Herald R. Cox

Propagation of Canine Distemper Virus on the Chorio-Allantolc Membrane of Embryonated Hen Eggs

Summary Studies are reported in which the Lederle stock strain of canine distemper virus was adapted and maintained through 74 serial passages on the chorio-allantoic membranes of developing chick embryos. The chick embryo adapted strain retained its capacity to produce lethal infection in ferrets through the 24th egg passage level but apparently lost its ability to do so between the 24th and 28th egg passages. Ferrets inoculated with chick embryo adapted virus of passage levels higher than the 28th transfer failed to show any signs of disease, but were found immune on rechallenge with fully virulent ferret spleen virus. Cross neutralization tests carried out in chick embryos and in ferrets indicate that the chick embryo adapted strain and the parent strain of ferret spleen virus are immunologically related.

Human laboratory infection with Venezuelan equine encephalomyelitis virus; report of four cases.

THE occurrence of human cases of encephalomyelitis due to laboratory infection with Venezuelan equine encephalomyelitis virus was reported for the first time by Casals et al.1 and by Lennette and K...

Adaptation of Colorado Tick Fever Virus to Mouse and Developing Chick Embryo.

Colorado tick fever is a human infection, probably of tick-borne viral etiology, which runs a mild course and is characterized by intermittent nonexanthematic fever. The disease has been recognized as a separate clinical entity by Becker under the name of Colorado tick fever 1 and by Toomey under the name of American Mountain Fever. 2 Topping. Cully-ford and Davis 3 conducting a clinical and epidemiological survey of Colorado tick fever were unsuccessful in their attempts to establish the causative agent of the disease in laboratory animals or developing chick embryo. Recently, however, Florio and his colleagues 4 reported transmission of Colorado tick fever virus to the Syrian hamster and found that the virus passes gradocol collodion membrane filters with calculated pore diameters of 24 milli-microns. 5 The present observation deals with adaptation of Colorado tick fever virus to the dba (dilute brown agouti) mouse, the white mouse and developing chick embryo. Experimental. Infected hamster serum representing the 30th hamster passage of the virus was obtained through the courtesy of Dr. Lloyd Florio, of the School of Medicine and Hospitals, University of Colorado. The serum was diluted 5 times in saline and inoculated, respectively into: 5 hamsters by intraabdominal route, 6 dba mice by intra-nasal route and 12 dba mice by intracerebral route. The hamsters weighed 90-120 g and the mice 10-12 g. Two hamsters were found dead and 2 sick on the fourth day after inoculation. A low WBC count, characteristic for Colorado tick fever 4 was observed on blood smears obtained from the 2 sick animals. None of the intranasally-inoculated mice showed any signs of illness. Neverthelessone animal was sacrificed on the fifth day after inoculation and a bacteriologically sterile suq)ension of its lungs was instilled intranasally into a second group of 6 dba mice.

Persistence of Antibody after Oral Trivalent Poliovirus Vaccine (Sabin Strains).

BECAUSE of the favorable rates of serum-antibody conversion obtained in a pilot study in Elkins, West Virginia, with two feedings of a trivalent oral poliovirus vaccine,1 the same vaccine was used ...

Cumulative Testing Experience with Consecutive Lots of Oral Poliomyelitis Vaccine

Anderson, J. R., Goudie, R. B., and Gray, K. G. (1959a). Scot. med. J., 4, 64. -(1959b). Lancet, 1, 644. Balfour, B. M., Doniach, D., and Roitt, I. M. (1959). In preparation. Beierwaltes, W. H., Dodson, V. N., and Wheeler, A. H. (1959). J. clin. Endocr., 19, 179. Belyavin, G., and Trotter, W. R. (1959). Lancet, 1, 648. Blizzard, R. M., Hamwi, G. J., Skillman, T. G., and Wheeler, W. E. (1959). New Engl. J. Med., 260, 112. Briinger, H. (1914). Mitt. Grenzgeb. Med. Chir., 28, 213. Buchanan, W. W., Anderson, J. R., Goudie, R. B., and Gray, K. G. (1958). Lancet, 2, 928. Cline, M. J., Selenkow, H. A., and Brooke, M. S. (1959). New Engl. J. Med., 260, 117. Crile, G., jun. (1948). Ann. Surg., 127, 640 and Rumsey, E. W. (1950). J. Amer. med. Ass., 142, 458. Donia:h, D., and Hudson, R. V. (1957). Brit. med. J., 1, 672. (1(959). Proc. roy. Soc. Med., 52, 178. and Roitt, I. M. (1957). J. clin. Endocr., 17, 1293. and Hudson, R. V. (1959). Rev. Maladies du Foie. In press. Druez, G., Hainaut, J., Parmentier, R., and Musin, L. (1958). Acta clin. belg., 13, 280. Eason, J. (1928). Edinb. med. J., 35, Trans. med.-chir. Soc. Edinb., p. 169. Eylan, E., Zmucky, R., and Sheba, C. (1957). Lancet, 1, 1062. Felix-Davies, D. (1958). Ibid., 1, 880. Goudie, R. B., Anderson, J. R., and Gray, K. G. (1959). J. Path. Bact., 77, 389. Clark, D. H., Murray, I. P. C., and McNicol, G. P. (1957). Lancet, 2, 976. Greene, R. (1953). Memoirs Soc. Endocr., 1, 16. Hudson, R. V. (1959). Irish J. med. Sci., p. 149. Joplin, G. F., and Fraser, R. (1959). Proc. roy. Soc. Med., 52, 177. Lee, J. M., and Schneider, H. A. (1957). Proc. 6th int. Congr. Neurol. Brussels, p. 22. McConahey, W. M., and Keating, F. R. (1951). J. clin. Endocr., 11, 1116. Mackay, I. R., and Larkin, Lois (1958). Aust. Annz. Med., 7, 251. Mahaux, J. (1959). Rapports de la Se reunion des endocrinologistes de langue frangaise, Paris. -and Pirart, J. (1959). Acta clin. belg., 14, 59. Morgans, M. E., and Trotter, W. R. (1957). Lancet, 1, 553. (1958). Ibid., 1, 607. Owen, C. A. (1958). J. clin. Endocr., 18, 1015. Owen, S G., and Smart, G. A. (1958). Lancet, 2, 1034. Pasteur, W. (1900). Trans. clin. Soc. Lond., 33, 189. Pulvertaft, R. J. V., Doniach, D., Roitt, I. M., and Hudson, R. V. (1959). Lancet, 2, 214. Robertson, J. D., and Reid, D. D. (1952). Ibid., 1, 940. Roitt, I. M., Campbell, P. N., and Doniach, D. (1958). Biochem. J., 69, 248. and Doniach, D. (1958). Lancet, 2, 1027. -(1959). In Henry Ford Symposium on Mechanisms of Hypersensitivity, p. 325., Little, Brown, Boston. Campbell, P. N., and Hudson, R. V. (1956). Lancet, 2, 820. Trotter, W. R., Belyavin, G., and Waddams, A. (1957). Proc. roy. Soc. Med., 50, 961. Volpe, R., and Johnston, McW. (1957). Canad. med. Ass. J., 77, 297. Waksman, B. (1959). Int. Arch. Allergy, Suppl. to vol. 14. Werner, S. C., and Spooner, M. (1955). Bull. N.Y. Acad. Med., 31, 137. White, R. G. (1957). Proc. roy. Soc. Med., 50, 953. Whitesell, F. B., and Black, B. M. (1949). J. clin. Endocr., 9, 1202. Witebsky, E. (1959). In Seelisberg Symposium on Immunopathology, p. 182. Benno Schwabe, Basel.

Failure of Phenosulfazole (Darvisul) in Treating Experimental Viral Infections

Conclusion Phenosulfazole (Darvisul) showed no beneficial effect in the treatment of mice infected with the Columbia SK virus and other neurotropic viruses, including a rodent adapted strain of poliomyelitis (MEF-1 strain). The lack of therapeutic effectiveness of the drug was also observed in monkeys infected intranasally with the Brunhilde strain of poliomyelitis virus. In other viral and rickettsia1 infections of mice and of developing chick embryos treatment with pheno-sulfazole was of no therapeutic value, with the possible exception of psittacosis in which a slight inhibition of growth in the developing chick embryos was observed. However, even in the latter case the effectiveness of phenosulfazole therapy was of much lower magnitude than that observed with other sulfonamides,4-6 or antibiotics.7

Ultrafiltration of the Virus of Equine Encephalomyelitis

Summary The virus of equine encephalomyelitis, both the Eastern and the Western strains, was found to pass collodion membranes with an average pore diameter of 66 mμ, but was completely held back by those of 60 mμ. The results indicate that this virus has the same filtration end-point, and consequently the same particle size, as that of St. Louis encephalitis, which was shown by Bauer, Fite, and Webster, 8 and Elford and Perdrau 9 to be 20 to 30mμ.

Absorption through the Nasal Mucosa of Tannic-Acid Treated Mice

Reports have been given of the absorption of Prussian blue through the nasal mucosa of rabbits 1 and mice. 2 , 3 A study by Olitsky and Cox 2 of the absorption of the pigment 1 to 3 hours after its instillation in mice previously treated intranasally with tannic-acid shows that absorption at this time is less evident than in untreated mice. Subsequently Rake 3 demonstrated that absorption through the nasal mucosa, chiefly the olfactory, is very rapid and has reached its maximum in mice killed at 2 minutes; furthermore, that much of the absorption occurs by way of the olfactory sensory cells. It seemed advisable to study the absorption of Prussian blue from the nasal cavities of tannic-acid treated mice during these early periods. Nine Swiss mice each received eight preliminary intranasal instillations over 3 days. Each dose, consisting of 0.03 cc. of 0.8 percent tannic-acid in 1% glycerine, was dropped on the outside of the nostrils from whence it was gradually breathed into the nose. Four hours after the last dose each mouse received 0.02 cc. of a mixture of equal parts of 10% iron ammonium citrate and 10% potassium ferrocyanide intranasally by the same technique. Two minutes after commencement of intranasal inoculation 5 mice were sacrificed by decapitation and 13 minutes later the other 4 were so sacrificed. Fixation, sectioning, and staining with Mayers carmine were carried out as indicated elsewhere. 3 A few sections were stained with hematoxylin and eosin to facilitate study of the inflammation. Following intranasal treatment with tannic-acid as given above, it is very rarely that any mouse shows exudate from the nostrils. Nevertheless, it was found in the present study that there is to be observed, microscopically, marked exudate into the nasal cavity, especially over the olfactory mucosa and around the turbinates. This exudate is composed of mucus, leucocytes—chiefly polymorphonuclears—and some red cells. The olfactory mucosa and submucosa show swelling and infiltration with oedema fluid and leucocytes. The capillaries of the submucosa are dilated and contain many polymorphonuclear leucocytes. Changes in the respiratory mucosa are slight. Six mice show intense inflammation. In 3 the inflammation is less. Preliminary treatment with 1% tannic acid in normal saline, instead of 0.8% in 1% glycerine, is found to produce similar inflammatory changes.

Effect of Tannic Acid on Intranasal Infection with Pneumococci

It has been shown 1 , 2 that certain viruses are infective for mice when instilled into the nares and that this capacity to invade the body of the mouse through the mucosa of the nose can be greatly diminished by preliminary intranasal treatments with tannic acid or alum-solutions. It has also been shown 3 that certain strains of pneumococci are infective for mice by the nasal route and that, under the conditions of the experiment, invasion takes place both through the nasal mucosa and through the pulmonary alveolar walls. 4 The present experiments were planned to test whether this nasal infectivity of pneumococcus strains is modified by preliminary tannic-acid treatments. For this purpose, 5 experiments were undertaken using groups of young (3 to 6 weeks old) Swiss mice. In each experiment 15 mice were treated with weak solutions of tannic acid for 3 or 4 days before the infecting inoculation was made. In 2 cases, 0.7 or 1.0% solution of tannic acid in normal saline was used and in the other 3, 0.8% solutions of tannic acid in 1.0% glycerol. In the first test, 7 preliminary treatments, each of 0.03 cc. of the tannic acid, were given intranasally; in the second, 9, in the third, 6, and in the last 2, 8 similar treatments. Fifteen mice remained untreated as controls in each test. Cultures of strains of pneumococci of both Type III and Type XIX, known to be intranasally virulent, were used. In the past the use of intranasally virulent strains of pneumococci has been attended with the difficulty that this virulence or invasiveness, which is independent of the intraäbdominal virulence is apt to be lost suddenly and without apparent reason during subculture or on animal passage 3 and cannot be regained by any of the means so far adopted.

Serological Distinctions Between the Viruses of Encephalitis in St. Louis, 1933, Equine Encephalomyelitis, and Vesicular Stomatitis.

The problem of the immunological relationship of the viruses of the encephalitis prevailing in St. Louis and Kansas City in 1933 and the viruses of equine encephalomyelitis and vesicular stomatitis has been studied by a series of cross-protection tests with the viruses associated with these diseases and their respective immune sera. A mouse strain of encephalitis virus obtained from Webster and Fite, 1 a neurotropic guinea pig strain of equine encephalomyelitis virus of the western type, 2 as well as similar mouse strains of the New Jersey and Indiana types 3 of vesicular stomatitis virus were employed, together with antisera from a monkey convalescent from experimental encephalitis, antisera from a rabbit and a horse convalescent from encephalomyelitis of the eastern type, 4 serum from a rabbit immunized against the western type of encephalomyelitis virus, and similar sera from rabbits immunized against the 2 strains of vesicular stomatitis virus. Hormone broth suspensions of the stomatitis and encephalitis materials were mixed with equal volumes of antisera so as to make final dilutions of 10-3, 10-4, 10-5, and 10−6. In the case of encephalomyelitis material, final dilutions of only 10-3 and 10-4 were used. Normal rabbit serum was substituted for antisera in similar series as controls. The mixtures were incubated at 37.5°C. for 2 hours, placed in the icechest overnight, and then examined for the presence of virus by intracerebral injection into animals.† For the stomatitis and the encephalitis virus mice were employed, and for the encephalomyelitis virus guinea pigs. The results as tabulated showed that a particular virus was neutralized only by its homologous antiserum. Hence no cross-immunity reactions occur between any of the viruses studied. Webster and Fite 1 have stated that the results of animal inoculation tests indicate that encephalitis virus is not related to equine encephalomyelitis or vesicular stomatitis viruses. The results of the serological tests here reported are in accord with their observations.