Chorioallantoic Membrane Of Embryonated Eggs Research Articles

STUDIES OF VACCINIA HEMAGGLUTININ OBTAINED FROM VARIOUS VACCINIA INFECTED TISSUES. DENSITY GRADIENT CENTRIFUGATION AND ELECTRON MICROSCOPY.

SummarySerologically specific vaccinial hemagglutinin (VHA) was obtained from vaccinia infected HeLa cells, chorioallantoic membranes of embryonated eggs and mouse fibroblast cell cultures. When concentrated and partially purified preparations were examined by density gradient centrifugation, the preparations varied in density according to the tissue of origin. The host-associated differences in peak density were accompanied by a marked degree of poly-dispersion not resolvable by a second density gradient centrifugation or by prolonged centrifugation. Electron micrographs showed that the particle size was in the range of 45–54 mμ. Substructure was detected in some particles which suggested the coiling or folding of a strand.

Separation and characterization of hemagglutinins from chorioallantoic membranes of embryonated eggs infected with vaccinia virus

A process is described which separates two hemagglutinins from vaccinia-infected chorioallantoic membrane suspensions by centrifugation in a suspending medium of density 1.063. The properties of these hemagglutinins have been studied and the results indicate that one hemagglutinin is virus specific and the other hemagglutinin is a normal tissue agglutinin also present in uninfected membrane suspensions. Criteria are presented for the differentiation of the two hemagglutinins and for the determination of the virus specificity of one of the hemagglutinins.

Linear and parabolic estimates of the titers of herpes simplex from pock counts on the chorioallantoic membrane of embryonated eggs

Analysis of 33 randomly selected titrations of herpes simplex on chorioallantoic membranes of embryonated hens' eggs revealed that the variance of pock counts among replicate eggs was considerably higher than expected and that the mean pock counts at high concentrations of virus were less than predicted from counts obtained following infection at low concentration of virus. The curve relating mean pock count to virus concentration leveled off in 26 titrations. The deviation was significant in 4 of the titrations. The upper limit of the linear portion of the dose response curve as defined in the text varied from 11 to 1100 mean pocks per membrane. Methods for estimating virus titers based on linear and parabolic approximations of the pock count-dose relationship were described and compared. The parabolic approximation was recommended for studies in which a more precise estimate of virus titer was desired.

孵化鶏卵に馴化せしめたHSTウイルスの免疫学的並びに形態学的研究に就て

HST virus is one of filtrable agents successfully isolated from Yoshida tumor in 1950 by Hamazaki passing through the different species of animals, and it has already been verified that immunoserologically it is specific to this tumor and that it is also an indispensable factor for the growth and transplantation of the tumor. This time we carried out immunological and morphological studies on this virus adapted to embryonated eggs.The HST virus successively inoculated into the yolk sack and chorioallantoic membrane of embryonated eggs in the microbiological department of our school can well become adaptation to embryonated eggs, and consequently the chick embryos usually die 4-6 days after the HST virus inoculation. In the liver of chick embryos dissociation, fatty degeneration, and necrobiosis of parenchymal cells, and the infiltration of monocytes and lymphoid cells can patho-anatomically be recognized.It has been found that this virus adapted to the embryonated eggs maintains the properties essentially the same as the original HST virus in that when it is inoculated successively back to its host animals, guinea pigs, inflammatory tissue reaction occur successively, though slightly milder than in the case of original virus.Next, in order to determined whether the virus successfully replaced from the embryonated eggs back to guinea pigs to be the same as the original HST virus, the cross-neutralization of two viruses by antisera were performed and it was proven that the tests were positive. Therefore the former was serologically quite identified with the latter, original HST virus.On the other hand, after purifying the agent in the allantoic fluid and examining it under an electron-microscope, round particles of 30-60mμ in size were obtained.

The Immunising Properties of An Egg-Adapted Strain of Fowl Pox Virus

Summary The adaptation of a Kenya strain of fowl pox virus to the chorioallantoic membrane of embryonated eggs is described. When used as a vaccine, consisting of infected chorio-allantoic membranes suspended in glycerine and broth, the egg-adapted virus provoked a solid immunity, lasting at least twelve months against challenge with homologous scab virus. The egg-adapted vaccine remained viable for 6 to 12 weeks at room temperature and for 6 to 12 months at refrigerator temperature.

Human Skin Grafted Upon the Chorioallantois of the Chick Embryo for Virus Cultivation

Summary and Conclusions(1) Human skin may readily be grafted onto the chorioallantoic membrane of embryonated eggs. (2) The prepuce removed by usual surgical circumcision is a satisfactory source of adequate amounts of skin. (3) Penicillin and streptomycin obviate most bacterial contamination. (4) Grafted human skin may be passed serially from egg to egg at weekly intervals and remain viable. (5) Herpes simplex and vaccinia grow readily on grafted human skin, with the formation of characteristic inclusion bodies. (6) A successful inoculation with herpes zoster confirms the previous single successful cultivation of this virus outside of the human body.

CULTIVATION OF THE HOG CHOLERA VIRUS

The work of Hecke on the cultivation, of hog cholera virus was confirmed with ease. Virus was grown in the presence of fresh minced swine testicle in flasks containing Tyrode solution, on the chorioallantoic membrane of embryonated eggs, and on the surface of swine serum agar. In flasks it was grown for 14 transfers; while on eggs it was grown for 13 transfers, followed by an equal number of transfers on agar, making 26 transfers in all. Only one strain of virus was used and we do not know whether all strains can be cultivated so readily or whether we were particularly fortunate in the selection of the strain used. Neither do we know whether swine testicle is better than other tissues for growth. The cultured virus produces characteristic hog cholera when injected into swine, and its effect can be neutralized with commercial anti-hog cholera serum. No evidence of attenuation of the virus was obtained, the last culture being highly virulent when small amounts were injected. No evidence for the adaptation to the egg could be secured, since passages without swine testicle on the membrane or intravenously for 2 transfers resulted in a loss of the virus. No contaminating virus that might favor the cultivation could be detected by animal or egg inoculation. Not only has the virus been cultivated but it has been demonstrated in large amounts in the culture. Four suspensions containing slightly over 0.5 mg. of protein nitrogen produced typical hog cholera when 1 x 10(-6) cc. was injected, and one suspension made in the same way was active in one-tenth this amount. Few titrations on what is commonly known as hog cholera virus, i.e. the serum from acutely ill pigs, are available. We made one such titration and produced a delayed disease with 1 x 10(-5) cc. of infectious serum. It seems probable that the culture virus is more active than the commonly used virus and that its practical use in hog cholera vaccination and hyperimmunization would result in a considerable saving. All of the methods used yielded active cultures, but the serum agar method is the one of choice since larger amounts of suspension can be obtained with less labor.

Development of Vaccinia and Variola Viruses in Embryonated Eggs at 28 C

SummaryVaccinia and variola viruses were established and maintained on transfer in the chorioallantoic membrane of embry-onated eggs incubated at a temperature sufficiently low to prevent embryonic development (28°C). The number of elementary bodies present in the membrane on the 3rd day approximated that in eggs incubated at a temperature which favored growth of the embryo (37°C). At 28°C, however, the effect of the respective viruses on the membrane and the embryo as well as the cellular response of the former to them was significantly retarded.