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The Control of Molds on Shell Treated Eggs
The Control of Molds on Shell Treated Eggs PATRICIA CLEMENTS AND A.
R.
WINTER
Ohio Agricultural Experiment Station, Columbus, Ohio (Received for publication May 4, 1956)
T IS a common practice to shell treat eggs that are to be held in cold storage, as shown by the reviews of the literature by Funk (1950, 1955). It may also be used as a means of preserving the original quality of eggs during movement in the normal channels of trade as shown by Winter et at. (1954). The dipping solution, usually paraffin base mineral oil, may become contaminated from microorganisms on the shells as shown by Rosser et al. (1942), Bart (1949) and McFarland (1950). Repeated use of the shell treating solution, without sterilization, may actually contaminate or seed successive lots of eggs with mold spores and bacteria. Molds will grow on and/or inside of the shells of treated eggs as shown by Mailman and Davidson (1944), Bart (1949), Yushok and Romanoff (1949, 1950), McFarland (1950), Puffenbarger (1951), Brown and Gibbons (1954), and Lorah, Funk and Forward (1954). Rosser et al. (1942) prevented mold growth on plastic (polyvinal alcohol) coated eggs by incorporating 2.8 percent dimethyl urea in the dipping solution. Mailman and Davidson (1944) prevented mold growth on oil-treated eggs by adding 0.25 percent Dowicide 7 (pentachlorophenol) to the oil. Young (1948) prevented mold growth on eggs by dipping them in a 0.5 percent solution of salicylanilide. Preliminary studies on the work to be reported here were made from our department in theses by Bart (1949) and McFarland (1950). They found that Zerlate (zinc dimethyl dithiocarbomate), Shirlan Extra (salicylanilide), Isothan DL-1 (dialkyl dimethyl ammonium bromide), Ceepryn (cetyl pyridinium chloride), Cetab (cetyl
trimethyl ammonium bromide) and Hadbac (higher alkyl dimethyl benezylammonium chloride) were effective fungicides for use in egg treating oils in low concentration (0.1 to 0.25 percent). Yushok and Romanoff (1950) have confirmed the efficiency of Ceepryn, and Lorah et al. (1954) have confirmed the value of Zerlate and Hadbac. The latter have also reported that BTC (alkyl dimethyl benzyl ammonium chloride) and thiourea (thiocarbamate) are effective fungicides, when used in low concentration. The objectives of this study were: (1) To obtain additional data on the efficiency of fungicides for use in egg treating paraffin base mineral oil, (2) To obtain data on fungicides suitable for use in Foamrex (a wettable wax emulsion), (3) To obtain data on the germicidal value of fungicides when used, in low concentration in egg treating oil and Foamrex. EXPERIMENTAL
Eggs. The eggs were obtained from the university farm. They consisted of both white and brown eggs. The eggs were held in an egg room, maintained at 55°F., one day before beginning of the test. All eggs, except the controls, were washed in a small amount of water slightly warmer than the eggs, and dried without rinsing, in front of a fan. The eggs were candled after washing and drying and only those with sound shells and grade A quality used. They were randomized into as many lots of two eggs each as required for the particular trial to be conducted. The eggs were dipped momentarily into the treating solution, maintained at room temperature. They were then placed on wire
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I
CONTROL OF MOLDS ON SHELL EGGS
eral oil and a wettable wax emulsion. Oil is widely used at present for shell treating eggs. It must be applied when the eggs are dry. The wettable wax emulsion may be applied when the eggs are wet, immediately after washing. The oil used in this experiment was Sohio No. 72. This is a paraffin base mineral oil with a viscosity of 64 to 75 Saybolt seconds at 100°F., pour point 25°F. and specific gravity of 0.845 to 0.858. This is a slightly heavier oil than generally used for treating eggs but provides slightly better protection as shown by Cotterill (1948), Winter and Cotterill (1949) and Puffenbarger (1951). Foamrex is the trade name of an alkaline wettable wax emulsion manufactured and distributed by the Socony Vacuum Oil Company. It contains small (1-2 micron) particles of wax dispersed in water by means of an emulsifying agent. It contains 47 percent solids (melting point 125°F.) and has a pH of 9.7. Foamrex was diluted with an equal volume of wated before use in the tests. Fungicides. The fungicides were obtained direct from the manufacturers. In most instances they had not been tested for use in egg treating solutions. Most fungicides are soluble in water, but not in oil. Nearly all of the fungicides tested were those formerly screened by Bart (1949) or McFarland (1950) and found to be most promising for use in oil. The fungicides were tested at 0.25 percent level. The earlier work of Bart (1949) and McFarland (1950), showed that some fungicides were effective at this or greater dilution. , RESULTS
Six fungicides were tested for their fungicidal and bactericidal properties on shell treated eggs. They were tested at 0.25 percent concentration in water for dipping eggs before shell treating and for use in Foamrex and oil egg dipping solutions.
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racks in front of a fan, to drain and dry. The eggs were then transferred to covered staining jars the bottoms of which were kept covered with water. They were supported on glass rings above the water. The eggs were held at room temperature (65°F.9S°F.) and 95 to 100 percent relative humidity for 15 days. . The eggs were examined for visible signs of mold growth at the end of 5, 10 and 15 days. They were examined under a normal day light and fluorescent (black) candling lamp. Mold counts. Mold counts were made, to obtain a more quantitative measure of viable molds on eggs than was possible by observing visible appearance. The eggs were washed by placing them in a wire basket suspended above the blades in a waring blender bowl. The washing solution consisted of 500 ml. of sterile water, containing emulsifying agents, 0.1 percent Tween 80 and 0.15 percent G-2800. These compounds improve washing and sampling efficiency in the presence of oil as shown by McFarland (1950). The wash solution was examined for molds by the dilution plate method using Bacto-potato dextrose agar acidified to pH 3.5. The plates were incubated 5 days at room temperature before the mold counts were made. Bacterial counts. Bacterial counts were made on the shell washing to determine the germicidal, as well as fungicidal value of the compounds tested. Dilution plates were prepared using Bacto tryptone glucose extract agar and incubated 48 hours at room temperature. Interior quality examination. At the end of the 15-day holding period, the eggs were candled, broken out and examined under ordinary white light and fluorescent light for mold development, rot, off color, odor and fluorescence. Shell treating solutions. The fungicides were tested for use in a paraffin base min-
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C L E M E N T S AND A.
WINTER
trials. No special seeding of egg shells with microflora was necessary. The microflora present on the washed eggs was probably typical of that likely to be found under practical conditions. Dipping the eggs in Foamrex or oil made little difference on visible mold growth on the shells, occurrence of rots or mold and bacterial counts in the shell washings at the end of the holding period. Data on the fungicidal and bactericidal properties of the fungicides tested with Foamrex and oil are summarized in Table 3. Cetab was an effective fungicide when used alone or in oil. It was not effective in preventing mold growth on egg shells when used with Foamrex. Cetab also had some germicidal value at 0.25 percent concentration when used alone or in connection with oil. It was not effective when used with Foamrex in the same concentration. The findings that Cetab is an effective fungicide for treating shell eggs is in agreement with the earlier reports by Bart (1949) and McFarland (1950). B 104 was an effective fungicide when used alone or in combination with Foamrex or oil in 0.25 percent concentration. It had
TABLE 1.—Fungicides tested for use in egg treating oil and Foamrex Fungicides Trade names
Solubility Water
Oil
Arquad S
1
1
B 104
+ +
1
Ceepryn Cetab * Duomeen T Shirlan Extra
2
3
2
3
3
4
2
Alkyl trimethyl ammonium chloride (Armour and Company) Di-duodecnyl dimethyl ammonium chloride (Rohm and Haas Company) Cetyl pyridinium chloride (National Biochemical Company) Cetyl trimethyl ammonium bromide (National Biochemical Company) Duomeen T mono Acetate (Armour and Company) Salicylanilide (Du Pont Company)
1. Dissolved in isopropanol and then in water or oil. 2. One part fungicide dissolved in 4 parts of a mixture consisting of dibutyl phthalate 75% and ethyl alcohol 25% and this in turn mixed with oil. 3. With addition of heat. 4. Dissolved in ammonium hydroxide and then in water. -+- Without addition of heat.
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The trade names of the fungicides tested, their active ingredients and their solubility properties are summarized in Table 1. The compounds had a tendency to settle out when dispersed in oil. Data on the growth of molds and bacteria on clean, soiled and washed eggs are summarized in Table 2. At the end of the IS-day holding period, in the warm moist chamber, the microbial growth was about the same on the clean soiled and washed eggs. The mold count in the shell washings ranged from 0 in 4 of the 54 analysis made to a high of 800,000,000 in one trial with an average of 1,000,000 per shell. The bacterial count ranged from 64,000 to 1,600,000,000 with an average of 16,000,000. The microbial count, even on clean appearing eggs is likely to be high as shown by the review of the literature by Winter et al. (1955). The fact that the viable mold and bacterial populations were about the same on the clean, soiled and washed eggs may have been due to the biological saturation point being reached during the holding period. The washed eggs, used for testing fungicides, had a heavy heterogenous microflora in 17 of the 18
R.
1119
CONTROL OF MOLDS ON S H E L L EGGS T A B L E 2.—Mold and bacterial growth on clean, soiled and washed eggs Trials Kind . of eggs
Egg treatment
•vr
No. microorganisms per shell (log)
No. showing mold
made
Growth Rots
Molds Av.
High
Bacteria Low
Av.
High
Low
None Dipped in Foamrex Dipped in oil
6 6 6
5 6 5
4 5 2
5.1 6.8 6.3
7.1 8.9 7.0
0 5.5 5.1
7.3 7.4 6.9
9.2 8.3 7.9
5.7 6.1 5.1
Soiled
None Dipped in Foamrex Dipped in oil
6 6 6
5 6 5
5 5 4
5.2 6.6 5.3
6.8 8.3 7.8
0 5.6 0
7.5 7.3 6.7
9.1 9.0 8.7
5.7 6.1 4.8
Washed
None Dipped in Foamrex Dipped in oil
6 6 6
5 5 5
4 4 4
5.6 6.7 6.5
7.3 8.3 7.2
0 4.0 5.6
7.3 7.7 6.9
8.7 8.2 8.1
4.8 6.8 5.2
TABLE 3.—Fungicidal and bactericidal properties of some fungicides in Foamrex and oil Percent of trials showing Fungicides >
Eggs a n d treatments
Mi croorgamsms per shell (lc3g of count) Bacteria
Molds Mold
Rot Ave.
High
Low
Ave.
High
Low
7.2 8.2
4.8
2 2 2 2 2 2 2 2
50 50 50 0 0 50 0 0
50 50 50 0 50 0 50 0
3.5 7.0 6.7 0 6.4 0 2.5 0
7.1 7.2 7.2 0 7.0 0 4.9 0
0 6.9 6.1 0 5.9 0 0 0
6.0 8.2 2.7 5.9 2.3 6.6 0
5.4 5.9 4.7 7.2 0
0 5.8 0 5.9 0
100 100 50 0 0 0 0
6.3 6.5 6.6 0 0 0 0 5.4
6.2 6.4 5.6 0 0 0 0 0
7.5 8.0 5.7 4.8 5.4 3.4 3.9 2.2
7.8 8.0 6.2 4.9 6.0 6.8 7.8 4.4
5.2 4.7 3.8 0.0 0.0 0.0
Cetab
Control Control Control Dipped Dipped Dipped Dipped Dipped
B104
Control Control dipped in Foamrex Control dipped in oil Dipped in B 104 Dipped in B 104 then Foamrex Dipped in B 104 then oil Dipped in Foamrex containing B 104 Dipped in oil containing B 104
2 2 2 2 2 2 2 2
100 100 100 0 0 0 0 0
• o
6.25 6.4 6.1 0 0 0 0 2.7
Control Control dipped in Foamrex Control dipped in oil Dipped in Duomeen T Dipped in Duomeen then in Foamrex Dipped in Duomeen then in oil Dipped in Foamrex containing D . T . Dipped in oil containing D . T .
4 4 4 4 4 4 4 4
100 100 100 0 0 0 25 0
100 100 75 75 50 75 50 50
6.7 7.0 6.5 2.9 2.4 3.0 3.2 3.0
7.3 8.3 7.1 6.0 4.9 7.0 8.4 6.3
6.2 6.4 5.6 0.0 0.0 0.0 0.0 0.0
8.0 7.6 6.9 5.8 5.8 6.4 4.8 4.5
8.7 8.0 8.2 7.3 6.2 6.8 7.7 7.3
7.2 6.8 5.2 4.9 5.5 6.2 0.0 0.0
2 2 2 2 2 2 2 2
50 50 50 0 0 0 0 0
50 50 50 0 50 0 0 50
3.5 7.0 6.7 0 3.9 0 0 3.0
7.1 7.2 7.2 0 6.7 0 0 5.9
0.0 6.9 6.1 0 5.9 0 0 0
6.0 8.2
7.2 8.2
4.8
Ceeprin
Control Control dipped in Foamrex Control dipped in oil Dipped in fungicide Dipped in fungicide then Foamrex Dipped in fungicide then oil Dipped in Foamrex containing fungicide Dipped in oil containing fungicide
0 6.3 2.4 0 3.1
0 6.4 4.7 0 6.1
0 6.2 0 0 0
Arquad
Control Control in Foamrex Control in oil Fungicide Fungicide then Foamrex Fungicide then oil Fungicide in Foamrex Fungicide in oil
4 4 4 4 4 4 4 4
100 100 100 0 25 0 50 0
100 100 75 SO 75 25 75 25
6.7 7.0 6.5 2.9 3.3 2.7 3.2 3.7
7.3 8.3 7.1 6.1 7.3 5.8 7.2 5.8
6.2 6.4 5.6 0 0 0 0 0
8.0 7.6 6.9 6.8 7.9 5.6 6.6 4.8
8.7 8.0 8.2 7.4 8.4 7.1 7.2 7.7
7.2 6.8 5.2 6.2 7.5 4.7 5.9 0
Shirlan Extra
Control Control in Foamrex Control in oil Fungicide Fungicide then Foamrex Fungicide then oil Fungicide in Foamrex Fungicide in oil
4 4 4 4 4 4 4 4
75 100 100 0 25 25 0 50
75 75 75 25 50 75 0 25
5.3 7.3 6.8 2.9 5.2 2.9 1.6 4.3
7.3 8.3 7.2 6.0 7.2 5.8 6.5 6.5
0 6.7 6.1 0 0 0 0 0
7.3 7.6 8.1 6.3 6.9 7.0 4.6 3.2
8.7 8.2 8.2 7.9 7.7 7.8 .8.4 6.6
4.8 6.9 8.1 5.2 6.5 0 0 0
Duomeen
untreated dipped in Foamrex dipped in oil in fungicide in fungicide then Foamrex in fungicide then oil in Foamrex containing fungicide in oil containing fungicide
No. of trials
—
—
—
—
— —
7.2
—
— —
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Clean
1120
P. CLEMENTS AND A. R. WINTER
SUMMARY
Molds grow readily on eggs treated with oil or Foamrex, within a period of 15 days, when held under favorable conditions. Some of the fungicides tested have bactericidal properties but are not efficient germicides for use in oil or Foamrex.
Some fungicides, incorporated in oil or Foamrex at 0.25 percent concentration, inhibit mold growth on shell-treated eggs. Cetab (cetyl trimethyl ammonium bromide), Ceepryn (cetyl pyridinium chloride) and Shirlan Extra (salicylanilide) were efficient fungicides for use in oil for treating shell eggs. B 104 (di-duodecnyl dimethyl ammonium chloride) was an effective fungicide for use in both oil and Foamrex for treating shell eggs. REFERENCES Bart, G., 1949. The control of fungi found on poultry eggs in storage. M. A. Thesis, The Ohio State University. Brown, H. J., and N. E. Gibbons, 1954. Air cell mold in oiled eggs. Food Tech. 8:307-11. Cotterill, O. J., 1948. The preservation of shell egg quality by oil treatment. M.S. Thesis, The Ohio State University. Funk, E. M., 1950. Maintenance of quality in shell eggs by thermostabilization. Missouri Agr. Exp. Sta. Res. Bull. 467. Funk, E. M., 1955. Treating shell eggs to maintain quality. North Central Regional Publication No. 62 Mo. Bull. 659. Lorah, M. E., E. M. Funk and J. Forward, 1954. Reducing Spoilage in shell eggs by the use of fungicides. Missouri Agr. Exp. Sta. Res. Bull. 573. Mailman, W. L., and J. A. Davidson, 1944. Oilprotected shell eggs. U.S. Egg and Poultry Mag. 50:113. McFarland, C. R., 1950. The effects of germicides in mineral oil used to shell-treat storage eggs. M.S. Thesis, The Ohio State University. Puffenbarger, D. W., 1951. Preservation of egg quality by shell treatment. M.S. Thesis, The Ohio State University. Rosser, F. T., W. H. White, A. H. Woodcock and D. A. Fletcher, 1942, Preservation of eggs. I. Treatment for maintenance of quality in shell eggs at ordinary temperatures. Can. J. Res. 20 D : 57-70. Yushok, W. D., and A. L. Romanoff, 1949. Studies on preservation of shell eggs by coating with plastics. Food Res. 14:113-122. Yushok, W. D. and A. L. Romanoff, 1950. Prevention of mold growth in intact eggs by treat-
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some bactericidal properties, reducing the bacterial count in the washings about 40 percent. Duomeen T prevented visible mold growth on the shells, reduced spoilage and mold counts in the shell washings about 55 percent and bacteria about 27 percent. Ceepryn eliminated mold growth on egg shells when used alone or in combination with oil in 0.25 percent concentration. It reduced bacterial counts when used as above. Ceepryn was ineffective as a fungicide or germicide when used in connection with Foamrex. Bart (1949), McFarland (1950) and Yushok and Romanoff (1950) have also reported that Ceepryn is an effective fungicide for treating shell eggs. Arquad practically eliminated visible mold growth on egg shells, reducing spoilage and the mold count about 50 percent. It had little bactericidal effect. Arauad was more effective when used in connection with oil than with Foamrex. Shirlan Extra eliminated visible mold growth on egg shells when used alone or in combination with oil in 0.25 percent concentration. It eliminated mold counts 100 percent from shell washings in 9 of 18 trials and bacteria in 6 to 18 trials. Shirlan Extra was effective in most trials when used with either oil or Foamrex. The fact that it was not completely effective in all trials may have been due to the type of microflora present. The findings, that Shirlan Extra is a good fungicide for treating shell eggs, is in agreement with the earlier reports by Young (1948), Bart (1949) and McFarland (1950).
CONTROL OF MOLDS ON SHELL EGGS ment with a cationic detergent. Food Res. IS: 1-7. Winter, A. R., and O. J. Cotterilll, 1949. The preservation of shell egg quality by oil dipping right on the farm. Amer. Egg and Poultry Rev. 10: (3) 14-16. Winter, A. R., P. Shields, L. MacDonald, and I. Prudent, 1954. Improving the keeping quality of eggs during marketing by hot oil treatment.
1121
Food Tech. 8:515-18. i Winter, A. R., B. Burkart, P. Clements and L. MacDonald, 1955. Cleaning eggs with detergents and detergent-sanitizers. Ohio Agr. Exp. Sta. Res. Bull. 762. Young, H. E., 1948, Blue case timber and mold rots in hens eggs, Queensland J. Agric. Sci. 5:125-40.
(Continued from page 1099) physiology, effects of antibiotics on blood formation, pharmacology of antibiotics, effects on growth of human subjects, miscellaneous nutritional effects, effects on guinea pigs, public health aspects of feeding antibiotics to animals, and conclusions.
This book is an excellent presentation of this field. It presents and weighs the evidence with judicial choice of facts and figures. This is not easy to accomplish, for the literature is full of contradictions and opposing theories, but it has been well done in this book.—H.D.B.
NEWS AND NOTES NOMINATIONS—PENB ACHIEVEMENT AWARD The Poultry and Egg National Board Research Achievement Award is given annually for outstanding research leading to the increased consumption of poultry and eggs. The recipient will receive an appropriate plaque and will be honored at the annual meeting of PENB, January 31-February 1, 1957. Selection of the winner is based on the importance of poultry and/or egg research contributions concerning processing, quality preservation, nutritive value, and new and improved product development, leading to the increased use of poultry and eggs in the diet. Nominations should be supported by biographical sketches of the nominees, including enumeration of contributions pertinent to the Award objectives, a list of publications during the past ten years, and, if available, endorsement or seconds. Nominations should be sent to Hans Lineweaver, Chairman of the PENB Technical Advisory Committee, Western Utilization Research Laboratory, Albany 10, California, not later than December 3, 1956. NEWMAN MEMORIAL AWARD Dr. B. R. Burmester, Biologist of the Regional Poultry Research Laboratory, Animal and Poultry Husbandry Research Branch, Agricultural Research Service, U. S. Department of Agriculture, East (Continued
Lansing, Michigan, received the Tom Newman Memorial International Award for Poultry Husbandry Research for 1956. This Award consists of a medal and i50. sterling. It is given for the most important contribution to poultry husbandry resarch work published in the previous year. The Award was made by the Committee, to Dr. Burmester, primarily for his poultry disease research work, with particular reference to lymphomatosis. The papers cited were: Immunity to Visceral Lymphomatosis in Chicks Following Injection of Virus into Dams, published in the Proceedings of the Society for Experimental Biology and Medicine; In Vitro and in Vivo Neutralization of the Virus of Visceral Lymphomatosis, published in the Proceedings of the Society for Experimental Biology and Medicine; The Presence of the Virus of Visceral Lymphomatosis in Embryonated Eggs of Normal Appearing Hens, published in Poultry Science; Tumor Incidence in the Progeny of Hens Repeatedly Injected as Adults with Visceral Lymphomatosis Virus, published in Poultry Science; The Propagation of the Virus of Visceral Lymphomatosis in Embryonated Eggs, published in Poultry Science; Plasma Phosphatase Activities of Normal and Lymphomatous Chickens, published in Cancer Research; and The Role of the Infected Egg in the Transmission of Visceral Lymphomatosis, published in Poultry Science. Dr. Burmester was born in Petaluma, California, and obtained a B.S. degree, majoring in Poulon page 1124)
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BOOK REVIEWS
R.
WINTER
Ohio Agricultural Experiment Station, Columbus, Ohio (Received for publication May 4, 1956)
T IS a common practice to shell treat eggs that are to be held in cold storage, as shown by the reviews of the literature by Funk (1950, 1955). It may also be used as a means of preserving the original quality of eggs during movement in the normal channels of trade as shown by Winter et at. (1954). The dipping solution, usually paraffin base mineral oil, may become contaminated from microorganisms on the shells as shown by Rosser et al. (1942), Bart (1949) and McFarland (1950). Repeated use of the shell treating solution, without sterilization, may actually contaminate or seed successive lots of eggs with mold spores and bacteria. Molds will grow on and/or inside of the shells of treated eggs as shown by Mailman and Davidson (1944), Bart (1949), Yushok and Romanoff (1949, 1950), McFarland (1950), Puffenbarger (1951), Brown and Gibbons (1954), and Lorah, Funk and Forward (1954). Rosser et al. (1942) prevented mold growth on plastic (polyvinal alcohol) coated eggs by incorporating 2.8 percent dimethyl urea in the dipping solution. Mailman and Davidson (1944) prevented mold growth on oil-treated eggs by adding 0.25 percent Dowicide 7 (pentachlorophenol) to the oil. Young (1948) prevented mold growth on eggs by dipping them in a 0.5 percent solution of salicylanilide. Preliminary studies on the work to be reported here were made from our department in theses by Bart (1949) and McFarland (1950). They found that Zerlate (zinc dimethyl dithiocarbomate), Shirlan Extra (salicylanilide), Isothan DL-1 (dialkyl dimethyl ammonium bromide), Ceepryn (cetyl pyridinium chloride), Cetab (cetyl
trimethyl ammonium bromide) and Hadbac (higher alkyl dimethyl benezylammonium chloride) were effective fungicides for use in egg treating oils in low concentration (0.1 to 0.25 percent). Yushok and Romanoff (1950) have confirmed the efficiency of Ceepryn, and Lorah et al. (1954) have confirmed the value of Zerlate and Hadbac. The latter have also reported that BTC (alkyl dimethyl benzyl ammonium chloride) and thiourea (thiocarbamate) are effective fungicides, when used in low concentration. The objectives of this study were: (1) To obtain additional data on the efficiency of fungicides for use in egg treating paraffin base mineral oil, (2) To obtain data on fungicides suitable for use in Foamrex (a wettable wax emulsion), (3) To obtain data on the germicidal value of fungicides when used, in low concentration in egg treating oil and Foamrex. EXPERIMENTAL
Eggs. The eggs were obtained from the university farm. They consisted of both white and brown eggs. The eggs were held in an egg room, maintained at 55°F., one day before beginning of the test. All eggs, except the controls, were washed in a small amount of water slightly warmer than the eggs, and dried without rinsing, in front of a fan. The eggs were candled after washing and drying and only those with sound shells and grade A quality used. They were randomized into as many lots of two eggs each as required for the particular trial to be conducted. The eggs were dipped momentarily into the treating solution, maintained at room temperature. They were then placed on wire
1116
Downloaded from http://ps.oxfordjournals.org/ at West Virginia University, Health Science Libraryy on April 11, 2015
I
CONTROL OF MOLDS ON SHELL EGGS
eral oil and a wettable wax emulsion. Oil is widely used at present for shell treating eggs. It must be applied when the eggs are dry. The wettable wax emulsion may be applied when the eggs are wet, immediately after washing. The oil used in this experiment was Sohio No. 72. This is a paraffin base mineral oil with a viscosity of 64 to 75 Saybolt seconds at 100°F., pour point 25°F. and specific gravity of 0.845 to 0.858. This is a slightly heavier oil than generally used for treating eggs but provides slightly better protection as shown by Cotterill (1948), Winter and Cotterill (1949) and Puffenbarger (1951). Foamrex is the trade name of an alkaline wettable wax emulsion manufactured and distributed by the Socony Vacuum Oil Company. It contains small (1-2 micron) particles of wax dispersed in water by means of an emulsifying agent. It contains 47 percent solids (melting point 125°F.) and has a pH of 9.7. Foamrex was diluted with an equal volume of wated before use in the tests. Fungicides. The fungicides were obtained direct from the manufacturers. In most instances they had not been tested for use in egg treating solutions. Most fungicides are soluble in water, but not in oil. Nearly all of the fungicides tested were those formerly screened by Bart (1949) or McFarland (1950) and found to be most promising for use in oil. The fungicides were tested at 0.25 percent level. The earlier work of Bart (1949) and McFarland (1950), showed that some fungicides were effective at this or greater dilution. , RESULTS
Six fungicides were tested for their fungicidal and bactericidal properties on shell treated eggs. They were tested at 0.25 percent concentration in water for dipping eggs before shell treating and for use in Foamrex and oil egg dipping solutions.
Downloaded from http://ps.oxfordjournals.org/ at West Virginia University, Health Science Libraryy on April 11, 2015
racks in front of a fan, to drain and dry. The eggs were then transferred to covered staining jars the bottoms of which were kept covered with water. They were supported on glass rings above the water. The eggs were held at room temperature (65°F.9S°F.) and 95 to 100 percent relative humidity for 15 days. . The eggs were examined for visible signs of mold growth at the end of 5, 10 and 15 days. They were examined under a normal day light and fluorescent (black) candling lamp. Mold counts. Mold counts were made, to obtain a more quantitative measure of viable molds on eggs than was possible by observing visible appearance. The eggs were washed by placing them in a wire basket suspended above the blades in a waring blender bowl. The washing solution consisted of 500 ml. of sterile water, containing emulsifying agents, 0.1 percent Tween 80 and 0.15 percent G-2800. These compounds improve washing and sampling efficiency in the presence of oil as shown by McFarland (1950). The wash solution was examined for molds by the dilution plate method using Bacto-potato dextrose agar acidified to pH 3.5. The plates were incubated 5 days at room temperature before the mold counts were made. Bacterial counts. Bacterial counts were made on the shell washing to determine the germicidal, as well as fungicidal value of the compounds tested. Dilution plates were prepared using Bacto tryptone glucose extract agar and incubated 48 hours at room temperature. Interior quality examination. At the end of the 15-day holding period, the eggs were candled, broken out and examined under ordinary white light and fluorescent light for mold development, rot, off color, odor and fluorescence. Shell treating solutions. The fungicides were tested for use in a paraffin base min-
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C L E M E N T S AND A.
WINTER
trials. No special seeding of egg shells with microflora was necessary. The microflora present on the washed eggs was probably typical of that likely to be found under practical conditions. Dipping the eggs in Foamrex or oil made little difference on visible mold growth on the shells, occurrence of rots or mold and bacterial counts in the shell washings at the end of the holding period. Data on the fungicidal and bactericidal properties of the fungicides tested with Foamrex and oil are summarized in Table 3. Cetab was an effective fungicide when used alone or in oil. It was not effective in preventing mold growth on egg shells when used with Foamrex. Cetab also had some germicidal value at 0.25 percent concentration when used alone or in connection with oil. It was not effective when used with Foamrex in the same concentration. The findings that Cetab is an effective fungicide for treating shell eggs is in agreement with the earlier reports by Bart (1949) and McFarland (1950). B 104 was an effective fungicide when used alone or in combination with Foamrex or oil in 0.25 percent concentration. It had
TABLE 1.—Fungicides tested for use in egg treating oil and Foamrex Fungicides Trade names
Solubility Water
Oil
Arquad S
1
1
B 104
+ +
1
Ceepryn Cetab * Duomeen T Shirlan Extra
2
3
2
3
3
4
2
Alkyl trimethyl ammonium chloride (Armour and Company) Di-duodecnyl dimethyl ammonium chloride (Rohm and Haas Company) Cetyl pyridinium chloride (National Biochemical Company) Cetyl trimethyl ammonium bromide (National Biochemical Company) Duomeen T mono Acetate (Armour and Company) Salicylanilide (Du Pont Company)
1. Dissolved in isopropanol and then in water or oil. 2. One part fungicide dissolved in 4 parts of a mixture consisting of dibutyl phthalate 75% and ethyl alcohol 25% and this in turn mixed with oil. 3. With addition of heat. 4. Dissolved in ammonium hydroxide and then in water. -+- Without addition of heat.
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The trade names of the fungicides tested, their active ingredients and their solubility properties are summarized in Table 1. The compounds had a tendency to settle out when dispersed in oil. Data on the growth of molds and bacteria on clean, soiled and washed eggs are summarized in Table 2. At the end of the IS-day holding period, in the warm moist chamber, the microbial growth was about the same on the clean soiled and washed eggs. The mold count in the shell washings ranged from 0 in 4 of the 54 analysis made to a high of 800,000,000 in one trial with an average of 1,000,000 per shell. The bacterial count ranged from 64,000 to 1,600,000,000 with an average of 16,000,000. The microbial count, even on clean appearing eggs is likely to be high as shown by the review of the literature by Winter et al. (1955). The fact that the viable mold and bacterial populations were about the same on the clean, soiled and washed eggs may have been due to the biological saturation point being reached during the holding period. The washed eggs, used for testing fungicides, had a heavy heterogenous microflora in 17 of the 18
R.
1119
CONTROL OF MOLDS ON S H E L L EGGS T A B L E 2.—Mold and bacterial growth on clean, soiled and washed eggs Trials Kind . of eggs
Egg treatment
•vr
No. microorganisms per shell (log)
No. showing mold
made
Growth Rots
Molds Av.
High
Bacteria Low
Av.
High
Low
None Dipped in Foamrex Dipped in oil
6 6 6
5 6 5
4 5 2
5.1 6.8 6.3
7.1 8.9 7.0
0 5.5 5.1
7.3 7.4 6.9
9.2 8.3 7.9
5.7 6.1 5.1
Soiled
None Dipped in Foamrex Dipped in oil
6 6 6
5 6 5
5 5 4
5.2 6.6 5.3
6.8 8.3 7.8
0 5.6 0
7.5 7.3 6.7
9.1 9.0 8.7
5.7 6.1 4.8
Washed
None Dipped in Foamrex Dipped in oil
6 6 6
5 5 5
4 4 4
5.6 6.7 6.5
7.3 8.3 7.2
0 4.0 5.6
7.3 7.7 6.9
8.7 8.2 8.1
4.8 6.8 5.2
TABLE 3.—Fungicidal and bactericidal properties of some fungicides in Foamrex and oil Percent of trials showing Fungicides >
Eggs a n d treatments
Mi croorgamsms per shell (lc3g of count) Bacteria
Molds Mold
Rot Ave.
High
Low
Ave.
High
Low
7.2 8.2
4.8
2 2 2 2 2 2 2 2
50 50 50 0 0 50 0 0
50 50 50 0 50 0 50 0
3.5 7.0 6.7 0 6.4 0 2.5 0
7.1 7.2 7.2 0 7.0 0 4.9 0
0 6.9 6.1 0 5.9 0 0 0
6.0 8.2 2.7 5.9 2.3 6.6 0
5.4 5.9 4.7 7.2 0
0 5.8 0 5.9 0
100 100 50 0 0 0 0
6.3 6.5 6.6 0 0 0 0 5.4
6.2 6.4 5.6 0 0 0 0 0
7.5 8.0 5.7 4.8 5.4 3.4 3.9 2.2
7.8 8.0 6.2 4.9 6.0 6.8 7.8 4.4
5.2 4.7 3.8 0.0 0.0 0.0
Cetab
Control Control Control Dipped Dipped Dipped Dipped Dipped
B104
Control Control dipped in Foamrex Control dipped in oil Dipped in B 104 Dipped in B 104 then Foamrex Dipped in B 104 then oil Dipped in Foamrex containing B 104 Dipped in oil containing B 104
2 2 2 2 2 2 2 2
100 100 100 0 0 0 0 0
• o
6.25 6.4 6.1 0 0 0 0 2.7
Control Control dipped in Foamrex Control dipped in oil Dipped in Duomeen T Dipped in Duomeen then in Foamrex Dipped in Duomeen then in oil Dipped in Foamrex containing D . T . Dipped in oil containing D . T .
4 4 4 4 4 4 4 4
100 100 100 0 0 0 25 0
100 100 75 75 50 75 50 50
6.7 7.0 6.5 2.9 2.4 3.0 3.2 3.0
7.3 8.3 7.1 6.0 4.9 7.0 8.4 6.3
6.2 6.4 5.6 0.0 0.0 0.0 0.0 0.0
8.0 7.6 6.9 5.8 5.8 6.4 4.8 4.5
8.7 8.0 8.2 7.3 6.2 6.8 7.7 7.3
7.2 6.8 5.2 4.9 5.5 6.2 0.0 0.0
2 2 2 2 2 2 2 2
50 50 50 0 0 0 0 0
50 50 50 0 50 0 0 50
3.5 7.0 6.7 0 3.9 0 0 3.0
7.1 7.2 7.2 0 6.7 0 0 5.9
0.0 6.9 6.1 0 5.9 0 0 0
6.0 8.2
7.2 8.2
4.8
Ceeprin
Control Control dipped in Foamrex Control dipped in oil Dipped in fungicide Dipped in fungicide then Foamrex Dipped in fungicide then oil Dipped in Foamrex containing fungicide Dipped in oil containing fungicide
0 6.3 2.4 0 3.1
0 6.4 4.7 0 6.1
0 6.2 0 0 0
Arquad
Control Control in Foamrex Control in oil Fungicide Fungicide then Foamrex Fungicide then oil Fungicide in Foamrex Fungicide in oil
4 4 4 4 4 4 4 4
100 100 100 0 25 0 50 0
100 100 75 SO 75 25 75 25
6.7 7.0 6.5 2.9 3.3 2.7 3.2 3.7
7.3 8.3 7.1 6.1 7.3 5.8 7.2 5.8
6.2 6.4 5.6 0 0 0 0 0
8.0 7.6 6.9 6.8 7.9 5.6 6.6 4.8
8.7 8.0 8.2 7.4 8.4 7.1 7.2 7.7
7.2 6.8 5.2 6.2 7.5 4.7 5.9 0
Shirlan Extra
Control Control in Foamrex Control in oil Fungicide Fungicide then Foamrex Fungicide then oil Fungicide in Foamrex Fungicide in oil
4 4 4 4 4 4 4 4
75 100 100 0 25 25 0 50
75 75 75 25 50 75 0 25
5.3 7.3 6.8 2.9 5.2 2.9 1.6 4.3
7.3 8.3 7.2 6.0 7.2 5.8 6.5 6.5
0 6.7 6.1 0 0 0 0 0
7.3 7.6 8.1 6.3 6.9 7.0 4.6 3.2
8.7 8.2 8.2 7.9 7.7 7.8 .8.4 6.6
4.8 6.9 8.1 5.2 6.5 0 0 0
Duomeen
untreated dipped in Foamrex dipped in oil in fungicide in fungicide then Foamrex in fungicide then oil in Foamrex containing fungicide in oil containing fungicide
No. of trials
—
—
—
—
— —
7.2
—
— —
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Clean
1120
P. CLEMENTS AND A. R. WINTER
SUMMARY
Molds grow readily on eggs treated with oil or Foamrex, within a period of 15 days, when held under favorable conditions. Some of the fungicides tested have bactericidal properties but are not efficient germicides for use in oil or Foamrex.
Some fungicides, incorporated in oil or Foamrex at 0.25 percent concentration, inhibit mold growth on shell-treated eggs. Cetab (cetyl trimethyl ammonium bromide), Ceepryn (cetyl pyridinium chloride) and Shirlan Extra (salicylanilide) were efficient fungicides for use in oil for treating shell eggs. B 104 (di-duodecnyl dimethyl ammonium chloride) was an effective fungicide for use in both oil and Foamrex for treating shell eggs. REFERENCES Bart, G., 1949. The control of fungi found on poultry eggs in storage. M. A. Thesis, The Ohio State University. Brown, H. J., and N. E. Gibbons, 1954. Air cell mold in oiled eggs. Food Tech. 8:307-11. Cotterill, O. J., 1948. The preservation of shell egg quality by oil treatment. M.S. Thesis, The Ohio State University. Funk, E. M., 1950. Maintenance of quality in shell eggs by thermostabilization. Missouri Agr. Exp. Sta. Res. Bull. 467. Funk, E. M., 1955. Treating shell eggs to maintain quality. North Central Regional Publication No. 62 Mo. Bull. 659. Lorah, M. E., E. M. Funk and J. Forward, 1954. Reducing Spoilage in shell eggs by the use of fungicides. Missouri Agr. Exp. Sta. Res. Bull. 573. Mailman, W. L., and J. A. Davidson, 1944. Oilprotected shell eggs. U.S. Egg and Poultry Mag. 50:113. McFarland, C. R., 1950. The effects of germicides in mineral oil used to shell-treat storage eggs. M.S. Thesis, The Ohio State University. Puffenbarger, D. W., 1951. Preservation of egg quality by shell treatment. M.S. Thesis, The Ohio State University. Rosser, F. T., W. H. White, A. H. Woodcock and D. A. Fletcher, 1942, Preservation of eggs. I. Treatment for maintenance of quality in shell eggs at ordinary temperatures. Can. J. Res. 20 D : 57-70. Yushok, W. D., and A. L. Romanoff, 1949. Studies on preservation of shell eggs by coating with plastics. Food Res. 14:113-122. Yushok, W. D. and A. L. Romanoff, 1950. Prevention of mold growth in intact eggs by treat-
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some bactericidal properties, reducing the bacterial count in the washings about 40 percent. Duomeen T prevented visible mold growth on the shells, reduced spoilage and mold counts in the shell washings about 55 percent and bacteria about 27 percent. Ceepryn eliminated mold growth on egg shells when used alone or in combination with oil in 0.25 percent concentration. It reduced bacterial counts when used as above. Ceepryn was ineffective as a fungicide or germicide when used in connection with Foamrex. Bart (1949), McFarland (1950) and Yushok and Romanoff (1950) have also reported that Ceepryn is an effective fungicide for treating shell eggs. Arquad practically eliminated visible mold growth on egg shells, reducing spoilage and the mold count about 50 percent. It had little bactericidal effect. Arauad was more effective when used in connection with oil than with Foamrex. Shirlan Extra eliminated visible mold growth on egg shells when used alone or in combination with oil in 0.25 percent concentration. It eliminated mold counts 100 percent from shell washings in 9 of 18 trials and bacteria in 6 to 18 trials. Shirlan Extra was effective in most trials when used with either oil or Foamrex. The fact that it was not completely effective in all trials may have been due to the type of microflora present. The findings, that Shirlan Extra is a good fungicide for treating shell eggs, is in agreement with the earlier reports by Young (1948), Bart (1949) and McFarland (1950).
CONTROL OF MOLDS ON SHELL EGGS ment with a cationic detergent. Food Res. IS: 1-7. Winter, A. R., and O. J. Cotterilll, 1949. The preservation of shell egg quality by oil dipping right on the farm. Amer. Egg and Poultry Rev. 10: (3) 14-16. Winter, A. R., P. Shields, L. MacDonald, and I. Prudent, 1954. Improving the keeping quality of eggs during marketing by hot oil treatment.
1121
Food Tech. 8:515-18. i Winter, A. R., B. Burkart, P. Clements and L. MacDonald, 1955. Cleaning eggs with detergents and detergent-sanitizers. Ohio Agr. Exp. Sta. Res. Bull. 762. Young, H. E., 1948, Blue case timber and mold rots in hens eggs, Queensland J. Agric. Sci. 5:125-40.
(Continued from page 1099) physiology, effects of antibiotics on blood formation, pharmacology of antibiotics, effects on growth of human subjects, miscellaneous nutritional effects, effects on guinea pigs, public health aspects of feeding antibiotics to animals, and conclusions.
This book is an excellent presentation of this field. It presents and weighs the evidence with judicial choice of facts and figures. This is not easy to accomplish, for the literature is full of contradictions and opposing theories, but it has been well done in this book.—H.D.B.
NEWS AND NOTES NOMINATIONS—PENB ACHIEVEMENT AWARD The Poultry and Egg National Board Research Achievement Award is given annually for outstanding research leading to the increased consumption of poultry and eggs. The recipient will receive an appropriate plaque and will be honored at the annual meeting of PENB, January 31-February 1, 1957. Selection of the winner is based on the importance of poultry and/or egg research contributions concerning processing, quality preservation, nutritive value, and new and improved product development, leading to the increased use of poultry and eggs in the diet. Nominations should be supported by biographical sketches of the nominees, including enumeration of contributions pertinent to the Award objectives, a list of publications during the past ten years, and, if available, endorsement or seconds. Nominations should be sent to Hans Lineweaver, Chairman of the PENB Technical Advisory Committee, Western Utilization Research Laboratory, Albany 10, California, not later than December 3, 1956. NEWMAN MEMORIAL AWARD Dr. B. R. Burmester, Biologist of the Regional Poultry Research Laboratory, Animal and Poultry Husbandry Research Branch, Agricultural Research Service, U. S. Department of Agriculture, East (Continued
Lansing, Michigan, received the Tom Newman Memorial International Award for Poultry Husbandry Research for 1956. This Award consists of a medal and i50. sterling. It is given for the most important contribution to poultry husbandry resarch work published in the previous year. The Award was made by the Committee, to Dr. Burmester, primarily for his poultry disease research work, with particular reference to lymphomatosis. The papers cited were: Immunity to Visceral Lymphomatosis in Chicks Following Injection of Virus into Dams, published in the Proceedings of the Society for Experimental Biology and Medicine; In Vitro and in Vivo Neutralization of the Virus of Visceral Lymphomatosis, published in the Proceedings of the Society for Experimental Biology and Medicine; The Presence of the Virus of Visceral Lymphomatosis in Embryonated Eggs of Normal Appearing Hens, published in Poultry Science; Tumor Incidence in the Progeny of Hens Repeatedly Injected as Adults with Visceral Lymphomatosis Virus, published in Poultry Science; The Propagation of the Virus of Visceral Lymphomatosis in Embryonated Eggs, published in Poultry Science; Plasma Phosphatase Activities of Normal and Lymphomatous Chickens, published in Cancer Research; and The Role of the Infected Egg in the Transmission of Visceral Lymphomatosis, published in Poultry Science. Dr. Burmester was born in Petaluma, California, and obtained a B.S. degree, majoring in Poulon page 1124)
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