The Project Gutenberg EBook of Outlines of dairy bacteriology, by H. L. Russell and E. G. Hastings This eBook is for the use of anyone anywhere at no cost and with almost no restrictions whatsoever. You may copy it, give it away or re-use it under the terms of the Project Gutenberg License included with this eBook or online at www.gutenberg.org Title: Outlines of dairy bacteriology A concise manual for the use of students in dairying Author: H. L. Russell E. G. Hastings Release Date: May 14, 2010 [EBook #32367] Language: English Character set encoding: ISO-8859-1 *** START OF THIS PROJECT GUTENBERG EBOOK OUTLINES OF DAIRY BACTERIOLOGY *** Produced by Stacy Brown, Peter Vachuska, Julia Miller and the Online Distributed Proofreading Team at https://www.pgdp.net OUTLINES OF DAIRY BACTERIOLOGY A CONCISE MANUAL FOR THE USE OF STUDENTS IN DAIRYING BY H. L. RUSSELL DEAN OF THE COLLEGE OF AGRICULTURE UNIVERSITY OF WISCONSIN AND E. G. HASTINGS PROFESSOR OF AGRICULTURAL BACTERIOLOGY UNIVERSITY OF WISCONSIN TENTH EDITION MADISON, WISCONSIN H. L. RUSSELL [Pg 1] 1914 Copyright 1914 BY H. L. RUSSELL and E. G. HASTINGS PREFACE TO THE TENTH EDITION. This text was originally the outgrowth of a series of lectures on the subject of dairy bacteriology to practical students in the winter Dairy Course in the University of Wisconsin. The importance of bacteriology in dairy processes has now come to be so widely recognized that no student of dairying regards his training as complete until he has had the fundamental principles of this subject. The aim of this volume is not to furnish an exhaustive treatise of the subject, but an outline and sufficient detail to enable the general student of dairying to obtain as comprehensive an idea of the bacteria and their effects on milk and other dairy products as may be possible without the aid of laboratory practice. When possible the dairy student is urged to secure a laboratory knowledge of these organisms, but lacking this, the student and general reader should secure a general survey of the field of bacteriology in relation to dairying. In this, the tenth edition, the effort has been made to include all of the recent developments of the subject. Especially is this true in regard to the subject of market milk, a phase of dairying that has gained greatly in importance in the last few years. The changes in the methods of handling market milk have been marked. The results of these changes in influencing the quality of milk offered to the consumer are fully discussed. H. L. R. E. G. H. CONTENTS Structure, Growth and Distribution of Bacteria 7 Methods of Studying Bacteria 20 Contamination of Milk 28 Infection of Milk with Pathogenic Bacteria 62 Fermentations of Milk 82 Preservation of Milk 113 Bacteria and Butter Making 136 Bacteria and Cheese Making 161 Bacteria in Market Milk 189 CHAPTER I. STRUCTURE, GROWTH AND DISTRIBUTION. Relation of bacteriology to dairying. The arts which have been developed by mankind have been the outgrowth of experience. Man first learned by doing, how to perform these various activities, and a scientific knowledge of the underlying principles which govern these processes was later developed. The art of dairying has been practiced from time immemorial, but a correct understanding of the fundamental principles on which the practice of dairying rests is of recent origin. In working out these principles, chemistry has been of great service, but in later years, bacteriology has also been most successfully applied to the problems of modern dairying. Indeed, it may be said that the science of dairying, as related to the problems of dairy manufacture is, in large degree, dependent upon an understanding of bacteriological principles. It is therefore essential that the student of dairying, even though he is concerned in large measure with the practical aspects of the subject, should acquire as complete an understanding of these principles as possible. While bacteriology is concerned primarily with the activities of those microscopic forms of plant life known as the [Pg 2] [Pg 3] [Pg 4] [Pg 7] bacteria, yet the general principles governing the life of this particular class of organisms are sufficiently similar to those governing the molds and other types of microscopic life that affect milk and its products to make it possible to include all of these types in a general consideration of the subject. Nature of bacteria. The vegetable kingdom to which the bacteria belong consists of plants of the most varying size and nature. Those of most common acquaintance are the green plants varying in size from those not visible to the naked eye to the largest trees. Another class of plants known as fungi or fungous plants do not contain chlorophyll, the green coloring matter, but are usually colorless and, as a rule, of small size; among them are included such forms as the mushrooms, smuts, rusts and mildews, as well as the molds and yeasts. The bacteria are closely allied to this latter class. When first discovered they were thought to be animals because of the ability of some forms to move about in liquids. The bacteria, like other kinds of living organisms, possess a definite form and shape. They are the simplest in structure of all the plants, the individual organism consisting of a single cell. The larger and more highly organized forms of life are made up of many microscopic cells, and the life of the individual consists of the work of all the cells. The bacteria are very comparable to the single cells of the higher plants and animals, but in the case of the bacteria the single cell is able to exist apart from all other cells and to carry out all of its life processes including reproduction. Forms of bacteria. With the multicellular organisms much variation in form is possible, but with these single-celled organisms the possible variation in form is greatly limited. Three well marked types occur among the bacteria: the round or coccus form (plural cocci); the rod-shaped or bacillus (plural bacilli); and the twisted or spirillum type (plural spirilla). Most organisms of special significance in dairying belong to the coccus or bacillus group. Size of bacteria. The bacteria, as a class, are among the smallest of living objects. None of them are individually visible to the naked eye, and they can be so seen only when clumps or masses are formed in the process of growth. Fig. 1.—Forms of Bacteria. A, coccus; B, bacillus; C, spirillum. While there is considerable relative variation in size, yet in actual dimensions, this difference is so small as to make careful microscopic determinations necessary. An average diameter may be taken as about one thirty-thousandth of an inch, while the length varies naturally several fold, depending upon whether the type under observation is a coccus or a bacillus. It is very difficult to conceive of the minuteness of the bacteria; the following may give some idea of their size. In a drop of cream ready for churning may be found as many as 10,000,000 and in a piece of fresh cheese as large as a cherry there may be as many living bacteria as there are people on our earth. While the bacteria are very minute, the effect which they exert in milk and other dairy products is great on account of their enormous numbers. Manner of growth. The cells of which all plants and animals consist increase in numbers by the division of each cell into two cells through the formation of a division wall across the cell. The new cells divide and the plant or animal continues to grow. The same cell division occurs in the bacteria but since the bacteria are single celled, division of the cells means an increase in numbers rather than growth as in the higher forms of life. Fig. 2.—Division of Bacteria. The bacteria increase in numbers by the division of each cell into two cells. (After Novy.) In the case of those bacteria that have a greater length than diameter, the new wall is formed at right angles to the long axis of the cell. As soon as the division is complete each cell is a complete individual, capable of carrying on all of its life processes. The cells may, however, cohere and thus form distinctive groupings that may serve to identify certain types. Some of the cocci form long chains and the term streptococcus is applied to such. Other groupings may be similar to a bale of twine or they may be massed in clusters with no regularity distinguishable. Spores. Just as ordinary plants form resistant structures, known as seeds, capable of retaining vitality under conditions unfavorable for growth thereby perpetuating the species, so with certain of the bacteria, definite structures, known as spores, that are analogous in some respects to the seeds of the higher plants, are produced within the mother cell. The spores are exceedingly resistant to the influence of an unfavorable environment, such as heat, cold, drying, and even chemical agents. It is this property of the spores which makes it so difficult to destroy the bacterial life in the process of [Pg 8] [Pg 9] [Pg 10] [Pg 11] sterilizing milk. The property of spore-formation is fortunately confined to a comparatively small number of different species of bacilli. Movement. Many of the bacteria are provided with vibratory organs of locomotion, known as cilia (singular cilium) which are variously distributed on the surface of the cell. By the movement of these relatively long, thread-like appendages the individual cell is able to move in liquids. It must be remembered, when these moving cells are observed under the microscope, that their apparent rate of movement is magnified relatively as much as their size. Conditions for growth. All kinds of living things need certain conditions for growth such as food, moisture, air and a favorable temperature. The bacteria prefer as food such organic matter as milk, meat, and vegetable infusions. Those living on dead organic matter are known as saprophytes, while those which are capable of thriving in the tissues of the living plant or animal are known as parasites. Certain of the parasitic forms are capable of causing disease in plants and animals. In the first group are embraced most of the bacteria that are able to develop in milk or its products, such as those forms concerned in the spoiling of milk or its fermentation. It is true that milk may contain disease-producing bacteria coming either from a diseased animal or from a diseased human being. It is also true that some of such harmful forms are able to grow in milk, such as the organisms causing typhoid fever and diphtheria. Food. The bacteria like all other plants must have their food in solution. Where they apparently live on solids, such as meats, fruits, etc., they dissolve the food substances before utilizing the same. If the solutions are highly concentrated, as in the case of syrups, preserves and condensed milk, the bacteria cannot readily grow, although all of the necessary food ingredients are present. When such concentrated solutions are diluted, bacterial growth will take place and the solutions will spoil. Fig. 3.—Photomicrograph of Lactic Acid Bacteria. Each cell is an individual organism, magnified 1250 diameters. Generally speaking the bacteria grow best in a neutral or slightly alkaline solution rather than in acid liquids. Temperature. One of the most important conditions influencing the rate of growth of bacteria is the temperature. Each form has a minimum temperature below which growth can not take place; also a maximum above which growth is again impossible. For the majority of species the minimum temperature ranges from 40 to 45° F. the maximum from 105 to 110° F. Growth takes place most rapidly at the optimum temperature, which, for each species, lies close to the maximum temperature at which growth can occur. Most of the bacteria of importance in the dairy grow well at from 70 to 100° F. There are forms that can grow below the freezing point of water when they are in solutions that do not freeze at this temperature. There are still other bacteria that can grow at 140° F. a temperature that is quickly fatal to most forms. These are of importance in the dairy since they limit the temperatures at which milk can be stored for long periods of time. Air supply. Living organisms, both plant and animal, require air or oxygen for the combustion of their food and for the production of energy. Most bacteria use, as do the green plants and animals, the free oxygen of the air for their respiration. Such organisms are called aerobic or air-living. A much smaller group possess the power of taking oxygen from organic compounds such as sugar and the like and therefore are able to live under conditions where air is excluded. These are called anaerobic bacteria. A large number of bacteria are able to live either in the presence or in the absence of free oxygen. Most of the bacteria of importance in the dairy are of this nature. Rate of growth. When there is an abundant supply of food and when the temperature conditions are favorable, the bacteria increase in numbers with astounding rapidity. It has been determined by actual experiment that the process of [Pg 12] [Pg 13] [Pg 14] cell division under favorable conditions takes place in a few moments. Barber has shown that one of the forms of bacteria constantly found in milk will divide in 17 minutes at 98° F. and that a single organism kept at this temperature for ten hours would increase to 1,240,000,000. If the temperature is reduced to 50° F., the time required for division is increased to several hours. The explanation for the rapid spoiling of milk that is not well cooled is thus apparent. The initial rapid rate of increase cannot be maintained for any length of time as the conditions become more and more unfavorable as growth continues, due to the accumulation of the by-products of the cell activity. Thus, the growth of acid-forming organisms in milk becomes checked by the formation of acid from the fermentation of the sugar. Detrimental effect of external conditions. Environmental conditions of a detrimental character are constantly at work tending to repress the activity of bacteria or to destroy them. These act more readily on the vegetating cells than on the more resistant spores. It is of the utmost importance that those engaged in dairy work be familiar with these antagonistic forces since it is constantly necessary to repress or to kill outright the bacteria in milk and other dairy products. In many lines of dairy work it is likewise important to be familiar with the conditions favorable for bacterial growth. Effect of cold. While it is true that chilling largely prevents fermentative action, and actual freezing stops all growth processes, still it does not follow that exposure to low temperatures will effectually destroy the vitality of bacteria, even in the growing condition. Numerous non-spore-bearing species remain alive in ice for a prolonged period, and experiments with liquid air show that even a temperature of-310° F. maintained for hours does not kill all exposed cells. Effect of heat. High temperatures, on the other hand, will destroy any form of life, whether in the vegetative or latent spore stage. The temperature at which the vitality of the cell is lost is known as the thermal death point. This limit is dependent not only upon the nature of the organism, but upon the time of exposure and the condition in which the heat is applied. In a moist atmosphere, the penetrating power of heat is great, consequently cell death occurs at a lower temperature than in a dry atmosphere. An increase in time of exposure lowers the temperature point at which death occurs. For growing organisms, the thermal death point of most species ranges from 130° to 140° F. for ten minutes. When spores are present, resistance is greatly increased, some forms being able to withstand steam at 212° F. from one to three hours. In the sterilization of milk, it is often necessary to heat for several hours, where a single exposure is made, to destroy the resistant spores, that seem to be more abundant under summer than winter conditions. Steam under pressure is a much more effective agent, as the temperature is thus raised considerably beyond 212° F. An exposure of twenty minutes, at a temperature of 230° to 240° F. will kill all spores. Where heat is used in a dry state, it is much less effective, a baking temperature of 260° to 300° F. for an hour being necessary to kill spores. This condition is of the utmost importance in the destruction of bacteria in the dairy and creamery. Effect of drying. The spore-bearing bacteria withstand effects of desiccation without serious injury, and many of the non-spore-producing types retain their vitality for some months. The bacteria found in the air are practically all derived from the soil, and exist in the air in a dried condition, in which they are able to remain alive for considerable periods of time. In a dried condition, active cell growth is not possible, but when other conditions, such as moisture and food supply are present, resumption of growth quickly begins. This property is also of importance in the dairy as in the preparation of dry starters for creameries and cheese factories. Effect of light. Bright sunlight exerts a markedly injurious effect on bacterial life, both in a spore and in a growing condition. Where the direct sunlight strikes, more or less complete disinfection results in the course of a few hours, the effect being produced by the chemical or violet rays, and not by the heat or red rays of the spectrum. This action, however, does not penetrate opaque objects, and is therefore confined to the surface. In diffused light, the effect is much lessened, although it is exerted to some extent. Sunlight exerts a beneficial effect on the general health and well- being of animal life, and is a matter of importance to be taken into consideration in the erection of buildings for animals as well as for people. Effect of chemicals. A great many chemical substances exert a more or less powerful toxic action on various kinds of life. Many of these are of great service in destroying bacteria or holding them in check. Those that are toxic and result in the death of the cell are known as disinfectants; those that merely inhibit, or retard growth are known as antiseptics. All disinfectants must of necessity be antiseptic in their action, but not all antiseptics are disinfectants, even when used in large amounts. Disinfectants have no place in dairy work, except to destroy disease-producing bacteria, or to preserve milk for analytical purposes. The so-called chemical preservatives used to "keep" milk depend for their effect on the inhibition of bacterial growth. In this country, most states prohibit the use of these substances in milk. Their only function in the dairy should be to check fermentative and putrefactive processes outside of milk and so keep the air free from taints. Products of growth. All bacteria, as a result of their growth in food substances, form more or less characteristic compounds that are known as by-products. The changes brought about are those of decomposition and are collectively known as fermentations; they are characterized by the production of a large amount of by-products as the result of the development of a relatively small amount of cell life. The souring of milk, the rotting of eggs, the spoiling of meats, the making of vinegar from cider are examples of fermentations caused by different bacteria. If the substances decomposed contain but little sugar, as do animal tissues, the conditions are favorable for the growth of the putrefactive bacteria, and foul-smelling gases are formed. When sugars are present, as in milk, the environmental [Pg 15] [Pg 16] [Pg 17] conditions are most favorable for the acid-forming bacteria that do not as a rule produce offensive odors. Many of the bacteria form substances known as enzymes which are able to produce certain decomposition changes in the absence of the living cells, and it is by virtue of these enzymes that the organisms are able to break down such enormous quantities of organic matter. Most of these enzymes react toward heat, cold, and chemical poisons in a manner quite similar to the living cells. In one respect, they are readily differentiated, and that is, that practically all of them are capable of producing their characteristic chemical transformations under conditions where the activity of the cell is wholly suspended as in a saturated ether or chloroform atmosphere. The production of enzymes is not confined to bacteria, but they are found throughout the animal and plant world, especially in those processes that are concerned in digestion. Rennet, used in cheese making, is an example of an animal enzyme. Distribution of bacteria. As bacteria possess greater powers of resistance than almost any other form of life, they are found very widely distributed over the surface of the earth. In soil they are abundant, because of the fact that all of the conditions necessary for growth are here best satisfied. They are, however, distributed with reference to the layers of the soil; the soil proper, i.e., that turned over by the plow, is extremely rich in them on account of the abundance of organic matter. But at the depth of a few feet they decrease rapidly in numbers, and in the deeper layers, from six to ten feet, or more, they are normally not present, because of the lack of proper food supply and oxygen. The fertility of the soil is closely associated with their presence. The bacteria are found in the air because of their development in the soil below. They are unable to grow even in a moist atmosphere, but are so readily dislodged by wind currents from the soil that over land areas the lower strata of the air always contain them. They are more numerous in summer than in winter; city air contains larger numbers than country air. Wherever dried fecal matter is present, as in barns, the air contains many forms. Water generally contains enough organic matter in solution, so that certain types of bacterial life find favorable growth conditions. Water in contact with the soil surface takes up many impurities, and is of necessity rich in bacteria. As the rain water percolates into the soil, it loses its germ content, so that the normal ground water, like the deeper soil layers, contains practically no bacterial life. Springs, therefore, are relatively deficient in germ life, except as they become contaminated with soil organisms, as the water issues from the ground. Wells vary in their germ content, depending upon manner of construction, ease of contamination at surface, etc. Wells are too frequently insufficiently protected from surface leachings, and consequently may contain all kinds of organisms found in the surface soil. Typhoid fever is very frequently disseminated in this way, as is cholera and a number of animal maladies. While the inner tissues of healthy animals are free from bacteria, the natural passages, as the respiratory and digestive tracts, being in more direct contact with the exterior, become readily infected. This is particularly true with reference to the intestinal tract, and in the undigested residue of the food, bacterial activity is at a maximum. The result is that fecal matter of all kinds contains enormous numbers of organisms so that the pollution of any food medium, such as milk, with such material is sure to introduce elements that seriously affect its quality. CHAPTER II. METHODS OF STUDYING BACTERIA. Necessity of artificial cultivation. The bacteria are so extremely small, that it is impossible to study individual germs separately without the aid of powerful microscopes. Little advance was made in the knowledge of these lower forms of plant life until the introduction of culture methods, whereby a single organism could be cultivated, and the progeny of this cell increased to such an extent in a short course of time that the resulting mass of cells would be visible to the unaided eye. This is done by growing the bacteria on various kinds of nutrient media that are prepared for the purpose, but inasmuch as bacteria are so universally distributed, it becomes an impossibility to cultivate any special form alone, unless the medium in which they are grown is first freed from all pre-existing forms of germ life. Food materials. Many kinds of food substances are used for the cultivation of bacteria in the laboratory. In fact, bacteria will grow on almost any organic substance, whether it is solid or liquid, provided the other essential conditions of growth are furnished. The food substances that are used for culture purposes are divided into two classes,—solids and liquids. Solid culture media may be either permanently solid, like potatoes and coagulated egg, or they may retain their solid properties only at certain temperatures, like gelatin or agar. The latter two, which were devised by Robert Koch, are of utmost importance in bacteriological research, for their use permits the separation of the different forms of bacteria that may happen to be in any mixture. Gelatin is advantageously used, because the majority of bacteria present wider differences, due to growth upon this medium, than upon any other. It remains solid at ordinary temperatures, becoming liquid at about 80° F. Agar, a gelatinous product derived from a Japanese seaweed, has a much higher melting point, and is used especially with those organisms whose optimum temperature for growth is above the melting point of gelatin. Besides these solid culture media, different liquid substances are extensively used, such as beef broth, milk and infusions of various vegetable and animal tissues. Skim milk is of especial value in studying the milk bacteria, and may be used in [Pg 18] [Pg 19] [Pg 20] [Pg 21] its natural condition, or a few drops of litmus solution may be added, in order to detect any change in its chemical reaction due to the bacteria. Sterilization. The various ingredients that are used in the preparation of culture media are not free from micro- organisms, hence the media would soon spoil if they were not destroyed, and the media subsequently protected from contamination from the air, etc. The process of rendering the media free from living micro-organisms is known as sterilization. It may be accomplished in a number of ways, but most often is done by the use of heat. For culture material, which is always organic in character, moist heat is employed. The various culture media, in appropriate containers, are subjected to a thorough steaming in a steam cooker. This destroys all of the vegetating cells but not the resistant spores that may be present. The media are then stored, for twenty-four hours, at temperatures favorable for the germination of the spores and are then again heated. Three such applications on successive days are usually sufficient to free the media from all living germs, since between the heating periods the spores germinate and the resulting vegetative cells are more easily destroyed. The sterile media will keep for an indefinite period in a moist place. The media are usually placed in glass containers which may be sterilized before use by heating them in an oven, it being possible to thus secure a much higher temperature than with streaming steam. All glass or metal articles may be sterilized by the use of dry heat but for organic media, to avoid burning, moist heat must be used. All kinds of materials may be sterilized by treatment with steam under pressure. An exposure for a few moments at 250° F., a temperature attained with 15 pounds steam pressure, will destroy all kinds of bacteria and their spores. This method of sterilization is used in the canning of meats and vegetables and in the preparation of evaporated milk. To avoid contamination of the media after sterilization, the flasks and tubes are, after being filled, stoppered with plugs of cotton-wool, which effectually filter out all bacteria and mold spores from the air, and yet allow the air to pass freely in and out of the containers. Methods of determining the number of bacteria. The method of determining the number and kinds of bacteria in any substance can be illustrated by the process as applied to milk. For this purpose the method of procedure is as follows: Sterile gelatin in glass tubes is melted and then cooled until it is barely warm. To this melted gelatin a definite quantity of milk is added. The medium is gently shaken, so as to thoroughly mix the milk and gelatine, and the mixture then poured into a sterile, flat, glass dish, and quickly covered, where it is allowed to cool until the gelatin hardens. After the culture plate has been left for twenty-four to thirty-six hours at the proper temperature, tiny spots will begin to appear on the surface, or in the depth of the culture-medium. These spots are called colonies, and are composed of an almost infinite number of individual cells, the result of the continued growth of a single organism that was in the drop of milk and which was firmly held in place when the gelatin solidified. The number of these colonies represents approximately the number of living bacteria that were present in the amount of milk added to the tube of gelatin. If the plate is not too thickly sown with the bacteria, the colonies will continue to grow and increase in size, and as they do, minute differences will begin to appear. These differences may be in the color, the contour, and the texture of the colony, or the manner in which it acts toward gelatin. Fig. 4.—Plate Culture. Each of the dots is a colony that has been formed by the growth of an organism embedded in the solid culture- medium. By counting the colonies, the number of living bacteria in the amount of milk added to the culture is determined. In order to make sure that the number of colonies is not so numerous as to prevent counting and further study of their characteristics, a series of plate cultures is usually made in which varying amounts of milk are added to the tubes of gelatine. This is attained by adding a definite amount of the milk or other substance to be examined to a measured [Pg 22] [Pg 23] [Pg 24] amount of sterile water, e.g., one cubic centimeter of milk to ninety-nine cubic centimeters of water. One cubic centimeter of this mixture may be used for the inoculation of the plate culture. This dilution may be carried on to any desired extent; in the examination of many dairy products, it is necessary to use very minute quantities of material, often only one one-millionth of a cubic centimeter. To study further the peculiarities of the different bacteria, small portions of the individual colonies are transferred to tubes of sterile culture-media. In order to do this the colony is touched with a piece of platinum wire; the minute amount of growth that adheres to the wire is sufficient to seed the tube of fresh culture-medium. The inoculating needle must always be sterilized before use by passing it through a gas flame. A culture thus obtained is called a pure culture since it contains but a single kind of an organism, as the colony is the result of the growth of a single cell. These cultures then serve as a basis for continued study, and must be planted and grown upon the different kinds of media that are obtainable. In this way the slightest variations in the growth of different forms are detected, and the peculiar characteristics are determined, so that the student is able to recognize this form when he meets it again. Fig. 5.—Different Kinds of Bacteria Growing in Gelatin. A, meager growth, no liquefaction or surface growth; B, profuse surface growth, radiating filaments from the growth below the surface; C, a rapid liquefying form; D, a gas producer that grows equally well in the presence or absence of air; E, form that grows only in the absence of air, an anaerob. These culture methods are of essential importance in bacteriology, as it is the only way in which it is possible to secure a quantity of germs in a pure state. The microscope in bacterial investigations. In order to verify the purity of the cultures, the microscope is in constant demand throughout all the different stages of the isolating process. For this purpose it is essential that the instrument used shall be one of high magnifying powers (600 to 800 diameters), combined with sharp definition. The microscopical examination of any germ is quite as essential as the determination of culture characteristics, in fact, the two must go hand in hand. The examination reveals not only the form and size of the individual germs but the manner in which they are united with each other, as well as any peculiarities of movement that they may possess. In carrying out the microscopical part of the work, not only is the organism examined in a living condition, but colored preparations are made by using solutions of anilin dyes as staining agents. These are of great service in bringing out almost imperceptible differences. The art of staining has been carried to the highest degree of perfection in bacteriology, especially in the detection of germs that are found in diseased tissues in the animal or human body. In studying the peculiarities of any special organism, not only is it necessary that these cultural and microscopical characters should be closely observed, but special experiments must be made in different ways, in order to determine any special properties that the germ may possess. Thus, the ability of any form to act as a fermentative organism can be tested by fermentation experiments; the property of causing disease, studied by the inoculation of pure cultures into experimental animals, like rabbits, guinea pigs and white mice. The methods of the bacteriologist in his laboratory are in their effect not dissimilar to those which the farmer employs in securing his crop of pure-bred grain. The laboratory farmer kills the weed seeds in his culture field by the application of heat. His field, which is embraced in his culture dish, has been fertilized and prepared by the addition of certain favorable ingredients. When he has garnered his crop, he maintains its purity by keeping his selected seed, the pure culture, free from all contamination. The dairyman, even though he may not expect to carry on the detailed operations of the laboratory, will understand the reason for the directions which he is often required to follow much better if he knows how the simple operations of the laboratory are carried out. For a fuller knowledge of these matters, the reader is [Pg 25] [Pg 26] [Pg 27] referred to the special texts on bacteriology. CHAPTER III. CONTAMINATION OF MILK. Spoiling of milk. Materials of animal origin are peculiarly prone to undergo changes, rendering them unfit for use, and of these, milk is exceedingly susceptible to such changes. This is due to the fact that the composition of milk is especially adapted to bacterial growth, and that the opportunity for entrance of such organisms is likewise such as to permit of abundant contamination. The consequence is that milk readily undergoes fermentative changes, due to the development of one or another type of micro-organism. Milk, a suitable bacterial food. While milk is designed by nature for the nourishment of mammalian life, it is, curiously enough, equally well adapted to the growth of these lowest forms of vegetable life. The nutritive substances required by bacteria are here sufficiently dilute to make possible rapid growth. Milk also contains all the necessary chemical substances to make a suitable bacterial food supply. Of the nitrogenous compounds, albumen is in a readily assimilable form. Casein, the principal nitrogenous constituent of milk, exists in an insoluble condition, and cannot be directly utilized, until it is acted upon by digesting enzymes. The fat in milk does not readily decompose, and while there are a few bacteria capable of splitting this substance, the majority of organisms are unable to utilize it. Milk sugar, on the other hand, is an excellent food for most species. Fig. 6.—Fat Globules and Bacteria. Note the relative size of the fat globules of milk and the lactic acid bacteria. Sources of contamination. Inasmuch as milk is especially exposed to the inroads of bacterial growth, and because of the fact that much of the contamination can easily be prevented, it is highly important that the milk producer and dealer should be thoroughly cognizant of the various sources of contamination. The different factors concerned in contamination may be grouped as follows: the interior of the udder; utensils, including all apparatus with which the milk is brought in contact subsequent to withdrawal from the animal; infection coming from the animal herself, from the milker, and the surrounding air. Condition of milk when secreted. Immediately after withdrawal from the udder, milk always contains bacteria, yet in the secreting cells of the udder of a healthy cow, germ life does not seem to be present. Only when the gland is diseased are bacteria found in any abundance. In the passage of the milk from the secreting cells to the outside, it receives its first infection, so that when drawn from the animal it generally contains a considerable number of organisms. A study of the structure of the udder shows the manner in which such infection occurs. Structure of the udder. The udder is composed of secreting tissue (gland cells) that is supported by fibrous connective tissue. The milk is elaborated in these cells and is discharged into microscopic cavities, from whence it flows through the numerous channels (milk sinuses) that ramify through the substance of the udder, until finally it is conveyed into the milk cistern, a common receptacle holding about one half pint that is located just above the teat. This cavity is connected with the outside by a direct opening (milk duct) through the teat. During the process of milking, the milk is elaborated rapidly in the gland cells, and their contents upon rupture of the milk cells, flow down into the cistern. The normal contraction of the muscles at the lower opening of the outer duct prevents the milk from passing out except when pressure is applied, as in milking. The inner walls of the milk duct and cistern are always more or less moist, and [Pg 28] [Pg 29] [Pg 30] therefore afford a suitable place for bacteria to develop, if infection once occurs, and conditions are favorable for growth. Manner of invasion. Two possible sources of invasion of the udder by bacteria may exist. If bacteria are present in the circulating blood, there is the possibility of organisms passing directly through the tissues into the milk-secreting cells. The other alternative is the possible direct contamination from the outside by organisms passing up through the milk duct, and so spreading through the open channels in the udder. Fig. 7.—Sectional View of Udder. Teat with milk duct connecting the exterior with the milk cistern. Milk sinuses which conduct the milk from the secreting tissue to the milk cistern. (After Moore & Ward.) Number of bacteria in fore-milk. If a bacteriological examination is made of the milk drawn from each teat at different periods during the milking process, it will be found that the fore-milk, i.e., the first few streams, contains, as a rule, many more organisms per cubic centimeter than that removed later. Not infrequently thousands of organisms per cubic centimeter may be found in the first streams while the middle milk, or strippings, will contain much smaller numbers. Distribution and nature of bacteria in udder. If the udder itself is carefully examined as to its bacterial content, it appears that the majority of organisms found is confined to the lower portion of this organ, in the teat, milk-cistern and large milk-ducts; while bacteria occur in contact with the secreting tissue, they are relatively less abundant. This would seem to indicate that the more probable mode of infection is through the open teat. While there is no constant type of bacteria found in the fore-milk, yet it is noteworthy that nearly all observers agree that the organisms most commonly found are not usually the acid-producing, or gas-generating type, so abundant on the skin or hairy coat of the udder and which predominate in ordinary milks. Coccus forms, belonging to both liquefying and non-liquefying types are most generally present. Many of these produce acid slowly and in small quantities. The bacteria coming from the interior of the udder are of small practical significance since they do not grow rapidly at the temperatures at which milk is stored. If the milk is protected from contamination from other sources, the bacteria from the udder will ultimately cause it to spoil, but under ordinary conditions other forms are present in such greater numbers, and grow so much more rapidly in milk, that the udder forms have small opportunity to exert any effect. It is interesting to note that the bacteria found in the udder are similar to those that seem to be most abundant in such glandular tissues as the liver and spleen. This fact increases the probability that these comparatively inert coccus forms of the udder may originate directly from the blood stream. The organisms that normally are found in the udder exert no [Pg 31] [Pg 32] [Pg 33] harmful effects on the gland. It might be thought that due to the presence of abundant food and a favorable temperature that growth would be abundant, but such is not the case. At times the udder may be invaded by forms that are not held in check by the natural factors and an inflammation of the udder is likely to result. Germicidal property of milk. It has been claimed that freshly drawn milk, like other body fluids, possesses germicidal properties, i.e., the power of destroying bacteria with which it may be brought in contact. If milk is carefully examined bacteriologically, hour by hour, after it is withdrawn from the udder, it will generally be found that there is at first not only no increase in number of organisms during a longer or shorter period when it is kept at temperatures varying from 40° to 70° F., but that an actual reduction not infrequently takes place. When cultures of bacteria, such as B. prodigiosus, a red organism, lactic acid organisms, and even the yellow, liquefying coccus, so commonly found in the fore-milk, are artificially introduced into the udder, it has been found that no growth occurs and that in the course of a few days the introduced organisms actually disappear. Whether this failure to colonize can be regarded as evidence of a germicidal property or not is questionable. In fact, this question is a matter of but little practical importance in the handling of milk since, under the best of conditions, the keeping quality of the milk is not materially enhanced. It may be of importance in inhibiting growth in the udder. Rejection of fore-milk. The fact that the fore-milk contains per cubic centimeter so much more germ life than the remainder of the milk has led some to advocate its rejection when a sanitary milk supply is under consideration. While from a purely quantitative point of view, this custom may be considered advantageous, in practice, however, it is hardly worth while since it is not at all certain that the rejection will have any effect on the keeping quality or healthfulness of milk. This is especially true if the ends of the teats are thoroughly cleaned before milking. It is true that the fore-milk is relatively deficient in fat so that the loss of butter fat occasioned by the rejection of the first few streams is comparatively slight. Contamination from utensils. One of the most important phases of contamination is that which comes from the utensils used to hold the milk from the time it is drawn until it is utilized. Not only is this important because it is a leading factor in the infection of milk, but because much improvement can be secured with but little trouble, and it is especially necessary that the dairy student should be made familiar with the various conditions that obtain. Pails and cans used to hold milk may be apparently clean to the eye, and yet contribute materially to the germ content of the milk placed in them. Not only does much depend upon their condition, but it is equally important to take into consideration their manner of construction. Dairy utensils should be simple in construction, rather than complex. They should be made so that they can be readily and easily cleaned, or otherwise the cleaning process is apt to be neglected. Of first importance are those utensils that are used to collect the milk and in which it is handled while on the farm. The warm milk is first received in pails, and unless these are scrupulously cleaned, an important initial contamination then occurs. As ordinarily washed, the process falls far short of ridding the utensils of the bacterial life that is adherent to the inner surface of the pail. Then, too, all angles or crevices afford an excellent hiding place for bacteria, and it is very important to see that all seams are well soldered. Round corners and angles flushed with solder greatly facilitate thorough cleaning of utensils. Tin utensils are recognized as most satisfactory. Shipping cans are likely to serve as greater infecting agents than pails for they are subject to more wear and tear and are harder to clean. As long as the surface is bright and smooth, it may be easily cleaned, but large utensils, such as cans, are likely to become dented and rusty in spots on the inner side. The storage of milk in such utensils results in its rapid deterioration. The action of rennet has been found to be greatly retarded where milk comes in contact with a rusty iron surface. It is also probable that some of the abnormal flavors in butter are due to the action of acid cream on iron or copper surfaces from which the tin has been worn. It is equally important that attention be paid to the care of strainers, coolers, and the small utensils. Cloth strainers are more or less of a hotbed for bacterial growth, for unless they are boiled, and then dried quickly and thoroughly, germ growth will continue apace in them, as long as they contain any moisture. Milking machines and farm separators. The introduction of these special types of dairy machinery in the handling of milk on the farm has materially complicated the question of the care of milk. Both of these types of apparatus are much more complicated than the usual milk utensil; consequently, the danger of imperfect cleaning is thereby increased. This is still further accentuated by the fact that cleansing of utensils on the farm can never be done so well as at the factory or milk depot where steam is available. The milking machine may be easily kept in a comparatively germ-free condition, but unless this is done, it contributes its quota of germ life to the milk. The farm separator is more widely used than the milking machine and in actual practice the grossest carelessness prevails in the matter of its care. Frequently it is not taken apart and thoroughly cleansed, but is rinsed out by passing water through the machine. It is impossible by such a treatment to remove the slime that collects on the wall of the bowl; the machine remains moist and bacterial growth can go on. Such a machine represents a most important source of contamination of milk and cream and it is probable that the widespread introduction of the hand separator has contributed more to lower the quality of cream delivered at the factory than any other single factor. Contamination from factory by-products. The custom of returning factory by-products in the same set of cans that is used to bring fresh milk is a prominent cause of bad milk. Whey and skim milk are rich in bacterial life, and not infrequently are so handled as to become a foul, fermenting mass. If the cans used to transport this material are not scrupulously cleaned on the farm, transfer of harmful bacteria to the milk is made possible. In this way the carelessness [Pg 34] [Pg 35] [Pg 36] [Pg 37] of a single patron may be the means of seeding the whole factory supply. This custom is not only liable to produce a poor quality of milk, but it is more or less of a menace to all the patrons of a factory, inasmuch as the opportunity always obtains that disease-producing organisms may thus be introduced into the supply. Not infrequently is tuberculosis thus spread through the medium of factory by-products. Fig. 8.—Whey Disposal. Whey barrels at a Wisconsin Swiss cheese factory. Each patron's share is placed in a barrel which is so situated that it is impossible to empty it completely; thus it is not cleaned during the season. The manufacture of Swiss cheese presents a striking example of the disregard which factory operators show toward the employment of bacteriological principles. In these factories, the custom is widely practiced of apportioning the patrons' allotment of whey into individual barrels which are supposed to be emptied each day. As these barrels are, however, rarely ever cleaned from the beginning to the end of the season, they become very foul, and the whey placed in them from day to day highly polluted. It is this material which is taken back to the farms in the same set of cans that is used for the fresh milk. When one recalls that the very best type of milk is essential for the making of a prime quality of Swiss cheese, and that to secure such, the maker insists that the patron bring the product to the factory twice daily, the before mentioned practice appea...