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The Bacteriology of Tuberculosis

The Bacteriology of Tuberculosis

Egons Darzins
Copyright Date: 1958
Edition: NED - New edition
Pages: 500
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  • Book Info
    The Bacteriology of Tuberculosis
    Book Description:

    The Bacteriology of Tuberculosis was first published in 1958. Although tuberculosis is one disease against which, it may be said with little argument, medical science has scored tremendous victories, the goal of complete conquest still lies ahead. To fix accurate sights for that goal, a thorough understanding of the present status of knowledge about the disease is needed. This volume is published in response to that need. Through an exhaustive study of the literature on tuberculosis bacteriology from the late nineteenth century to the present, Dr. Darzins presents a comprehensive account of the knowledge and practices which have developed in this field. An important aspect is the discussion of the relatively new problems raised in bacteriological science by recent advances in the use of chemotherapeutics, antibiotics, and surgery for the treatment of tuberculosis. The first section is devoted to the morphology and cytology of the tubercle bacillus. Here Dr. Darzins outlines the physical, biological, and chemical methods of identifying cell structures. In the next section he considers the sources of energy and growth of the bacillus. He proceeds in the following section to a discussion of the methods of isolating and identifying the bacillus. The fourth part deals with a major problem of tuberculosis bacteriology, that of distinguishing the types of bacilli and determining their pathogenicity. In the final section he considers the problems of experimental work and points out the hazards and the need for precautions in laboratory work.

    eISBN: 978-0-8166-6203-6
    Subjects: Health Sciences

Table of Contents

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  1. Front Matter
    (pp. i-viii)
  2. Table of Contents
    (pp. ix-xii)
  3. Part One. Morphology and Cytology of the Tubercle Bacillus

    • I Tubercle Bacillus as a Unicellular Organism
      (pp. 3-7)

      ROBERT KOCH (1882), in his paper on the discovery of the cause of tuberculosis, described the tubercle bacillus, stained with methylene blue and counterstained with vesuvin, in the following terms (p. 222): “The bacteria made visible through this procedure are somewhat peculiar in aspect. They have the shape of a rod and, because of this form, belong to the group of bacilli. They are very thin and one-fourth to one-half as long as the diameter of a red blood cell; sometimes they reach a greater length and are as long as the whole diameter of a red cell. Under certain...

    • II Physical Methods for Identifying Cell Structures
      (pp. 8-41)

      THE resolving power of a microscope is inversely related to the wave length of the illuminant used; that is, the resolving power increases with decreasing wave length. For this reason, short-wave microscopy could be a considerable advance over earlier methods of penetrating into fine structures of the cellular world. But the practical realization of short-wave microscopy presented difficulties with respect to the source of the short-wave light and the material to be used for the lenses of the microscope and the slides. Light with a wave length of less than 300 mμ is ultraviolet, and

      ultraviolet light cannot penetrate glass....

    • III Biological Methods for Identifying Cell Structures
      (pp. 42-53)

      THE young bacterial cell, when treated with a basic dye, is uniformly and deeply stained. The stain prevents the observer from looking into the interior of the cell. On the other hand, a stain with a specific affinity to chromatin, which would stain cytoplasm in a color different from that of the nucleus, would be capable of indicating the presence or absence of a nuclear substance in the cell.

      Romanowsky (1891) started his study of malaria with the intention of finding a combination of dyes that would stain the nuclei and the cytoplasm in different colors. A mixture of basic...

    • IV Chemical Methods for Identifying Cell Structures
      (pp. 54-65)

      THE high affinity of chromatin to basic dyes was attributed to the acid compounds of chromatin, and chiefly to its large nucleic acid content. The staining process of the nucleus itself was regarded as a process of salt formation between the acid and the base. If this were the case, the staining properties of the nucleus would not necessarily point out chromatin. Indeed, the cartilage and the mucus are equal to chromatin in their Romanowsky staining properties. The notion of chromatin was morphologic rather than chemical. The discovery of a reaction which would specifically point out chromatin would be of...

    • V The Bacterial Cell and the Metazoan Cell
      (pp. 66-74)

      THE nucleus is an isolated body in the cytoplasm of a cell. The presence of an organized nucleus composed of nucleolus, chromosomes, and genes will indicate the appurtenance of a cell to a higher form of life. Although the notion of nucleus is more morphologic than chemical, some general criteria of distinction between the organized nucleus, nuclear bodies, and other cell organelles should be indicated.

      The nucleus of higher forms of life is present in the cytoplasm at all conditions of nutrition, at any age, and at any stage of cell development. The disappearance of the nucleus means the death...

  4. Part Two. Sources of Energy and Growth of the Tubercle Bacillus

    • VI Oxidation, Fermentation, and Growth of the Tubercle Bacillus
      (pp. 77-98)

      TO LIVE means to consume energy. The living organism does not create energy but uses the energy of the sun (as found in green plants) or gets it by transforming the chemically bound energy of foodstuffs. The tubercle bacillus, like all other living cells devoid of chlorophyll, is unable to utilize the radiant energy of the sun. The process of breaking down complex compounds into simple ones permits the organism to use the energy thus liberated. The breakdown of foodstuffs is performed by means of oxidation or fermentation. As Lavoisier recognized in 1770, respiration, like combustion, is a process of...

    • VII The Action of Fatty Acids, Salicylates, and Benzoates on Tubercle Bacilli
      (pp. 99-114)

      RESPIRATION, growth, and reproduction are different phases of cellular life and are of different significance to it. Respiration may go down to a very low level, as occurs in die bacterial spores, but its total cessation would mean death to the cell. These findings led to the erroneous conception that the contrary is also true, that is, that any increase in respiration signifies increased growth or reproduction of the cell. There is no growth and reproduction without respiration, but increased respiration of the cell does not necessarily entail better growth and greater reproduction. The reproduction of a cell, which follows...

    • VIII Sources of Carbon
      (pp. 115-145)

      NOCARD and Roux (1887) discovered that the addition of glycerol to the agar medium stimulates the growth of tubercle bacilli highly. Th. Smith (1904—1905) observed that there exists some difference between the ways the human and bovine types of tubercle bacilli react to glycerol. Both types consume glycerol, but most human strains, after several weeks of incubation, acidify the glycerol broth, whereas most bovine bacilli alkalize it. If bacilli are grown on broth without glycerol, these differences are not revealed; the reaction is alkaline in both cases.

      Siebert (1909), like Th. Smith, attributed the acidification of glycerol broth under...

    • IX Sources of Nitrogen and Phosphorus
      (pp. 146-155)

      NITROGEN is indispensable to the growth of tubercle bacilli. As tubercle bacilli do not utilize the nitrogen of the air (Proskauer and Beck, 1894), it must be supplied to them in the form of soluble compounds.

      The nitrogen content of tubercle bacilli ranges from 7.0 to 7.8 per cent (Bance, 1942) and there is no relation between the level of nitrogen in the bacilli and its concentration in the medium (Henley and LeDuc, 1939). Tubercle bacilli can for a short time proliferate in a medium lacking a source of nitrogen but containing a source of carbon. Such a process of...

    • X The Mineral Requirements of Tubercle Bacilli
      (pp. 156-159)

      THE good growth of tubercle bacilli on mineral media or on purely vegetable media, such as potatoes and carrots, reversed the conception of the growth requirements of tubercle bacilli, based upon the first successful attempt at tubercle bacilli cultivation on a coagulated serum by Koch. The opinion that the tubercle bacillus is a highly pretentious parasite which, in order to grow, requires complex animal proteins as nutrients, has proved to be erroneous. It was early found that this bacillus lives on relatively simple nitrogen, carbon, and mineral compounds.

      The recognition that the tubercle bacillus grows on solutions of relatively simple...

    • XI Growth Factors and Trace Elements
      (pp. 160-168)

      THE greatest part of the ingredients of egg media used for the cultivation of tubercle bacilli remains unknown and the preparation of these media stereotyped (van Niel, 1944; Dubos and Noufflard, 1950; Middlebrook, 1950). The basic elements needed for the growth of a cell, carbon, nitrogen, phosphorus, potassium, iron, and others, are supplied by egg yolk, or may be obtained from the added chemicals. From this primary building material the cell manufactures its constituents. Besides these ingredients, derived directly from the foodstuffs or produced from the nutrients by the cell, there are substances, necessary for the growth, which the cell...

  5. Part Three. The Isolation and Identification of the Tubercle Bacillus

    • XII The Collection of Tuberculous Material
      (pp. 171-184)

      WHEN tubercle bacilli invade an organism previously affected by tuberculosis, they meet with a highly sensitized tissue which reacts to the renewed infection with the death of local tissue or caseation necrosis. This defense mechanism of the organism is a result of the contact between the allergic tissue and the tuberculous antigens and is a by-product of the infection. This defense reaction will, to a great extent, determine the issue of the infection itself. From the moment of the caseation of the tissue on, the organism is combating not only the invasion of tubercle bacilli but also its own dead...

    • XIII Diagnostic Staining of Tubercle Bacilli
      (pp. 185-190)

      THE early discovery of a simple and highly effective method for staining tubercle bacilli secured for staining a place of great importance among the diagnostic procedures of tuberculosis, a place which it has kept for long years.

      The staining method of Koch (1882) required a long time (24 hours), although Koch indicated that by heating the specimen to 40°C. the staining procedure could be shortened to half an hour. Koch’s method was not sufficiently sensitive and failed in many instances (Ehrlich, 1913).

      Ehrlich (1882), instead of using potassium hydroxide and methylene blue as Koch had, introduced the alkaline aniline oil...

    • XIV Fluorescence and Phase Contrast Microscopy as Diagnostic Procedures
      (pp. 191-193)

      HAGEMANN (1938) investigated tubercle bacilli in pathologic material with a fluorescence microscope and compared the results with those obtained with the Ziehl-Neelsen staining method. His results were extremely encouraging. In sputa confirmed by cultivation as tuberculous, the Ziehl-Neelsen staining method revealed 35 per cent of this material as positive, whereas fluoresence microscopy showed 70 per cent as positive; in the case of tuberculous urine, pus, and feces, fluorescence microscopy revealed 200 per cent more positive cases than the Ziehl-Neelsen method.

      The publications of Hagemann and the creation by the Reichert Optical Company at Vienna of a simple and inexpensive fluorescence...

    • XV Cultivation of Tubercle Bacilli
      (pp. 194-212)

      ROBERT KOCH (1882) was the first to succeed in cultivating tubercle bacilli. He describes this achievement as follows. “The principle involved in cultivating tubercle bacilli is the use of a solid, transparent medium, which even at incubator temperature does not lose its solid consistency . . . Serum of cattle or sheep blood, which is collected as clean as possible, is distributed in test tubes which are closed with a plug of cotton and heated at 58°C. one hour daily for six days. Through this procedure we are able, if not always, at least in most cases, to sterilize the...

    • XVI The Shaking-Precipitation (SP) Method
      (pp. 213-228)

      THE cultivation results of tubercle bacilli on an artificial medium depend upon (a) the viable properties of the bacilli inoculated; (b) the number of bacilli inoculated; (c) the composition of the medium used.

      Both acids and alkalies may be used to eliminate contaminants from pathologic material, and in a material rich in tubercle bacilli the cultivation results will be successful with all these substances. The results will be different if there is a smaller number of bacilli in the material, as in cases where the material has been collected from incipient or healing tuberculosis. The disastrous effect on tubercle bacilli...

    • XVII Submerged Growth
      (pp. 229-235)

      EXPERIENCE derived from the preparation of tuberculin has taught that pellicles that have sunk to the bottom of the vessel will not produce a culture. It was deduced from these observations that tubercle bacilli are only able to grow on the surface of a liquid medium.

      The adequate aeration of the surface of liquid media inoculated with pellicles of tubercle bacilli has always been of concern to investigators, and the death of submerged cultures was attributed to the asphyxia of the bacilli, although the first culture of tubercle bacilli in a liquid medium, which was accomplished by Nocard and Roux...

    • XVIII Dispersed Growth
      (pp. 236-254)

      S. ARLOING (1898) like all other workers of his time, met with little success in his attempts to obtain a homogeneous suspension of human tubercle bacilli grown on solid media. The occasional observation of the presence of diffuse growth in the liquid on the bottom of potato slants inoculated with tubercle bacilli gave S. Arloing the idea of creating a homogeneous culture of tubercle bacilli and of using such a culture instead of a suspension of tubercle bacilli. To facilitate the dispersed growth of bacilli in the liquid, the culture in glycerol broth was agitated daily. Growth throughout the broth...

    • XIX Efficacy of Microscopic Examination, Culture, and Animal Inoculation
      (pp. 255-263)

      A TREMENDOUS amount of work was devoted to the problem of direct microscopic examination and culturing or animal inoculation as a means of revealing the presence of tubercle bacilli in pathologic material. The avalanche of publications about this problem has brought very few new data, but it keeps on rolling.

      The microscopic examination of pathologic material was for a long time, and in many countries still is, the limit beyond which it is impossible to go in detecting tubercle bacilli. The history of the cultivation of tubercle bacilli shows that for many years the cultivation of tubercle bacilli was the...

    • XX Quantitative Aspects of Tubercle Bacilli in Sputum
      (pp. 264-270)

      THE idea of a classification and prognosis of tuberculosis on the basis of the number and the morphology of bacilli in sputum provoked extensive works and discussions in the early days of tuberculosis bacteriology, and the scale of classification of Gaffky (1884) particularly attracted the attention of physicians. The following years were poor in results pointing toward a cure for active tuberculosis, so that physicians gradually abandoned Gaffky’s classifications as worthless for the estimation of the results of the treatment and the prognosis. An easy way of dealing with the problem was adopted. In the presence of few or thousands...

  6. Part Four. The Types and Pathogenicity of the Tubercle Bacillus

    • XXI Do Fixed Types of Tubercle Bacilli Exist?
      (pp. 273-286)

      THE identity of human and bovine tuberculosis was assumed by the early investigators, even before the tubercle bacillus itself was discovered. Villemin’s (1865, 1868) classical experiments on the transmission of the human and bovine tuberculous virus to animals led him to this conclusion. His observations were confirmed by Chauveau’s (1868), Klebs’ (1870), and Gerlach’s (1870) successful attempts at infecting animals with tuberculosis by feeding them with tuberculous material of human and bovine origin. Koch (1882) held thatPerlsucht(the pearly disease of cattle) is identical to human tuberculosis and, consequently, transmissible to humans. The identity of tubercle bacilli derived from...

    • XXII Methods of Determining Virulence
      (pp. 287-314)

      UNDER the influence of the hypothesis of the stability of types, the study of the basic property of tubercle bacilli—their virulence—was overshadowed by the conventional typing of bacilli as a means of determining the presumed sources of infection. It is evident that the degree of pathogenicity or virulence of bacilli must play a decisive role in an attempt to determine the types of bacilli. To cause the disease, tubercle bacilli must establish and multiply in the host, although the rate of multiplication alone does not explain the degree of their virulence. The slowly evolving tuberculous lesions of a...

  7. Part Five. Experimenting with the Tubercle Bacillus

    • XXIII Problems in Experimenting with Tubercle Bacilli
      (pp. 317-321)

      EXPERIMENTS with tubercle bacilli bring up some problems which are common to general microbiology and others that are restricted to the bacteriology of mycobacteria.

      Experiment, observation, and reasoning, which were recognized by Francis Bacon and Claude Bernard as the only sources of human knowledge, has its limitations because our senses and the procedures we employ to collect scientific data (weighing, diluting, injecting, etc.) are imperfect and subject to variations which lead to errors. To overcome this, a large number of average data are often collected. These created averages do not exist in nature (Claude Bernard). In an account of an...

    • XXIV Estimating the Antituberculous Activity of a New Drug
      (pp. 322-326)

      CAREFULLY collected and critically selected data will produce lasting achievement. The investigation of a new drug must always be started with the study of the action of this drugin vitro.The results obtained may give important indications about the mode of its action as well as save time and work. However, the actions of drugsin vitroandin vivorarely run parallel. Generally, the action of a drug is the result of its concentration. High concentrations have bactericidal or bacteriolytic, low ones bacteriostatic, action.

      The chemotherapeutic or serotherapeutic action of an unknown compound must be compared with the...

    • XXV The Guinea Pig in Experimental Tuberculosis
      (pp. 327-332)

      THE guinea pig is the animal most sensitive to infection with tubercle bacilli. It is universally used to detect tuberculous infection, to screen the pathogenicity of acid-fast bacilli, and to test the efficiency of antituberculous drugs.

      A guinea pig infected with pathogenic tubercle bacilli develops exudative, progressive, and lethal tuberculosis which resembles but slightly the pathology of the torpid cavernous phthisis of human beings. These peculiarities of guinea pig tuberculosis forced investigators to look for animals whose tuberculosis is more similar to that of men (mice, rabbits, hamsters).

      In tuberculosis experiments young and healthy guinea pigs not under 300 g....

    • XXVI The Rabbit in Experimental Tuberculosis
      (pp. 333-339)

      RABBITS are used for the differentiation of human and bovine types of tubercle bacilli and in the study of the problems of the pathogenesis and therapy of tuberculosis. Lurie (1949b) showed that rabbits uniformly develop ulcerative chronic pulmonary phthisis when exposed to the inhalation of controlled numbers of virulent bovine tubercle bacilli. Tubercle bacilli multiply in the walls of the lung cavities, and the tuberculous process in the animal closely resembles human chronic pulmonary tuberculosis. The testing of chemotherapeutic substances under these experimental conditions may be of importance in the evaluation of new antituberculous drugs.

      In the experiments of Lurie,...

    • XXVII The Mouse in Experimental Tuberculosis
      (pp. 340-356)

      THE susceptibility of the white mouse and the field mouse(Arvicola arvalis)to infection with the tubercle bacillus was demonstrated by Koch (1884). Tubercle bacilli, injected subcutaneously or intraperitoneally into mice, produced a slowly progressing infection. Koch concluded that the mouse possesses high resistance to tuberculosis. This view was to be shared by most later workers in this field.

      Straus (1895) reports his successful attempts to infect mice with tubercle bacilli. Römer (1903) was the first to use large numbers of white mice in a comparative study of the biology of tubercle bacilli, although mice at that time were regarded...

    • XXVIII The Hamster in Experimental Tuberculosis
      (pp. 357-362)

      ONE of the many different animals infected with tubercle bacilli by Koch (1882) was the hamster. It was probably the EuropeanCricetus cricetus.

      The striped hamster,Cricetus griseusThomas and Milne Edwards, is a small rodent, weighing about 30 g. It is abundant in China and was first used in experiments with tubercle bacilli by Korns and Lu (1927) of the Peking Union Medical College. These investigators inoculated the striped hamster, as well as white mice and guinea pigs, with tubercle bacilli of human, bovine, and avian types. The hamsters and mice were injected subcutaneously with 0.1 mg., the guinea...

    • XXIX Chorioallantoic Membrane and Chick Embryo in Experimental Tuberculosis
      (pp. 363-381)

      SINCE it has been demonstrated that membranes of chick embryo are susceptible to infection with microorganisms, the developing egg has been much used in the studies of bacteriologic, chemotherapeutic, and histologic problems. (See Fig. 43.)

      The most outstanding feature of this method is the use of living developing embryonic tissue as the medium of culture. A large number of organisms which can be brought to development only in a living organism or in tissue culture, multiply readily in a developing chick embryo.

      Like every great discovery, the chick embryo method opened wide possibilities for research. It rendered unrivaled service to...

    • XXX Hazards and Precautions in the Laboratory
      (pp. 382-402)

      THE greatest of all dangers is perhaps becoming acclimatized to a danger. Human nature is endowed with the power of adaptation, and daily work with tubercle bacilli may lead to laxness in the routine. Remaining aware of a persisting danger during long years of work, often in the same place and with the same microorganism, is one of the peculiarities of the true scientific mind.

      Laboratory accidents and the infection of bacteriologists and pathologists have considerably increased with the growing number of persons working in diagnostic and research laboratories. The hazards of laboratory infection are far beyond those met in...

  8. Bibliography
    (pp. 403-461)
  9. Author Index
    (pp. 462-473)
  10. Subject Index
    (pp. 474-488)