'
`\i
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`‘.9-
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`January-February 1975
`
`Volume 40 : Volume 1
`
`9
`JOURNAL
`of FOOD SCIEl\LCE
`
`
`
`Director of Publications
`John B. Klis
`
`Publisher
`Calvert L. Willey
`
`Managing Editor
`Bernard Schukraft
`
`Scientific Editor
`Bernard J. Liska
`
`Director of Advertising
`Edward H. Hoffman
`
`Asst. to Scientific Editor
`Anna May Schenck
`
`Board of Editors
`R. Berry (77)
`W. Clark (76)
`R. Cassens (75)
`G. Bookwalter (77)
`Ft. Eiserle (76)
`D. Goll (75)
`O. Fennema (77)
`G. Giddings (76)
`H. Hultin (75)
`E. Gullett (77)
`S. Kazeniac (75)- D. Heldman (76)
`E. Laramond (77)
`T. Labuza (75)
`P. Hopper (76)
`R. Maxcv (75)
`Y. Pomeranz (76) T. Richardson (77)
`P. Nelson (75)
`M. Solberg (76)
`B. Stillings (77)
`
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`© Copyright 1975 by Institute of Food Technologists. All rights reserved. JOURNAL ofFOOD SCIENCE (formerly Food
`Research) is published six times a year (bimonthly) by Institute of Food Technologists, Suite 2120, 221 N. LaSalle Street,
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`138—JOURNAL OF FOOD SCIENCE— Volume 40 (7.975)
`
`including
`alcohols,
`The polyhydric
`propylene glycol, were
`inhibitory at
`lower concentrations compared to the
`
`amount allowed for various uses by the
`FDA. The minimum inhibitory concen-
`tration determined for 1,3-butanediol
`
`the mold spores was omitted from the formula-
`tion'of the systems to maintain the desired aW
`of 0.85.
`After 3 min mixing in the Brabender bowl,
`samples of the systems were plated to deter-
`mine the initial viable mold count. 5g of the
`food was blended with 45 m1 of sterile de-
`ionized water for 1 min and TSY agar plates
`were used ih duplicate at 23"C for 3 days.
`The pH of the systems was determined by
`two methods. A direct reading was taken by
`pressing a .nonaqueous Beckman electrode
`(#39142) into the squares of food. The gran
`plot method of Labuza (1974a, b) was also
`used. To 3.0g of food either 1, 2 or 3 m1 of
`distilled deionized water was added and stirred
`in to make a slurry. The pH was read after 5
`min equilibration. The pH was plotted against
`the grams of H20 added on gran plot paper
`(100% volume—corrected, Orion
`eat. no
`900093). The value at zero addition is the pH.
`This method is useful for IMF systems and was
`found more reliable than the method recom-
`mended in. the AOAC book of standard meth-
`ods (AOAC, 1970). The two methods used in
`this study were found to give the same pH value
`within 1 0.05 pH units which is the probable
`variation in composition.
`The water activity (aw) was measured by a
`manometer technique (Labuza, 1974a, b). The
`technique has an accuracy of t 0.005 at an aw
`of 0.85. Storage of the samples over the satu-
`rated salt solution made certain that this aw
`was constant throughout storage.
`The moisture content of representative du—
`plicatesamples’ofvthe systems with and without
`citric acid- was determined by the vacuum oven
`method at 29 in. Hg and 60°C for 24 hr.
`
`RESULTS & DISCUSSION
`THE PARAMETERS and results of this
`study are shown in Table 2.’The criterion
`for no inhibition was when mold became
`visible. This could indicate a consumer ac-
`ceptance criterion. As
`should be ex—
`pected, all the acid-type inhibitors were
`completely effective at pH 4.2 showing
`no growth for over 9 months in this inter-
`mediate moisture food. With a pH in the
`normal
`range for meat products, 0.3%
`K—sorbate is an effective mold inhibitor
`without
`the added effect of propylene
`glycol.
`If the food were higher in pH,
`more K-sorbate than the FDA allowance
`would be necessary. A similar trend is
`found for the propionate. Benzoic acid is
`not effective in the amount allowed by
`FDA restriction (0.1%) at the higher pH.
`The parabens inhibited the mold at all
`levels tested. As seen, a lower concentra-
`tion than found for the acid-type inhibi-
`tors is effective. The antibiotic, pimaricin,
`is effective at 0.002% (or 20 ppm) at
`both pH 5.7 and pH 4.2. Klis et
`a1.
`(1959) found inhibition at 5 ppm in agar
`at pH 5.6. However, they only incubated
`for 2 wk. It is possible growth might have
`occurred after that time. From a shelf—life
`testing standpoint, a longer time should
`be used. This study found 10 ppm to be
`ineffective. Most likely the antibiotic was
`not distributed as well in the heterogene-
`“‘ ous food of this study.
`
`
`PEANUTS
`
`FREEZE—‘DRIED CHICKEN
`
`BLEND ALL DRY
`COMPONENTS
`
`NON-FAT
`DRY MILK
`
`
`
`
`TEST
`ORGANISM
`
`-NH|B|TOR
`
`
`
`
`PEANUT BUTTER
`HONEY
`
`H20
`
`Shape Subdivide'50,1.-
`
`o a
`a 0 ‘3
`
`BRABENDER
`BOWL
`
`Fig. 1—Cald—mixing procedure used to prepare Hennican, the chicken-
`based IMF used.
`
`Table 2—Microbial inhibitors in Hennican, aw 0.85
`
`Time for 1st appearance
`of A. niger3 (wkl
`
` Inhibitor %wlw pH 5.7 pH 4.2
`
`
`
`
`
`
`
`Potassium sorbate
`
`Calcium propionate
`
`Benzoic acid
`
`Methyl paraben
`
`Propyl paraben
`
`Parabens Me/Pro
`i2:1)
`Fimaricin
`
`1,3 Butanediol
`
`Propylene glycol
`
`Mannitol
`
`Sorbirol
`
`Glycerol
`
`Control
`
`ng
`2
`0.15
`ng
`ng
`0.30
`ng
`2
`0.1
`ng
`19
`0.2
`ng
`ng
`0.3
`ng
`7
`0.2
`ng
`ng
`0.3
`ng
`ng
`0.03
`ng
`ng
`0.05
`ng
`ng
`0.10
`ng
`ng
`0.01
`ng
`ng
`0.03
`ng
`ng
`0.04
`ng
`ng
`0.05
`ng
`ng
`0.10
`4.5
`1
`0.001
`ng
`ng
`0.002
`ng
`ng
`0.005
`22
`1
`1.0
`ng
`ng
`2.0
`ng
`ng
`4.0
`ng
`ng
`1.0
`ng
`ng
`2.0
`"Q
`ng
`4.0
`ng
`ng
`1.0
`ng
`ng
`2.0
`ng
`ng
`1.0
`n9
`n9
`2.0
`ng
`ng
`1.0
`ng
`ng
`1.0
`1 4.5
`
`3- 9 months storage at 23°C: ng = no mold growth during the period of
`storage
`
`-
`
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`(2.0%) is below the inhibitory concentra-
`tion found by Frankenfeld et a1. (1973)
`in studies of A. niger on food systems of
`higher aw, and similar pH.
`The interaction of aw, solute used and
`pH in their effect on microorganisms has
`been reported by many workers (Troller,
`1973). The solutes used to lower aw are
`often polyols, such as glycerol, propylene
`glycol, 1,3-butanediol, and it
`is certain
`that their inhibitory effect is not entirely
`related to their water binding capacity;
`however, the reason for their toxicity is
`not known. Working with Neurospora sp. ,
`Charlang and Horowitz (1971) found that
`glycerol was less inhibitory as compared
`to NaCl or sucrose at the same aw. They
`suggest the difference is due to the sol-
`ute’s electrolytic properties. They found
`that at low aw, a substance essential for
`spore germination was lost to the medium
`and when the substance was isolated and
`supplied to the spores, germination oc-
`curred. They suggest that' the release of
`this substance is due to Osmotic effects
`which are related to the permeability of
`the cell to a solute. Solutes such as glyc-
`erol, which easily entered the cell pre-
`venting osmotic imbalance, did not in-
`hibit germination as much.
`Webb (1960) suggested that death at
`lowered aw was due to the dehydration
`of an essential macromolecule. He sug-
`gested that if the solute had a hydrogen
`bonding ability, it may bind on the mac-
`romolecule
`and
`prevent denaturation
`from loss of the hydration shell as aw
`decreases. This could explain why glyc-
`erol was less toxic than NaCl in the Char-
`lang and Horowitz (1971) study, how-
`ever, it does not explain the toxicity in
`this study.
`Homer and Anagnostopoulos (1973)
`studied the growth rate of several molds
`as a function of pH, aw, temperature and
`
`INHIBITION OF A. niger IN AN IMF SYSTEM—139
`
`the solute used to adjust aw. They found
`glycerol to be more inhibitory to A. niger
`than sucrose at the same pH and aw. 0n
`agar at aw 0.86 and pH 3.7, growth ofA
`niger was visible on media containing
`glycerol as the humectant after 5 days at
`25°C. This is a very short induction time
`compared to the present study in which
`the control Hennican (no glycerol added)
`at aw 0.85, pH 4.2 didn't show growth of
`the mold for 4.5 wk. Under the stress
`presented by this food system as com-
`pared to nutrient agar,
`the additional
`adverse effects of only 1% of glycerol was
`enough to completely inhibit
`the mold
`for over 9 months.
`The mode of action of these inhibitors
`is not known, but they are effective in-
`hibitors of the test organism in this study
`at suboptimal pH and aw. This study will
`be extended to other molds commonly
`found as contaminants at low aw and to
`the pathogenic bacteria Staphylococcus
`aureus.
`
`REFERENCES
`
`AOAC. 1970. “Official Methods of Analysis.”
`Ed. Horowitz. W.. 11th ed. p. 214. Assoc.
`D.
`.
`Oféicial Analytical Chemists. Washington.
`Charlang. G.W. and Horowitz. N.I-I. 1971. Get-
`mination and growth of neurospora at low
`2 0.
`wgter activities. Proc. Nat. Acad. Sci. 68:
`Chichester. D.F. and Tanner. F.W. Jr. 1968.
`Antimicrobial food additives. In “Handbook
`of Food Additives." Ed. Furia. T.E.. p. 137.
`Chemical Rubber 00.. Cleveland. Ohio.
`Christian. J.H.B. 1963. Water activity and the
`growth of microorganisms. In “Recent Ad-
`vances in Food Science.” Ed. Leitch. J.M.
`and Rhodes. D.N.. Vol 3. p. 248. Butter-
`worths 8: Co., London.
`.
`Clark. W.. Shirk, R. and Kline. E. 1964. Pimari-
`cin. a new food fungistat.
`In “4th Int’l
`Symp. on Food Microbiology." p. 167. SIK
`Goteborg. Sweden.
`Frankenfeld. A.J.W.. Karel. M. and Labuza.
`”LP. 1973. Intermediate moisture food com-
`position containing aliphatic 1. 3- diols. U.S.
`Patent No. 3.732.112.
`
`Hollis. F.. Kaplow. M.. Halik. J. and Nord-
`strom. H. 1969. Parameters for moisture
`content for stabilization of food products.
`Phase 2. U.S. Army Natick Labs. Contract
`DAAG-17-67-C-0098.
`Homer. K.J. and Anagnostopoulos. GD. 1973.
`Combined effects of water activity. PK and
`temperature on the growth and spoilage
`potential of fungi. J. Appl Bacterial. 36:
`427.
`Kaplow. M. 1970. Commercial development of
`intermediate moisture foods. Food Technol.
`24: 889.
`Klis. J.B.. Witter. L.D. and Ordal. Z.J. 1959.
`The effect of several antifungi antibiotics on
`the growth of common food spoilage fungi.
`Food Technol. 13: 124.
`Labuza. T.P. 1974a. Sorption theory and meas-
`urement. In “Physical Properties of Food.”
`Ed. Rha. C.. p. 119. Reidel Press. Dor-
`drecht. Holland.
`Labuza. T.P. 1974b. Storage stability and im-
`provement of intermediate moisture foods.
`Phase 2. Contract #NAS 9-12560. National
`Aeronautics 8: Space Administration. Hous-
`ton. Texas.
`Labuza. T.P.. Cassil. S. and Sinskey. AJ. 1972.
`Stability of intermediate moisture foods. 2.
`Microbiology. J. Food Sci. 37: 160.
`Plitman. M.. Park. Y.. Gomez. R. and Sinskey.
`A.J. 1973. Viability of Staphylococcus
`aureus in intermediate moisture meats. J.
`Food Sci. 38: 1004.
`Sauer. F. 1972. Control of mold by chemical
`preservatives. Fungi and Food. 7th Annual
`Symposium. Western New York Section.
`IFT. Oct. 19.
`Scott. W.J. 1957. Water relations of food spoil-
`age microorganisms. Adv. Food Research 7:
`84.
`Troller. J.A. 1973. The water relations of food-
`borne bacterial pathogens: A review. J. Milk
`Food Technol. 36: 276.
`Webb. S.J. 1960. Factors affecting the viability
`of air-borne bacteria. 3. The role of bound
`water and protein structure in the death of
`air-borne cells. Can. J. Microbiol. 6: 89.
`Ms received 6/8/74; revised 8/24/74: accepted
`8/26/74.
`
`Presented at the 34th Annual Meeting of the
`Institute of Food Technologists in New Or-
`leans.
`Paper no. 8722 from the University of Min-
`nesota Agric. Experiment Station. This study
`was supported in part by the University of Min-
`nesota Agric. Expt. Station Project No.
`18—52HM and Contract
`#NAS
`9-12560.
`Lyndon B. Johnson Space Center. Houston.
`Texas.
`Reference to any product mentioned does
`not mean endorsement.
`
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