may be inactivated but infec- tivity is still demonstrated in the cell or animal system because of the low level infectivity of the free nucleic acid.
The thermal process used to inactivate enzymes in foods, not less than 73 C or more than 77 C, probably will be greater than that re- quired to inactivate viruses. In Figure 1, a 3D virus inactivation pro- cess is depicted. If a 12D process is required, the slope will approach the USPHS pasteurization standard for ice cream mix when high temperatures are used. At low temperatures («60 C) longer time-temperature processes will be needed to inactivate the viruses than is required by the pasteuri- zation standard.
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Table 1. Some characteristics of animal viruses.
Family Density*0*
(Cesium chloride) Nucle_ic_acid (Type-stranded)
Surface characteristics
(Lipid) (Envelope)
1. Arenavlridae 1.18(1) RNA, SS +(b) +
2. Bunyaviridae 1.20-1.23 RNA, SS .(c) +
3. Coronavirldae 1.19-1.23 RNA, SS - +
4. Herpetoviridae 1.27-1.29 DNA, DS + +
5. Orthomyxoviridae 1.17-1.20 RNA, SS - +
6. Paramyxoviridae 1.21-1.24 RNA, SS + +
7. Poxvlridae DNA, DS + -
8. Retroviridae 1.16-1.18(1) RNA, SS + +
9. Rhabdoviridae 1.20 R*4A, SS - +
10. Togav.1 ridae 1.25 RNA, SS + +
11. Adenovlridae 1.33-1.35 DNA, DS - -
12. Papovaviridae 1.34 DNA, DS - -
13. Parvoviridae 1.38-1.46 DNA, SS - -
14. Picornaviridae 1.32-1.41 RNA, SS - -
15. Reoviridae 1.31-1.38 RNA, DS - -
g/cm ; + present; - absent
(1) In sucrose gradient
Abbreviations: RNA-ribonucleic acid, DNA-deoxyribonucleic acid, SS-single stranded and DS-double stranded.
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Bibliography
Some Characteristics of Animal Viruses
Andrewes, S. C, and H. G. Pereira. 1972. Viruses of vertebrates (3rd ed.) The Williams and Wilkins Co., Baltimore, Maryland, William Clowes & Sons, Ltd., London, Beccles and Colchester.
Fenner. F. 1976. Classification and nomenclature of viruses. Second report of the International Committee on Taxonomy of Viruses. Intervirol. 7(1-2):1-116.
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Bibliography
Viral Thermal Inactivation
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miJVW**Up!' M"l L»'.IP ■ .1 .»■Lin.iPlili'v.i^ll^"»"11.11 ^" ' "lu « ' ■! ' " »■"' M,nwi_nm i|i«upi
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23
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Bibliography
Thermrl Inactivation of Viruses in Foods
Aizen, M. S., and E. R. Pille. 1972. An experimental study of the survival of certain enteroviruses in meat products (In Russian; brief English summary). Gigiyena i Sanitariya 3:28-30.
Bauragartener, L., C. Olson, and II. Onuma. 1976. Effect of pasteuri- zation and heat treatment on bovine leukemia virus. JAVMA 169(11): 1189-1191.
Blackwell, J. H. 1976. Survival of foot-and-mouth disease virus in cheese. J. Dairy Sei. 59(9):1574-1579.
Blackwell, J. H., and J. L. Hyde. 1976. Effect of heat on foot-and- mouth disease virus (FMDV) in the components of milk from FMDV- infected cows. J. Hyg. Camb. 77:77-83.
Cliver, D. 0. 1973. Cheddar cheese as a vehicle for viruses. J. Dairy Sei. 56:1329-1331.
DiGirolamo, R., J. Liston, and J. R. Matches. 1970. Survival of virus in chilled, frozen and processed oysters. Appl. Microbiol. 20:58-63.
Demopoullos, G. T., 0. N. Fellowes, J. J. Collis, G. C. Poppensiek, A. G. Edwards, and J. H. Graves. 1959. Thermal inactivation and antigenicity studies of heated tissue suspensions containing foot- and-mouth disease virus. Amer. J. Vet. Res. 20:510-521.
Dupre', M. V., and M. Frobisher. 1966. Thermal inactivation: Animal viruses. In Environmental Biology (ed.) Altman, P. L., and D. S. Dittmer, FASEB, Bethesda, Maryland.
Felkai, V. T., F. Solyom, M. Szent-Ivanyi, and A. Wagner. 1970. The heat tolerance of foot-and-mouth disease virus in milk. Magyar Allatorv. Lapja 25^378-384,
Filppi, J. A., and G. J. Banwart. 1974. Effect of the fat content of ground beef on the heat inactivation of poliovirus. J. Food Sei. 39:865-866.
Foster, B. S., and C. H. Thompson. 1957. Report of the 60th Annual Meeting, U. S. Livestock Sanitary Association, Chicago, Illinois. Vet. Med. 52:118-121.
Gough, R. E. 1973. Thermostability of Newcastle disease virus in liquid whole egg- The Vet. Record 93(24):632-633.
31
:'.*-•:
Gresikova-Kohutova, M. 1959. The effect of heat on infectivity of the tick-borne encephalitis virus. Acta Virologica 3:215-221.
Heidelbaugh, N. D., and J. H. Graves. 1968. Effects of some tech- niques applicable in food processing on the infectivity of foot- and-mouth disease virus. Food Technol. 22(2):120-124.
Herniman, K. A. J., P. M. Hedhurst, J. N. Wilson, and R. F. Sellers. 1973. The action of heat, chemicals and disinfectants on swine vesicular disease virus. The Vet. Record, 93:620-624.
Hermann, J. E., and D. 0. Cliver. 1973. Enterovirus persistence in sausage and ground beef. J. Milk and Food Technol. 36(0):426-423.
Hyde, J. L., J. H. Blackwell, and J. J. Callis. 1975. Effect of pas- teurization and evaporation on foot-and-mouth disease virus in whole milk from infected cows. Can. J. Comp. M.ed. 39:305-309.
Kantor, II. A., and N. N. Potter. 1975. Persistence of echovirus and poliovirus in fermented sausages. Effects of sodium nitrite and processing variables. J. Food Sei. 40:960-972.
Kästli, P., and G. A. Moosbrugger. 1960. Destruction of foot-and-mouth disease virus by heat in milk products (in French; brief summary in English). Schweizer Archiv für Tierheill.urde 110:09-94.
Kaplan, A. S., and J. L. Melnick. 1954. Effect of milk and other daily products on the thermal inactivation of coxsackievirus. Am. J. Pub. Health 44:1174-1134.
K <lan, A. S., and J. L. llelnick. 1954. Differences in thermostability of antigenically related strains of poliomyelitis virus. Proc. Soc. 7,xptl. Biol. tied. 06:301-304.
Kavlan, A. S.» and J. L. llelnick. 1952. Effect of milk and cream on the thermal inactivation of human poliomyelitis virus. Am. J. Pub. Health 42:525-534.
Kelly, 3). F. 1964. Studies on onteroviruses of the pig. VII. Sone properties of a porcine enterovirus (F ). ?.es. Vet. Sei. 5:56-69.
Kostenko, An. N., and N. E. Botsman. 1971. Transmission of entero- viruses by milk and milk products (in Russian). Ilaterialakh U ShC'yezda gigiyenistov Ukrainskoy SSR v 1971 g. v g. Kieve 165-160.
Lawson, R. B., and J. L. Melnick. 1947. Inactivation of raurine polio- myslitis viruses by heat. J. Infec. Dis. 00:201-205.
Moosbrugger, P. K. 1968. La destruction du virus aphteux par la chaleur dans les produits laitiers. Extrait du Schweizer Archiv fUr Tierheilleundex fasc. 2, 110:09-94.
32
,-»^.-'*.-l ■ v-1: mai»nua.iiia«iii riMiririli) llll'J> iiViitrtfflftr tiri
fp^i'-wy,!', '^t^"*-.^i •idüN.^»-
Sellers, R, F. 1969. Inactivatlon of foot-and-mouth disease virus in milk. Br. Vet. J. 125:163-167.
Strock, N. R., and N. N. Potter. 1972. Survival of poliovirus and echo- virus during simulated commercial egg pasteurization treatments. J,* Milk and Food Tech. 35:247-251.
Sullivan, R., R. M. Marnell, E. P. Larkin, and R. B. Read, Jr. 1975. Inacti- vatlon of poliovirus 1 and coxsackievirus B-2 in broiled hamburgers. J. Milk and Food Technol. 38(8):473-475.
Sullivan, R., J. T. Tierney, E. P. Larkin, and ?w. B. Read, Jr. 1971. Thermal resistance of certain oncogenic viruses suspended in milk and milk products. Appl. Microbiol. 22(3):315-320.
Terpstra, C, and B. Krol. 1976. Effect of heating on the survival of swine fever virus in pasteurised canned ham from experimentally infected animal3. Tijdschr. Diergeneesk., deel 101, afl. 22:1237-1241.
33 <4
s::** I» :'jats«
Table A. Agents modifying virus thernal inactivation.
Virus Remarks Reference
Poliovirus
Poliovirus 1
Polioviruses 1, 2, 3
Polioviruses 1+2 Echo 6 and 7 Coxsackie B-5
Poliovirus 1
Poliovirus 1
ItgH-h stabilized virus at 45 C
Cations stabilized virus at 65 C
Ackerman
Fulioka
UgCl, stabilized virus during storage at 4 C Me.lnick
Reduced sulfhydryl groups Stabilized viruses to inactivation At less than 50° C
L-cystine stabilized, 5 different stocks of virus to temperatures below 50 C
Elemental sulfides (tetrasulfide) Stabilized virus against inactivation at 50° C
Halsted
Pohjanpelto
?ons
Poliovirus 1, 2, 9, 3 Coxaackie A-9, B~3 and Echo-1 Echo-3, 6 and 19
Echo-32
Poliovirus 2
Rhinoviruses and Enteroviruses
Simian virus SV-2
Herpe8virus (JES)
50 ug/nl of L-cystine stabilized viruses 500-2500 UR/ml stabilized of the viruses
No stabilization
Pch.lanpelto
MgClj stabilized virus at 50 C Branche
Soeir 0.1 M NaCl at 56 C for 1 hr * 6 log drop in infectivity 2.0 M NaCl at 56° C for 1 hr ^ 2 log drop in infectivity
MgCl« less effectively stabilized Dimmock Rhinoviruses at 50° C
MgCl0 stabilized virus at 50° C Heberling
Ka.SO, and Na-HPO. stabilized at 50° J Wallis 2 4 2 4
!IgCl2, MgSO , KH PO , 2 MKC1 or NaCl did not
stabilize virus Very thermosensitive in i9otonic salt solution
34
Table 4. Agents modifying virus thar: ia! inactivat?on (contd.)
Virus Remarks Aeferenee
7\b.abdoviruses EDTA and serum denonstrated MichalsL.i protective effect on virus at 37 and 5f° G
Hewcastle disease Casein protected virus at 45 C Ballestero- HgSO, did not protect virus
) ' Tick-borne I'.onovalent Metallic cations Mayer encephalitis stabilize'', virus at .'JO C
. /V.TA viruses '.Tot stabilized by (,iva1ent cations "7allii;
I'.eOTirus Stabilized by divalent cations Wallis
Vesicular Ho catn'onie stabilisation at 50 C 7<e.e. exantchoraa characteristic of liuman c.nteroviruses only of swine
•
■
35
.^."ü:., r-MUt... -J
Bibliography
Agents Modifying Virus Thermal Inactivation
Ackerman, W. W., R. S. Fujioka, and H. B. Kurtz. 1970. Cationic modulation of the inactivation of poliovirus by heat. Arch. Environ. Hlth. 21(3): 377-381.
Ballesteros, S., and B. Lucio. 1972. Effect of magnesium sulfate and casein hydrolysate on thermoinactivation of Newcastle disease virus vaccine. Avian Dis. 16(4):724-728.
Branche, W, C, Jr., V. M. Young, F. M. Houston, and L. W. Koontz. 1965. Characterization of prototype virus Echo-32. Proc. Soc. Exptl. Biol. Med. 118(1):186-190.
Dimmock, N. J., and D. A. J. Tyrrell. 1964. Some physiochemical properties of rhinoviruses. Brit. J. Exptl. Pathol. 45:271-280.
Fujioka, R., H. Kurtz, and W. W. Ackermann. 1969. Effects of cations and organic compounds on inactivation of poliovirus with urea, guanidine, and heat. Proc. Soc. Exp. Biol. Med. 132(3):825-829.
Halsted, C. C, D. S. Y. Seto, J. Simkins, and D. H. Carver. 1970. Protec- tion of enteroviruses against heat inactivation by sulfhydryl-reduclng substances. Virol. 40(3):751-754.
Heberling, R. L., and F. S. Cheever. 1965, Simian enterovirus SV-2 heraag- glutination studies. I. Relationship of infectivity to hemagglutination. Proc. Soc. Exptl. Biol. Med. 151-154.
Mayer, V., and I. Slavik. 1966. Thermostabilization of the tick-borne encephalitis virus. Virol. 29:492-493. ♦
Melnick, J. L., and C. Wallis. 1963. Effect of pH on thermal stabilization oral poliovirus vaccine by magnesium chloride. Proc. Soc. Exptl. Biol. Med. 112:394-397.
Michalak!, F., II. F. Parks, F. Sokol, and !I. F. Clark. 1976. Thermal inactivation of rabies and other rhabrtoviruses: stabilization of the chelating agent Ethylenediaminetetraacetic acid at physiological temperatures. Infec. and Immun. 14(1):135-143.
Pohjanpelto, P. 1961. Response of enteroviruses to cystine. Virol. 15:225-230.
Pohjanpelto, P. 1958. Stabilization of poliovirus by cystine. Virol. 6:472-487.
36
-JSüS/W.'/E;iSKsasyer^z™rL:_ - :.n.l.\r::^r_:-.. ■■_ .. ...:,-.^,, „. . :;—-i-: la^^—u .ü,"*" "T.;..;:^:;;,..-..: if^rat-u.»
Pona, M. 1964. Stabilization of poliovlrus by sodium tetrasulfide. Virol. 22(2):253-261.
Speir, R. W. 1961. Thermal stability of raengo and poliovirus in Hypertonie aalt. Virol. 14:382-383.
Wallis, C, and J. L. Me In Ick. 1965. Thermos tab ilizat ion and thermosensitization of herpesvirus. J. Bacterlol. 90:1632-1637.
Wallis, C, and J. L. Melnick. 1964. Reovirus activation by heating and inactivation by cooling in MgCl_ solutions. Virol. 22:608-619.
Wallis, C, C. S. Yang, and J. L. Melnick. 1962. Effect of cations on thermal inactivation of vaccinia, herpes simplex, and adenoviruses J. Immunol. 89:41-46.
Zee, Y. C, and A. J. Hackett. 1967. The influence of cations on the thermal inactivation of vesicular exanthema of swine virus. Arch. Gesamte Virusforsch 20(4):473-475.
37
ESE3ÜZ.: '—....i"L. .':>•' l'-^VT .f '.'jjigfa«!
Table 5. Naturally occurring virus concentrations In Food Animal Products
Virus Tlter Remarks Reference
Foot-and-mouth
Disease virus
(FMDV)
FMDV
FMDV
FMDV
Tickborne encephalitis
10*
10
10
10
A.5-5.3
3.3-5.2
10 4.5
2.0 10
10A.0-5.0
100-7 to 102-1
101-5 to IQ2*2
10 1.7
2.A 10
IG0'7 to 102-8
ID1'8 to IQ2'3
* 10 3.7
Enterovlruses < 10
Swine fever virus 10A-7 to IQ6'0
In serum, bovine
Pharynx
Milk
Saliva
Bone marrow, bovine
Muscle, brain
Blood
Beef, bovine
Liver
Kidney
Blood
Rumen
Lymph node
Goat milk
Shellfish
Blood
Burrows
Cox
Cottral
Henderson
Gresikova
Denis
Terpstra
38
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i
Bibliography
Naturally Occurring Virus Concentrations in Food Animal Products
Burrows, R., J. A. Mann, A. Greig, W. G. Chapman, and D. Goodridge. 1971. The growth and persistence of foot-and-moutb disease virus in the bovine mammary gland. J. Hyg. Camb. 69:307-321.
Cottral, G. E., B. F. Con, and D. E. Baldwin. 1960. The survival of foot-and-mouth disease virus in cured and uncured meat. Am. J. Vet. Ras. 21:288-297.
Cox, B. F., G. E. Cottral, and D. E. Baldwin- 1961. Further studies on the survival of foot-and-mouth disease virus in meat. Am. J. Vet. Res. 22:224-226.
Denis, F., and J. F. Brisou. 1976. Contamination virale des fruits de raer: etude portant sur Panalyse de 15,000 coquillages. Bull, de L'Academie Nationale de Medecine, Tome 160(1):18-22.
Gresikova, !i., I. Havranek, and F. Gorner. 1961. The effect of pasteuri- zation on the infectivity of tick-borne encephalitis virus. Acta virol. Prague 5:31-36.
Henderson, W. M. , and J. B. Brooksby. 1943. The survival of foot-and- "jiouth disease virus in meat and offal. J. Hyg. 46:394-402.
Terpstra, C, en B. Krol. 1976. Effect of heating on the survival of swine fever virus in pasteurised canned ham from experimentally infected animals. Tijdschr. Diergeneesk., deel 101, afl. 22:1237-1241.
39
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Table 6. The effect of temperature on nucleic acids.
Virus Remarks Reference
Foot-and-mouth disease (FMDV)
Virus heated to 61° and 85° C in RNase free medium produced infectious nucleic equivalent to that produced by phenol extraction
Bachrach
FMDV Prolonged incubation at 25 C for Brown 24 hrs or 37 C for 8 hrs resulted in a loss of ^ 3 logs of infectious nucleic acid
Tobacco mosaic virus (TMV)
Poliovirus 1 and TMV
Poliovirus 1
Polioviruses
Loss of RNA biological activity oc- Ginoza curs at moderate heat in marked con- trast to DNA which is inactivated with a high-temperature coefficient
* 90% loss of RNA infectivity in Gordon 80-120 min at 65° C
~ 99% loss of RNA infectivity in 40-45 min at 80° C
* 99% loss of RNA infectivity in 3-5 min at 100° C
Virus rapidly degraded at 56 C Jordan liberating viral RNA
Protein denaturation precedes Meitens dissociation of viral RNA at 50° C
EEE and VEE
Poliovirus
RNA inactivated «v 90% in 4 hrs Mika at 50° C
Heating RNA for 5 min at 60 C results Gorman in loss of 1 log of infectivity - virus heated same procedure results in 7 log loss of infectivity
Poliovirus mutants Variation in sensitivity to heat mutants
Pspaevangelou
Bacteriophaga Heat (50-60 C) inactivation is accompanied by release of native DNA. The molecules are intact - 90% removed from virus in 60 min at 60° C
Ritchie
40
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Bibliography
The Effect of Temperature on Nucleic Acids
Bachrach, H. L. 1961. Thermal degradation of foot-and-mouth disease virus into infectious ribonucleic acid. Proc. Soc. exp. Biol. lied. 107:610-613.
Brown, F., B. Cartwright, and D. L. Stewart. 1963. The effect of various inactivating agents on the viral and ribonucleic acid infectivities of foot-and-mouth disease and on its attachment to susceptible cells. J. Gen Microbiol. 31:179-186.
Ginoza, W. 1958. Kinetics of heat inactivation of ribonucleic acid of tobacco mosaic virus. 1958. Nature 181:958-961.
Gordon, M. P., J. W. Huff, and J. J. Holland. 1963. Heat inactivation of the infectious ribonucleic acids of polio and tobacco mosaic viruses. Virol. 19(3):416-418.
Jordan, L., and H. D. Maylor. 1974. Studies on the degradation of polio- virus by heat. Microbos 9:51-60.
Mietens, C, and K. Koschel. 1971. RNA content and entigenicity of polio- virua before and after inactivation by heating. Z. Naturforsch. 26 b:945-950.
Mika, L. A., J. E. Officer, and A. Brown. 1963. Inactivation of two arbo- viruses and their associated infectious nucleic acids. J. Infect. Dis. 113:195-203.
Norman, A., and R. C. Veomett. 1960. Heat inactivation of poliovirus ribonucleic acid. Virol. 12(1):136-139.
Papaevangelou, G. J., and J. S. Youngner. 1961. Thermal stability of ribonucleic acid froö poliovirus mutants. Virol. 15:509-511.
Ritchie, D. A. 1970. Physical characterization of the DNA released from phage particles by heat inactivation. Fed. Eur. Btochera. Soc. Lett. ll(l):257-260.
41
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Bibliography
Nucleic Acids
Alexander, H. E., G. Koch, I. lt. Mountain, and 0. Van Damme. 1958. Inactivity of ribonucleic acid from poliovirus in human cell monolavers. J. Exotl. Tied. 103:493-506.
Colter, J. S., H. II. Bird, A. W. Moyer, and R. A. 3rown. 1957. Infectivity of ribonucleic acid isolated from virus-infected tissues. Virol. A:522-532.
Dimmock, U, J. 1966. Biophysical studies of a rhinovirus: extraction and assay of infectious ribonucleic acid. Nature 209:792-794.
Gierer, A., and G. Schramm. 1956. Infectivity of ribonucleic acid from tobacco mosaic virus. Nature 177:702-703.
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