Читать книгу: «Experiments on the Spoilage of Tomato Ketchup», страница 4
STERILITY OF KETCHUP
To determine the sterility of ketchup, cultures were made from 77 of the bottles. The method used was to wipe the bottles and cork stoppers with a damp towel and then remove the cork. The cork puller which was used grasps the neck of the bottle in such a way as to cover the opening and remove the cork without the inrush of air that occurs when the ordinary corkscrew is used. A flame was then passed over the mouth of the bottle, after which the upper layer of ketchup was poured out, so as to discard any material which might have been contaminated in handling. Tomato gelatin was used as a medium and cultures were made in petri dishes.
There were 17 plates on which no organisms developed, indicating that the ketchup was sterile. Of the 60 plates having organisms, 54 had molds, 22 of these having molds alone; 21 plates had yeast-like organisms, 3 plates having these only; 29 plates had bacteria, 4 having bacteria alone. Sometimes a plate would have only one form of organism, but more often there was a mixture present. Of 15 plates having only one form of organism, 3 had yeast alone, 2 bacteria alone, and 10 had mold alone. Of the 77 bottles of ketchup from which the inoculations were made, 41 were without and 36 with preservative, and of the 17 sterile ketchups, 8 contained sodium benzoate and 9 were without preservative.
A considerable part of the experimental ketchup proved not to be sterile. The organisms present were of the class which require oxygen for their growth and therefore they had only been arrested in their activity. No growth could take place so long as the air was excluded and therefore no spoilage could occur. When the cork was drawn, the organisms could grow and cause spoilage, and this is a much more potent factor than the entrance of germs from without. Bottling and sealing the ketchup quickly while hot so completely excludes the air that only a few colonies of yeast or mold may be found on subsequent microscopical examination. Filling at a low temperature and corking while cool allows sufficient air to remain incorporated in the ketchup and neck of the bottle to permit a considerable growth of the organisms and a product derived from good stock may thus acquire the appearance of ketchup derived from partially decayed material. A ketchup in which bubbles of air are incorporated in filling may show a growth of mold at each bubble throughout the mass. The foregoing statements apply to ketchup containing sodium benzoate as well as to the non-preservative goods of the character used in these experiments.
EXPERIMENTS WITH PRESERVATIVES
SODIUM BENZOATE
The preservative in general use in ketchup is sodium benzoate. Salicylic acid is used, but only to a limited extent. The amount of sodium benzoate used, according to the labels, varies from one-sixteenth to one-tenth of 1 per cent; but on some labels the amount is not stated. Experiments were made to determine the amount necessary to check the spoilage of ketchup.
Two organisms, a mold and a yeast, were selected on which to make the tests. The mold was the ordinary blue mold, Penicillium, which was present in many of the brands of ketchup and is found commonly on acid foods. It was selected on account of its prevalence and resistive power. The yeast was obtained from ketchup and was also a vigorous grower, forming a thick, wrinkled film on various media. Any effect on the growth of the yeast could be seen readily in its manner of forming the film.
Portions of tomato gelatin to which 0.1, 0.5, 1, and 2 per cent, respectively, of sodium benzoate were added, were first inoculated with the mold. There was no development in those containing 1 and 2 per cent; a retarded development resulted in that containing 0.5 per cent, and the growth when 0.1 per cent was used was nearly normal, showing very little difference from that in the gelatin without sodium benzoate.
Ketchup was next tried as a medium, but the amount of benzoate was reduced to one-sixteenth, one-twelfth, and one-tenth of 1 per cent, as it was thought that some of the other constituents of the ketchup were antiseptic to a slight degree. The growth in the ketchup was irregular, though the benzoate checked development in all. Equal amounts of benzoate were used in tomato bouillon, with practically the same results as in the ketchup. The development was checked in all, and in some plates one-sixteenth of 1 per cent seemed to be fully as efficacious as one-tenth of 1 per cent. When the mold was examined under the microscope, the filaments were found to be much swollen and distorted in shape, and filled with a coarsely granular protoplasm, containing much fat, as indicated by the blackening with osmic acid. The culture containing the mold which gave the least development seemed to show the least distortion and swelling of the filaments.
The results indicated that in using sodium benzoate as a preservative there is uncertainty as to results, even when using the maximum amount allowed – one-tenth of 1 per cent. They also indicated that this preservative had an injurious effect on the living matter of the mold. (See Pl. II; compare with normal growth, Pl. I.)
SALT
The effect of salt in checking development was tested by using tomato bouillon as a medium and adding 5, 10, 15, 20, 25, and 30 grams of salt, respectively, to 100 cc. These were inoculated with the mold. The 5-gram solution seemed to have no effect on development. When 10 grams were used growth appeared as soon as in the bouillon without salt, but was not so extensive. In the 15-gram solution growth was retarded four days, and most of that which did develop remained submerged, the mold growing normally on the surface. With 20 grams the growth was five days slower than the normal in starting, and after that there was only a slight development. In the 25-gram solution, the growth started at the same time as when 20 grams were employed, but remained stationary, while with the 30-gram solution, no development occurred.
The yeast was checked slightly by 5 grams, and very materially by the 10-gram solution, as it required two days for a thin, delicate film to form, whereas in ordinary solutions a rather thick film is formed within twenty-four hours or even in less time. There was no development in the 15-gram solution.
SUGAR
The effect of sugar was tested on both the mold and the yeast by adding it to tomato bouillon. It was supposed that a low percentage of sugar like the salt would plasmolyze the cells, and in this way check growth, but it seemed to have no effect until the amount was increased to 25 grams per 100 cc of bouillon. In this solution growth appeared as soon as with the weaker solutions, but there was a smaller amount. In the 25 to 40 gram solutions there was less development as the amount of sugar increased. In the 70 and 75 gram solutions growth was delayed one day in its appearance. In the 80, 85, and 90 gram solutions growth was delayed two days, the colonies growing submerged at first, but after a time forming on the surface. The mycelium remained very thin, but a thick layer of spores formed. From this point on the amounts were increased by 10 grams up to 200. The development became slower and less successively until 170 grams were added. In this case a small colony appeared on the surface in seven days, but seemed to grow less after that. The solutions were held, and in time crystals separated from the thick sirups. After two months dry-looking colonies developed along the edges, forming a ring, and some formed on the surface, these occurring also in the flasks containing 170, 180, 190, and 200 grams of sugar per 100 cc. The colonies were a dull greenish drab in spots, the remainder being white.
For the yeast the 80-gram solution of sugar was the strongest in which any development took place.
SPICES
Experiments to determine the value of the spices as antiseptics were made, using water infusions, acetic-acid extracts, and oil extracts.
WATER INFUSIONS
In making the water infusions 20 grams of the whole spices, with 200 cc of water, were boiled for forty-five minutes. This is approximately the length of time that the spices are cooked in the ketchup in the factory. The liquid was then filtered and from 0.1 to 5 cc of the filtrate was used in 10 cc of tomato bouillon. The same organisms were used as in the former experiments.
The tests showed that cinnamon and cloves were the strongest antiseptically. These checked growth when used in small amounts, but it required 3 cc of the cinnamon and 1 cc of the cloves to inhibit the growth of the mold. Mustard, paprika, and cayenne pepper checked growth also, but 5 cc, the highest strength used, did not inhibit growth. The ginger, mace, and black pepper had no apparent effect in the quantities used.
The effect of the spices on the development of the yeast was somewhat different from their effect on Penicillium. The cinnamon showed the strongest action, 3 cc being effective, whereas 5 cc of the cloves was required. The cayenne pepper came next in effectiveness, and after that the black pepper. The ginger, mace, and mustard solutions had no effect in the strengths used.
The remainder of the spice infusions were kept in glass-stoppered bottles in the laboratory, and in a few weeks’ time there was a coating of mold formed over the surface of the mace, the mustard, and the black and cayenne peppers. The paprika had small, stunted colonies dotting the surface.
At the time that these experiments were made a quantity of the ground spices were placed in large petri dishes and water was added to make a heavy paste. One set of these was inoculated with the mold, and another set with the yeast, and all were kept in a warm place. No development of either organism appeared on the cinnamon, cloves, or mustard; on the others a growth first showed in three days. On a normal medium growth appears in twenty-four hours. On the mace, paprika, and cayenne pepper the Penicillium and yeast with which the pastes were inoculated were overgrown in a few days with black mold (Rhizopus nigricans).
