Constantin Levaditi and the Veterinary Medicine Constantin Levaditi Şi Medicina Veterinară

Rev Rom Med Vet (2016) 26 | 1: 67-72 67 THE HISTORY OF VETERINARY MEDICINE | ISTORIA MEDICINII VETERINARE

CONSTANTIN LEVADITI AND THE VETERINARY MEDICINE CONSTANTIN LEVADITI ŞI MEDICINA VETERINARĂ

R. IFTIMOVICI1)

This article is the fruit of bibliographical research I Acest articol este rezultatul unei cercetări biblio- conducted in the archives of the Pasteur Institute in grafice efectuată în arhiva Institului Pasteur din Paris. Paris. The text is based on unpublished documents, Textul se bazează pe documente inedite, în special especially letters sent by professor Constantin Levaditi scrisori trimise de către profesorul Constantin Levaditi (1874-1953) to prominent figures of veterinary medi- (1874–1953) unor personalităţi marcante ale medici- cine. Constantin Levaditi, a Romanian physician who nei veterinare. Constantin Levaditi, medic român, care worked in France, at the Pasteur Institute, is consi- a lucrat în Franţa la Institutul Pasteur, este considerat dered to be one of the founders of virology and anti- a fi fost unul dintre fondatorii virusologiei şi chimiote- infectious chemotherapy. He was initially (1896-1903) rapiei antiinfecţioase pe plan mondial. a disciple of Victor Babeş, Paul Ehrlich (in Frankfurt A fost iniţial (1896–1903) discipol a lui Victor Ba- /M.), I.I. Metchnikov and Emile Roux (in Paris). beş, Paul Ehrlich la Frankfurt /M., I.I Mecinikov şi Emile With Nobel Prize winner Karl Landsteiner (who dis- Roux la Paris. tinguished the four blood groups and the Rh factor in Împreună cu Karl Landsteiner, laureat Nobel (cel man), Levaditi discovered the polio virus in 1909. care a evidenţiat cele 4 grupe sanguine şi factorul Rh la Three years later he described (with C. Kling) the om), Levaditi a descoperit în 1909 virusul poliomielitei. ways the virus is transmitted, and introduced the first Ulterior, în 1912, a precizat căile de răspândire ale laboratory diagnosis of infantile paralysis (1913). acestui virus (cu C. Kling) şi a introdus primul diagnos- Levaditi, a Pasteurian, was the first to cultivate the tic de laborator al paraliziei infantile (1913). polio virus in vitro (1913), paving the way for vaccina- Pasteurianul român a reuşit să cultive în premieră, tion against this terrible disease. in vitro, virusul polio (1913) deschizând, principial, At a time when syphilis treatment with arsenic- drumul vaccinării împotriva acestei teribile maladii. based drugs (Salvarsan, Neosalvarsan) was outdated, Într-o epocă în care terapia sifilisului cu preparate Levaditi, R. Sazerac and Şt. S. Nicolau extrapolated arsenicale (Salvarsan, Neosalvarsan) era depăşită, the bismuth-salt therapy for syphilis patients (starting Levaditi, R. Sazerac şi Şt. S. Nicolau, pornind de la stu- from chemotherapy studies on chicken spirillosis) and dii de chimioterapie a spirilozei găinilor au extrapolat obtained remarkable results (1925-1931). tratamentul cu săruri de bismut la bolnavii de sifilis, After discovering with yet another Romanian Pas- obţinând rezultate excelente (1925–1931). teurian, Ştefan S. Nicolau, two of the general charac- Descoperind împreună cu un alt pasteurian ro- teristics of viruses (ultrafiltrability and predominantly mân, Ştefan S. Nicolau, două dintre caracterele gene- cell-mediated immunity in viral infections), Levaditi rale ale virusurilor (ultrafiltrabilitatea şi imunitatea also described the special tropism of certain viruses for predominat celulară în viroze) Levaditi a semnalat şi cancerised tissues (viral oncolysis) which, developed tropismul special al unor virusuri pentru ţesuturile by modern research, stands out as a major promise for cancerizate (oncoliza virală), proprietate care rafinată cancer treatment in the next 10 to 15 years. de cercetările moderne, reprezintă o promisiune pen- Levaditi's relationship with veterinary physicians, tru tratamentul cancerului în următorii 10–15 ani. which we are going to highlight, first of all consists of a Relaţiile lui C. Levaditi cu medici veterinari, rele- number of letters setting forth research ideas and ex- vate de noi, constau în primul rând dintr-o serie de perimental protocols. One of the main addresses was scrisori, nepublicate până în prezent, în care sunt ex- veterinary professor Camille Guérin (1872-1961), co- primate idei de lucru şi chiar protocoale experimen- author with A. Calmette of the first vaccine for tuber- tale. Unul din principalii adresanţi este profesorul ve- culosis (BCG). Levaditi in his turn received letters from terinar Camille Guérin (1872-1961) coautor alături de Gaston Ramon, while Bucharest professor Paul Rie- A. Calmette al primului vaccin împotriva tuberculozei gler, the founder of Romanian Veterinary Microbiology, (BCG). Scrisori primite de Levaditi poartă semnătura is the one who used to send Levaditi “The Veterinary lui Gaston Ramon, iar “Arhiva veterinară”, revistă ce Archives”, a magazine published in Bucharest. apărea la Bucureşti, îi este cerută de către Levaditi profesorului bucureştean Paul Riegler, fondatorul mi- 1) Academy of Medical Sciences, Bucharest, Romania crobiologiei veterinare româneşti. E-mail: [email protected] O pleiadă de tineri medici veterinari din mai multe 68 Rev Rom Med Vet (2016) 26 | 1

Levaditi's pupils and associates at the Pasteur In- ţări ale Europei au fost elevi şi colaboratori ai lui Leva- stitute included a number of young vets from several diti în Institutul Pasteur din Paris: englezul Ian Gallo- European countries. Among them we should mention way, francezii J. Verge şi P. Goret, românii Alexandru Ian Galloway (UK), J. Verge et P. Goret (France), Al. Ciucă, Nicolae Stamatin, Ioan Borcilă ş.a. Ciucă, N. Stamatin, I. Borcilă (Romania). Interesul lui C. Levaditi pentru medicina veteri- Levaditi's interest in veterinary medicine showed nară apare pregnant mai ales atunci când a coordonat out when he coordinated and wrote articles devoted to şi elaborat texte de patologie specifică în monumenta- specific pathology for his monumental treaty "Les ul- lul său tratat „Les ultravirus des maladies animales”, travirus des maladies animales", published in Paris in apărut la Paris în 1943, lucrare care, cel puţin 30 de 1943. For at least 30 years, this was the reference ani, a fost cartea de căpătâi a medicilor veterinari in- book for infectionist veterinarians on all continents. fecţionişti de pe toate continentele. Keywords: Constantin Levaditi, virology, Cuvinte cheie: Constantin Levaditi, virusologie, anti-infectious chemotherapy, polio virus chimioterapie antiinfecţioasă, virusul polio

The links between the Pasteurian school and ve- Gaston Ramon (1866-1963), who discovered ana- terinary medicine were intense and steadfast. toxine therapy and in the 1940's became the general The current, unanimously-accepted saying "there director of the Pasteur Institute, is another example of is only one medicine" has old roots in Pasteur's thin- a vet promoted to the top ranks of the scientific world of king. In his famous address to the Medical Academy in the time. (Fig. 1) Paris (1878) titled "La théorie des germes et ses appli- Constantin Levaditi (1874-1953), a graduate of cations à la médecine et à la chirurgie" (Germ Theory the Bucharest Faculty of Medicine (1898) and an MD and Its Applications in Medicine and Surgery), the with a degree obtained in Paris (1902), is considered most convincing arguments, which blew away the as one of the main founders of virology and chemothe- naive theory according to which infectious diseases rapy (Fig. 2). Initially an assistant of Victor Babeş in are the nefarious work of miasmas, originated for the Bucharest (1898-1900), then of Paul Ehrlich in Frank- most part in animal pathology or zoonoses. furt /Main (1900-1902), Levaditi settled down in Paris The examples he used referred to the diseases of where he worked first at the Collège de France with silkworms, to avian cholera, rabies, and anthrax. One physiopathologist Albert Charrin, and then with I.I. single example, based on the isolation of a particular Metchnikov and Emile Roux (Pasteur's right hand) in microbe - a streptococcus - referred to a specifically the world-famous Institute. human disease: puerperal fever. Levaditi was an illustrious figure of the Pasteur In- If Pasteur was able to bring forward arguments stitute for nearly 40 years. His major breakthroughs documenting the microbes' "guilt", it was mainly be- include the discovery of infantile paralysis virus (1909) cause he had read articles written by researchers of with Karl Landsteiner (15) (who in his turn distin- animal pathology and because he used to work toge- guished the four blood groups and the Rh factor in ther with them. Among the works he had read featured man) and of the the way poliomyelitis is transmitted articles published between 1877 and 1883 by veteri- (10), the first laboratory diagnosis of poliomyelitis nary physicians Henri Toussaint of Toulouse (on avian (with Al. Netter), based on the seroneutralised anti- cholera), Pierre Victor Galtier of Lyon (on rabies), Hen- body titre in bovine blood and healthy virus carriers ri Boulle of Paris (on microbian infections), and others. blood, as well as the study of neuro-viroses such as Understanding the importance of veterinary pa- (epidemic) von Economo encephalitis, neurotropic thology for medicine generally, Pasteur and later on herpes, viral encephalitis, etc. (19) Emile Duclaux and Emile Roux, who succeeded him at At a time when viruses and especially the dyna- the head of the Pasteur Institute after 1889, backed mics of viral multiplication were very little known the research conducted by veterinary physicians such (1909-1915), Levaditi was the first in the world to cul- as Edmond Etienne Nocard (1850-1903), unanimous- tivate viruses in tissue culture by using the novel me- ly known for his studies on bovine and zoonosical tu- thod of the "suspended drop" invented by American berculosis and for having laid the basis, with E. Roux, physician Harrison and improved by French Nobel for industrially producing therapeutical serum on hor- Prize winner (1912) Alexis Carrel. What he did was to ses, at Garches (antidiphteric, antitetanic, antistrep- cultivate the polio virus on ape non-neuronal spinal tococcic, a.o.). ganglions (16). His work in this field paved the way for Rev Rom Med Vet (2016) 26 | 1 69

the preparation, after 1950, of vaccines for infantile paralysis (the Albert Sabin, Jonas Salk, a.o. formulae). Also in virology, Levaditi and his Romanian pupil Ştefan S. Nicolau (1896-1967) defined two of the general characteristics of viruses: their ultrafiltability through calodium membranes and the fact that anti- viral immunity is predominantly cell-mediated, while the humoral one is derivative and less frequent than the one in bacterial diseases. Finally, Levaditi and Nicolau in 1922 found that many virus types have a special electivity for young tissues, for fast multiplying cells (embryonary and cancerised tissue). This discovery led to the concept of viral oncolysis. Obviously, the hopes pinned on anti-cancer therapy based on oncolytical viruses did not materialise in the initial form thought out by Levaditi. But intense research is fectious agents and thus discovered Herpetomonas currently under way on the use of certain viruses bombycis (12). (cowpox, for instance) as vehicles carrying lithic fac- Going back to this Romanian Pasteurian's work as tors to be directed precisely towards the tumoral tis- a chemotherapist, we should recall that after 1915 the sue where they should generate the apoptosis of the civilised world lived a great medical disillusion. After neoplastic cells. the burst of enthusiasm sparked in the early 20th cen- Levaditi was the coordinator and author of major tury by the introduction of arsenic-based Salvarsan parts of the monumental treatises (counting more (1907) and Neosalvarsan (1912), general despair gra- than 3,000 pages each) "Les ultravirus des maladies dually set in: as time went by, the treponemes would humaines" (The Ultra-Viruses of Human Diseases, select arsenic-resistant strains. This is when the bis- Paris, 1938 and 1944) and "Les ultravirus des mala- muth-based antiluetic drugs invented by Levaditi and dies animales" (The Ultra-Viruses of Animal Diseases, Sazerac came in. And so syphilis could be checked, Paris-Montpellier, 1943). On the volume dealing with until the introduction in 1944 of the penicillin-based animal viral diseases, Levaditi coordinated a team of therapy (Mc Mahoney). experts in veterinary infectious-contagious pathology Undoubtedly, a major part of Levaditi's scientific including Jean Verge, Pierre Goret and others. work served human medicine. But at a closer look we Levaditi's work as a chemotherapist is no less im- see the scientist had not only an inerest in animal pa- portant. His master in Frankfurt/M., Paul Ehrlich, was thology but also close ties with veterinary physicians. searching for a way to cure syphilis. His team made up Striving to learn more about these ties, I spent of German and Japanese researchers (Shiga, Hata) in several months in various Paris libraries and above all in 1907 managaed to synthesise the first "magic bullet" the Pasteur Institute's which killed treponemes (preparation 606 or Salvar- archives, which shelter san). In 1912, Ehrlich's team produced Neosalvarsan. an important collection Very few people know however that the year when Sal- of unpublished docu- varsan was introduced in clinics, Levaditi and his Bri- ments drawn up by C. tish pupil K. McIntosh were focusing on veterinary pa- Levaditi. He had a re- thology, studying the parasiticide action of Atoxil on markable exchange of avian spirochetosis (13). Their aim was obviously to views with Jean-Marie extrapolate it to the treatment of human syphilis tre- Camille Guérin (1872- ponema. 1961) (Fig. 3). As we Sometimes between 1865 and 1868, Pasteur was know, Guérin's name is gathering arguments in favour of the microbian theory linked to the creation including from the field of silkworm diseases. Follo- between 1905-1908 of wing the advice from his veterinarian associates, Le- the BCG strain of atte- vaditi did not hesitate (starting in 1905) to conduct nuated bovine tubercu- himself research on the parasitism of Bombyx mori in- losis bacills (3), initially 70 Rev Rom Med Vet (2016) 26 | 1

used in the vaccination of cattle (1907-1908) and sub- As results from the letters Levaditi sent to Camille sequently of newborns (first human vaccinations, B. Guérin (Fig. 4), it was the latter who constantly de- Weill-Halle, 1922). Working both at the Pasteur Insti- livered the vaccine pulp that was used on the sub- tute and at the National Veterinary School of Alfort, sequent passages on rabbit and mouse brain. It was a Guérin was an authority in microbiology and immuno- material specially prepared by Guérin under strict ste- logy. This is proved, among others, by the letters he rility conditions. Levaditi sent Guérin the detailed pro- received from Jules Bordet (Nobel Prize winner in tocol of his experiments and, more than once, asked 1919), Albert Calmette, Charles Nicole (director of the for advice about it. One can clearly see from these Pasteur Institute in Tunis, Nobel Prize winner in 1928), letters that Levaditi considered Guérin as a magister Edmond Sergent, director of the Pasteur Institute in dixit in matters of bacteriology and immunology. This Algiers, and, of course from Levaditi (20). The scien- is proved by a letter (Fig. 5) addressed to Guérin on tific collaboration Guérin - Levaditi spanned 3 years May 7, 1923, in which he asked for his opinion on cer- (1922-1925). The main object was the preparation of tain experiments, adding at the end: "Ecrivez-moi un neurovaccine by Levaditi and Nicolau. mot à ce sujet" (Send me a word on this topic) (21).

What was neurovaccine ? An attempt by the two Romanian Pasteurians to improve smallpox vaccine for children. Indeed, although cowpox pustulas had been used by prophylactic medicine for more than a century (Edward Jenner had introduced cowpox vaccination as early as 1797), the gross vaccine pulp obtained by scarifying the veals' skin was usually infected with ubiquitous, often pathogenic germs (Staphylococcus aureus, pathogenic fungi, etc).The sterilisation method of vaccinal (and vaccinating) material in the first two decades of the 20th century was complicated, as the antibiotic treatment of suspensions didn’t exist at the time. Besides, titration of the vaccine virus was not yet possible and thus the doses administered to children varied, often generating undesired local reactions. Levaditi and Nicolau thought that, by adapting the vaccine virus to rabbits' brains they would create a neurovaccine free of pathogenic bacteria, hence In the archives of the Pasteur Institute we can "pure" from a bacteriological point of view and whose also find a letter from C. Levaditi addressed to Paul titration on a white mouse brain was feasible (18). Riegler (Fig. 6), a professor of bacteriology at the Ve- Rev Rom Med Vet (2016) 26 | 1 71

terinary Faculty in Bucharest.Levaditi informed Riegler level. Obviously, at the beginning of the 1940, mole- in it that he was preparing a paper on anaphilaxis and cular genetics was barely at its beginnings. The only asked him to send him several copies of “The Veteri- phenomenon observed by the time in this field, though nary Archives” dating from 1906, 1907 and 1908 (the completely unexplained, was the discovery in 1928 by magazine was published by the Higher School of Vete- British bacteriologist Frederick Griffith (1878-1941) of rinary Medicine in Bucharest). After Riegler's death, a "mysterious factor". This "factor" turned type-II non the letter became the property of Nicolae Stamatin, a -pathogenic pneumococci into type-III, very aggres- disciple of Riegler's and subsequently a professor of sive ones (6). It was only 16 years later, in 1944, that microbiology at the Veterinary Faculty in Bucharest. the American Oswald Th. Avery and his assciates Colin Professor Stamatin thought it his duty to send the M. McLead and Maclyin McCarty demonstrated that letter back to the Levaditi’s son (28). Griffith's enigmatic factor was a nucleoprotein (1). A letter (dated October 11, 1951), received by C. A year before, in the summer of 1943, "Les ultra- Levaditi from Gaston Ramon, the inventor of ana- virus des maladies animales" had been published in toxines and the general director of the Pasteur Insti- Paris, actually anticipating the experimental conclu- tute, announced him that professor E. Leclainche sions of the American researchers. (1861-1953) from the National Veterinary School of Alfort, one of the founders of the OIE (International Office of Epizootics) and author of the erudite and ex- haustive "Histoire de la médecine vétérinaire" (1936) had proposed C. Levaditi to become a member of the French Academy of Sciences in Paris (27) (Fig. 7). Professor Pierre Hyacinthe Goret (1907-1944) of Lyon, just as professor Louis Armand Jean Verge (1892-1964) of Alfort, one of the most valuable ana- tomo-pathologists of interwar French veterinary medicine, intensely worked together with Levaditi, both on animal research protocols and on the"Les ultravirus des maladies animales" (Paris-Montpellier, 1943). Among the veterinary physicians who worked in Levaditi's Parisian laboratory featured British professor Ian A. Galloway, who was a prominent figure of the Institute of Animal Health in Pirbright (Surrey) after 1950. The young Briton worked with Levaditi and Nicolau in the 1930s and 1940s, subsequently beco- ming an expert in cultivating animal viruses (aphthous fever, swine pest, etc) in vitro. Levaditi's appreciation for veterinary medicine also shows in the drawing up and coordination of the vast treatise devoted to animal viral diseases "Les ultravirus des maladies animales" (1943). In achieving this masterpiece Levaditi appealed to the knowledge of well-known veterinarian virologists, actually materialising the concept "human medicine and animal medicine, just one medicine". Besides the fact that in 1943, when it was published, the treatise made pubic throughout the world (still engulfed in war) the most recent breakthroughs of virology in general and of the medical-veterinary one in particular, we should stress that the work con- tained essential thoeries concerning the relationship That was a time when the overwhelming majority between the virus and the victim cells at a molecular of bacteriologists and virologists believed that viruses 72 Rev Rom Med Vet (2016) 26 | 1

multiply the same way as bacteria do, through scissi- 7. Iftimovici R. (1979), C. Levaditi, A forerunner of parity. Levaditi and his associates, both human and modern research and interpretation of virus multi- veterinary physicians, proved to have another opinion, plication, Rev.Roumaine de Virologie, 30,237-242 so much the more so as they had noticed that viruses 8. Iftimovici R. (2015), Louis Pasteur şi medicina ve- do not have a cell structure. This treatise of veterinary terinară, Rev Rom Med Vet, vol. 25, nr. 1 medicine thus sets forth the following hypothesis con- 9. Iftimovici R. (2015), Istoria universală a medicinii cerning viral multiplication: "The infected receptive şi farmaciei, Ed. Academiei Române, vol.II cell produces a virus-nucleoprotein identical to that of 10. Kling C., Levaditi C. (1913), Etudes sur la Polio- the agent that caused the disease". Further on, de- myélite aiguë épidémique, Monographies de fining with extreme accuracy the mechanism of viral l'Institut Pasteur, Paris multiplication, Levaditi, Lepine and Verge for the first 11. Lepine P. (1953), Constantin Levaditi, La Presse time stated that: "Thus, the ultra-virus appears to be a médicale, 71, Nov 7, p. 1455 producer of specific nucleoproteins, whose synthesis it 12. Levaditi C. (1905), Sur un nouveau flagelle para- generates, on the basis of the cell's normal nucleopro- site du Bombyx mori (Herpetomonas bombycis), teins. The ultra-virus thus intervenes in cell meta- C.R. de l'Académie des Sciences, Paris, octobre bolism, diverting its rhythm" (23). 13. Levaditi C., McIntosh K., (1907) L'influence de We no longer need to stress that this viral multi- l'atoxyl sur la spirillose provoquée par le Spirillum plication mechanism was subsequently proven to be gallinam, C.R. de la Société de Biologie, Paris, juin accurate by both Oswald Th. Avery's team in 1944 and 14. Levaditi C., Landsteiner K. (1909), La transmission by another team of US researchers made up of Alfred de la paralysie infantile au chimpanzé, C.R. de Hershey and Marta Chase (1952), who demonstrated l'Académie des Sciences Paris,149, pp.1014-1016 that in the case of the bacteriophage virus of the 15. Levaditi C., Landsteiner K. (1910), Sur la paralysie colibacillus, the virus does not need to completely pe- infantile expérimentale, C.R. de l'Académie des netrate the body of the bacteria but can merely inject Sciences, Paris, 150, pp.55-57 its specific nucleoproteins carrying a viral reproductive 16. Levaditi C. (1913), Symbiose entre le virus de la message into the victim's cytoplasma (23). poliomyélite et les cellules des ganglions spinaux à With this demonstration Alfred Hershey earned l'état de vie prolongé in vitro, C.R. de la Société de the Nobel Prize (1944). It was exactly what Levaditi, Biologie, Paris, mai Lepine and Verge had discovered as early as 1943. 17. Levaditi C. (1921), Unpublished letter to C.Guérin, Our conclusion can only be that, by assuming the Nov.12, Levaditi Archives, Pasteur Institute, Paris "Pasteurian spirit", C. Levaditi showed his appreciaton 18. Levaditi C., Nicolau S. (1922), Vaccine pure céré- for his veterinary colleagues with whom he worked brale, C.R. de l'Académie des Sciences, Paris and exchanged opinions still valid in today's medicine. 19. Levaditi C. (1922), Ectodermoses neurotropes, Masson, Paris REFERENCES 20. Levaditi C. (1922), Unpublished letter to C.Guérin, March 9, Levaditi Archives, Pasteur Institute, Paris 1. Avery T.O., McLead C.M., McCarty M. (1944), Stu- 21. Levaditi C. (1923), Unpublished letter to C.Guérin, dies on the chemical nature of the substance indu- May 7, Levaditi Archives, Pasteur Institute, Paris cing transformation of penumococcal types, The 22. Levaditi C., Lépine P., Verge P. (1943), Les ultra- Journal of Experimental Medicine, 62(2), 137-158 virus des maladies animales, Montpellier, p.78 2. Athanasiu P. (1953), Le professeur Levaditi, Presse 23. Navarro-Martin A. (1954), Constantino Levaditi, Médicale, octobre, document 26990, Levaditi Ar- Atlas Dermo-syphiligraficas,Buenos Aires, 2, 46, chives of the Pasteur Institute, Paris doc. 26991, Levaditi Arch., Pasteur Institute, Paris 3. Calmette A., Guérin C. (1908), Nouvelles contribu- 24. Nicol L. (1974), L'épopée pastorienne et la mé- tions à l'étude de la vaccination des bovidés contre decine vétérinaire, chez l'auteur, 92380, Garches la tuberculose, Annales de l'Institut Pasteur, Paris 25. Nicolau S. St., Iftimovici R. (1967), Constantin 4. Delaunay A. (1962), L'Institut Pasteur des origines Levaditi, Ed. Ştiinţifică, Bucharest à aujourd'hui, Paris, pp.179-180 26. Ramon G. (1951), Letter to C. Levaditi, Oct. 11, 5. Girond M. (1953), C. Levaditi (obituary), Bull. de Levaditi Arch., Pasteur Institute, Paris, no. 37509 l'Acad.Nationale de Médecine, Paris, 34, 35, p. 556 27. Stamatin N. (1968), Letter to Jean Levaditi (C. 6. Griffith F.(1928), J.Hygiene, Cambridge,27, p.113 Levaditi's son), Bucharest, June 7