The Wonderful Development of Modern Bacteriology

The Wonderful Development of Modern Bacteriology

BY J. E. GREAVES, PH. D., PROFESSOR OF BACTERIOLOGY, STATE AGRI- CULTURAL COLLEGE.

Probably in no field of human endeavor has research been crowned with such glorious achievements, at least in so far as the welfare of the human race is concerned, as in the field of bac- teriology and this, in spite of the fact that it had the most humble ;

and recent beginning. Even the very dawn of its development dates back only to 1675 when Leeuwenhoek, a Dutch linen-draper, spent his leisure time in the grinding of lenses. In this he became so proficient that he perfected a lens superior to any that had been made up to his time. With his crude lens he examined raindrops, his own saliva, and many putrefying substances. In these he ob- served many living, moving forms, which, prior to this time, had been unseen. We can imagine his joy and surprise from his statement

"I saw with wonder that my material contained many tiny animals which moved about in a most amusing fashion; the largest of these showed the liveliest and most active motion, moving through the water or saliva as a fish of prey darts through the sea. They we're found everywhere, although in no large numbers. A second kind sometimes spun around in a circle like a top. They were present in larger quan- tities. A third kind could not be distinguished so clearly; now they appeared oblong, now quite round. They looked like a swarm of gnats or flies dancing about together. I had the impression that I was looking at several thousand in a given part of the material from the teeth no larger than a grain of sand, when only one part of the mate- rial was added to nine parts of water or saliva."

We find this patient worker giving a very accurate description of these minute forms of life, but even this did not awaken the world to a full realization of the wonderful, invisible forms of life which were present in everything, and always working for good or evil. It did, however, stimulate discussion as to whether these micro-organisms were the result of spontaneous generation due to vegetative changes in the substances in which the organisms were found, or whether they were the descendants of pre-existing or- ganisms. Back in the sixteenth century, we find the famous physicist and chemist, Van Helmont, stating that mice can be spontaneously generated by merely placing some dirty rags in a receptacle to- gether with a few grains of wheat or a piece of cheese. The same

308 IMPROVEMENT ERA philosopher's method of engendering scorpions appears to us very amusing "Scoop out a hole in a brick. Put into it some sweet basil. Lay a second brick upon the first, so that the hole may be perfectly covered. Expose the two bricks to the sun, and at the end of a few days the smell of the sweet basil, acting as a ferment, will change the herb into a real scorpion."

An Italian, Buonami, tells of a wonderful metamorphosis which he had witnessed. Rotten timber, which he rescued from the sea, produced worms these gave rise to butterflies, and the ;

butterflies, strangest of all, became birds. These false notions were overthrown by the Italian poet and physician, Redi, who clearly demonstrated that larvae were not spontaneously generated in decomposing meat. He merely took the precaution of placing the meat in a wide-mouthed bottle and covered the mouth of the bottle with gauze. Flies, attracted by the odor, deposited their eggs on the gauze and it was shown that it was from these, and not through spontaneous generation, that the so-called worms arose. The theory of the spontaneous generation of maggots and the like had been proven to be untenable but how about these ;

microscopic organisms ? Could they develop directly from organic material? Now anyone provided with this new instrument, the microscope, could easily demonstrate for himself the spontaneous generation of microscopic eels in vinegar or produce myriads of different and interesting living creatures in a simple infusion of hay or other organic material. Needham and the great naturalist, Buffon, evolved the theory that a force called Productive of vegetative force existed which was responsible for the production of organized beings. Buffon elaborated the theory that there were certain unchangeable parts common to all living things. These ultimate organic constituents he supposed capable of taking various molds or shapes which constituted the various living creatures. After death these ultimate constituents were supposed to be set free and become very active. Uniting with one another and other particles they formed swarms of microscopic creatures. Needham took decaying organic matter and enclosed it in vessels which he placed upon hot ashes to destroy any existing animalculae. Yet later in these fluids he found micro-organisms which were not there in the beginning. In 1769 Spallanzani repeated the work, using hermetically sealed flasks, and sterilized by boiling for one hour. He writes "I used hermetically sealed vessels. I kept them for one hour in boiling water and, after opening and examining their contents after a reasonable interval. I found not the slightsst trace of animalculae, though I had examined the infusion from nineteen different vessels ''

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But the reply of Needham was that the boiling had altered the character of the infusion so that it was unable to produce life. Voltaire, with his characteristic satire, took up the fight at this point and ridiculed the operations of the English clergy who had engendered eels in the gravy of boiled mutton, and he wittily re- marks, "It is strange that men should deny a Creator and yet attribute to themselves the power of creating eels." But this was a controversy to be settled not by ridicule but by experimental evidence. So we find Spallanzani answering this by cracking one of the flasks so that air could enter. Decay soon set in. Even this was not sufficient to overthrow a popular belief, for the claim was made that the hermetically sealing of the flasks excluded the air and this was essential to the normal development of these forms of life. This objection was answered by the work of many an ingenious investigator, some by passing the air through tubes con- taining acid and then into the infusion, the acid removing the micro-organisms, others by passing the air through a red hot tube. But the final proof came when it was shown that it was sufficient to place cotton plugs in the bottles so that, as the air passed in, the microscopic organisms would be held back by the cotton. Every now and then the contents of a flask would spoil, even after it had been carefully stoppered and then boiled. This remained a stum- bling block in the way of those who maintained that life sprang only from life, until in the year 1865, when Pasteur demonstrated the fact that many bacteria may pass into a resting stage, and while in this condition they will withstand conditions which kill them quickly while in the vegetation stage. Eleven years later Conn, of Breslau, investigated very carefully organisms when in this resting or spore stage, and today we know forms of micro- organisms which will withstand boiling water for sixteen hours without killing, and others are even resistant enough to endure for many hours a ten per cent solution of carbolic acid. Since the dawn of history man has been interested in the won- derful process known as fermentation, and many an ingenious theory has been formulated to explain it, but nothing more than theory existed until the classic works of Louis Pasteur appeared on fermentation, about 1837. He claimed that all forms of fer- mentation were due to the action of microscopic organized cells. An idea such as this, at this late date, did not go unchallenged, for we find no less illustrious workers than Helholtz and Liebig opposing him. Liebig even scoffed at such an idea for we find him writing: "Those who pretend to explain the putrefaction of animal sub- stances by the presence of micro-organisms reason very much like a child who would explain the rapidity of the Rhine by attributing it to the violent motions imparted to it in the direction of Burgen by the numerous wheels of the mills of Venence."

310 IMPROVEMENT ERA Pasteur's carefully planned experiments, however, soon showed that without these micro-organisms there would be no fermentation, no putrefaction, no decay of any tissues, except by the slow process of oxidation. The care with which his experi- ments were planned and executed are well shown in the experi- ments with grape sugar, concerning which he writes

"I prepared forty flasks of a capacity of from two hundred and fifty to three hundred cubic centimeters and filled them half full with filtered grapemust, perfectly clear, and which, as is the case of all acidulated liquids that have been boiled for a few seconds, remains uncontaminated, although the curved neck of the flask containing them remains constantly open during several months or years. "In a small quantity of water, I washed a part of a bunch of grapes, the grapes and the stalks together, and the stalks separately. This washing was easily done by means of a small barber's hair brush. The washing-water collected the dust upon the surface of the grapes and the stalks, and it was easily shown under the microscope that this water held in suspension a multitude of minute organisms closely re- sembling either fungoid spores or those of alcoholic yeast, or those of Mycoderma vini, etc. This being done, ten of the forty flasks were preserved for reference; in ten of the remainder, through the straight tube attached to each, some drops of the washing-water were intro- duced; in a third series of ten flasks a few drops of the same liquid were placed after it had been boiled; and finally in the ten remaining flasks were placed some drops of grape juice taken from the inside of a perfect fruit. In order to carry out this experiment the straight tube of each flask was drawn out into a fine and firm point in the lamp, and then curved. This fine and closed point was filed round near the end and inserted into the grape while resting upon some hard substance. When the point was felt to touch the support of the grape it was by a slight pressure broken off at the file mark. Then if care had been taken to create a slight vacuum in the flask, a drop of the juice of the grape got into it, the filed/ point was withdrawn, and the aperture immediately closed in the alcohol lamp. This decreased pressure of the atmosphere in the flask was obtained by the following means: After warming the sides of the flask, either in the hands or in the lamp flame, thus causing a small quantity of air to be driven out of the end of the curved neck, this end was closed in the lamp. After the flask was cooled, there was a tendency to suck in the drop of grape-juice in the manner just described. "The drop of grape-juice which enters into the flask by this suction ordinarily remains in the curved part of the tube, so that to mix it with the must it was necessary to incline the flask so as to bring the must into contact with the juice and then replace the flask in its normal position. The four series of comparative experiments produced the following results: "The first ten flasks containing the grape-must boiled in pure air did not show the production of any organism. The grape-must could possibly remain in them for an indefinite number of years. Those in the second series, containing the water in which the grapes had been washed separately and together, showed without exception an alcoholic fermentation which in several cases began to appear at the end of forty-eight hours when the experiment took place at ordinary summer temperature. At the same timethat the yeast appeared, in the form o c white traces, which little by little united themselves in the form of a deposit on the sides of all the flasks, there were seen to form little

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flakes of Mycellium, often as a single fungoid growth or in combina- tion, these fungoid growths being quite independent of the must or of any alcoholic yeast. Often, also, the Mycoderma vini appeared after some days upon the surface of the liquid. The Vibria and the lactic ferments properly so-called did not appear on account of the nature of the liquid. "The third series of flasks, the washing-water in which had been previously boiled, remained unchanged, as in the first series. Those of the fourth series, in which was the juice of the interior of the grapes, remained equally free from change, although I was not always able, on account of the delicacy of the experiment, to eliminate every chance of error. These experiments cannot leave the least doubt in the mind as to the following facts: "Grape-must, after heating, never ferments on contact with air, when the air has been deprived of the germs which it ordinarily holds in a state of suspension. "The boiled grape-must ferments when there is introduced into it a very small quantity of water in which the surface of the grapes or their stalks have been washed. "The grape-must does not ferment when there is added to it a small quantity of the juice of the inside of the grape. "The yeast, therefore, which causes the fermentation of the grapes in the vintage-tub comes from the outside and not from the inside of the grapes. Thus it destroyed the hypothesis of MM. Trecol and Fremy, who surmised that the albuminous matter transformed itself into yeast on account of the vital germs which were natural to it. With greater reason, therefore, there is no longef any question of the theory of Liebig of the transformation of albuminoid matter into ferments on account of the oxidation."

Pasteur's work did not stop here, for he soon proved that a disease that was attacking the silk worm was caused by bacteria. And from this there developed the idea that disease in general is due to bacteria. The profound importance of his work is well summarized by Lord Lister, when he said of him

"Truly there does not exist in the entire world any individual to whom the medical science owes more than they do to you" [referring to Pasteur]. "Your researches on fermentation have thrown a power- ful beam, which has lightened the baleful darkness of surgery, and has transformed the treatment of wounds from a matter of uncertain and too often disastrous empiricism into a scientific art of sure beneficence. Thanks to you; surgery has undergone a complete revolution which has deprived it of its terrors and has extended almost without limit its efficacious power."

And we find Tyndall stating: "We have been scourged by miserable throngs, attacked from im- penetrable ambuscades, and it is only today that the light of science is being let in upon the murderous dominion of our foes."

If there was any doubt in the mind of the scientific world as to the fallacy of the theory of spontaneous generation, after the work of Pasteur, it was dispelled by the work of Tyndall. He _

proved that in an atmosphere aboslutely devoid of dust, as is the

312 IMPROVEMENT ERA case on the top of mountains, and in some ingeniously constructed boxes used by him, perishable substances such as beef-tea will keep for an indefinite time. As early as 1863, Davanie had seen in the blood of some animals that had died of a disease known as anthrax, a very smajl rod-like organism which permeated all the capillaries. Experi- ments showed that blood from such an animal when injected into the veins of a second animal caused it to die of the same disease. But he found that there were times when the organism could not be found in the blood of the animal after death, yet blood from this animal would cause the death of another animal when injected into its veins. This left a doubt in the minds of thinking men as to whether this rod-shaped organism was causing the death of the animal, or- was it some invisible element in the blood itself. This was not fully settled until thirteen years later, when the work of Robert Koch appeared. He had not only seen the organism but he obtained it free from all other substances and had proven that it was the specific cause of the disease. This was followed by many other discoveries, until today we know that practically all diseases are due to these invisible foes. Yes, even many of the changes taking place in the body and associated with old age are probably caused by the products generated by bacteria. The workers in this field are not satisfied with knowing the cause of a disease, but they want to know how they can ward off disease and how to cure it when it once gains access to the body of an animal. Pasteur soon announced that he had found a pre- vention for anthrax, and his statement was immediately challenged by the president of an agricultural society in such a way that it was brought to the attention of the entire civilized world. He suggested that the subject be submitted to a decisive public test and offered to furnish fifty sheep, half of which should be pro- tected by the attenuated virus prepared by Pasteur. Later they were all to be infected by the disease-producing organism and if the vaccine be a success the protected ones were to remain healthy, the unprotected ones to die of the disease. Pasteur accepted the challenge and suggested that for two of the sheep there should be substituted two goats, and that there be added to the herd ten cows, but he stated that these latter animals should not be considered as falling rigidly within the test, for his experiments had not yet been extended to cattle. Before this time the fame of Pasteur had been considered firmly established but now all the world looked on with doubt to think that any man should make such a prepos- terous claim. On May 5th the animals to be protected received their first treatment with the vaccine and a second two weeks later. On May 31st all the animals were inoculated with an extremely virulent culture of the disease-producing organism. The results of the test were dramatic indeed for

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"Two days later, June 2nd, at the appointed hour of rendezvous, a vast crowd, composed of veterinary surgeons, newspaper correspond- ents, and farmers from far and near, gathered to witness the closing scenes of this scientific tourney. What they saw was one of the most dramatic scenes in the history of peaceful science, a scene which Pasteur declared afterwards, 'amazed the assembly.' Scattered about the enclosure, dead, dying or manifestly sick unto death, lay the un- protected animals, one and all, while each and every protected animal stalked unconcernedly about with every appearance of perfect health. Twenty of the sheep and the one goat were already dead; two 'other sheep expired under the eyes of the spectators; the remaining victims lingered but a few hours longer. Thus in a manner theatrical enough, not to say tragic, was proclaimed the unequivocal victory of science."

In 1885, he announced his cure for hydrophobia, the disease following the bite of a mad dog, and since that time thousands have been rescued from this terrible disease. This was followed by other great advances, until today diph- theria, in place of being a disease in which the death rate is 30 per cent, it is now cut to less than three. Typhoid fever is nearly conquered, as is shown by the fact that during the first five months of the year 1913, there was not a single case of typhoid fever occurred in the United States army, while in 1909 there were 173 cases and 16 deaths. In the last quarter of 1911, anti-typhoid vaccination of all persons in military service under forty-five years of age was made compulsory, and since that time there has not been a single vaccinated soldier die of this disease. Nor is it fol- lowed by bad after-effects for, during the past four years, over ;

200,000 persons, mostly in the military or naval service, have been immunized without any fatalities or serious complications. Surely this is a great triumph. The Asiatic cholera and the yellow fever have been nearly wiped from the face of the earth. And thanks to the wonderful work of Lord Lister, of whom it has been written, "He saved more lives, due to his work in antiseptic surgery, than were lost by Napoleon in all of his wars," the hospitals are no longer filled with the harrows that were once to be seen there. The abdomen has become the playground of the surgeon, nearly all parts of the body may be entered and it is not followed by that awful infection which used to follow even the slightest operation. The organism which causes lockjaw following the wound with the toy pistol has been ferreted out, and today this kind of wound if prop- erly treated is only slightly more dangerous than a cut. The stage to which antiseptic surgery has been developed is well shown by the record of the Mayo Brothers who, during the last ten years, have performed over 3,000 difficult operations with a death rate of less than three per cent. Yes, greater advances than this have been made, for we find the death rate amongst their patients dur- ing the last year to be only slightly greater than one per cent. The investigators in some of these fields have gone into it not

314 IMPROVEMENT ERA only with a knowledge of the fact that failure may be their lot, but they must even risk their lives in the work, as is shown in the fight against yellow fever. Dr. Lazear, an American army sur- geon, allowed himself to be bitten by a mosquito in an infected ward. He soon acquired the disease in its most terrible form and died a martyr to science and a true hero. Truly can we say of him, "Greater love hath no man than this, that a man lay down his life for his friends." Dr. Lazear lost his life, but due to him and his comrade's work, yellow fever has been conquered. Throughout this summary we have been tracing man's fight with these invisible foes, but there is another side to the story. But it has not been developed to the extent to which this has, for here man has not been driven on by the struggle against an in- visible enemy. Only a very small fraction of these invisible or- ganisms are man's enemies. Most are his friends. Some of them toil for him night and day. We find them helping him in the dairy, on the farm, in every avocation of life. Their true signifi- cance is probably partly understood, when we realize that without them life upon this planet would be impossible. The whole world would become filled with never-changing dead matter. One of the essential elements of the soil would soon be exhausted, and plants and animals would soon perish from starvation. So we find them increasing the fertility of the farmer's soil and helping him in a thousand and one ways. We find them directed by man's will today in many a large industry, not in the haphazard way as was the case in the past, but with quantitative precision. One may think from the preceding that in this field of science there is little yet to be done, but this is not the case, for there are diseases still unconquered. The great "White Plague" still claims its millions each year. There are diseases which are sapping the very life blood of the nation, yet they go unchecked. Science as yet has not come to the aid of the unfortunate victims. As regards the beneficial organisms, we have only just started to realize their great possibilities. In the soil we have five great classes of organisms which deal with the transformation of nitro- gen in the soil. One class carries on purification, changing the insoluble proteins into ammonia, another picks the ammonia up as formed, transforming it into nitrites, and even this must be changed into nitrates before the farmer's plants can use it. Under what condition are these changes carried on at a maximum rate ? What influence has moisture, temperature, crop and methods of tillage on this change? Some of these questions are being an- swered by the work being carried on by the Utah Experiment Station, but there are many yet unanswered. Still they are vital questions, for in many cases the yield of the farmer's crop will be determined by the skill with which he controls these various changes. We have another set of organisms in the soil, the func-

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tion ofwhich is to take the practically valueless nitrogen of the atmosphere and change it into forms such that the higher plants can feed upon. How may we control them for maximum yields ? for if treated right they will never tire, but toil on forever. Then again, it- is possible that we may become so familiar with these minute plants that we may use them to measure the requirements of the soil, and it is quite possible that they will be more sensitive than any tests we yet have. Truly can we say that in this field great things have been accomplished, but there remains yet to conquer fields richer than the workers in the past have ever dreamed of. LOGAN, UTAH

M. I. A. CLASS IN LEADERSHIP

This class, in which Supt. Heber J. Grant appears, held a three- days' session in the Latter-day Saints University, Salt Lake City, under the guidance of Oscar A. Kirkham and Emily C. Adams. In addition to the six-weeks' course given in the Brigham Young College, in Logan, which was well attended, a three-days' separate course was '

given there on January 11, 12, and 13, at which some 200 or more were in attendance. The three-day courses in nearly all parts of the Church have been very successful, and the thanks of the General Superin- tendencies and the Boards are due to the managements and teachers of the different Church schools, for favors and accommodations ex- tended. This applies not only to one but to all of them, and thanks are. hereby expressed to all who have given aid to the officers and members of the M. I. A. in these splendid three-day schools in leader- ship. The General Officers are hopeful that good results will follow for the entire membership, from the training given to M. I. A. workers in these gatherings.