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The Story Of Germ Life
by H. W. Conn
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Occasionally bacteria are of some value in food products. The gamy flavour of meats is nothing more than incipient decomposition. Sauer Kraut is a food mass intentionally allowed to ferment and sour. The value of bacteria in producing butter and cheese flavours is noticed elsewhere. But commonly our aim must be to prevent the growth of bacteria in foods. Foods must be dried or cooked or kept on ice, or some other means adopted for preventing bacterial growth in them. It is their presence that forces us to keep our ice box, thus founding the ice business, as well as that of the manufacture of refrigerators. It is their presence, again, that forces us to smoke hams, to salt mackerel, to dry fish or other meats, to keep pork in brine, and to introduce numerous other details in the methods of food preparation and preservation.



CHAPTER III.

RELATION OF BACTERIA TO THE DAIRY INDUSTRY.

Dairying is one of the most primitive of our industries. From the very earliest period, ever since man began to keep domestic cattle, he has been familiar with dairying. During these many centuries certain methods of procedure have been developed which produce desired results. These methods, however, have been devised simply from the accumulation of experience, with very little knowledge as to the reasons underlying them. The methods of past centuries are, however, ceasing to be satisfactory. The advance of our civilization during the last half century has seen a marked expansion in the extent of the dairy industry. With this expansion has appeared the necessity for new methods, and dairymen have for years been looking for them. The last few years have been teaching us that the new methods are to be found along the line of the application of the discoveries of modern bacteriology. We have been learning that the dairyman is more closely related to bacteria and their activities than almost any other class of persons. Modern dairying, apart from the matter of keeping the cow, consists largely in trying to prevent bacteria from growing in milk or in stimulating their growth in cream, butter, and cheese. These chief products of the dairy will be considered separately.

SOURCES OF BACTERIA IN MILK.

The first fact that claims our attention is, that milk at the time it is secreted from the udder of the healthy cow contains no bacteria. Although bacteria are almost ubiquitous, they are not found in the circulating fluids of healthy animals, and are not secreted by their glands. Milk when first secreted by the milk gland is therefore free from bacteria. It has taken a long time to demonstrate this fact, but it has been finally satisfactorily proved. Secondly, it has been demonstrated that practically all of the normal changes which occur in milk after its secretion are caused by the growth of bacteria. This, too, was long denied, and for quite a number of years after putrefactions and fermentations were generally acknowledged to be caused by the growth of micro- organisms, the changes which occurred in milk were excepted from the rule. The uniformity with which milk will sour, and the difficulty, or seeming impossibility, of preventing this change, led to the belief that the souring of milk was a normal change characteristic of milk, just as clotting is characteristic of blood. This was, however, eventually disproved, and it was finally demonstrated that, beyond a few physical changes connected with evaporation and a slight oxidation of the fat, milk, if kept free from bacteria, will undergo no change. If bacteria are not present, it will remain sweet indefinitely.

But it is impossible to draw milk from the cow in such a manner that it will be free from bacteria except by the use of precautions absolutely impracticable in ordinary dairying. As milk is commonly drawn, it is sure to be contaminated by bacteria, and by the time it has entered the milk pail it contains frequently as many as half a million, or even a million, bacteria in every cubic inch of the milk. This seems almost incredible, but it has been demonstrated in many cases and is beyond question. Since these bacteria are not in the secreted milk, they must come from some external sources, and these sources are the following:

The first in importance is the cow herself; for while her milk when secreted is sterile, and while there are no bacteria in her blood, nevertheless the cow is the most prolific source of bacterial contamination. In the first place, the milk ducts are full of them. After each milking a little milk is always left in the duct, and this furnishes an ideal place for bacteria to grow. Some bacteria from the air or elsewhere are sure to get into these ducts after the milking, and they begin at once to multiply rapidly. By the next milking they become very abundant in the ducts, and the first milk drawn washes most of them at once into the milk pail, where they can continue their growth in the milk. Again, the exterior of the cow's body contains them in abundance. Every hair, every particle of dirt, every bit of dried manure, is a lurking place for millions of bacteria. The hind quarters of a cow are commonly in a condition of much filth, for the farmer rarely grooms his cow, and during the milking, by her movements, by the switching of her tail, and by the rubbing she gets from the milker, no inconsiderable amount of this dirt and filth is brushed off and falls into the milk pail The farmer understands this source of dirt and usually feels it necessary to strain the milk after the milking. But the straining it receives through a coarse cloth, while it will remove the coarser particles of dirt, has no effect upon the bacteria, for these pass through any strainer unimpeded. Again, the milk vessels themselves contain bacteria, for they are never washed absolutely clean. After the most thorough washing which the milk pail receives from the kitchen, there will always be left many bacteria clinging in the cracks of the tin or in the wood, ready to begin to grow as soon as the milk once more fills the pail The milker himself contributes to the supply, for he goes to the milking with unclean hands, unclean clothes, and not a few bacteria get from him to his milk pail. Lastly, we find the air of the milking stall furnishing its quota of milk bacteria. This source of bacteria is, how ever, not so great as was formerly believed. That the air may contain many bacteria in its dust is certain, and doubtless these fall in some quantity into the milk, especially if the cattle are allowed to feed upon dusty hay before and during the milking. But unless the air is thus full of dust this source of bacteria is not very great, and compared with the bacteria from the other sources the air bacteria are unimportant.

The milk thus gets filled with bacteria, and since it furnishes an excellent food these bacteria begin at once to grow. The milk when drawn is warm and at a temperature which especially stimulates bacterial growth. They multiply with great rapidity, and in the course of a few hours increase perhaps a thousandfold. The numbers which may be found after twenty-four hours are sometimes inconceivable; market milk may contain as many as five hundred millions per cubic inch; and while this is a decidedly extreme number, milk that is a day old will almost always contain many millions in each cubic inch, the number depending upon the age of the milk and its temperature. During this growth the bacteria have, of course, not been without their effect. Recognising as we do that bacteria are agents for chemical change, we are prepared to see the milk undergoing some modifications during this rapid multiplication of bacteria. The changes which these bacteria produce in the milk and its products are numerous, and decidedly affect its value. They are both advantageous and disadvantageous to the dairyman. They are nuisances so far as concerns the milk producer, but allies of the butter and cheese maker.

THE EFFECT OF BACTERIA ON MILK.

The first and most universal change effected in milk is its SOURING. So universal is this phenomenon that it is generally regarded as an inevitable change which can not be avoided, and, as already pointed out, has in the past been regarded as a normal property of milk. To-day, however, the phenomenon is well understood. It is due to the action of certain of the milk bacteria upon the milk sugar which converts it into lactic acid, and this acid gives the sour taste and curdles the milk. After this acid is produced in small quantity its presence proves deleterious to the growth of the bacteria, and further bacterial growth is checked. After souring, therefore, the milk for some time does not ordinarily undergo any further changes.

Milk souring has been commonly regarded as a single phenomenon, alike in all cases. When it was first studied by bacteriologists it was thought to be due in all cases to a single species of micro-organism which was discovered to be commonly present and named Bacillus acidi lactici (Fig. 19). This bacterium has certainly the power of souring milk rapidly, and is found to be very common in dairies in Europe. As soon as bacteriologists turned their attention more closely to the subject it was found that the spontaneous souring of milk was not always caused by the same species of bacterium. Instead of finding this Bacillus acidi lactici always present, they found that quite a number of different species of bacteria have the power of souring milk, and are found in different specimens of soured milk. The number of species of bacteria which have been found to sour milk has increased until something over a hundred are known to have this power. These different species do not affect the milk in the same way. All produce some acid, but they differ in the kind and the amount of acid, and especially in the other changes which are effected at the same time that the milk is soured, so that the resulting soured milk is quite variable. In spite of this variety, however, the most recent work tends to show that the majority of cases of spontaneous souring of milk are produced by bacteria which, though somewhat variable, probably constitute a single species, and are identical with the Bacillus acidi lactici (Fig. 19). This species, found common in the dairies of Europe, according to recent investigations occurs in this country as well. We may say, then, that while there are many species of bacteria infesting the dairy which can sour the milk, there is one which is more common and more universally found than others, and this is the ordinary cause of milk souring.

When we study more carefully the effect upon the milk of the different species of bacteria found in the dairy, we find that there is a great variety of changes which they produce when they are allowed to grow in milk. The dairyman experiences many troubles with his milk. It sometimes curdles without becoming acid. Sometimes it becomes bitter, or acquires an unpleasant "tainted" taste, or, again, a "soapy" taste. Occasionally a dairyman finds his milk becoming slimy, instead of souring and curdling in the normal fashion. At such times, after a number of hours, the milk becomes so slimy that it can be drawn into long threads. Such an infection proves very troublesome, for many a time it persists in spite of all attempts made to remedy it. Again, in other cases the milk will turn blue, acquiring about the time it becomes sour a beautiful sky-blue colour. Or it may become red, or occasionally yellow. All of these troubles the dairyman owes to the presence in his milk of unusual species of bacteria which grow there abundantly.

Bacteriologists have been able to make out satisfactorily the connection of all these infections with different species of the bacteria. A large number of species have been found to curdle milk without rendering it acid, several render it bitter, and a number produce a "tainted" and one a "soapy" taste. A score or more have been found which have the power of rendering the milk slimy. Two different species at least have the power of turning the milk to sky-blue colour; two or three produce red pigments (Fig. 20), and one or two have been found which produce a yellow colour. In short, it has been determined beyond question that all these infections, which are more or less troublesome to dairymen, are due to the growth of unusual bacteria in the milk.

These various infections are all troublesome, and indeed it may be said that, so far as concerns the milk producer and the milk consumer, bacteria are from beginning to end a source of trouble. It is the desire of the milk producer to avoid them as far as possible—a desire which is shared also by everyone who has anything to do with milk as milk. Having recognised that the various troubles, which occasionally occur even in the better class of dairies, are due to bacteria, the dairyman is, at least in a measure, prepared to avoid them. The avoiding of these troubles is moderately easy as soon as dairymen recognise the source from which the infectious organisms come, and also the fact that low temperatures will in all cases remedy the evil to a large extent. With this knowledge in hand the avoidance of all these troubles is only a question of care in handling the dairy. It must be recognised that most of these troublesome bacteria come from some unusual sources of infection. By unusual sources are meant those which the exercise of care will avoid. It is true that the souring bacteria appear to be so universally distributed that they can not be avoided by any ordinary means. But all other troublesome bacteria appear to be within control. The milkman must remember that the sources of the troubles which are liable to arise in his milk are in some form of filth: either filth on the cow, or dust in the hay which is scattered through the barn, or dirt on cows' udders, or some other unusual and avoidable source. These sources, from what we have already noticed, will always furnish the milk with bacteria; but under common conditions, and when the cow is kept in conditions of ordinary cleanliness, and frequently even when not cleanly, will only furnish bacteria that produce the universal souring. Recognising this, the dairyman at once learns that his remedies for the troublesome infections are cleanliness and low temperatures. If he is careful to keep his milk vessels scrupulously clean; if he will keep his cow as cleanly as he does his horse; and if he will use care in and around the barn and dairy, and then apply low temperatures to the milk, he need never be disturbed by slimy or tainted milk, or any of these other troubles; or he can remove such infections speedily should they once appear. Pure sweet milk is only a question of sufficient care. But care means labour and expense. As long as we demand cheap milk, so long will we be supplied with milk procured under conditions of filth. But when we learn that cheap milk is poor milk, and when we are willing to pay a little more for it, then only may we expect the use of greater care in the handling of the milk, resulting in a purer product.

Bacteriology has therefore taught us that the whole question of the milk supply in our communities is one of avoiding the too rapid growth of bacteria. These organisms are uniformly a nuisance to the milkman. To avoid their evil influence have been designed all the methods of caring for the dairy and the barn, all the methods of distributing milk in ice cars. Moreover, all the special devices connected with the great industry of milk supply have for their foundation the attempt to avoid, in the first place, the presence of too great a number of bacteria, and. in the second place, the growth of these bacteria.

BACTERIA IN BUTTER MAKING.

CREAM RIPENING.—Passing from milk to butter, we find a somewhat different story, inasmuch as here bacteria are direct allies to the dairyman rather than his enemies. Without being aware of it, butter makers have for years been making use of bacteria in their butter making and have been profiting by the products which the bacteria have furnished them. Cream, as it is obtained from milk, will always contain bacteria in large quantity, and these bacteria will grow as readily in the cream as they will in the milk. The butter maker seldom churns his cream when it is freshly obtained from the milk. There are, it is true, some places where sweet cream butter is made and is in demand, but in the majority of butter-consuming countries a different quality of butter is desired, and the cream is subjected to a process known as "ripening" or "souring" before it is churned. In ripening, the cream is simply allowed to stand in a vat for a period varying from twelve hours to two or three days, according to circumstances. During this period certain changes take place therein. The bacteria which were in the cream originally, get an opportunity to grow, and by the time the ripening is complete they become extremely numerous. As a result, the character of the cream changes just as the milk is changed under similar circumstances. It becomes somewhat soured; it becomes slightly curdled, and acquires a peculiarly pleasant taste and an aroma which was not present in the original fresh cream. After this ripening the cream is churned. It is during the ripening that the bacteria produce their effect, for after the churning they are of less importance. Part of them collect in the butter, part of them are washed off from the butter in the buttermilk and the subsequent processes. Most of the bacteria that are left in the butter soon die, not finding there a favourable condition for growth; some of them, however, live and grow for some time and are prominent agents in the changes by which butter becomes rancid. The butter maker is concerned with the ripening rather than with later processes.

The object of the ripening of cream is to render it in a better condition for butter making. The butter maker has learned by long- experience that ripened cream churns more rapidly than sweet cream, and that he obtains a larger yield of butter therefrom. The great object of the ripening, however, is to develop in the butter the peculiar flavour and aroma which is characteristic of the highest product. Sweet cream butter lacks flavour and aroma, having indeed a taste almost identically the same as cream. Butter, however, that is made from ripened cream has a peculiar delicate flavour and aroma which is well known to lovers of butter, and which is developed during the ripening process.

Bacteriologists have been able to explain with a considerable degree of accuracy the object of this ripening. The process is really a fermentation comparable to the fermentation that takes place in a brewer's malt. The growth of bacteria during the ripening produces chemical changes of a somewhat complicated character, and concerns each of the ingredients of the milk. The lactic-acid organisms affect the milk sugar and produce lactic acid; others act upon the fat, producing slight changes therein; while others act upon the casein and the albumens of the milk. As a result, various biproducts of decomposition arise, and it is these biproducts of decomposition that make the difference between the ripened and the unripened cream. They render it sour and curdle it, and they also produce the flavours and aromas that characterize it. Products of decomposition are generally looked upon as undesirable for food, and this is equally true of these products that arise in cream if the decomposition is allowed to continue long enough. If the ripening, instead of being stopped at the end of a day or two, is allowed to continue several days, the cream becomes decayed and the butter made therefrom is decidedly offensive. But under the conditions of ordinary ripening, when the process is stopped at the right moment, the decomposition products are pleasant rather than unpleasant, and the flavours and aromas which they impart to the cream and to the subsequent butter are those that are desired. It is these decomposition products that give the peculiar character to a high quality of butter, and this peculiar quality is a matter that determines the price which the butter maker can obtain for his product.

But, unfortunately, the butter maker is not always able to depend upon the ripening. While commonly it progresses in a satisfactory manner, sometimes, for no reason that he can assign, the ripening does not progress normally. Instead of developing the pleasant aroma and flavour of the properly ripened cream, the cream develops unpleasant tastes. It may be bitter or somewhat tainted, and just as sure as these flavours develop in the cream, so sure does the quality of the butter suffer. Moreover, it has been learned by experience that some creameries are incapable of obtaining an equally good ripening of their cream. While some of them will obtain favourable results, others, with equal care, will obtain a far less favourable flavour and aroma in their butter. The reason for all this has been explained by modern bacteriology. In the milk, and consequently in the cream, there are always found many bacteria, but these are not always of the same kinds. There are scores, and probably hundreds, of species of bacteria common in and around our barns and dairies, and the bacteria that are abundant and that grow in different lots of cream will not be always the same. It makes a decided difference in the character of the ripening, and in the consequent flavours and aromas, whether one or another species of bacteria has been growing in the cream. Some species are found to produce good results with desired flavours, while others, under identical conditions, produce decidedly poor results with undesired flavours. If the butter maker obtains cream which is filled with a large number of bacteria capable of producing good flavours, then the ripening of his cream will be satisfactory and his butter will be of high quality. If, however, it chances that his cream contains only the species which produce unpleasant flavours, then the character of the ripening will be decidedly inferior and the butter will be of a poorer grade. Fortunately the majority of the kinds of bacteria liable to get into the cream from ordinary sources are such as produce either good effects upon the cream or do not materially influence the flavour or aroma. Hence it is that the ripening of cream will commonly produce good results. Bacteriologists have learned that there are some species of bacteria more or less common around our barns which produce undesirable effects upon flavour, and should these become especially abundant in the cream, then the character of the ripening and the quality of the subsequent butter will suffer. These malign species of bacteria, however, are not very common in properly kept barns and dairies. Hence the process that is so widely used, of simply allowing cream to ripen under the influence of any bacteria that happen to be in it, ordinarily produces good results. But our butter makers sometimes find, at the times when the cattle change from winter to summer or from summer to winter feed, that the ripening is abnormal. The reason appears to be that the cream has become infested with an abundance of malign species. The ripening that they produce is therefore an undesirable one, and the quality of the butter is sure to suffer.

So long as butter was made only in private dairies it was a matter of comparatively little importance if there was an occasional falling off in quality of this sort. When it was made a few pounds at a time, and only once or twice a week, it was not a very serious matter if a few churnings of butter did suffer in quality. But to-day the butter-making industries are becoming more and more concentrated into large creameries, and it is a matter of a good deal more importance to discover some means by which a uniformly high quality can be insured. If a creamery which makes five hundred pounds of butter per day suffers from such an injurious ripening, the quality of its butter will fall off to such an extent as to command a lower price, and the creamery suffers materially. Perhaps the continuation of such a trouble for two or three weeks would make a difference between financial success and failure in the creamery. With our concentration of the butter- making industries it is becoming thus desirable to discover some means of regulating this process more accurately.

The remedy of these occasional ill effects in cream ripening has not been within the reach of the butter maker. The butter maker must make butter with the cream that is furnished him, and if that cream is already impregnated with malign species of bacteria he is helpless. It is true that much can be done to remedy these difficulties by the exercise of especial care in the barns of the patrons of the creamery. If the barns, the cows, the dairies, the milk vessels, etc., are all kept in condition of strict cleanliness, if especial care is taken particularly at the seasons of the year when trouble is likely to arise, and if some attention is paid to the kind of food which the cattle eat, as a rule the cream will not become infected with injurious bacteria. It may be taken as a demonstrated fact that these malign bacteria come from sources of filth, and the careful avoidance of all such sources of filth will in a very large measure prevent their occurrence in the cream. Such measures as these have been found to be practicable in many creameries. Creameries which make the highest priced and the most uniform quality of butter are those in which the greatest care is taken in the barns and dairies to insure cleanliness and in the handling of the milk and cream. With such attention a large portion of the trouble which arises in the creameries from malign bacteria may be avoided.

But these methods furnish no sure remedy against evils of improper species of bacteria in cream ripening, and do not furnish any sure means of obtaining uniform flavour in butter. Even under the very best conditions the flavour of the butter will vary with the season of the year. Butter made in the winter is inferior to that made in the summer months; and while this is doubtless due in part to the different food which the cattle have and to the character of the cream resulting therefrom, these differences in the flavour of the butter are also in part dependent upon the different species of bacteria which are present in the ripening of cream at different seasons. The species of bacteria in June cream are different from those that are commonly present in January cream, and this is certainly a factor in determining the difference between winter and summer butter.

USE OF ARTIFICIAL BACTERIA CULTURES FOR CREAM RIPENING.

Bacteriologists have been for some time endeavouring to aid butter makers in this direction by furnishing them with the bacteria needful for the best results in cream ripening. The method of doing this is extremely simple in principle, but proves to be somewhat difficult in practice. It is only necessary to obtain the species of bacteria that produce the highest results, and then to furnish these in pure culture and in large quantity to the butter makers, to enable them to inoculate their cream with the species of bacteria which will produce the results that they desire. For this purpose bacteriologists have been for several years searching for the proper species of bacteria to produce the best results, and there have been put upon the market for sale several distinct "pure cultures" for this purpose. These have been obtained by different bacteriologists and dairymen in the northern European countries and also in the United States. These pure cultures are furnished to the dairymen in various forms, but they always consist of great quantities of certain kinds of bacteria which experience has found to be advantageous for the purpose of cream ripening.

There have hitherto appeared a number of difficulties in the way of reaching complete success in these directions. The most prominent arises in devising a method of using pure cultures in the creamery. The cream which the butter makers desire to ripen is, as we have seen, already impregnated with bacteria, and would ripen in a fashion of its own even if no pure culture of bacteria were added thereto. Pure cultures can not therefore be used as simply as can yeast in bread dough. It is plain that the simple addition of a pure culture to a mass of cream would not produce the desired effects, because the cream would be ripened then, not by the pure culture alone, but by the pure culture plus all of the bacteria that were originally present. It would, of course, be something of a question as to whether under these conditions the results would be favourable, and it would seem that this method would not furnish any means of getting rid of bad tastes and flavours which have come from the presence of malign species of bacteria. It is plainly desirable to get rid of the cream bacteria before the pure culture is added. This can be readily done by heating it to a temperature of 69 degrees C. (155 degrees F.) for a short time, this temperature being sufficient to destroy most of the bacteria. The subsequent addition of the pure culture of cream-ripening bacteria will cause the cream to ripen under the influence of the added culture alone. This method proves to be successful, and in the butter making countries in Europe it is becoming rapidly adopted.

In this country, however, this process has not as yet become very popular, inasmuch as the heating of the cream is a matter of considerable expense and trouble, and our butter makers have not been very ready to adopt it. For this reason, and also for the purpose of familiarizing butter makers with the use of pure cultures, it has been attempted to produce somewhat similar though less uniform results by the use of pure cultures in cream without previous healing. In the use of pure cultures in this way, the butter maker is directed to add to his cream a large amount of a prepared culture of certain species of bacteria, upon the principle that the addition of such a large number of bacteria to the cream, even though the cream is already inoculated with certain bacteria, will produce a ripening of the cream chiefly influenced by the artificially added culture. The culture thus added, being present in very much greater quantity than the other "wild" species, will have a much greater effect than any of them. This method, of course, cannot insure uniformity. While it may work satisfactorily in many cases, it is very evident that in others, when the cream is already filled with a large number of malign species of bacteria, such an artificial culture would not produce the desired results. This appears to be not only the theoretical but the actual experience. The addition of such pure cultures in many cases produces favourable results, but it does not always do so, and the result is not uniform. While the use of pure cultures in this way is an advantage over the method of simply allowing the cream to ripen normally without such additions, it is a method that is decidedly inferior to that which first pasteurizes the cream and subsequently adds a starter.

There is still another method of adding bacteria to cream to insure a more advantageous ripening, which is frequently used, and, being simpler, is in many cases a decided advantage. This method is by the use of what is called a natural starter. A natural starter consists simply of a lot of cream which has been taken from the most favourable source possible—that is, from the cleanest and best dairy, or from the herd producing the best quality of cream—and allowing this cream to stand in a warm place for a couple of days until it becomes sour. The cream will by that time be filled with large numbers of bacteria, and this is then put as a starter into the vat of cream to be ripened. Of course, in the use of this method the butter maker has no control over the kinds of bacteria that will grow in the starter, but it is found, practically, that if the cream is taken from a good source the results are extremely favourable, and there is produced in this way almost always an improvement in the butter.

The use of pure cultures is still quite new, particularly in this country. In the European butter-making countries they have been used for a longer period and have become very much better known. What the future may develop along this line it is difficult to say; but it seems at least probable that as the difficulties in the details are mastered the time will come when starters will be used by our butter makers for their cream ripening, just as yeast is used by housewives for raising bread, or by brewers for fermenting malt. These starters will probably in time be furnished by bacteriologists. Bacteriology, in other words, is offering in the near future to our butter makers a method of controlling the ripening of the cream in such a way as to insure the obtaining of a high and uniform quality of butter, so far, at least, as concerns flavour and aroma.

BACTERIA IN CHEESE.

Cheese ripening.—The third great product of the dairy industry is cheese, and in connection with this product the dairyman is even more dependent upon bacteria than he is in the production of butter. In the manufacture of cheese the casein of the milk is separated from the other products by the use of rennet, and is collected in large masses and pressed, forming the fresh cheese. This cheese is then set aside for several weeks, and sometimes for months, to undergo a process that is known as ripening. During the ripening there are developed in the cheese the peculiar flavours which are characteristic of the completed product. The taste of freshly made cheese is extremely unlike that of the ripened product. While butter made from unripened cream has a pleasant flavour, and one which is in many places particularly enjoyed, there is nowhere a demand for unripened cheese, for the freshly made cheese has a taste that scarce any one regards as pleasant. Indeed, the whole value of the cheese is dependent upon the flavour of the product, and this flavour is developed during the ripening.

The cheese maker finds in the ripening of his cheese the most difficult part of his manufacture. It is indeed a process over which he has very little control. Even when all conditions seem to be correct, when cheese is made in the most careful manner, it not infrequently occurs that the ripening takes place in a manner that is entirely abnormal, and the resulting cheese becomes worthless. The cheese maker has been at an entire loss to understand these irregularities, nor has he possessed any means of removing them. The abnormal ripening that occurs takes on various types. Sometimes the cheese will become extraordinarily porous, filled with large holes which cause the cheese to swell out of proper shape and become worthless. At other times various spots of red or blue appear in the manufactured cheese; while again unpleasant tastes and flavours develop which render the product of no value. Sometimes a considerable portion of the product of the cheese factory undergoes such irregular ripening, and the product for a long time will thus be worthless. If some means could be discovered of removing these irregularities it would be a great boon to the cheese manufacturer; and very many attempts have been made in one way or another to furnish the cheese maker with some details in the manufacture which will enable him in a measure to control the ripening.

The ripening of the cheese has been subjected to a large amount of study on the part of bacteriologists who have been interested in dairy products. That the ripening of cheese is the result of bacterial growth therein appears to be probable from a priori grounds. Like the ripening of cream, it is a process that occurs somewhat slowly. It is a chemical change which is accompanied by the destruction of proteid matter; it takes place best at certain temperatures, and temperatures which we know are favourable to the growth of micro-organisms, all of which phenomena suggest to us the action of bacteria. Moreover, the flavours and the tastes that arise have a decided resemblance in many cases to the decomposition products of bacteria, strikingly so in Limburger cheese. When we come to study the matter of cheese ripening carefully we learn beyond question that this a priori conclusion is correct. The ripening of any cheese is dependent upon several different factors. The method of preparation, the amount of water left in the curd, the temperature of ripening, and other miscellaneous factors connected with the mechanical process of cheese manufacture, affect its character. But, in addition to all these factors, there is undoubtedly another one, and that is the number and the character of the bacteria that chance to be in the curd when the cheese is made. While it is found that cheeses which are treated by different processes will ripen in a different manner, it is also found that two cheeses which have been made under similar conditions and treated in identically the same way may also ripen in a different manner, so that the resulting flavour will vary. The variations between cheeses thus made may be slight or they may be considerable, but variations certainly do occur. Every one knows the great difference in flavours of different cheeses, and these flavours are due in considerable measure to factors other than the simple mechanical process of making the cheese. The general similarity of the whole process to a bacterial fermentation leads us to believe at the outset that some of the differences in character are due to different kinds of bacteria that multiply in the cheese and produce decomposition therein.

When the matter comes to be studied by bacteriology, the demonstration of this position becomes easy. That the ripening of cheese is due to growth of bacteria is very easily proved by manufacturing cheeses from milk which is deprived of bacteria. For instance, cheeses have been made from milk that has been either sterilized or pasteurized—which processes destroy most of the bacteria therein—and, treated otherwise in a normal manner, are set aside to ripen. These cheeses do NOT ripen, but remain for months with practically the same taste that they had originally. In other experiments the cheese has been treated with a small amount of disinfective, which is sufficient to prevent bacteria from growing, and again ripening is found to be absolutely prevented. Furthermore, if the cheese under ordinary conditions is studied during the ripening process, it is found that bacteria are growing during the whole time. These facts all taken together plainly prove that the ripening of cheese is a fermentation due to bacteria. It will be noticed, however, that the conditions in the cheese are not favourable for very rapid bacterial growth. It is true that there is plenty of food in the cheese for bacterial life, but the cheese is not very moist; it is extremely dense, being subjected in all cases to more or less pressure. The penetration of oxygen into the centre of the mass must be extremely slight. The density, the lack of a great amount of moisture, and the lack of oxygen furnish conditions in which bacteria will not grow very rapidly. The conditions are far less favourable than those of ripening cream, and the bacteria do not grow with anything like the rapidity that they grow in cream. Indeed, the growth of these organisms during the ripening is extremely slow compared to the possibilities of bacterial growth that we have already noticed. Nevertheless, the bacteria do multiply in the cheese, and as the ripening goes on they become more and more abundant, although the number fluctuates, rising and falling under different conditions.

When the attempt is made to determine the relation of the different kinds of ripening to different kinds of bacteria, it has thus far met with extremely little success. That different flavours are due to the ripening produced by different kinds of bacteria would appear to be almost certain when we remember, as we have already noticed, the different kinds of decomposition produced by different species of bacteria. It would seem, moreover, that it ought not to be very difficult to separate from the ripened cheese the bacteria which are present, and thus obtain the kind of bacteria necessary to produce the desired ripening. But for some reason this does not prove to be so easy in practice as it seems to be in theory. Many different species of bacteria have been separated from cheeses. One bacteriologist, studying several cheeses, separated about eighty different species therefrom, and others have found perhaps as many more from different sources. Moreover, experiments have been made with a considerable number of these different kinds of bacteria to determine whether they are capable of producing normal ripening. These experiments consist of making cheese out of milk that has been deprived of its bacteria, and which has been inoculated with large quantities of the species in question. Hitherto these experiments have not been very satisfactory. In some cases the cheese appears to ripen scarcely at all; in other cases the ripening occurs, but the resulting cheese is of a peculiar character, entirely unlike the cheese that it is desired to imitate. There have been one or two experiments in recent times that give a little more promise of success than the earlier ones, for a few species of bacteria have been used in ripening with what the authors have thought to be promising success. The cheese made from the milk artificially inoculated with these species ripens in a satisfactory manner and gives some of the character desired, though up to the present time in no case has the typical normal ripening been produced in any of these experiments.

But these experiments have demonstrated beyond question that the abnormal ripening which is common in cheese factories is due to the presence of undesirable species of bacteria in the milk. Many of the experiments in making cheeses by means of artificial cultures of bacteria have resulted in decidedly abnormal cheeses. Many of the cheeses thus manufactured have shown imperfections in ripening which are identical with those actually occurring in the cheese factory. Several different species of bacteria have been found which, when artificially used thus for ripening cheese, will give rise to the porosity and the abnormal swelling of the cheese already referred to (Fig. 24). Others produced bad tastes and flavours, and enough has been done in this line to demonstrate beyond peradventure that the abnormal ripening of cheese is due primarily to the growth of improper species therein. Quite a long list of species of bacteria which produce abnormal ripening have been isolated from cheeses, and have been studied and experimented with by bacteriologists. As a result of this study of abnormal ripening, there has been suggested a method of partially controlling these—remedying them. The method consists simply in testing the fermenting qualities of the milk used. A small sample of milk from different dairies is allowed to stand in the cheese factory by itself until it undergoes its normal souring. If the fermentation or souring that thus occurs is of a normal character, the milk is regarded as proper for cheese making. But if the fermentation that occurs in any particular sample of milk is unusual; if an extraordinary amount of gas bubbles are produced, or if unpleasant smells and tastes arise, the sample is regarded as unfavourable for cheese making, and as likely to produce abnormal ripening in the cheeses. Milk from this source would therefore be excluded from the milk that is to be used in cheese making. This, of course, is a tentative and an unsatisfactory method of controlling the ripening, and yet it is one of some practical value to cheese makers. It is the only method that has yet been suggested of controlling the ripening.

Our bacteriologists, of course, are quite confident that in the future more practical results will be obtained along this line than in the past. If it is true that cheeses are ripened by bacteria; if it is true that different qualities in the cheese are due to the growth of different species of bacteria during the ripening, it would seem to be possible to obtain the proper kind of bacteria and to furnish them to the cheese maker for artificially inoculating his cheese, just as it has been possible to furnish artificially cultivated yeasts to the brewer, and as it has become possible to furnish artificially cultivated bacteria to the butter maker. We must, however, recognise this to be a matter for the future. Up to the present time no practical results along the lines of bacteria have been obtained which our cheese manufacturers can make use of in the way of controlling with any accuracy this process of cheese ripening.

Thus it will be seen that in this last dairy product bacteria play even a more important part than in any of the others. The food value of cheese is dependent upon the casein which is present. The market price, however, is controlled entirely by the flavour, and this flavour is a product of bacterial growth. Upon the action of bacteria, then, the cheese maker is absolutely dependent; and when our bacteriologists are able in the future to investigate this matter further, it seems to be at least possible that they may obtain some means of enabling the cheese maker to control the ripening accurately. Not only so, but recognising the great variety in the flavours of cheese, and recognising that different kinds of bacteria undoubtedly produce different kinds of decomposition products, it seems to be at least possible that a time will come when the cheese maker will be able to produce at— will any particularly desired flavour in his cheese by the addition to it of particular species of bacteria, or particular mixtures of species of bacteria which have been discovered to produce the desired effects.



CHAPTER IV.

BACTERIA IN NATURAL PROCESSES.—AGRICULTURE.

Thus far, in considering the relations of bacteria to mankind, we have taken into account only the arts and manufactures, and have found bacteria playing no unimportant part in many of the industries of our modern civilized life. So important are they that there is no one who is not directly affected by them. There is hardly a moment in our life when we are not using some of the direct or indirect products of bacterial action. We turn now, however, to the consideration of a matter of even more fundamental importance; for when we come to study bacteria in Nature, we find that there are certain natural processes connected with the life of animals and plants that are fundamentally based upon their powers. Living Nature appears limitless, for life processes have been going on in the world through countless centuries with seemingly unimpaired vigour. At the very bottom we find this never-ending exhibition of vital power dependent upon certain activities of micro-organisms. So thoroughly is this true that, as we shall find after a short consideration, the continuance of life upon the surface of the world would be impossible if bacterial action were checked for any considerable length of time. The life of the globe is, in short, dependent upon these micro-organisms.

BACTERIA AS SCAVENGERS.

In the first place, we may notice the value of these organisms simply as scavengers, keeping the surface of the earth in the proper condition for the growth of animals and plants. A large tree in the forest dies and falls to the ground. For a while the tree trunk lies there a massive structure, but in the course of months a slow change takes place in it. The bark becomes softened and falls from the wood. The wood also becomes more or less softened; it is preyed upon then by insect life; its density decreases more and more, until finally it crumbles into a soft, brownish, powdery mass, and eventually the whole sinks into the soil, is overgrown by mosses and other vegetation, and the tree trunk has disappeared from view. In the same way the body of the dead animal undergoes the process of the softening of its tissues by decay. The softer parts of the body rapidly dissipate, and even the bones themselves eventually are covered with the soil and disintegrated, until in time they, too, disappear from any visible existence. This whole process is one of decay, and the result is that the solid mass of the body of the tree or of the animal has been decomposed. What has become of it? The answer holds the secret of Nature's eternal freshness. Part of it has dissipated into the air in the form of gases and water vapour; part of it has changed its composition and has become incorporated into the soil, the final result being that the body of the plant or animal disappears as such, and its substance is converted into gaseous form, which is dissipated in the air or into simple compounds which sink into the earth.

This whole process of decay of organic life is one in which bacteria play the most important part. In the case of the decomposition of the woody matter of the tree trunk, the process is begun by the agency of moulds, for this group of organisms alone appears to be capable of attacking such hard woody structure. The later part of the decay, however, is largely carried on by bacterial life. In the decomposition of the animal tissues, bacteria alone are the agents. Thus the process by which organic matter is dissipated into the air or incorporated into the soil is one which is primarily presided over by bacterial life.

Viewing this matter in a purely mechanical light, the importance of bacteria in thus acting as scavengers can hardly be overestimated. If we think for a moment of the condition of the world were there no such decomposing agents to rid the earth's surface of the dead bodies of animals and plants, we shall see that long since the earth would have been uninhabitable. If the dead bodies of plants and animals of past ages simply accumulated on the surface of the ground without any forces to reduce them into simple compounds for dissipation, by their very bulk they would have long since completely covered the surface of the earth so as to afford no possible room for further growth of plants and animals. In a purely mechanical way, then, bacteria as decomposition agents are necessary to keep the surface of the earth fresh and unencumbered so that life can continue.

BACTERIA AS AGENTS IN NATURE'S FOOD CYCLE.

But the matter by no means ends here. When we come to think of it, it is a matter of considerable surprise that the surface of the earth has been able to continue producing animals and plants for the many millions of years during which life has been in existence. Plants and animals both require food, animals depending wholly upon plants therefor. Plants, however, equally with animals, require food, and although they obtain a considerable portion of their food from the air, yet no inconsiderable part of it is obtained from the soil. The question is forced upon us, therefore, as to why the soil has not long since become exhausted of food. How could the soil continue to support plants year after year for millions of years, and yet remain as fertile as ever?

The explanation of this phenomenon is in the simple fact that the processes of Nature are such that the same food is used over and over again, first by the plant, then by the animal, and then again by the plant, and there is no necessity for any end of the process so long as the sun furnishes energy to keep the circulation continuous. One phase of this transference of food from animal to plant and from plant to animal is familiar to nearly every one. It is a well-known fact that animals in their respiration consume oxygen, but exhale it again in combination with carbon as carbonic dioxide. On the other hand, plants in their life consume the carbonic dioxide and exhale the oxygen again as free oxygen. Thus each of these kingdoms makes use of the excreted product of the other, and this process can go on indefinitely, the animals furnishing our atmosphere with plenty of carbonic acid for plant life, and the plants excreting into the atmosphere at the same time an abundant sufficiency of oxygen for animal life. The oxygen thus passes in an endless round from animal to plant and from plant to animal.

A similar cycle is true of all the other foods of animal and plant life, though in regard to the others the operation is more complex and more members are required to complete the chain. The transference of matter through a series of changes by which it is brought from a condition in which it is proper food for plants back again into a condition when it is once more a proper food for plants, is one of the interesting discoveries of modern science, and one in which, as we shall see, bacteria play a most important part. This food cycle is illustrated roughly by the accompanying diagram; but in order to understand it, an explanation of the various steps in this cycle is necessary.

It will be noticed that at the bottom of the circle represented in Fig. 25, at A, are given various ingredients which are found in the soil and which form plant foods. Plant foods, as may be seen there, are obtained partly from the air as carbonic dioxide and water; but another portion comes from the soil. Among the soil ingredients the most prominent are nitrates, which are the forms of nitrogen compounds most easily made use of by plants as a source of this important element. It should be stated also that there are other compounds in the soil which furnish plants with part of their food—compounds containing potassium, phosphorus, and some other elements. For simplicity's sake, however, these will be left out of consideration. Beginning at the bottom of the cycle (Fig. 25 A), plant life seizes the gases from the air and these foods from the soil, and by means of the energy furnished it by the sun's rays builds these simple chemical compounds into more complex ones. This gives us the second step, as shown in Fig. 25 B, the products of plant life. These products of plant life consist of such materials as sugar, starches, fats, and proteids, all of which have been manufactured by the plant from the ingredients furnished it from the soil and air, and through the agency of the sun's rays. These products of plant life now form foods for the animal kingdom. Starches, fats, and proteids are animal foods, and upon such complex bodies alone can the animal kingdom be fed. Animal life, standing high up in the circle, is not capable of extracting its nutriment from the soil, but must take the more complex foods which have been manufactured by plant life. These complex foods enter now into the animal and take their place in the animal body. By the animal activities, some of the foods are at once decomposed into carbonic acid and water, which, being dissipated into the air, are brought back at once into the condition in which they can serve again as plant food. This part of the food is thus brought back again to the bottom of the circle (Fig. 25, dotted lines). But while it is true that animals do thus reduce some of their foods to the simple condition of carbonic acid and water, this is not true of most of the foods which contain nitrogen. The nitrogenous foods are as necessary for the life as the carbon foods, and animals do not reduce their nitrogenous foods to the condition in which plants can prey upon them. While plants furnish them with nitrogenous food, they can not give it back to the plants. Part of the nitrogenous foods animals build into new albumins (Fig. 25 C); but a part of them they reduce at once into a somewhat simpler condition known as urea. Urea is the form in which the nitrogen is commonly excreted from the animal body. But urea is not a plant food; for ordinary plants are entirely unable to make use of it. Part of the nitrogen eaten by the animal is stored up in its body, and thus the body of the animal, after it has died, contains these nitrogen compounds of high complexity. But plants are not able to use these compounds. A plant can not be fed upon muscle tissue, nor upon fats, nor bones, for these are compounds so complex that the simple plant is unable to use them at all. So far, then, in the food cycle the compounds taken from the soil have been built up into compounds of greater and greater complexity; they have reached the top of this circle, and no part of them, except part of the carbon and oxygen, has become reduced again to plant food. In order that this material should again become capable of entering into the life of plants so as to go over the circle again, it is necessary for it to be once more reduced from its highly complex condition into a simpler one.

Now come into play these decomposition agencies which we have been studying under the head of scavengers. It will be noticed that the next step in the food cycle is taken by the decomposition bacteria. These organisms, existing, as we have already seen, in the air, in the soil, in the water, and always ready to seize hold of any organic substance that may furnish them with food, feed upon the products of animal life, whether they are such products as muscle tissue, or fat, or sugar, or whether they are the excreted products of animal life, such as urea, and produce therein the chemical decomposition changes already noticed. As a result of this chemical decomposition, the complex bodies are broken into simpler and simpler compounds, and the final result is a very thorough destruction of the animal body or the material excreted by animal life, and its reduction into forms simple enough for plants to use again as foods. Thus the bacteria come in as a necessary link to connect the animal body, or the excretion from the animal body, with the soil again, and therefore with that part of the circle in which the material can once more serve as plant food.

But in the decomposition that thus occurs through the agency of the putrefactive bacteria it very commonly happens that some of the food material is broken down into compounds too simple for use as plant food. As will be seen by a glance at the diagram (Fig. 25 D), a portion of the cleavage products resulting from the destruction of these animal foods takes the form of carbonic-acid gas and water. These ingredients are at once in condition for plant life, as shown by the dotted lines. They pass off into the air, and the green leaves of vegetation everywhere again seize them, assimilate them, and use them as food. Thus it is that the carbon and the oxygen have completed the cycle, and have come back again to the position in the circle where they started. In regard to the nitrogen portion of the food, however, it very commonly happens that the products which arise as the result of the decomposition processes are not yet in proper condition for plant food. They are reduced into a condition actually too simple for the use of plants. As a result of these putrefactive changes, the nitrogen products of animal life are broken frequently into compounds as simple as ammonia (NH3), or into compounds which the chemists speak of as nitrites (Fig. 25 at D). Now these compounds are not ordinarily within the reach of plant life. The luxuriant vegetation of the globe extracts its nitrogen from the soil in a form more complex than either of the compounds here mentioned; for, as we have seen, it is nitrates chiefly that furnish plants with their nitrogen food factor. But nitrates contain considerable oxygen. Ammonia, which is one of the products of putrefactive de- composition, contains no oxygen, and nitrites, another factor, contains less oxygen than nitrates. These bodies are thus too simple for plants to make use of as a source of nitrogen. The chemical destruction of the food material which results from the action of the putrefactive bacteria is too thorough, and the nitrogen foods are not yet in condition to be used by plants.

Now comes in the agency of still another class of micro-organisms, the existence of which has been demonstrated to us during the last few years. In the soil everywhere, especially in fertile soil, is a class of bacteria which has received the name of nitrifying bacteria (Fig. 26). These organisms grow in the soil and feed upon the soil ingredients. In the course of their life they have somewhat the same action upon the simple nitrogen cleavage products just mentioned as we have already noticed the vinegar- producing species have upon alcohol, viz., the bringing about a union with oxygen. There are apparently several different kinds of nitrifying bacteria with different powers. Some of them cause an oxidation of the nitrogen products by means of which the ammonia is united with oxygen and built up into a series of products finally resulting in nitrates (Fig. 26). By the action of other species still higher nitrogen compounds, including the nitrites, are further oxidized and built up into the form of nitrates. Thus these nitrifying organisms form the last link in the chain that binds the animal kingdom to the vegetable kingdom (Fig. 25 at 4). For after the nitrifying organisms have oxidized nitrogen cleavage products, the results of the oxidation in the form of nitrates or nitric acid are left in the soil, and may now be seized upon by the roots of plants, and begin once more their journey around the food cycle. In this way it will be seen that while plants, by building up compounds, form the connecting link between the soil and animal life, bacteria in the other half of the cycle, by reducing them again, give us the connecting link between animal life and the soil. The food cycle would be as incomplete without the agency of bacterial life as it would be without the agency of plant life.

But even yet the food cycle is not complete. Some of the processes of decomposition appear to cause a portion of the nitrogen to fly out of the circle at a tangent. In the process of decomposition which is going on through the agency of micro-organisms, a considerable part of the nitrogen is dissipated into the air in the form of free nitrogen. When a bit of meat decays, part of the meat is, indeed, converted into ammonia or other nitrogen compounds, but if the putrefaction is allowed to go on, in the end a considerable portion of it will be broken into still simpler forms, and the nitrogen will finally be dissipated into the air in the form of free nitrogen. This dissipation of free nitrogen into the air is going on in the world wherever putrefaction takes place. Wherever decomposition of nitrogen products occurs some free nitrogen is eliminated. Now, this part of the nitrogen has passed beyond the reach of plants, for plants can not extract free nitrogen from the air. In the diagram this is represented as a portion of the material which, through the agency of the decomposition bacteria, has been thrown out of the cycle at a tangent (Fig. 25 E). It will, of course, be plain from this that the store of nitrogen food must be constantly diminishing. The soil may have been originally supplied with a given quantity of nitrogen compound, but if the decomposition products are causing considerable quantities of this nitrogen to be dissipated in the air, it plainly follows that the total amount of nitrogen food upon which the animal and vegetable kingdoms can depend is becoming constantly reduced by such dissipation.

There are still other methods by which nitrogen is being lost from the food cycle. First, we may notice that the ordinary processes of vegetation result in a gradual draining of the soil and a throwing of its nitrogen into the ocean. The body of any animal or any plant that chances to fall into a brook or river is eventually carried to the sea, and the products of its decomposition pass into the ocean and are, of course, lost to the soil. Now, while this gradual extraction of nitrogen from the soil by drainage is a slow one, it is nevertheless a sure one. It is far more rapid in these years of civilized life than in former times, since the products of the soil are given to the city, and then are thrown into its sewage Our cities, then, with our present system of disposing of sewage, are draining from the soil the nitrogen compounds and throwing them away.

In yet another direction must it be noticed that our nitrogen compounds are being lost to plant life—viz., by the use of various nitrogen compounds to form explosives. Gunpowder, nitro-glycerine, dynamite, in fact, nearly all the explosives that are used the world over for all sorts of purposes, are nitrogen compounds. When they are exploded the nitrogen of the compound is dissipated into the air in the form of gas, much of it in the form of free nitrogen. The basis from which explosive compounds are made contains nitrogen in the form in which it can be used by plants. Saltpetre, for example, is equally good as a fertilizer and as a basis for gunpowder. The products of the explosion are gases no longer capable of use by plants, and thus every explosion of nitrogen compounds aids in this gradual dissipation of nitrogen products, taking them from the store of plant foods and throwing them away.

All of these agencies contribute to reduce the amount of material circulating in the food cycle of Nature, and thus seem to tend inevitably in the end toward a termination of the processes of life; for as soon as the soil becomes exhausted of its nitrogen compounds, so soon will plant life cease from lack of nutrition, and the disappearance of animal life will follow rapidly. It is this loss of nitrogen in large measure that is forcing our agriculturists to purchase fertilizers. The last fifteen years have shown us, however, that here again we may look upon our friends, the bacteria, as agents for counteracting this dissipating tendency in the general processes of Nature. Bacterial life in at least two different ways appears to have the function of reclaiming from the atmosphere more or less of this dissipated free nitrogen.

In the first place, it has been found in the last few years that soil entirely free from all common plants, but containing certain kinds of bacteria, if allowed to stand in contact with the air, will slowly but surely gain in the amount of nitrogen compounds that it contains. These nitrogen compounds are plainly manufactured by the bacteria in the soil; for unless the bacteria are present they do not accumulate, and they do accumulate inevitably if the bacteria are present in the proper quantity and the proper species. It appears that, as a rule, this fixation of nitrogen is not performed by any one species of microorganisms, but by two or three of them acting together. Certain combinations of bacteria have been found which, when inoculated in the soil, will bring about this fixation of nitrogen, but no one of the species is capable of producing this result alone. We do not know to what extent these organisms are distributed in the soil, nor how widely this nitrogen fixation through bacterial life is going on. It is only within a short time that it has been demonstrated to exist, but we must look upon bacteria in the soil as one of the factors in reclaiming from the atmosphere the dissipated free nitrogen.

The second method by which bacteria aid in the reclaiming of this lost nitrogen is by a combined action of certain species of bacteria and some of the higher plants. Ordinary green plants, as already noted, are unable to make use of the free nitrogen of the atmosphere It was found, however, some fifteen years ago that some species of plants, chiefly the great family of legumes, which contains the pea plant, the bean, the clover, etc, are able, when growing in soil that is poor in nitrogen, to obtain nitrogen from some source other than the soil in which they grow. A pea plant in soil that contains no nitrogen products and watered with water that contains no nitrogen, will, after sprouting and growing for a length of time, be found to have accumulated a considerable quantity of fixed nitrogen in its tissues The only source of this nitrogen has been evidently from the air which bathes the leaves of the plant or permeates the soil and bathes its roots This fact was at first disputed, but subsequently demonstrated to be true, and was found later to be associated with the combined action of these legumes and certain soil bacteria. When a legume thus gains nitrogen from the air, it develops upon its roots little bunches known as root nodules or root tubercles. The nodules are sometimes the size of the head of a pm, and sometimes much larger than this, occasionally reaching the size of a large pea, or even larger. Upon microscopic examination they are found to be little nests of bacteria In some way the soil organisms (Fig 27) make their way into the roots of the sprouting plant, and finding there congenial environment, develop in considerable quantities and produce root tubercles in the root. Now, by some entirely unknown process, the legume and the bacteria growing together succeed in extracting the nitrogen from the atmosphere which permeates the soil, and fixing this nitrogen in the tubercles and the roots in the form of nitrogen compounds. The result is that, after a proper period of growth, the amount of fixed nitrogen in the plant is found to have very decidedly increased (Fig 25 E).

This, of course, furnishes a starting point for the reclaiming of the lost atmospheric nitrogen. The legume continues to live its usual life, perhaps increasing the store of nitrogen in its roots and stems and leaves during the whole of its normal growth. Subsequently, after having finished its ordinary life, the plant will die, and then the roots and stems and leaves, falling upon the ground and becoming buried, will be seized upon by the decomposition bacteria already mentioned. The nitrogen which has thus become fixed in their tissues will undergo the destructive changes already described. This will result eventually in the production of nitrates. Thus some of the lost nitrogen is restored again to the soil in the form of nitrates, and may now start on its route once more around the cycle of food.

It will be seen, then, that the food cycle is a complete one. Beginning with the mineral ingredients in the soil, the food matter may start on its circulation from the soil to the plant, from the plant to the animal, from the animal to the bacterium and from the bacterium through a series of other bacteria back again to the soil in the condition in which it started. If, perchance, in this progress around the circle some of the nitrogen is thrown off at a tangent, this, too, is brought back again to the circle through the agency of bacterial life. And so the food material of animals and plants continues in this never-ceasing circulation. It is the sunlight that furnishes the energy for the motion. It is the sunlight that forces the food around the circle and keeps up the endless change; and so long as, the sun continues to shine upon the earth there seems to be no reason why the process should ever cease. It is this repeated circulation that has made the continuation of life possible for the millions and millions of years of the earth's history. It is this continued circulation that makes life possible still, and it is only this fact that the food is thus capable of ever circulating from animal to plant and from plant to animal that makes it possible for the living world to continue its existence. But, ah we have seen, one half of this great circle of food change is dependent upon bacterial life. Without the bacterial life the animal body and the animal excretion could never be brought back again within the reach of the plant; and thus, were it not for the action of these micro- organisms the food cycle would be incomplete and life could not continue indefinitely upon the surface of the earth. At the very foundation, the continuation of the present condition of Nature and the existence of life during the past history of the world has been fundamentally based upon the ubiquitous presence of bacteria and upon their continual action in connection with both destructive and constructive processes.

RELATION OF BACTERIA TO AGRICULTURE.

We have already noticed that bacteria play an important part in some of the agricultural industries, particularly in the dairy. From the consideration of the matters just discussed, it is manifest that these organisms must have an even more intimate relation to the farmer's occupation. At the foundation, farming consists in the cultivation of plants and animals, and we have already seen how essential are the bacteria in the continuance of animal and plant life. But aside from these theoretical considerations, a little study shows that in a very practical manner the farmer is ever making use of bacteria, as a rule, quite unconsciously, but none the less positively.

SPROUTING OF SEEDS.

Even in the sprouting of seeds after they are sown in the soil bacterial life has its influence. When seeds are placed m moist soil they germinate under the influence of heat. The rich albuminous material in the seeds furnishes excellent food, and inasmuch as bacteria abound in the soil, it is inevitable that they should grow in and feed upon the seed. If the moisture is excessive and the heat considerable, they very frequently grow so rapidly in the seed as to destroy its life as a seedling. The seed rots in the ground as a result. This does not commonly occur, however, in ordinary soil. But even here bacteria do grow in the seed, though not so abundantly as to produce any injury. Indeed, it has been claimed that their presence in the seed in small quantities is a necessity for the proper sprouting of the seed. It has been claimed that their growth tends to soften the food material in the seed, so that the young seedling can more readily absorb it for its own food, and that without such a softening the seed remains too hard for the plant to use. This may well be doubted, however, for seeds can apparently sprout well enough without the aid of bacteria. But, nevertheless, bacteria do grow in the seed during its germination, and thus do aid the plant in the softening of the food material. We can not regard them as essential to seed germination. It may well be claimed that they ordinarily play at least an incidental part in this fundamental life process, although it is uncertain whether the growth of seedlings is to any considerable extent aided thereby.

THE SILO.

In the management of a silo the farmer has undoubtedly another great bacteriological problem. In the attempt to preserve his summer-grown food for the winter use of his animals, he is hindered by the activity of common bacteria. If the food is kept moist, it is sure to undergo decomposition and be ruined in a short time as animal food. The farmer finds it necessary, therefore, to dry some kinds of foods, like hay. While he can thus preserve some foods, others can not be so treated. Much of the rank growth of the farm, like cornstalks, is good food while it is fresh, but is of little value when dried. The farmer has from experience and observation discovered a method of managing bacterial growth which enables him to avoid their ordinary evil effects. This is by the use of the silo. The silo is a large, heavily built box, which is open only at the top. In the silo the green food is packed tightly, and when full all access of air is excluded, except at its surface. Under these conditions the food remains moist, but nevertheless does not undergo its ordinary fermentations and putrefactions, and may be preserved for months without being ruined. The food in such a silo may be taken out months after it is packed, and will still be found to be in good condition for food. It is true that it has changed its character somewhat, but it is not decayed, and is eagerly eaten by cattle.

We are yet very ignorant of the nature of the changes which occur m the food while in the silo. The food is not preserved from fermentation. When the siloxis packed slowly, a very decided fermentation occurs by which the mass is raised to a high temperature (140 degrees F. to 160 degrees F.). This heating is produced by certain species of bacteria which grow readily even at this high temperature. The fermentation uses up the air in the silo to a certain extent and produces a settling of the material which still further excludes air. The first fermentation soon ceases, and afterward only slow changes occur. Certain acid- producing bacteria after a little begin to grow slowly, and in time the silage is rendered somewhat sour by the production of acetic acid. But the exclusion of air, the close packing, and the small amount of moisture appear to prevent the growth of the common putrefactive bacteria, and the silage remains good for a long time. In other methods of filling the silo, the food is very quickly packed and densely crowded together so as to exclude as much air as possible from the beginning. Under these conditions the lack of moisture and air prevents fermentative action very largely. Only certain acid-producing organisms grow, and these very slowly. The essential result in either case is that the common putrefactive bacteria are prevented from growing, probably by lack of sufficient oxygen and moisture, and thus the decay is prevented. The closely packed food offers just the same unfavourable condition for the growth of common putrefactive bacteria that we have already seen offered by the hard-pressed cheese, and the bacteria growth is in the same way held in check. Our knowledge of the matter is as yet very slight, but we do know enough to understand that the successful management of a silo is dependent upon the manipulation of bacteria.

THE FERTILITY OF THE SOIL.

The farmer's sole duty is to extract food from the soil. This he does either directly by raising crops, or indirectly by raising animals which feed upon the products of the soil. In either case the fertility of the soil is the fundamental factor in his success. This fertility is a gift to him from the bacteria.

Even in the first formation of soil he is in a measure dependent upon bacteria. Soil, as is well known, is produced in large part by the crumbling of the rocks into powder. This crumbling we generally call weathering, and regard it as due to the effect of moisture and cold upon the rocks, together with the oxidizing action of the air. Doubtless this is true, and the weathering action is largely a physical and chemical one. Nevertheless, in this fundamental process of rock disintegration bacterial action plays a part, though perhaps a small one. Some species of bacteria, as we have seen, can live upon very simple foods, finding in free nitrogen and carbonates sufficiently highly complex material for their life. These organisms appear to grow on the bare surface of rocks, assimilating nitrogen from the air, and carbon from some widely diffused carbonates or from the CO2 in the air. Their secreted products of an acid nature help to soften the rocks, and thus aid in performing the first step in weathering.

The soil is not, however, all made up of disintegrated rocks. It contains, besides, various ingredients which combine to make it fertile. Among these are various sulphates which form important parts of plant foods. These sulphates appear to be formed, in part, at least, by bacterial agency. The decomposition of proteids gives rise, among other things, to hydrogen sulphide (H2S). This gas, which is of common occurrence in the atmosphere, is oxidized by bacterial growth into sulphuric acid, and this is the basis of part of the soil sulphates. The deposition of iron phosphates and iron silicates is probably also in a measure aided by bacterial action. All of these processes are factors in the formation of soil. Beyond much question the rock disintegration which occurs everywhere in Nature is chiefly the result of physical and chemical changes, but there is reason for believing that the physical and chemical processes are, to a slight extent at least, assisted by bacterial life.

A more important factor of soil fertility is its nitrogen content, without which it is completely barren. The origin of these nitrogen ingredients has been more or less of a puzzle. Fertile soil everywhere contains nitrates and other nitrogen compounds, and in certain parts of the world there are large accumulations of these compounds, like the nitrate beds of Chili. That they have come ultimately from the free atmospheric nitrogen seems certain, and various attempts have been made to explain a method of this nitrogen fixation. It has been suggested that electrical discharges in the air may form nitric acid, which would readily then unite with soil ingredients to form nitrates. There is little reason, however, for believing this to be a very important factor But in the soil bacteria we find undoubtedly an efficient agency m this nitrogen fixation. As already seen, the bacteria are able to seize the free atmospheric nitrogen, converting it into nitrite and nitrates. We have also learned that they can act in connection with legumes and some other plants, enabling them to fix atmospheric nitrogen and store it m their roots. By these two means the nitrogen ingredient in the soil is prevented from becoming exhausted by the processes of dissipation constantly going on. Further, by some such agency must we imagine the original nitrogen soil ingredient to have been derived. Such an organic agency is the only one yet discerned which appears to have been efficient in furnishing virgin soil with its nitrates, and we must therefore look upon bacteria as essential to the original fertility of the soil. But in another direction still does the farmer depend directly upon bacteria The most important factor in the fertility of the soil is the part of it called humus. This humus is very complex, and never alike in different soils It contains nitrogen compounds in abundance, together with sulphates, phosphates, sugar, and many other substances. It is this which makes the garden soil different from sand, or the rich soil different from the sterile soil. If the soil is cultivated year after year, its food ingredients are slowly but surely exhausted. Something is taken from the humus each year, and unless this be replaced the soil ceases to be able to support life. To keep up a constant yield from the soil the farmer understands that he must apply fertilizers more or less constantly.

This application of fertilizers is simply feeding the crops. Some of these fertilizers the farmer purchases, and knows little or nothing as to their origin. The most common method of feeding the crops is, however, by the use of ordinary barnyard manure. The reason why this material contains plant food we can understand, since it is made of the undigested part of food, together with all the urea and other excretions of animals, and contains, therefore, besides various minerals, all of the nitrogenous waste of animal life. These secretions are not at first fit for plant food. The farmer has learned by experience that such excretions, before they are of any use on his fields, must undergo a process of slow change, which is sometimes called ripening. Fresh manure is sometimes used on the fields, but it is only made use of by the plants after the ripening process has occurred. Fresh animal excretions are of little or no value as a fertilizer. The farmer, therefore, commonly allows it to remain in heaps for some time, and it undergoes a slow change, which gradually converts it into a condition in which it can be used by plants. This ripening is readily explained by the facts already considered The fresh animal secretions consist of various highly complex compounds of nitrogen, and the ripening is a process of their decomposition. The proteids are broken to pieces, and their nitrogen elements reduced to the form of nitrates, leucin, etc, or even to ammonia or free nitrogen. Further, a second process occurs, the process of oxidation of these nitrogen compounds already noticed, and the ammonia and nitrites resulting from the decomposition are built into nitrates. In short, in this ripening manure the processes noticed in the first part of this chapter are taking place, by which the complex nitrogenous bodies are first reduced and then oxidized to form plant food. The ripening of manure is both an analytical and a synthetical process. By the analysis, proteids and other bodies are broken into very simple compounds, some of them, indeed, being dissipated into the air, but other portions are retained and then oxidized, and these latter become the real fertilizing materials. Through the agency of bacteria the compost heap thus becomes the great source of plant food to the farmer. Into this compost heap he throws garbage, straw, vegetable and animal substances in general, or any organic refuse which may be at hand. The various bacteria seize it all, and cause the decomposition which converts it into plant food again. The rotting of the compost heap is thus a gigantic cultivation of bacteria.

This knowledge of the ripening process is further teaching the farmer how to prevent waste. In the ordinary decomposition of the compost heap not an inconsiderable portion of the nitrogen is lost in the air by dissipation as ammonia or free nitrogen. Even his nitrates may be thus lost by bacterial action. This portion is lost to the farmer completely, and he can only hope to replace it either by purchasing nitrates in the form of commercial fertilizers, or by reclaiming it from the air by the use of the bacterial agencies already noticed. With the knowledge now at his command he is learning to prevent this waste. In the decomposition one large factor of loss is the ammonia, which, being a gas, is readily dissipated into the air. Knowing this common result of bacterial action, the scientist has told the farmer that, by adding certain common chemicals to his decomposing manure heap, chemicals which will readily unite with ammonia, he may retain most of the nitrogen in this heap in the form of ammonia salts, which, once formed, no longer show a tendency to dissipate into the air. Ordinary gypsum, or superphosphates, or plaster will readily unite with ammonia, and these added to the manure heap largely counteract the tendency of the nitrogen to waste, thus enabling the farmer to put back into his soil most of the nitrogen which was extracted from it by his crops and then used by his stock. His vegetable crops raise the nitrates into proteids. His animals feed upon the proteids, and perform his work or furnish him with milk. Then his bacteria stock take the excreted or refuse nitrogen, and in his manure heap turn it back again into nitrates ready to begin the circle once more. This might go on almost indefinitely were it not for two facts, the farmer sends nitrogenous material off his farm in the milk or grains or other nitrogenous products, which he sells, and the decomposition processes, as we have seen, dissipate some of the nitrogen into the air as free nitrogen.

To meet this emergency and loss the farmer has another method of enriching the soil, again depending upon bacteria. This is the so- called green manuring. Here certain plants which seize nitrogen from the air are cultivated upon the field to be fertilized, and, instead of harvesting a crop, it is ploughed into the soil. Or perhaps the tops may be harvested, the rest being ploughed into the soil. The vegetable material thus ploughed in lies over a season and enriches the soil. Here the bacteria of the soil come into play in several directions. First, if the crop sowed be a legume, the soil bacteria assist it to seize the nitrogen from the air. The only plants which are of use in this green manuring are those which can, through the agency of bacteria, obtain nitrogen from the air and store it in their roots. Second, after the crop is ploughed into the soil various decomposing bacteria seize upon it, pulling the compounds to pieces. The carbon is largely dissipated into the air as carbonic dioxide, where the next generation of plants can get hold of it. The minerals and the nitrogen remain in the soil. The nitrogenous portions go through the same series of decomposition and synthetical changes already described, and thus eventually the nitrogen seized from the air by the combined action of the legumes and the bacteria is converted into nitrates, and will serve for food for the next set of plants grown on the same soil. Here is thus a practical method of using the nitrogen assimilation powers of bacteria, and reclaiming nitrogen from the air to replace that which has been lost. Thus it is that the farmer's nitrogen problem of the fertile soil appears to resolve itself into a proper handling of bacteria. These organisms have stocked his soil in the first place. They convert all of his compost heap wastes into simple bodies, some of which are changed into plant foods, while others are at the same time lost. Lastly, they may be made to reclaim this lost nitrogen, and the fanner, so soon as he has requisite knowledge of these facts, will be able to keep within his control the supply of this important element. The continued fertility of the soil is thus a gift from the bacteria.

BACTERIA AS SOURCES OF TROUBLE TO THE FARMER.

While the topics already considered comprise the most important factors in agricultural bacteriology, the farmer's relations to bacteria do not end here. These organisms come incidentally into his life in many ways. They are not always his aids as they are in most of the instances thus far cited. They produce disease in his cattle, as will be noticed in the next chapter. Bacteria are agents of decomposition, and they are just as likely to decompose material which the farmer wishes to preserve as they are to decompose material which the farmer desires to undergo the process of decay. They are as ready to attack his fruits and vegetables as to ripen his cream. The skin of fruits and vegetables is a moderately good protection of the interior from the attack of bacteria; but if the skin be broken in any place, bacteria get in and cause decay, and to prevent it the farmer uses a cold cellar. The bacteria prevent the farmer from preserving meats for any length of time unless he checks their growth in some way. They get into the eggs of his fowls and ruin them. Their troublesome nature in the dairy in preventing the keeping of milk has already been noticed. If he plants his seeds in very moist, damp weather, the soil bacteria cause too rapid a decomposition of the seeds and they rot in the ground instead of sprouting. They produce disagreeable odours, and are the cause of most of the peculiar smells, good and bad, around the barn. They attack the organic matter which gets into his well or brook or pond, decomposing it, filling the water with disagreeable and perhaps poisonous products which render it unfit to drink. They not only aid in the decay of the fallen tree in his forests; but in the same way attack the timber which he wishes to preserve, especially if it is kept in a moist condition. Thus they contribute largely to the gradual destruction of wooden structures. It is therefore the presence of these organisms which forces him to dry his hay, to smoke his hams, to corn his beef, to keep his fruits and vegetables cool and prevent skin bruises, to ice his dairy, to protect his timber from rain, to use stone instead of wooden foundations for buildings, etc. In general, when the farmer desires to get rid of any organic refuse, he depends upon bacteria, for they are his sole agents (aside from fire) for the final destruction of organic matter. When he wishes to convert waste organic refuse into fertilizing material, he uses the bacteria of his compost heap. On the other hand, whenever he desires to preserve organic material, the bacteria are the enemies against which he must carefully guard.

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