`
`”
`
`January-February 1975
`
`Volume 40 : Volume 1
`
`i
`
`N
`WeevsmYY
`N
`eed
`JOURNAL
`of FOOD SCIENCE
`
`
`
`Director of Publications
`John 8. 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)
`R. Ejiserle (76)
`D. Goll (75)
`0. Fennema (77)
`G. Giddings (76)
`H. Hultin (75)
`©. Gullett {77)
`S. Kazeniac (75)- D. Heldman (76}
`E. Laramond (77)
`T. Labuza (75)
`P. Hopper (76)
`R. Maxcy (75)
`Y. Pomeranz (76) T. Richardson (77)
`P. Netson (75)
`M. Solberg (76)
`B. Stillings (77)
`
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`IFT Scientific Editor
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`© Copyright 1975 by Institute of Food Technologists. All rights reserved. JOURNAL of FOOD 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 (1975)
`
`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
`
`NON ~ FAT
`DRY MILK
`
`the mold spores was omitted from the formula-
`tion’ of the systems to maintain the desired ay,
`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 ml of sterile de-
`ionized water for 1 min and TSY agar plates
`were used in 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 ml of
`distilled deionizeil water was added and stirred
`ORGANISM
`in to make a slurry. The pH was read after 5
`TEST
`min equilibration. The pH wasplotted against
`
`
`[INHIBITOR| PEANUT BUTTER
`HONEY
`the grams of H,O added on gran plat paper
`
`(100% volume-corrected, Orion
`cat.
`no.
`HeO
`
`900093). The value at zero addition is the pH.
`_[so04[s004an
`petC089)<Sebdvite
`This method is useful for IMF systems and was
`found more reliable than the method recom-
`mended in the AOAC bookof standard meth-
`a°o
`BOWL
`ods (AOAC, 1970). The two methods used in
`this study were found to give the same pH value
`within + 0.05 pH units which is the probable
`variation in composition.
`The water activity (ay) was measured by a
`manometer technique (Labuza, 1974a, b). The
`technique has an accuracy of + 0.005 at ana,
`of 0.85. Storage of the samples over the satu-
`rated sdlt solution made certain that this ay
`was constant throughoutstorage.
`The moisture content of representative du-
`plicate samples’of the systems with and without
`citric acid was determined by the vacuum oven
`method at 29 in. Hg and 60°C for 24 hr.
`
`
`
`
`
`PEANUTS
`
`FREEZE—DRIED CHICKEN
`
`
`BLEND ALL DRY
`COMPONENTS
`
`Fig. 1~Cold-mixing procedure used to prepare Hennican, the chicken-
`based IMF used.
`
`Table 2—Microbial inhibitors in Hennican, ay, 0.85
`
`Time for 1st appearance
`of A. niger (wk)
`
`RESULTS & DISCUSSION
`THE PARAMETERSandresults 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 monthsin 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
`FDArestriction (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 al.
`(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.
`
`
`
` Inhibitor %wiw pH 5.7 pH 4.2
`
`
`
`
`
`Potassium sorbate
`
`Calcium propionate
`
`Benzoic acid
`
`Methy! paraben
`
`Propy! paraben
`
`Parabens Me/Pro
`(2:1)
`Pimaricin
`
`1,3 Butanediol
`
`Propylene glyco!
`
`Mannitol
`
`Sorbitol
`
`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
`ng
`ng
`4.0
`ng
`ng
`1.0
`ng
`ng
`2.0
`ng
`ng
`1.0
`ng
`ng
`2.0
`ng
`ng
`1.0
`ng
`ng
`1.0
`1 4.5
`
`29 monthsstorage 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 al. (1973)
`in studies of A. niger on food systems of
`higher ay, and similar pH.
`The interaction of ay, 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 ay. 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 waslost 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 ay 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 ay
`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.
`Horner 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 ay. They found
`glycerol to be more inhibitory to A. niger
`than sucrose at the same pH and ay. On
`agar at aw 0.86 and pH 3.7, growth of A
`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 ay 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 modeofaction 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 ay and to
`the pathogenic bacteria Staphylococcus
`aureus,
`
`REFERENCES
`
`AOAC. 1970. “Official Methods of Analysis,”
`Ed. Horowitz, W., 11th ed, p. 214. Assoc.
`Official Analytical Chemists, Washington,
`D.c.
`Charlang, G.W. and Horowitz, N.H. 1971. Ger-
`mination and growth of neurospora at low
`water activities. Proc. Nat. Acad. Sci. 68:
`260.
`Chichester, D.F. and Tanner, F.W. dr. 1968.
`Antimicrobial food additives. In ‘(Handbook
`of Food Additives,” Ed. Furia, T.E., p. 137.
`Chemical Rubber Co., 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 & 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,
`T.P. 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, G.D. 1973.
`Combined effects of water activity, pH and
`temperature on the growth and spoilage
`potential of fungi. J. Appl Bacteriol. 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 & Space Administration, Hous-
`ton, Texas.
`Labuza, T.P., Cassil, S. and Sinskey, A.J. 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 Annua) 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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