T. J. Mackie

PART a GENERAL SECTION: Organization of the Clinical Bacteriology Laboratory: Quality Assurance. Computers in Medical Microbiology. Safety in the Microbiology Laboratory. Laboratory Strategy in the Diagnosis of Infective Syndromes. Specimen Collection, Culture Containers and Media. Culture of Bacteria. Tests for the Identification of Bacteria. Laboratory Control of Antimicrobial Therapy. Some Serological Techniques for Microbial and Viral Infections. Nucleic Acid Techniques in Diagnostic Microbiology PART B BACTERIA and RELATED ORGANISMS: Staphylococcus: Cluster Forming Gram-Positive Cocci. Streptococcus and Enterococcus. Streptococcus Pneumoniae. Neisseria, Moraxella, Acinetobacter. Corynebacterium. Listeria, Erysipelothrix. Bacillus. Mycobacterium. Actinomycetes: Actinomyces, Actinomadra, Nocardia, Streptomyces and Related Genera. Enterobacteriacea: Escherichia, Kelbsiella, Proteus and Other Genera. Salmonella. Shigella. Psuedomonas, Stenotrophomas, Burkholderia. Vibrio: Aeromonas: Plesiomonas: Campylobacter: Arcobacter: Hel Icobacter: Wolinella. Haemophilus, Gardnerella and Other Bacilli. Bordetella. Brucella. Yersinia, Pasteurella, Francisella. Legionellaceae. Bacteroides, Fusobacterium and Other Gram-Negative Anaerobic Rods: Anaerobic Cocci: Identification of Anaerobes. Clostridia of Wound Infection. Enteropathogenic Clostridia and Clostridium Botulinum. Treponema: Serological Tests for Syphilis. Leptospira, Borrelia, Spirillum. Coxiella Burnetii and Other Medically Important Members for the Family Rickettsiaceae. Mycoplasma Pneumonaie and Other Medically Important Members of the Family Mycoplasmataceae. Chlamydia PART C VIRUSES: Laboratory Diagnosis of Virus Infections. Rapid Diagnosis of Viral Infections. Cell and Virus Culture PART D FUNGI: Fungi PART E PROTOZOA: Protozoa PART F HELMINTHS: Helminths PART G GENERAL METHODS: Immunofluorescence and Immunoelectronmicroscopy: Some Selected Aspects of Light and Electronmicroscopy. Staining Methods. Sterilization and Disinfection in the Laboratory. Ph Meas

1. An analytical study has been made of the mechanism of natural bactericidal action by the serum of various animals (ox, sheep, horse, rabbit, guinea-pig, rat, man) towards certain organisms ( B. typhosus, B. dysenteriae Shiga, B. proteus, V. cholerae ) exhibiting the maximum reactivity to this effect. 2. Serum-complement has no bactericidal action per se , and an antibodylike agent invariably acts as an intermediary agent, “sensitising” the particular organism to the action of the complement and capable of being “absorbed” by it from serum at 0° C. 3. This sensitising agent is stable at 55° C. but labile at 60°–65° C. In this respect it resembles natural haemolysins and agglutinins, but contrasts with the more stable immune antibodies and the more labile natural complement-fixing antibodies (for bacterial antigens). It is resident mainly in the carbonic-acid-insoluble fraction of the serum. It is present in the serum of young animals before certain other natural antibodies have developed. 4. Absorption tests demonstrate the high degree of specificity of these natural bactericidal antibodies for particular bacteria. 5. A non-specific extracellular substance occurs in bacterial cultures which may neutralise or inhibit these antibodies, and interfere with their sensitising action even at 0° C. 6. This substance is liberated in large amount in cultures heated at high temperatures (120° C). It can be removed by repeated washing of growths in saline solution. It may inactivate a bactericidal antibody in heated serum, though not in fresh unheated serum, and may inactivate a particular antibody in the serum of one animal species but not in another. Strains of bacteria vary in their production of this substance. 7. The observations submitted in this paper, correlated with previous studies of natural antibodies by the authors and others, indicate that immune antibodies have their precursors specifically differentiated in the serum of normal animals and that, in general, immune antibodies are not substances formed de novo .

Since the early work of Nuttall (1888), Buchner (1889), and others first disclosed the bactericidal property of normal serum, a considerable amount of attention has been devoted to this important biological phenomenon. The literature (which has been reviewed recently by Knorr, 1929) lacks, however, a comprehensive and co-ordinated study of the subject, and in fact presents a good many confusing data in regard to the nature of the active principles concerned and their immunological significance.

(1) Brilliant green exerts an inhibitory effect on the growth of bacilli of the coli group commonly occuring in faeces, which is in general more marked than its action on B.typhosus and paratyphiod bacilli. (2) By taking advantage of this property of brilliant green a method has been devised is the inoculation B. typhosus from faeces. The procedure adopted is the inoculation of a series of tubes of peptone-water medium containing varying amounts of brilliant green, incubating for 20–24 hours, and then inoculation on a suitable solid medium from each tube. (3) The reason for employing a series of concentrations of brilliant green is that the optium concentration for the growth and isolation of B. typhosus varies from case to case, depending probably both on the proportion of typhoid bacilli present and on the number and character of the accompnying bacteria as well as on the organie faecal material. (4) The method is very easily and rapidly carried out.