EXPERIMENTS ON THE SPOILAGE OF TOMATO KETCHUP TEXT

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_). ACETIC-ACID EXTRACTS. In the manufacture of ketchup acetic-acid extracts of the spices are sometimes used instead of the whole spices, on account of their supposed antiseptic properties as well as their greater strength and convenience in handling. One minim of the standard acetic-acid extracts is equal in strength to 1 grain of the whole spices. The acid extracts obtained included allspice, celery, cloves, coriander, garlic, and black pepper. In the tests 0.1, 0.2, 0.3, 0.4, 0.5, and 1 cc, respectively, of the extract was added to 10 cc of tomato bouillon. One set was inoculated with the mold and another set with the yeast. In the case of the mold, no growth occurred with the allspice and cloves; the celery checked the growth materially, there being no indication of mold until the sixth day. Normally a fairly strong growth occurs in twenty-four hours. In the solution containing 0.3 cc there was only one small colony in thirteen days, and no further development. In the solution containing the coriander, the growth in the 0.5 cc solution did not appear for three days, the 1 cc solution showing no growth. The garlic had practically the same effect as the coriander, while the black pepper was stronger, no growth appearing in the solution containing 0.5 cc. The yeast was slightly stronger in resisting the effect of the extracts. No growth appeared with the allspice and cloves; 0.5 cc of the celery and 1 cc of the coriander were required to inhibit growth, and the garlic and black pepper gave similar results, a weak development occurring in the solutions containing 1 cc. OIL EXTRACTS. Oil extracts of the spices were tested in the same manner as the water infusions and the acetic-acid extracts. The oils were so strong that in order to handle them easily they were mixed with equal volumes of alcohol, except that the mace, which was in the form of a paste, was mixed with two-thirds its volume of alcohol. To 10 cc of tomato bouillon were added 0.1, 0.2, 0.3, 0.4, and 0.5 cc, respectively, of the oils of cinnamon, cloves, mace, mustard, and black pepper. In the case of the mold, there was no development in the solutions containing cinnamon, cloves, and mustard; in those containing mace and black pepper the development was slower than the normal, that in the black pepper being more pronounced. On the yeast the effect was similar, no development occurring in the cinnamon, cloves, and mustard, and a retarded development taking place in the mace and black pepper, that in the black pepper being the more pronounced. The experiments show that some of the spices, notably allspice, cinnamon, and cloves have decided antiseptic value, but that the peppers are not as valuable as is generally supposed. The oil extracts have been advocated for use in ketchup instead of the whole spices, but in quantities which would be useful antiseptically their use would be objectionable, for when present in approximately the same proportions as are the whole-spice infusions, the flavor is too strong and masks the more delicate flavor of the tomato. The acetic-acid extracts are more effective than are the water infusions, and they are not objectionable in the ketchup. VINEGAR AND ACETIC ACID. An experiment was made to determine the antiseptic value of vinegar and acetic acid. Commercial 50-grain distilled vinegar was used. It was found that when 30 per cent of this vinegar was added to the tomato bouillon the development of mold was checked and the extent to which it was checked increased with the increased amounts of vinegar. The development in the solution containing 30 per cent of the vinegar was two days later than the normal in starting, while the solution containing 100 per cent was eleven days delayed and showed but little growth. An 80 per cent solution of glacial acetic acid was used. One-half of 1 per cent added to the tomato bouillon checked growth to the same extent as 30 per cent of vinegar, and no development occurred when the quantity was increased to 2 per cent. Experiments were then made in which vinegar was added to the ketchup in proportions varying from 1 part in 32 to 1 part in 8, with the result of greatly delaying the appearance of the mold as the proportion increased. With the increase in vinegar it was necessary to add sugar and slightly more spices to overcome the pungency of the acid and thus insure good flavor. The addition of the vinegar to the pulp had the effect of arresting the action of the oxidase and thus the bright color was maintained. The usual custom in factory practice is to add the vinegar near the close of the cooking process otherwise a considerable portion of the acid will be driven off. This practice was followed in the experimental work, but it has since been found that continued heating in the presence of the acid has some effect upon sterilization, and therefore the increased amount of vinegar is effective not only because of the additional acid present, but also because the heating in the after process is thereby rendered more efficacious. This line of experiments gives promise of practical results in producing a ketchup which will not only keep while in the bottle, but will also keep longer after it is opened. Each manufacturer must work out the quantities that could be used with his formula and still retain the character of his goods. OIL. In ketchup manufacturing it is customary, if an agitator is not used, to put a small amount of fat in the kettle to check the ebullition during the reduction of the pulp. The amount used in this manner is not sufficient, however, to be apparent in the ketchup. Brannt[D] states that in some factories, where the trimmings are allowed to accumulate for the season, they are given liberal doses of oils and condiments when cooked, in order to disguise their defects, so that the product can be placed on the market as “fresh tomato catchup.” That the use of oils is increasing is evident from the comparison of the ketchup of the past season with that of former years. [D] Brannt, W. L., A Practical Treatise on the Manufacture of Vinegar, 1900, p. 455. When oil is used in ketchup, it is easily detected under the microscope, as it appears in the form of shining, yellow globules which blacken gradually when treated with osmic acid. Besides this, the oil comes to the surface of the ketchup, where it can be seen readily, and if considerable oil has been used a distinct layer is formed. When the ketchup has been made for some time, the oil changes so that the ketchup has a peculiar “greasy” odor, or the oil may be so changed as to give a decidedly rancid smell to the ketchup. Oil usually causes a deterioration in flavor and odor, though some of the ketchups to which it has been added do not spoil readily. Olive oil, cottonseed oil, and oleomargarine are used. That the oil is not considered one of the regular known ingredients of the ketchup is shown by the failure to declare its presence on the label. To test the antiseptic value of oils in ketchup, experiments were made, using olive oil, cottonseed oil, and oleomargarine in the proportions of 1 part of oil to 1,000, 750, and 500 parts of ketchup, respectively. The ketchup was made in small quantities, 2 gallons for each experiment. After bottling, all except the check bottles were inoculated with Penicillium and kept at kitchen temperature. All spoiled, and neither the quantity nor kind of oil used had any marked effect in preventing spoilage. That the oils affected the development of the mold was evident. The mold developed first at the junction of the ketchup with the bottle forming a ring which spread gradually over the surface developing a somewhat heavy mycelium. This remained white longer than usual, spores forming very gradually, as indicated by the change in color from white to a delicate blue. At the end of three weeks only spots of color appeared on the surface and these were still blue, though in ordinary development the blue color changes to green in two or three days. Another test was made, using olive oil only, and in the proportions of 1 part of oil to 500, 400, and 300 parts, respectively, of the ketchup. Reduction was made in a steam-jacketed kettle, the oil being added when the ebullition of the ketchup was the strongest, after which the boiling was continued for fifteen minutes. The ketchup was bottled, unsterilized bottles being used, then covered loosely with the metal caps. The time required for the ketchup to spoil was longer than in the first set, but there was not sufficient difference nor enough uniformity in the time to indicate that the use of oil in ketchup is desirable, even if the change of flavor and odor be not taken into consideration. The average number of days before spoilage for those containing 1 part of oil to 500 parts of ketchup, was thirteen and two-thirds days; one has not yet spoiled (a period of forty-five days), while the first bottle spoiled in four days. Those having 1 part of oil to 400 parts of ketchup had an average life of nine and three-fourths days, the minimum being three days, and the maximum twenty-six days. Those having 1 part to 300 parts of ketchup on an average did not spoil for six and three-fourths days, the minimum being four days, and the maximum eleven days. The failure of some of the bottles to spoil, though similar in every known respect to those which did spoil, is a feature peculiar to ketchup and is familiar to manufacturers who make careful tests before putting their product on the market. For this reason a rather large number of bottles should be used in a test in order that the results may be approximately accurate and represent general conditions. STUDY OF PENICILLIUM IN KETCHUP. Penicillium is a plant which is distributed widely and apparently is able to grow wherever organic matter is found, though flourishing best when the material contains acid. It causes loss in canneries, breweries, distilleries, etc., the only use made of it being in the manufacture of Roquefort cheese, the immature cheese being inoculated with the conidia for the effect the mold produces in the maturing process.