1852 - 1915
Loeffler, pupil and assistant of the great Robert Koch, is justly famed for his part in the discovery of the causative organism of diphtheria. One of the most able investigators in the golden age of bacteriological discovery, he also made many other important contributions: he was the discoverer of Loefflerella mallei (the causative organism of glanders) with Schutz; of the organisms responsible for swine erysipelas and swine plague; and of Salmonella typhi in mice. He was an early pioneer in the field of virology. Working with Paul Frosch he demonstrated that foot and mouth disease was due to a filterable virus - the first recognition of a virus as the cause of an animal disease.
Loeffler was a great technician and made many important technical advances. Some of the methods and procedures he evolved are still in daily laboratory use. Every medical laboratory scientist must be familiar with Loeffler's methylene blue stain, which is in widespread use in bacteriological laboratories. He was one of the earliest to appreciate the value of aniline dyes for staining bacteria and tissues, and studied them extensively. Loeffler was responsible for the introduction of one of the first flagella staining methods.
Perhaps it is in the field of culture media, however, that Loeffler's techniques were most important in his own day. His blood serum medium for the cultivation of the diphtheria bacillus, his malachite green selective medium for B typhosus (S typhi), and his meat-juice peptone-gelatine medium were widely used. Whilst these media have now largely been replaced by others and by new techniques, developed as a result of much greater knowledge, there is little doubt that they represented a real breakthrough at the time of their introduction.
Though Loeffler was a man of vigorous physique
and great energy - an indefatigable worker in the true Koch tradition -
he published relatively little by comparison with many of his contemporaries.
His published work was always prepared with great thoroughness, however,
every findings being carefully checked, and it has fully stood the tests
of time and confirmation by later workers.
Edwin Klebs had long searched for the causative organism of diphtheria. A professor at Zurich in 1883, he certainly saw the diphtheria bacillus but, although he saw the organism in diphtheritic membranes, he was unable to grow it by artificial culture or to prove its relationship to the disease. At the time Loeffler took up the search he was working in Koch's laboratories. He began his studies by making a complete histological study of material from a number of fatal cases of diphtheria. In this work he used methylene blue, evolving his famous alkaline methylene blue staining solution. His work confirmed the findings of Klebs and Corynebacterium diphtheriae was known for many years as the Klebs-Loeffler bacillus (often referred to as 'KLB').
Loeffler realised that he must do what Klebs had failed to do - find some method of isolating and growing the bacillus in pure culture. Only in this way could he hope to reproduce the disease in animals and prove the organism's true role. Using a peptone gelatine medium his first attempts at growing the bacillus failed completely. He then turned to a medium consisting of glucose broth and blood serum coagulated by heat. While this was not selective, the diphtheria organisms grew as distinct colonies and Loeffler was able to obtain pure subcultures. The bacilli were the injected into guinea pigs and produced a fatal disease which, on post mortem examination, showed characteristic changes.
One other very significant fact also emerged
from these animal experiments; not only did Loeffler show the organism
to be the cause of the disease, he also found that it remained localised
at the site of injection in the same way that it remained localised in
the diphtheritic membrane in human cases. this led him to attribute
the rapidly fatal effects of the disease to the ability of the bacilli
to liberate a powerful exotoxin - a theory which he failed to prove.
Emile Roux, working with Alexandre Yersin at the Pasteur Institute in Paris,
later showed that filtered broth cultures of the diphtheria bacillus, containing
no organisms, produced death with the characteristic lesions when injected
into guinea pigs. This was proof of Loeffler's theory of a powerful
poison produced by the bacilli and carried throughout the body and proved
to be the starting point of Behring and Kitasato's discovery of diphtheria
antitoxin. Loeffler continued his investigations into diphtheria
and discovered the existence of pseudo-diphtheria bacilli, sometimes found
in the human throat and nose. In 1888 and Austrian investigator,
Van Hoffmann-Wellenhof, published an account of an organism isolated from
a healthy human throat, which he was unable to distinguish from the diphtheria
bacillus. Further investigation soon proved, however, that 'Hoffman's
Bacillus' was quite distinct from the bacillus responsible for diphtheria
simply another member of the Corynebacterium group.
Friedrich August Johannes Loeffler, the son
of a military surgeon, was born at Frankurt-on-the-Oder on 24 June 1852.
He studied medicine at Wurzburg and Berlin, taking his MD in 1874.
During the Franco-Prussian War of 1870 he served as an hospital assistant.
After passing his stated examination in 1875 Loeffler served as a military
surgeon in Hanover and Potsdam. In 1879 he transferred to the Imperial
Health Department, coming under the influence of Robert Koch with whom
he worked on steam disinfection and other problems related to public health.
In 1886 he became Privatdocent in Hygiene at the University of Berlin and
two years later was appointed to the Chair of Hygiene at the Institute
of Hygiene at Greifswald, of which he later became Rector. In 1913
he followed Georg Gaffky as Director of the Institut fur Infectionskrankheiten
(Institute of Infectious Diseases) in Berlin. Loeffler was the author
of the famous history of bacteriology, Vorlesungen über die geschichtliche
Entwickelung der Lebre von den Bacterien. This work, the first on
the subject, covered the history of bacteriology from earliest times until
the 1880s. It was Loeffler's intention to write a second volume covering
later developments in bacteriology but, unfortunately, pressure of work
prevented him from doing so. With the outbreak of the First World
War in 1914 Loeffler returned to the Army Medical Corps with the rank of
General, and was largely concerned with matters of hygiene. He was
now over sixty years of age and the strains of his important office proved
too great for his health; early in 1915 he was invalided home, where in
April he died. He was buried in Greifswald.
1861 - 1931
The name of Dr MacConkey has been familiar to generations of bacteriologists through the widespread use of the well-known culture media, which bear his name. The valuable media for differentiating intestinal organisms of the coli-typhoid group, because of their effectiveness and relative simplicity of preparation, were long favoured by bacteriologists, particularly in the British Isles. MacConkey's media were a development of the test which came to be known as the Voges-Proskauer reaction which had been introduced in 1898 by two associates of Robert Koch in Berlin, O Voges and B Proskauer.
In the early 1900s, while working as Assistant Bacteriologist to the Royal Commission on Sewage Disposal, in the Thompson-Yates Laboratories of Liverpool University and under the direction of Sir Rupert Boyce, MacConkey evolved his famous media formulae. His object was to produce a medium, which favoured the growth of Gram negative organisms yet with an inhibiting effect on those which were Gram positive. At this period knowledge of the intestinal bacteria was far from complete, and the technical procedures for their investigation were still very much in the developmental stage. The use of fermentation reactions had been established and J E Durham had published his method for the demonstration of fermentation activity. Conradi and Drigalski were evolving their selective medium containing crystal violet.
MacConkey's first experiments were with the addition of one to 1000 parts of carbolic acid to the medium. This was not a new idea; various workers had published media formulae in which minute amounts of an antiseptic substance were included, and there was evidence that its presence inhibited the growth of Gram positive organisms more than the Gram negative ones. After a number of trials MacConkey discovered that a suitable concentration with C A hill he added the indicator, litmus. Later, glucose was replaced by lactose.
Acting on a suggestion made by A S Grunbaum
and E H Hume, MacConkey replaced litmus with neutral red, which stained
the colonies as well as being an indicator. (Dr Grunbaum was not
German, as his name might suggest, but an Englishman who during the 1914
- 1948 war, found it desirable to change his name to Leyton). Particularly
in the bacteriological investigation of water supplies, use of MacConkey's
medium in the form of a peptone water without the agar base, was much favoured
as a preliminary culture procedure.
The Lister Institute
Alfred Theodore MacConkey, born in1861 the son of a minister in West Derby, Liverpool, studied medicine at Cambridge and Guy's Hospital. After qualification he went into private practice in Beckenham, Kent, but soon decided to specialise in bacteriology. In 1897 he joined the Bacteriology Department at Guy's Hospital. This was followed by his employment as assistant bacteriologist to the Royal Commission on Sewage Disposal in Liverpool. When, in 1904, Charles Todd left the staff of the Lister Institute (then the Jenner Institute of Preventive Medicine) to join the Egyptian Health Service, MacConkey accepted an invitation to take his place in London. A year later he went to the Institute's branch laboratories at Elstree as bacteriologist in charge.
At the time of MacConkey's transfer the Lister Institute was going through a period of financial stress and, though the antitoxin preparations produced at Elstree were recognised s among the best available, the department steadily lost money each year. On his appointment to Elstree, the chairman of the governing body (Sir Henry Roscoe) gave MacConkey a clear directive. While the high quality of the laboratory products must be maintained, the department must not only be made to pay its way but must also make a substantial contribution to the expenses of the Institute as a whole. This was certainly a difficult assignment but MacConkey, a strong personality, accepted the challenge with enthusiasm and immediately instituted a policy of stringent economy. Research not directly relevant to the production of therapeutic sera was discontinued, staff reductions were made, heating of the laboratories was reduced to a bare minimum, and even participation by any member of the staff in sporting activities was strongly discouraged as wasteful of energy. It is hardly surprising that such measures did little to win him universal popularity, and he acquired the reputation of being 'something of a character'. Nevertheless, MacConkey turned the department into a profitable undertaking, and during the 1914 - 1918 war it had a splendid record in satisfying the enormous demands of the armed forces for therapeutic sera.
In 1926 MacConkey retired and went to live
at Brindley Heath, in Surrey, where he died in 1931.
1844 - 1922
The title 'Father of tropical Medicine' has on many occasions been rightly bestowed on this very remarkable Scotsman, who was a pioneer of organised teaching in the undeveloped - but rewarding - field of tropical medicine. He was responsible for the foundation and subsequent development of two famous medical schools, the Hong Kong Medical School (which became the University of Hong Kong) and the London School of Tropical Medicine at the Albert Dock Hospital, now transformed into the London School of Hygiene and Tropical Medicine and with an international reputation. In the 50 or so years of his busy professional life Sir Patrick Manson made a series of important discoveries in tropical medicine, but perhaps his greatest contribution was the concept of the role of winged insects as the agents responsible for the spread of certain tropical diseases - a concept which paved the way for Sir Ronald Ross's subsequent discoveries about malaria.
Although many other investigators - including Joseph Bancroft in Australia, Wucherer in Brazil, and Lewis in India - played essential parts in the unravelling of the mystery of elephantiasis and its allied conditions, it was Manson in 1877 who showed the manner of development of the filarial worm Wucheria bancrofti and the role of the Culex mosquito in its spread. This was the first occasion on which it was proved that an infective disease was spread by animal vectors.
Manson's investigation was greatly helped by
his Chinese servant, Huito. It had been known for some time that
Huito's blood harboured filaria and, very courageously, he volunteered
to sleep in a mosquito cage containing specimens of the mosquito Culex
fatigans, the common mosquito of Amoy. It is to be feared that Huito
spent a far from peaceful night, but the experiment worked and in the morning
Manson was able to collect several of the engorged female insects and keep
them in captivity for some days. At intervals he killed specimens
and carried out a careful dissection. He quickly discovered that
the minute worms present in the blood ingested by the insect migrated from
the mosquito's stomach into the tissue of the thorax, and he noted the
remarkable change is size and shape undergone by the parasite. Manson
spent many months performing further experiments to confirm his findings.
The research work was carried out under great difficulties: Manson had
also to perform his ordinary duties, and his problems were increased by
the fact that in China at that time investigation of pathological changes
brought about in the human body by disease was very limited, it being almost
impossible to obtain permission to carry out autopsies. Furthermore,
the science of entomology was at an elementary stage of development; few
books on the subject existed, and knowledge of the mosquito's anatomy was
almost non-existent. An appeal to the British Museum for help brought
him a treatise on the anatomy of the cockroach - the best they could do.
Manson's laboratory facilities were limited to a microscope, a few slides, and simple stains, and he improvised insect dissection instruments using fine pen nibs. Manson also reported the nocturnal appearance of the parasite in patients' blood, noting that even patients with a heavy infestation rarely showed the presence of parasites in their blood during the day. Some of his critics regarded these observations on the nocturnal wanderings of the parasite with such scepticism that they inquired if the 'Filariae carried watches?' Some years later, when Manson had returned to London, he was able to demonstrate that during the day the parasite embryos collected in the capillaries of the lungs.
The lungs fluke
Manson's discovery of human infection by the oriental lung fluke Paragonimus westermani was something of a lucky chance. During a consultation with an important Chinese mandarin on a skin condition from which he was suffering, the patient suddenly expectorated onto the floor, right at Manson's feet. Disgusted by such objectionable behaviour, Manson was on the point of telling his patient to mind his manners when he noticed that the sputum contained a number of rusty brown coloured specks. An immediate examination of the specimen under the microscope revealed the presence of masses of tiny egg-like bodies. Some time late Manson discovered that these bodies hatched out into small embryos. It so happened that some months previously a Dr Ringer in Formosa had written to him about the finding at autopsy of a lung fluke in one of Manson's old patients. Ringer sent the specimen to Manson and further examination revealed more of the embryos, which were similar to those Manson had found in his Chinese patient. This simple observation formed the basis of the study of the parasite which followed in the next few years. When, in 1899, Manson returned to London he continued his studies in tropical medicine, including work on trypanosomiasis which brought him into contact with the tragic figure of Roger Casement, then a member of the Consular Service in Sierra Leone. He was also responsible for descriptions of several new species of blood filaria, and a demonstration of the left history of the parasite responsible for guinea worm infection. Manson's mosquito-malaria hypothesis was the basis of Ross's subsequent discoveries, and he constantly encouraged and advised Ross in his research. Posterity seems to have been less than just in the recognition it awarded Manson for his share in Ross's discovery: without Manson's vision the world might well have had to wait for this valuable knowledge.
Doctor, scientist and administrator
Sir Patrick Manson, who filled the roles of
physician, scientist and administrator was born on 3 October 1844, at Cromlet
Hill, Oldmeldrum, Aberdeenshire. He studied medicine at Aberdeen
University, and soon after qualification joined the Chinese Maritime Customs
Service. First he was posted to Formosa and later to Amoy, spending
twelve years in the service.
In 1883 he moved to Hong Kong where he established a large practice, took a leading part in the establishment of medical teaching in the colony, and founded a medical school - now the University of Hong Kong. When Manson returned to London he opened consulting rooms in Queen Anne Street and, despite the demands of a large practice, continued his research. In his house he equipped a small room as a laboratory which, from its chronically disorganised state, was referred to by his family as the 'muck room'. With the support of the Colonial Secretary, Joseph Chamberlain, Manson proposed to the Seaman's Hospital Society (of which he was physician) a scheme for the organisation of a School of Tropical Medicine at their Albert Dock Hospital. This important project was successfully launched in 1899, when the first session opened with 27 students. During the three quarters of a century of its existence the school has grown both in size and reputation, and is now the London School of Hygiene and Tropical Medicine. Sir Patrick Manson's advice on the problems of tropical medicine was widely sought and for many years he was consultant on these matters to the Colonial Office. He died on 9 April 1922.
NOTE: This historical note has been abstracted
from 'Founders of Medical Laboratory Science' and is provided for educational
James McIntosh - 1882 - 1948
Paul Fildes - 1882 - 1971
It sometimes happens that the name of an individual is best remembered, and in fact becomes a household word in the laboratory, by a piece of apparatus or a technical method bearing his name, although his achievements in the fields of medical research and discovery may be of much greater importance. The history of the medical laboratory sciences provides many examples of this, including such famous figures as Pasteur, Koch and Ebrlich. McIntosh and Fildes also fall into this category. Although both made many important contributions to medical knowledge they are chiefly remembered for their development of a new and revolutionary method of achieving anaerobic culture.
Since the early days of bacteriology there had been methods of growing anaerobic bacteria, but none of them was wholly satisfactory as they rarely removed all traces of oxygen. The simplest of these methods was to cover the surface of the culture with a substance which was impermeable to air. Alternatively, heat could be used to drive off the oxygen, or a reducing agent such as sodium sulphide - or even a fragment of living tissue - could be added. Mechanical means (such as a pump) might be used or as in the case of Buchner and Bulloch's method, chemical means. Nowadays it is perhaps difficult to appreciate the relative inefficiency of the methods then available, and how difficult it was to obtain satisfactory cultures of anaerobic organisms. Only in exceptional circumstances was the procedure attempted and by no means could it be considered a routine measure.
In 1907 Pfuhl published a paper in which he described how he had made use of the catalytic properties of spongy platinum to obtain anaerobic conditions for the culture of organisms. In 1915 Laidlaw suggested the use if platinised charcoal and colloidal platinum for the same purpose. During the First World War the bacteriology of war wounds, and in particular the investigation of clostridial wound infection, became a matter of prime importance and it became a matter of urgency to develop better methods of culturing anaerobic organisms.
Starting from the previous work of Pfuhl and
Laidlaw, McIntosh and Fildes developed the highly successful 'McIntosh
and Fieldes Jar', which quickly became a standard item of bacteriology
McIntosh and Fildes anaerobic jar
McIntosh and Fildes published their first paper on the anaerobic jar in 1916, entitled A New Apparatus for the Isolation and Cultivation of Anaerobic Micro-organisms. Five years later a second paper described an improved form, virtually the McIntosh and Fildes jar of today. Though a leading laboratory supply house immediately made arrangements to manufacture this apparatus, the paper gave full details for making the jars in the laboratory. A 'do-it-yourself' approach was prevalent in laboratory practice in those days.
The use of a paint tin measuring seven inches by five inches, with a lever-off lid, was suggested by the authors. A small stopcock - the type used on model steam engines was found to be very suitable - was fitted in the lid and carefully made gas-tight with solder. to the thread of the stopcock was attached a strip of brass, bent to form a bracket. This carried the palladium capsule, which was made by impregnation asbestos wool with a solution of palladium chloride, and was enclosed in a small envelope made from wire gauze. After impregnating the capsule was heated in a smokey flame, which coated it with carbon; finally, complete reduction was brought about by further heating with a blowpipe.
After filling the tin with cultures, the method of use was to fill the gutter of the tin with a thick layer of plasticine, heat the capsule in a bunsen burner, and immediately close the tin and connect a hydrogen generator to the opened stopcock. In their first experiments McIntosh and Fildes had used glass vessels as container, which made it possible to judge whether the removal of molecular oxygen was complete by the bleaching of an indicator solution of broth and methylene blue inside the jar. When tins where used it was impossible to use an indicator in the same way; it was found, however, that provided an air-tight seal was obtained (and it was essential that the stopcock should be well greased) there was little risk of failure.
In 1921 Fildes and McIntosh published a paper describing an improved form of anaerobic jar. The main improvement was the introduction of electrical means of heating the palladium asbestos capsule. Not only was this a much more convenient arrangement but it had the great advantage that any trace of oxygen which might later diffuse out of the culture medium could be removed easily be reactivating the palladium capsule by simply passing current through the heater; if necessary the current could be allowed to run continuously.
Born in Aberdeen in 1882 James McIntosh graduated
MC ChB at Aberdeen University in 1905 and then spent some time at the Pasteur
Institute in Paris before returning to Aberdeen in 1908. In the same
year he moved south to The London Hospital, where he remained until becoming
Professor of Pathology and Director of the Bland-Sutton Institute at the
Middlesex Hospital in 1920. Among many important offices he held
was a term as President of what was later to become the Institute of Medical
McIntosh, alone or in collaboration with other, published well over a hundred papers. His work on syphilis contributed a great deal to the knowledge of the serological diagnosis and treatment of the disease. He did much work on the Wasserman reaction, which had been introduced in 1906. Among his many contributions on this subject was a paper published in 1912 which gave details of a formula for the preparation of a cholesterolised alcoholic heart extract, which became the standard Wasserman antigen. His work covered a very wide field including chemotherapy, the Rous sarcoma, tar tumours, dental caries, and viral disease. In his investigation of influenza he became convinced of the role played by a virus in this condition. During the First World War he turned hiss attention to bacteriological culture media and was particularly interested in the estimation of pH. McIntosh died at his birthplace, Aberdeen, in 1948 at the comparatively early age of 66.
Paul Gordon Fildes, the son of the painter Sir Luke Fildes, was born in London in 1882. He studied medicine at The London Hospital, where he subsequently worked as a bacteriologist. In the First World War he served in the Royal Navy and was in charge of the pathology laboratory at the Royal Naval Hospital, Haslar. In 1934 he went to the Middlesex Hospital as Director of the newly established Medical Research Council unit for the study of bacterial chemistry. During the Second World War he was leader of a team of scientists at Porton Down, who were concerned with counter measures against the possible use of bacteriological warfare. After the war the Bacterial Chemistry Unit was reconstituted at the Lister Institute of Preventive Medicine and Sir Paul Fildes remained as the Director for three years. He retired in 1949 and went to the Department of Pathology at Oxford University, working there for the next thirteen years.
While Sir Paul Fildes' work on bacterial chemistry earned him an international reputation, he did important work in many other fields of medicine. His published work included papers on haemophilia, cerebrospinal fever, tetanus, and influenza. His studies on Haemophilus influenzae led to the introduction of the special culture medium which carries his name. He was recipient of many honours: awarded an OBE in 1919, knighted in 1946, elected a Fellow of the Royal Society and presented with the Royal Medal of that Society in 1953. Honorary degrees were conferred on him by the Universities of Cambridge and Reading. He died at the age of 88 in 1971.
NOTE: This historical note has been abstracted
from 'Founders of Medical Laboratory Science' and is provided for educational
1863 – 1943
Of all the early pioneer bacteriologist's, the life of the shy, introverted, but adventurous and strong-willed Alexandre Yersin was one of the most colourful and exciting. An ardent disciple of the Pasteur School, Yersin's work with Emile Roux in the discovery of diphtheria toxin earned him an early reputation as a bacteriologist of international fame. Yersin spent a large part of his working in what was then French Indo-China, where he was able to combine the life of a working bacteriologist with that of an active explorer.
Although the discovery by Edwin Klebs in 1883 of the diphtheria bacillus, and its successful cultivation a year later by Friedrich Loeffler, were tremendous advances, there were still many important problems of diphtheria left unsolved. At the Pasteur Institute, in the late 1880s, Emile Roux decided to investigate some of these and he invited Yersin - then a young scientist working at the Institute – to collaborate with him in this project. The research proved to be particularly arduous and for three years Roux and Yersin spent long hours in the laboratory, often working far into the night. The results of these labours were made public in three classic papers published in 1888, 1891 and 1890.
In the first of these publications they confirmed Loeffler's methods of cultivation and made the important observation that, in experimentally infected rabbits, when death did not occur quickly the animal was often paralysed. Roux and Yersin came to the very significant conclusion that the diphtheria bacillus was capable of producing a poison. They were able to demonstrate this poison, or diphtheria toxin, by passing bouillon cultures of the diphtheria bacillus through Chamberland filters, the sterile filtrate being capable of causing the disease when injected into experimental animals.
In their second paper they described in detail their procedure of producing the toxin, and reported the results of an elaborate study of its pathogenic action on other types of experimental animals.
The technique for the laboratory diagnosis of diphtheria was outlined in their third paper.
The demonstration of diphtheria toxin was to prove of tremendous importance in the treatment of the disease, as it formed the starting point of von Behring's monumental work on development of an immunising serum.
Yersin's work on plague
In May 1894 plague spread from the mainland of China to the colony of Hon Kong, where it caused many deaths. Two medical research teams were sent tot investigate and to discover the cause of the epidemic. One of these, the Japanese plague mission, was well staffed and headed by the famous bacteriologist Shibasaburo Kitasato. The other was a small French mission under Yersin. The Japanese team arrived first and were at work by 14 June, a day before Yersin was able to arrive in Hong Kong. A laboratory was provided for the Japanese but Yersin had to be content with a straw hut. Yersin's team consisted only of himself and tow almost untrained assistants, on of whom promptly disappeared taking Yersin's modest cash reserves with him. A courtesy visit to Kitasato was a failure because of language difficulties, and it became clear that both missions would have to work independently. Yersin's problems were by no means at an end, however, and everything that could go wrong seemed to do so. He had great difficulty in getting plague autopsy material as it was reserved for the Japanese mission, and it was only after considerable delay that he was able to carry out a few autopsies.
The rapidity with which Kitasato reported discovery of the causative organism is well known, yet despite the difficulties he encountered Yersin arrived at the same conclusion within weeks.
The validity of Kitasato's original findings has been questioned and priority of discovery has been the subject of controversy. rightly or wrongly the Japanese have been credited with the discovery, the work of the unfortunate Yersin merely serving as confirmation of the Japanese findings. One cannot help feeling some sympathy for Yersin who, with grossly inadequate resources, successfully solved on of the world's greatest disease problems just a few weeks too late to claim priority of discovery.
Alexander Emile Jean Yersin was born in 1863 in the Swiss town of Aubonne, not far from the shore of Lake Geneva. his father, who had been a professor of natural history, died a few days before his birth. In early life young Yersin showed an interest in, and aptitude for, scientific studies. He began his medical education at Lausanne then moved to Marburg, and finally completed his training in Paris. Yersin had little liking for routine clinical work, and right from the start of his medical career he decided to specialise in pathology. He became an assistant to Andre Cornil, the pathologist at the Hotel-Dieu Hospital in Paris, and it was while he was here that he suffered an accident, which determined his future medical career.
While carrying out an autopsy on rabies victim Yersin cut himself and beacme a potential victim of the disease. fortunately a course of treatment at the Pasteur Clinic prevented the disease developing, and young Yersin – greatly impressed both by the treatment he received, and by the enthusiasm of Pasteur's band of workers – determined to join them. First, however, he decided to see something of the methods used in Koch's rival school of bacteriology in Berlin, and in 1887 spent some months in that famous department. Here he was able to learn and practice the new technical methods introduced by Koch, and he was responsible for the introduction to the Paris School of many of the methods evolved in Koch's laboratory. His first research at the Pasteur Institute was concerned with tuberculosis and the action of antiseptic and heat treatment on tubercle bacilli – further evidence of Koch's teaching. It was his next three years' work with Roux that were the most important, however:
Once the diphtheria project had been completed Yersin's restless nature asserted itself, and he gave up his post and set out to see something of the more distant parts of the world. He obtained a post as ship's doctor on a French vessel travelling in the Far East, making several voyages to Saigon and becoming fascinated by Indo-China, much of which was still completely unknown. Resigning his post as ship's doctor he set off on his first exploratory mission of the country's interiot. In the years 1892 to 1894 he made a number if explorations, usually with grossly inadequate resources, but somehow this determined little man survived all the misfortunes which befell him in the course of his adventurous journeys, and collected valuable information on the topography, flora and fauna of these regions.
In 1891 Albert Calmette had come to Saigon
to organise a branch of the Pasteur Institute and when, after two years,
he had to return to France he persuaded Yersin to join the French Colonial
Medical Service and left him in charge of the new Institute. This
position suited Yersin very well. It gave him an official position
in the country he had come to love and was determined to help. It
also permitted him to pursue both his scientific interests and his love
os exploration. In 1895 he was asked to found a second Pasteur Institute
in the country and he chose Nhatrang, a small fishing village in a beautiful
bay with a temperate climate and lush vegetaion, where he lived until his
death in 1943. In 1933 Yersin was appointed a member of the Scientific
Council of the Paris Pasteur Institute and once a year made the journey
by air to Paris to fulfil his official duties. The last of his visits
was in 1940, when he was on the last plane to leave Paris for Indo-China.
NOTE: This historical note has been abstracted
from 'Founders of Medical Laboratory Science' and is provided for educational
1863 - 1931
The striking success of modern drugs and methods of therapy in the control and treatment of tuberculosis has tended to dim memories of the devastation brought about by that disease less than a quarter of a century ago. The name of the Frenchman; Albert Calmette, is inextricably linked with one of the first successful measures to check the spread of this disease. Calmette, with his associate Guerin, produced the BCG vaccine (bacille de Calmette et Guerin) - after nearly 20 years' work and this vaccine, with the improvements subsequently made to it, gave millions of children protection from tuberculosis and saved countless young lives.
A vaccine to give protection from attacks of tuberculosis was not a new idea; following the successful outcome of von Behring's work in Germany on diphtheria, and Sir Almroth Wright's work in England on typhoid, it was the ambition of many research workers to produce equally effective protection against tuberculosis. In 1902 von Behring himself had experimented with measures to protect cattle from bovine tuberculosis by the administration of a strain of human tubercle of low virulence. Prior to von Behring's work Others had employed extracts of killed tubercle bacilli, or emulsions of tubercle bacilli cultures whose virulence they had attempted to diminish by growing the organisms in media containing formalin or similar reagents. The reputation of the German bacteriologist, Robert Koch, had been severely damaged by the failure of his ill-fated tuberculin as a therapeutic agent.
In 1894 Louis Pasteur, then director of the famous Pasteur Institute m Paris, was asked by the city of Lille, where he had started his career in bacteriology, to establish a branch of the Pasteur Institute in that city. Largely on the advice of his deputy, Emile Roux, Pasteur invited Calmette, who was working part-time at the Institute in Paris, to take charge of the new venture. Calmette was well suited to the post, having previously established a branch of the Pasteur Institute in Saigon, in what was then French Indo-China. Calmette accepted the post and in 1895 went to Lille as Director.
While suitable premises were being built he
started work in temporary laboratories. He was a gifted organiser and within
a short time had the laboratories producing sufficient smallpox and rabies
vaccine to meet all the needs of northern France. He also developed a new
commercial process for the conversion of starch into sugar and alcohol,
money from which greatly helped in financing the new project. When
the work of the laboratories was safely moved into the new building, and
vaccine production was in full swing, Calmette turned his attention to
what became his main interest - the discovery of some means of combating
Calmette soon found that the work of the Lille Insatute, and his research into the development of attenuated live vaccine in accordance with Pasteur's practice, required the services of a trained veterinarian for the maintenance of a stock of experimental animals, mostly bovine. Professor Nocard recommended his assistant, Camette Guerin, for the post and in 1897 Guerin joined Calmette in Lille. They then began the long series of cultures which culminated in the production of the successful vaccine, known as BCG.
Over a period of 13 years, using a bovine strain of tubercle which had been isolated by Nocard in 1902 they sub-cultured the organisms 231 times and the work continued even throughout the German occupation of Lille during the 1914-18 war. By 1908 Calmette and Guerin had succeeded in obtaining an attenuated strain of the bovine tubercle bacillus. The culture medium they used consisted of potato, glycerine, and bile salts. The addition of bile to the medium was to prove of considerable importance and came about quite accidentally. In the preparation of the finely divided emulsions of tubercle bacilli needed for the injection of the experimental animals, Calmette and Guerin discovered that sterile ox bile was a most effective agent for breaking up the clumps of organisms. Furthermore they discovered that, when bile was included in the culture medium, changes in the virulence and morphology of the organisms began to occur. Prolonged culture in media containing bile produced a strain of organisms from which a safe and effective vaccine could be made. The work continued for a number of years, until it was conclusively proved that the vaccine was harmless to the tuberculosis-susceptible guinea pig. Another ten years were to elapse before Calmette and Guerin were fully satisfied that it was equally suitable for children. These findings were duly confirmed by the Academy of Medicine in Paris and the first prophylactic tube of BCG on man was carried out in 1921, when a physician persuaded Calmette to allow his vaccine to be used to protect a child delivered of a tuberculous mother. This unplanned case was completely successful and Calmette, in collaboration with medical colleagues, then carried out extended - trials. t
In 1924 Calmette and Guerin published their historic paper, in which they showed that trials undertaken in many parts of the world had clearly demonstrated the success and safety of the vaccine as a protective measure. In 1928 a new building was erected at the Pasteur Institute to accommodate the Service du BCG.
The Lubeck disaster
In 1930, just at the time when BCG had gained
a large measure of acceptance and thousands of children had been successfully
treated, a tragic accident occurred. There had always been those
who doubted the safety of the method, fearing that the strain would not
always remain avirulent, and this disaster immediately brought a strong
reaction from its opponents.
The director of the public health laboratories at Lubeck, in Germany, had arranged for Calmette to send him a culture and for a vaccine to be prepared in the laboratories of the city hospital. The batch of vaccine was duly produced and given to some 251 children. Within a year 207 of these children had contracted tuberculosis, and 72 died. The whole world was profoundly shocked and Calmette was savagely attacked both by the medical and by the lay press. Doctors who had successfully used the vaccine rallied to his side, however, and from Germany there was particularly strong support. The German health authorities held a painstaking inquiry, which was conducted with scrupulous fairness, and their findings completely absolved Calmette of any blame. They found that the strain of organism supplied by Calmette had been contaminated in the Lubeck laboratories by a virulent strain, and the German doctors held responsible were given prison sentences.
Despite the inquiry's report completely clearing Calmette of any blame, the use of BCG suffered a serious setback for some years. In Great Britain the safety and value of the vaccine was clearly demonstrated as a result of a Medical Research Council investigation in 1959.
Pioneer of immunisation
Leon Charles Albert Calmette was born in Nice in 1863, the son of a lawyer. His first ambition was to follow a naval career, and in due course he went to Brest as a cadet. His studies were curtailed by an attack of typhoid fever which left him in such a poor state of health that he was rejected on medical grounds. Calmette did not abandon his ambition to enter the navy and after a period of recuperation was accepted for training as a naval physician. After qualification he saw service in various parts of the world. In 1890 he was transferred to the newly formed French Colonial Medical Service and given leave to undertake further study at the Pasteur Institute in Paris. Here he came to the notice of the great Pasteur, who was much impressed by his ability and nominated him for the job of organising the Institute in Saigon. Calmette soon had an efficient, if modest, organisation working on the production of smallpox and rabies vaccines and he undertook an extensive study of snake venoms and the production of anti-venom sera. Unfortunately, after two years he contracted a severe form of dysentery and had to return to France, leaving the Institute in the hands of Alexandre Yersin (Emile Roux's collaborator in his work on diphtheria) whom Calmette had persuaded to join the French Colonial Medical Service in Saigon.
In Paris Calmette recovered from his attack
of dysentery and was given a part-time administrative post at the Ministry
of the Colonies. The mornings, and most evenings, he spent working
in Roux's laboratory at the Pasteur Institute and it was while filling
these posts that the opening in Lille occurred.
During the 1914-18 war, when the German forces occupied Lille, all cattle were requisitioned and the bulk of Calmette's animal experiments had to cease. He continued, however, to keep some pigeons (in breach of the regulations) and this, and other acts, brought him into conflict with the occupying authorities. Madame Calmette was taken to Germany as a hostage, and Calmette himself was interned and only saved from a possible death sentence by the personal intervention of the highly respected German bacteriologist, Richard Pfeiffer, who at the time held the rank of Generalarzt in the German army.
On Metchnikoff's death in 1917 Calmette was named, in absentia, Associate Director of the Pasteur Institute in Paris and he took up this post at the end of the war. His death in 1933 was undoubtedly hastened by the worry brought about by the Lubeck disaster, and the long enquiry which followed.
His associate in the discovery and development
of BCG, Cermelle Guerin, was born in Paris in 1872 and trained as a veterinarian.
Almost all his professional life was spent in the service of the Pasteur
Institute in Lille, and later in Paris, where he continued to supervise
the production of BCG after Calmette's death. Before his death in
1961 Guerin had the satisfaction of seeing the final acceptance of BCG
as a safe and efficient prophylactic measure against tuberculosis.
Wilhelm Bunsen, the famous 19th century German chemist and physicist, made many important discoveries and contributed a great deal to the advancement of knowledge in his chosen field. He is, however, most widely known by the gas burner which bears his name. This burner, which was invented more than a hundred years ago, is still an indispensable piece of equipment in many widely different types of laboratory. Initially designed with the needs of the chemist and physicist in mind, it has proved essential in medical laboratories also, and especially in bacteriology. The pioneer bacteriologists were fortunate that the burner was available in the last quarter of the 19th century, when tremendous advances were made in that science.
While the Bunsen burner has found. universal application, and perpetuated the name of its inventor far outside the world of science, many of Bunsen's other discoveries were also of great importance. He was a prodigious worker and, although many of his researches were long-term projects, he published over a hundred scientific papers.
An explosive subject
Bunsen spent forty years as Professor of Chemistry
in the University of Heidelberg.
It was in the early years of his professorship that he invented his famous burner.
Many others before him had tried to devise an effective laboratory burner, using coal gas, but none of these efforts had proved satisfactory; in fact, some had been literally explosive failures. With the exception of the simple spirit lamp and the charcoal fire, the only available alternative was an oil burner (known as the Argand) which had originated in Switzerland in the 18th century. In the 1850 the laboratories of the
University of Heidelberg were moved from the rather ramshackle remains of a long-disused monastery to a new building in the Plockstrasse. This move coincided with the introduction of coal gas to the city and the university authorities decided that their new laboratories should be connected to the gas supply.
Bunsen was well versed in the conditions required
for the combustion of a co gas and air mixture and, despite the series
of failures experienced by other worker who had attempted to construct
a practical gas burner, he was fully convinced that such a device was possible.
Needing a hot, smokeless flame for some experiments on spectrum analysis
he resolved to design a burner which would give the kind of flame he required
without any danger of an explosion. In this enterprise he was assisted
by one of his pupils, Henry Roscoe, who later became Professor of Chemistry
in Manchester and was knighted. The team also included Peter Desaga,
a technician on the university staff.
Experimental work on the project began in 1853. Superficially the problem appeared a simple one; it was necessary merely to convert a bright, smoky flame into a hot, smokeless one. Yet it was only after two years of delicate and painstaking experiment that success was finally achieved, with the production of the familiar Bunsen burner There were two basic difficulties which had to be overcome. First, they had to determine the correct mixture of gas and air (they eventually settled on three volumes of air to one of gas): secondly, it was necessary to ensure complete burning of the gas and to minimise the amount of carbon produced. This was done by admitting air into the flame from below, so that there was air both inside and outside the flame.
The principle of the Bunsen burner was subsequently employed in the domestic gas stove, and indeed in any gas apparatus requiring this-kind of flame. Several modifications of Bunsen's original design appeared, mostly intended to give a larger area of hot flame, and a pilot light was also subsequently included.
Many people consider that Bunsen's most important findings arose from the long series of successful investigations in the field of spectrum analysis which he carried out in conjunction with his colleague at Heidelberg, Gustav Kirchoff. Yet these were not his only noteworthy discoveries. Before he went to Heidelberg, Bunsen had been employed at Cassel and as Professor of Chemistry at Marburg. During this phase of his life he invented the carbon-zinc battery (the now obsolete 'Bunsen cell'). He also conducted extensive research into the properties of the cacodyl compounds. In the course of this latter work an accident occurred which almost cost him his life, and which resulted in the loss of an eye.
Other areas of research which continued to interest Bunsen for many years were the development of methods for the accurate measurement of gaseous volume, and the investigation of gaseous diffusion and absorption. Unlike many of his contemporaries Bunsen was no compiler of manuals; the only book he ever published was one describing his gasometric researches. His years of work with his friend Kirchoff, who also held a chair at Heidelberg, established spectrum analysis as a science and led to Bunsen's discovery of the rare elements caesium andrubidium. He also observed the high luminosity displayed in the colourless flame of certain rare earth metals, a discovery which was to be of immense value in the incandescent gas mantle industry.
Liked and respected
Bunsen is said to have been a kindly and modest man, greatly respected by his contemporaries in the world of science and beloved by the students who came to him from all parts of the world. He spent 37 years as Professor of Chemistry at Heidelberg and during that time received many offers of more attractive appointments in larger university centres. All these invitations he refused, preferring the quiet and restful atmosphere of Heidelberg.
Born on 31 March 1811 in the city of Gottingen,
on the eastern side of the Prussian province of Westphalia, Bunsen was
the son of the chief librarian and Professor of Classical Philology at
the University of Gottingen. Young Bunsen was educated in his home
town and afterwards furthered his knowledge by periods of study in Berlin,
Paris and Vienna. At the age of 22 he returned to Gottingen to rake
up a post in the university. Three years later he moved to Cassel
and in 1838 he was appointed Professor of Chemistry at Marburg. In
1861 he moved to Breslau, and in the following year to his final appointment
at Heidelberg. Bunsen retired from active university life in 1889
and remained in Heidelberg until his death ten years later.
Sir David Bruce was one of the great pioneers of the 'Golden Age of Bacteriology, and although be is best remembered for bis outstanding discoveries in connection with two diseases-Malta fever and sleeping sickness-be also carried out important research in other fields. As well as being a great scientist he was a gifted medical administrator who had a distinguished military career.
Malta, or Mediterranean, fever was renamed undulant fever when its wider geographical distribution became appreciated; the latter title being derived from the character of the patients' temperature charts. The disease, probably first introduced into Malta during the Crimean war, quickly became a permanent scourge of the island with a high death rate-the result of large numbers of both the civilian and garrison populations suffering attacks of this lengthy and debilitating illness. Bruce's discovery in 1887 of the causative organism, and the part he played later in demonstrating the role of the goat in spreading the fever, eventually resulted in the eradication of the disease from Malta.
It was while serving in Malta as an army medical officer that Bruce became concerned with the problem of Malta fever. He was convinced that the disease was due to some form of bacterial infection and concentrated his efforts on a search for the causative organism. Within two years he was successful for, in the spleens of patients who had died of the disease, he discovered a 'micrococcus', and by careful experiment proved conclusively the aetiological significance of the organism, which he called Micrococcus melitensis. In 1920 Tensier and Meyer placed the organism in a separate genus, with the name Brucella in honour of its discoverer, and it became known as Brucella melitensis.
Bruce's discovery of this organism had little
immediate effect on the control of the disease, but the introduction in
1897, by Sir Almroth Wright and Frederick Smith of an agglutination test
for the diagnosis of undulant fever was the first step in bringing it under
control. In 1904, twenty years after his initial discovery, Bruce
returned to Malta and played an important part in the final conquest of
the disease. He headed the Royal Society's Malta Fever Commission,
which proved that the principal source of infection was contaminated goats'
While several workers contributed to the discovery of the cause of this dreaded tropical disease, Bruce's work was by far the most important. His earlier finding that a trypanosome was the cause of a disease in cattle and horses led directly to the discovery that sleeping sickness was also a form of trypanosomiasis. Later he played a leading role in the work that demonstrated the part played by the tsetse fly in the transmission of the disease.
Bruce's interest in trypanosomiasis began in 1895 when he was stationed in South Africa. At the special request of the Governor of Natal, Sir Walter Hely-Hutchinson who had been Lieutenant Governor of Malta at the time of the successful research into the cause of Malta fever - Bruce was sent to investigate a disease of domestic animals, called 'nagana', which caused a high mortality amongst horses and cattle. At first Bruce regarded the problem as bacteriological but, when this line of investigation proved fruitless, he switched his attention to the examination of blood films. Unfortunately his laboratory resources were limited. The only stain he had available was carbol-fuchsin - not a particularly suitable blood stain-and although he did find some curious objects between the blood cells he regarded them as artefacts. The search was, however, much more rewarding when he examined wet preparations of blood and found an organism which, at first, he thought might be a filaria. This organism was later named Trypanosoma brucei and, by experimental infection of test animals, Bruce proved it to be the cause not only of 'nagana', but also of another condition of domestic animals-tsetse fly disease; in fact, he established that they were the same disease. Recalled to military duties in January 1895 Bruce's research was interrupted, but later in the year he was able to return and complete his work on trypanosomiasis in domestic and wild animals, and on the part played by the tsetse fly, Glossina morsitans, in transmitting the infecting organism.
In 1902, the War Office seconded Bruce to the Royal Society's Commission on Sleeping Sickness, in Uganda. Working for this body he played a prominent role in proving that the disease was a form of trypanosomiasis, transmitted by the tsetse fly Glossina palpalis. This established the basic facts about the aetiology and epidemiology of sleeping sickness and was the foundation upon which all subsequent investigations were based.
Scientist and soldier
Sir David Bruce was born in Melbourne on 29 May 1855. His father, who was an engineer connected with the mining industry, was only a temporary resident in Australia and when David Bruce was five years old the family returned to Scotland and settled in Stirling.
Bruce entered Edinburgh University in 1876,
graduating in medicine in 1881. His first post was as an assistant
to a doctor in Reigate but within two years he decided to enter the Army
Medical Service and was commissioned in 1883.
Almost immediately he was posted to Malta where he did his famous work on Malta fever. His next appointment was that of assistant professor of pathology at the Army Medical College at Netley, where he stayed for the following five years. Before taking up this post, however, he spent his leave in Berlin working in Robert Koch's laboratories.
In 1894 he was posted to South Africa and commenced his investigations into trypanosomiasis. While in South Africa he was recalled to purely military duties during the Boer War. Shut up in Ladysmith during the siege, he took charge of a large military hospital and his wife, who accompanied him on all his travels, undertook the duties of sister-in-charge of the operating theatre. Sir David does not appear to have had a great liking for the finer details of laboratory technique and Lady Bruce, who became a highly-skilled bacteriologist and microscopist, was largely responsible for the technical work that his research required.
During the 1914-1918 war Bruce was commandant
of the Royal Army Medical College at Millbank and was responsible for research
into trench fever and tetanus. He was knighted in 1908 and received
many honours to this day his name is perpetuated by the David Bruce Laboratories
in Wiltshire, and the David Bruce Hospital in Malta (formerly the military
hospital). He died in London on 27 November 1931, four days after
the death of his wife.
1870 - 1961
The decade 1870-1880 is often referred to as 'The Golden Years of Bacteriologic Discovery'; one might, with equal justification, label the years 1890-1901 as 'The Golden Years of Immunological Discovery'. Famous figures such as Emil Von Behr his associate Shibasaburo Kitasato, Paul Ehrlich, Elie Metchnikoff, and Richard Pfeiffer are among the many who founded the new science of immunology.
It was a well-known fact that, following recovery from certain infections, individuals acquired protection from subsequent attacks, and that in some instances this protective power could be stimulated without an attack of the actual disease. The practice of vaccination against smallpox (introduced by Edward Jenner) and Louis Pasteur's work on anthrax and rabies clearly demonstrated the value of this knowledge. Understanding of the underlying mechanism of antigen-antibody reactions, and evaluation of methods of serological diagnosis, were the aims of the pioneers of immunology.
In this new field the Belgian Jules Bordet,
together with his fellow countryman Octave Gengou, played a leading role.
Their researches were on the properties of serum from immunised animals,
and the Bordet-Gengou complement fixation technique was later the basis
of a number of diagnostic serological tests. Bordet was not only
a great immunologist, he was also an eminent bacteriologist and, with Gengou,
was responsible for the discovery of the causative organism of whooping
The complement fixation reaction
Bordet's early interest in the field of immunity was shown in a paper published in 1895. In this he demonstrated that two quite different substances were involved in the phenomenon of bacteriolysis. These substances he called 'alexine' (from the Greek, I ward off) and 'substance sensibisatic'; terms which were replaced by Ehrlich's 'complement' and 'amboceptor'.
Bordet utilised the work carried out in the
preceding year by Richard Pfeiffer and Vasiliy Isayeff, who had demonstrated
the possibility of using immune serum as a diagnostic test for cholera.
Pfeiffer and Isayeff had found that cholera vibrios injected into the peritoneal
cavity of an immunised guinea pig quickly lost their motility and were
eventually phagocytosed by leucocytes. This discovery became known
as 'Pfeiffer's phenomenon', and Bordet and Gengou used the same principle
for their in-vitro tests.
In 1901 Bordet and Gengou published details of their complement fixation test. This method, based on their previous work on immune haemolysis and bacteriolysis of cholera vibrios, was applicable to the detection of antibodies to a variety of bacteria, and gave rise to a most important range of diagnostic serological tests. Bordet and Gengou's complement fixation method also had applications in the evolution of a technique for the detection of human blood in medico-legal work.
Causative organism of whooping cough
As a research team Bordet and Gengou proved an ideal combination and in 1906, with the publication of a paper on the causative organism of whooping cough, they made a further major research contribution. In this paper, Le Microbe de la Coquelusbe, they gave a full account of the properties and cultivation of the organism now known as Bordetella pertussis, together with strong evidence supporting its aetiological relationship to the disease. The organism they isolated was a minute coccobacillus which they found in sputum expectorated during paroxysms of coughing in acute whooping cough. For the cultivation of the organism they devised a special slope culture medium known as Bordet and Gengou's medium, the principal constituents are potato extract, agar, and blood.
In modern bacteriological nomenclature the genetic part of the organism's name-Bordetella-is a tribute to one of its discoverers. Although for many years Bord. pertussis was known as Haemophilus pertussis, it was originally called the 'Bordet-Gengou bacillus'
Born on 13 June 1870 in the Belgian town of
Soignies, some twenty miles from Brussels, Jules Bordet studied at the
University of Brussels and graduated as a doctor of medicine in 1892.
Two years later he moved to the Pasteur Institute in Paris, where he held
the post of 'preparateur' in Metchnikoff s laboratory. In 1901 he
was recalled to Brussels to found the Pasteur Institute and to become its
first director. and in 1907
he was appointed professor of bacteriology in the University of Brussels-a post he held until his retirement in 193 5.
During his long and distinguished career Bordet received many honours, including the Nobel Prize for medicine and physiology in 1919. Although Bordet and Ehrlich held very different views on the mechanism of the antigen-antibody reaction, this difference of opinion was never allowed to degenerate into bitter controversy, as sorry times arose between distinguished protagonists of opposing theories. Jules Bordet outlived most of his contemporaries: he was 90 years old when. m 1961, he died in home at Ixelles on the outskirts of Brussels.
NOTE: This historical note has been abstracted
out of 'Founders of Medical Laboratory Science" and is provided for educational
Emil von Behring
1854 - 1917
It is often held that the discovery of antitoxic immunity was one of the greatest achievements of medical science. Certainly the discovery of diphtheria antitoxin, offering dramatically successful treatment of a disease which caused thousands of deaths amongst young children each year, was an epoch-making event.
The nature of immunity had for a long time been the subject of deep controversy. There were two main schools of thought. The 'Solidists', led by the great Elie Metchnikoff, thought that phagocytes played the leading role. Supporters of the alternative theory, the 'Humoralists', considered that a substance evolved in the body was mainly responsible.
Von Behring's work gave tremendous support to the 'Humoralists'. He proved that the serum of an animal which had recovered from an attack of a certain disease could be injected into a second animal and give it immunity to that particular disease This work clearly established that some protective substance was present in the serum, which would neutralise toxins produced by certain infecting organisms. This substance Behring called antitoxin, and he was the first to use this word.
Many others made important observations that led to the discovery and development of diphtheria antitoxin- Loeffler, Roux, Yersin, Ehrlich, Kitasato, and probably Robert Koch-but the actual discovery was undoubtedly the work of Behring and his Japanese co-worker, Kitasato. What is perhaps equally remarkable is the rapidity with which it became a widely used practical measure. In l890 Behring and Kitasato published the result of their research into diphtheria and tetanus antitoxins, showing how they had cured infected animals and immunised healthy animals. Exactly one year later, on Christmas night 1891, a child was successfully treated in a Berlin clinic with diphtheria antitoxin.
Behring's search for an antidote to the toxic
products of bacteria started when he joined the team of workers in Robert
Koch's laboratories. As an army surgeon he had had considerable experience
of iodoform, an antiseptic dressing then used routinely in the German army.
He had formed the opinion that its beneficial action was not solely due
to any direct antibacterial power, but to other properties it had of neutralising
the toxic products of septic bacteria. Roux and Yersin, in Paris,
had already demonstrated that diphtheria brought about its lethal effects
by the liberation of a toxin. That it was possible to acquire immunity
to diphtheria had been shown by Loeffler, who had also worked in Koch's
Another worker in Koch's laboratories was Fracnkel, who at this time was attempting to discover a method of attenuating diphtheria bacilli to produce a vaccine. Behring's first experiments were on the lines of Fracnkel's work and were, no doubt, influenced by his experience of iodoform. To cultures of diphtheria he added very small amounts of iodine trichloride, and by this method he did have some success in protecting guinea pigs. His next step was to render guinea pigs immune to diphtheria by repeated injections of the exudate from an animal dead of the disease and containing no diphtheria bacilli, but only the toxins. He showed that sub-lethal doses of diphtheria toxin had an immunising effect..
At this time Shibasaburo Kitasato, a Japanese bacteriologist, was also working in Koch's laboratory. He had recently discovered the causative organism of tetanus an, had shown that it produced a lethal toxin, and was actively engaged in the search for tetanus antitoxin. Behring and Kitasato joined forces, and in December 1890 published a paper on their work on tetanus and diphtheria antitoxins. Almost concurrently Behring published a separate paper, giving details of his work on diphtheria antitoxin. Within a matter of months Behring succeeded in working out a method of rendering guinea pigs immune to diphtheria, or of curing infected animals by the intraperitoneal injection of blood from immunised animals, and at the Congress of Hygiene in 1891, in London, he made a public announcement of this work.
During the following three years much progress was made in methods of producing diphtheria antitoxin, and establishing the correct dosage and standardisation. In these measures Paul Ehrlich had a great deal of influence, and it was during this period that the horse was first used for the production of antitoxin. At the International Congress of Hygiene and Demography at Budapest in 1894 Roux fully confirmed all Behring's claims, and diphtheria antitoxin began to be readily available for general clinical use.
During the next fifty years many improvements in the specific treatment of diphtheria by antitoxin took place. In 1912 Behring was one of the first to explore the possibilities of active immunisation, and in some countries a policy of active immunisation of children was adopted. The almost complete eradication of diphtheria from Britain, however, had to await an official campaign in the 1940s.
Von Behring's life
Emil Von Behring was born in the small town
of Hansdorf in East Prussia on 15 March 1854; by a strange coincidence,
a day after the birth of his associate in later life, Paul Ehrlich. Behring
studied at the Army Medical School in Berlin and graduated in 1878.
The first ten years of his professional life were spent in the German Army
Medical Service, working at various military establishments, and in 1888
he became lecturer at the Army Medical College. In 1889 he was appointed
assistant to Robert Koch at the Institute of Hygiene, and within a year
moved to the Institute of Infectious Diseases as Koch's assistant.
In 1893 the title of professor was conferred on Von Behring in recognition of his work on diphtheria, and two years later he moved to Halle and occupied the Chair of Hygiene. This, however, was to be only a temporary post. Within a year he accepted the post of Professor of Hygiene and Director of the Hygiene Institute in the University of Marburg, when only 41 years of age.
Behring became interested in the commercial manufacture of antitoxin, and the German organisation Farbwerke Höchst built and equipped an extensive plant for this purpose.
The early diphtheria antitoxins had many serious disadvantages, and some dangers, which research overcame in the next fifty years. The commercial production of diphtheria antitoxin was quickly established, however, first in Germany and shortly afterwards in many other centres. In England production began in London in 1895 in what is now the Lister Institute.
Another large combine, the Behringwerk, developed a bovo-vaccine which Behring had devised for the immunisation of cattle against tuberculosis. Although this vaccine was at the time widely used in many parts of the world, the results of prolonged experience were disappointing. As a result of these commercial connections Von Behring became a rich man and a large landowner and on his extensive estate he maintained a big herd of cattle on which his tuberculosis immunisation process could be tested under his direct supervision.
In 1901 Von Behring was awarded the Nobel Prize
for Medicine and in Germany his work received official recognition by the
conferring of the title 'Excellency' on him. The Paris Academie de Medécine
awarded him a prize of £1000 and the Institute of France £2000.
Emil Von Behring died, following an attack of pneumonia. on 31 March 1917
at the age of 61, and was buried in Marburg.
NOTE: This historical note has been abstracted
out of 'Founders of Medical Laboratory Science" and is provided for educational
1840 - 1905
Improvement in the performance of the microscope proved to be of paramount importance in the emergence of the medical laboratory sciences in the last part of the 19th century. This in turn demanded tremendous progress in methods f or the preparation of specimens for microscopy, and new methods of staining tissues and bacteria. Many famous characters contributed to the evolution of the microscope from the first crude, single-lens instruments of the 17th century to the highly practical compound models, with achromatic lenses and much improved mechanisms, of the mid-19th century. It was, however, the German microscopist and mathematician, Ernst Abbe, who did most to lay the foundations for the development of the modern instrument. If he opened up new fields for microscopy by introducing the oil-immersion objective and the apochromatic lens, and initiated new manufacturing processes, replacing the manual methods of individual craftsmen with mass production of precision-built instruments.
Abbe was not the originator of the oil-immersion objective, but he was the first to produce a really practical system for its use. In 1873, six years before Abbe published his famous paper On Stephenson's system of homogeneous immersion of microscope objectives, an American instrument maker called Robert Tolles devised and produced a 1/10 inch immersion objective, using Canada balsam as the immersion medium. Tolles' invention did not meet with a great deal of success as the objective itself was a clumsy affair and Canada balsam a messy and inconvenient medium to work with. Also, as Tolles had an unfortunate manner of expressing himself in print his views were much misunderstood and this discouraged the further development of a promising project.
The immersion principle was not a new idea.
Robert Hooke, microscopist and curator of the Royal Society in the 17th
century, recorded having used little globules of liquid with his single-lens
microscope. The idea does not appear to have been developed any further,
however, until the Italian mathematician and physicist Giovanni Amica,
experimented in the 1850s with various immersion fluids before finally
deciding on water. Later the optician Edmund Hartnack fitted water
immersion objectives with correction collars, and in the 1860s these lenses
were produced in considerable numbers, particularly in continental Europe.
In 1878 an eminent English microscopist, J W Stephenson, suggested the use of an immersion fluid with similar refractive and dispersive powers to glass. Unfortunately Stephenson lacked the technical knowledge and the workshop facilities required to develop the idea, and he therefore wrote to Abbe suggesting that he might like to explore the possibilities. Abbe was very impressed by the scheme; he had considered such a project some years previously, but had not investigated the idea further. Stimulated by Stephenson's letter, and with the facilities of the Zeiss organisation at his disposal, Abbe developed the scheme and within a few years the oil-immersion high-power objective had become a standard item of equipment on all but the cheapest microscopes. The search for a suitable immersion medium presented some difficulty, and over 300 fluids were investigated before cedar-wood oil was selected.
Despite the usefulness of achromatic lenses, Abbe was well aware that it was not possible to correct completely for both chromatic and spherical aberrations using flint and crown glasses. While the achromatic lens gave a perfectly satisfactory performance in ordinary work, something very much better was required for work demanding highly critical high-power microscopy. Alternative types of glass with different optical properties were at that time simply not available.
Despite the impossibility of obtaining glasses with the special properties he needed, Abbe continued his experiments and was able to test his theory by using specially prepared fluid elements as lenses. Two such experimental objectives were made at the Zeiss works, and although they proved fully the correctness of Abbe's theory they were quite unsuitable for commercial production. There the whole matter might have ended had not a Dr Otto Schott learned of Abbe's failure to obtain the glass he needed. Schott was well suited to undertake the experimental work: he had a good theoretical background and had been trained m the practical side of glass manufacture in his father's glass factory, which also gave him ready access to the facilities he needed. In a short while he wrote to Abbe reporting his Success in formulating a new type of glass with optical properties quite different from those of ordinary glass. With Abbe's constant encouragement Schott continued his experiments and produced more new types of glass. Although he was only able to manufacture small quantities of the new glass there was enough available to make a prototype lens which demonstrated the possibilities of the new objectives.
With financial assistance from the Prussian
government, and the support of the Zeiss organisation, a glass research
laboratory-which later expanded into a small glass-making plant-was opened
in Jena with Schott in charge. The new Schott glass works and the
Zeiss organisation worked in close contact, and by the time Abbe had completed
his calculations for the new range of apochromatic objectives adequate
quantities of the new glass were available for commercial production to
Somewhat surprisingly, Abbe's name is perpetuated not by any of his original contributions, but by a piece of equipment which had been in use long before his time-the substage condenser. Abbe devised an efficient condenser with a special mounting, which is still known as the Abbe condenser.
Lifelong collaboration with Zeiss
Ernst Abbe, the son of a spinning-mill foreman in the small German town of Eisenach, was born on 23 January 1840. He went to the University of Jena and then to Gottingen, where in 1861 he obtained his degree. After posts at Gottingen and Frankfort-am-Main he returned in 1863 to a lectureship at Jena. Three years later he was invited by Carl Zeiss to join the Zeiss organisation as a scientific consultant, and so began one of the most fruitful of scientific partnerships. Abbe's duties with Zeiss allowed him to retain his university post, and in 1870 he became a professor. Abbe greatly valued his continued association with the university and was always ready to offer promising young scientists posts in the Zeiss organisation, and to encourage Zeiss scientists to lecture at the university. In 1875 his close collaboration with Carl Zeiss was further strengthened when he became a partner in the firm, and in 1888, on the death of Zeiss, he became its proprietor.
Abbe died on 14 January 1905, a few days before
his 65th birthday. A most impressive monument to his memory was erected
in Jena and each year is visited by scientists from all over the world.
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