Chorioallantoic Route Research Articles

In ovo inhibition of avian pox virus replication by mangosteen rind and red ginger ethanolic extracts.

Background and Aim:Avian pox is a contagious disease caused by the avian pox virus (APV). Mangostin and γ-mangostin in mangosteen rind (MR) and gingerol in red ginger (RG) exhibit antiviral activity. In this study, we evaluated the effect of MR and RG ethanolic extracts on APV based on pock lesions on the chorioallantoic membrane (CAM) of specific pathogen-free (SPF) embryonated chicken eggs (ECEs).Materials and Methods:Three APVs from chicken isolates (C1, C2, and C3), one APV from a pigeon isolate (P), 1.5% and 3% MR ethanolic extract, 5% and 10% RG ethanolic extract, and a combination of 1.5% MR and 5% RG at 0.1 mL/egg were inoculated in ovo (7th day incubation, chorioallantoic route) in SPF ECEs. A control group inoculated in ovo with APV alone was also established. Each treatment consisted of three replicates. Parameters including embryo survival, CAM lesions, and average number of pock lesions were determined.Results:In ovo inoculation of MR and RG ethanolic extracts was not harmful to the ECEs and did not induce CAM lesions. The average number of pock lesions in the control group (C1, C2, C3, and P) was 35, 14, 10, and 17, respectively, whereas in all treatment groups, the number was 0, except in the 5% RG group of C1, which had a value of 10.Conclusion:In ovo inoculation of 1.5% and 3% MR, 5% and 10% RG, and the combination of 1.5% MR plus 5% RG ethanolic extracts at 0.1 mL/egg inhibit APV by reducing the number of pock lesions on the CAM of the ECE.

MORPHOLOGICAL AND BIOLOGICAL STUDIES ON A VIRUS IN CULTURED LYMPHOBLASTS FROM BURKITT'S LYMPHOMA.

Lymphoblasts of two tissue culture strains (EB1 and EB2) from different biopsy specimens of Burkitt's lymphoma have been examined in thin sections by electron microscopy, and have each been found to carry a morphologically identical virus. The virus was observed in samples taken over many months, being present in about 1 to 2 per cent of the cells in two forms: Immature particles about 75 mmicro in diameter which were seen in both the nucleus and cytoplasm; and larger mature particles with a diameter of 110 to 115 mmicro, which were either within membrane-bounded cytoplasmic spaces or at the cell surface. There was some indication that the particles matured by budding through the cytoplasmic membranes. Both types of particle occurred in dead degenerating cells or, less frequently, in intact altered cells. The characteristic alterations of the latter included margination of the chromatin, fragmentation of the nuclear envelope, beaded opaque material in the mitochondria, and, with one of the cell strains (EB1), sheaves of altered spindle tubules. All attempts to isolate and identify the virus carried by the two strains of lymphoblasts failed. No pathological effects were caused in 8-day chick embryos inoculated either with whole lymphoblasts or extracts of disrupted lymphoblasts, using the intraallantoic, amniotic, and chorioallantoic routes, and the extraembryonic fluids of such chicks were without haemagglutinating activity for human, chicken, guinea pig, or monkey erythrocytes. Whole lymphoblasts or lymphoblast extracts were likewise without effect when inoculated intraperitoneally into newborn hamsters or two strains of newborn mice. Similar lymphoblast inocula did not cause detectable changes in 9 different test tissue culture systems even after 8 blind passages. The nature of the unknown, unidentified virus in the cultured lymphoblasts from Burkitt's lymphomas is considered and its possible relationship to the cells discussed.

STRUCTURE AND DEVELOPMENT OF VIRUSES AS OBSERVED IN THE ELECTRON MICROSCOPE

Chicken embryos were infected by the chorioallantoic route with influenza virus, PR8 strain, in the form of undiluted chorioallantoic fluid. Electron microscopic examination 24 hours after infection revealed that membrane-bound fragments of cytoplasm appeared to be in process of release from entodermal cells of the chorioallantois. The number of such fragments was greatly increased in proportion to the number of typical viral particles after the third serial passage, which was accompanied by a reduction of the infectivity-hemagglutinin ratio (von Magnus effect). The lack of recognizable internal components, together with the presence of surface structure which closely resembled that of the virus and frequently contained viral antigen, suggested that many of these fragments were incomplete viral particles. It is proposed that concentrated inocula damage the cells and interfere with differentiation of the virus, but do not inhibit formation and detachment of cytoplasmic processes. Under these circumstances the accumulation of viral antigen at the surface of the cell will result in the predominant formation of incomplete virus.

Studies with Live Attenuated Measles-Virus Vaccine

Introduction Control of measles through active immunization has been the object of numerous studies. Measles first was transmitted artificially to susceptible human beings by Home,1who exposed scarified skin of volunteers to blood from patients in the acute phase of this disease. In a similar attempt to produce active immunity, Hektoen2injected fresh infective blood subcutaneously into susceptible persons who subsequently developed measles. Failure to produce attenuated disease apparently discouraged further attempts to induce active immunity against measles. Adaptation of measles virus to growth in embryonated hen's egg and the subsequent use of this virus as an immunizing agent was reported by Rake and his associates between 1939 and 1943.3-8Virus passaged from 4 to 108 times via the chorioallantoic route did not induce in susceptible children a significant degree of protection from naturally occurring measles,9and interest in this problem again waned. In 1954, Enders

Research Note: A Rapid Macroscopic Plate Agglutination Test for Newcastle Disease. A Preliminary Report

RAPID and simple macroscopic test to detect antibodies against animal viruses would be of value in human and veterinary medicine. This paper (1) describes a plate agglutination test to detect antibodies for Newcastle disease (ND). The antigen was prepared from centrifuged chorioallantoic fluid of embryonating chicken eggs infected with the GB (Texas) strain1 of ND virus. Eighty 9-day-old embryonating eggs were inoculated via the chorio-allantoic route with one million ELD50 of the GB (Texas) strain of ND virus. The chorio-allantoic fluid (360 ml), harvested at death of the embryos 18 hours or more after inoculation, was differentially centrifuged. Extreme care was taken in harvesting asceptically only the chorio-allantoic fluid. If yolk material was aspirated by accident, the syringe was discarded. Another clean, sterile syringe and needle were used to continue the harvest. This was pooled into a 500 ml. sterile Erlenmayer flask, partially immersed in ice water. The temperature of the cubicle during harvesting was approximately 28 C. One cardboard flat of eggs (30 eggs) was harvested at a time. The remaining dead eggs were kept at 2 C until harvested. A suitable sample of the pooled chorio-allantoic fluid had an ELD,, of 10-9-0. Shortly after harvesting, the 360 ml of

ギラン・バレー症候群を呈する多発生神経根炎患児から分離された1 cytopathogenic Agent (柿木株) に関する研究

The attempts were made to isolate the agents into the tissue culture human embryonal skin muscle cells, from the fecal specimen of a case with typical Guillain-Barré syndrome, namely the dissociation of “cyto-albumique” of spinal fluid, paralysis of both legs and without fever, of age 5 years old boy. The cytopathogenicity was observed clearly at 5th tissue culture passage level following three days of incubation. The pattern of this changes were of similar nature to that of polio's. The maximum TCID50 was revealed to be of the vicinity of 10-4.5/ml. Neutralizing antibody was found in the patients sera against this agent, which was not apparently neutralized with 3 types of polio-antisera and their mixture sera. The agent was temporary designated as “Kakiki Strain”, of which neutralizing antibody titre exhibited 1:64 in both paired sera obtained at 12th and 21st day of illness. No discernible changes were observed in the tissue culture HeLa cells and chick embryo fibroblast cells No pathogenic effect was shown in suckling and adult mice as well as in the developing chick embryo inoculated through the chorioallantoic route and yolk sack route. The suggestion would seem to be made in the line that this agent could be of similar character to ECHO viruses.

Neutralization Tests with Eastern Equine Encephalomyelitis Virus in the Chick Embryo: A Statistical Treatment of the Results by the Probit Method

Summary The chick embryo has been successfully employed for the quantitative assay of residual equine encephalomyelitis virus in tests with immune serum. Under the conditions of these experiments maximum neutralization of equine encephalomyelitis virus by antiserum was attained within a few seconds of mixing the virus and the serum. Less neutralization was apparent if the virus-serum mixtures were kept at 4 C prior to inoculation. The amount of neutralization which was apparent was related directly to the susceptibility of the embryo or inversely to the number of virus particles necessary to initiate a lethal infection. The greatest neutralization was shown when 15 day eggs were inoculated by the yolk sac route and the least neutralization when 15 day eggs were inoculated by the chorio-allantoic route. Eleven day eggs inoculated by either route showed intermediate degrees of neutralization.

Studies on the Growth of Murine Encephalomyelitis Viruses in Fertile Eggs

Summary Theiler's virus of mouse encephalomyelitis, FA strain, was cultivated through ten generations in the developing chick embryo by the chorio-allantoic route of inoculation. Successful propagation was also obtained by inoculation into the yolk-sac and into the allantoic cavity. The virus was present throughout the egg with the highest concentration being found in the embryo, after twelve to fourteen days incubation at 35 C., following inoculation of 6–7 day old eggs. The ID50 titers of embryo suspensions ranged between 10−5.0 and 10−5.4 and no changes in the symptomatology of infection by this virus in mice were observed following these egg passages. Theiler's TO strain of mouse encephalomyelitis was also passed through four generations in the developing chick embryo following inoculation by the chorio-allantoic route. Virus was found to be present in the chorio-allantoic membrane and embryo and the maximal titer appeared to be found after twelve days incubation at 35 C. The ID50 titer of the embryo suspension after four egg passages was found to be 10−3.0. Successive passages in the chick embryo did not cause any apparent alteration in the symptomatology produced by this virus in mice. Efforts to cultivate in fertile eggs the Lansing, Y-SK and Ph strains of mouse-adapted poliomyelitis virus, two strains of poliomyelitis virus derived from monkey spinal cord and one strain of poliomyelitis virus derived from human spinal cord, were unsuccessful.

Resistance of Chicks to Infection with Influenza A Virus

SummaryVirus was not recovered after hatching from chicks infected as embryos with the Melbourne strain of influenza A virus. This virus survived and increased in cultures of 18- or 20-day embryo lung and lung tissue from 2-day-old chicks at 37° C, but not in cultures of embryo or chick lung at 41°C. Virus was not recovered from embryos inoculated by the chorio-allantoic route and kept at 41°C. Virus was demonstrated in the lungs of 1-day-old chicks at 19 but not at 48 hours after inoculation in the nares. No neutralizing antibodies for the virus were detected in sera of chicks following such inoculation nor in the sera of normal chicks or adult chickens.

OBSERVATIONS ON THE INFECTION OF CHICK EMBRYOS WITH BACTERIUM TULARENSE, BRUCELLA, AND PASTEURELLA PESTIS

1. Comparison of the infections of chick embryos by the chorio-allantoic route indicates that Bacterium tularense and Brucella suis, abortus, and melitensis exhibit varying degrees of facultative intracellular parasitism. Pasteurella pestis is adapted to rapid proliferation and spread in the intercellular fluids. 2. In the early stages of infection Bacterium tularense has a marked affinity for growth within ectodermal epithelial cells. Brucella suis and Brucella abortus differ in their selectivity for cells of mesodermal derivation and especially in their effect on vascular endothelium. The strain of Brucella melitensis studied is limited in its intracellular growth to ectodermal epithelium. 3. Many of the features characteristic of these infections in the natural hosts are reproduced in the chick embryo and its membranes. 4. The possible implications regarding the differences in behavior of these microorganisms in relation to the problem of infection and pathogenesis of these diseases are discussed.

A STUDY OF GENERALIZED VACCINIA IN THE CHICK EMBRYO

1. Chick embryos infected by the chorio-allantoic route with a bacteria-free strain of vaccinia virus develop a general dissemination of the virus throughout the entire organism with the exception of the central nervous system. 2. Quantitative estimation of the distribution of the virus in the various organs of the infected chick by cutaneous inoculation on the rabbit skin offers no evidence for a heightened affinity of the virus for special tissues. 3. Histological study of the lesions in the various organs demonstrates the focal character of the lesions which apparently originate as perivascular infiltrations around the smaller blood vessels. No lesions could be demonstrated in the central nervous system proper. 4. In the earlier stages of the disease Guarnieri bodies are clearly demonstrable in the cells of the epidermis and the squamous epithelium of the buccal mucosa. Inclusion bodies closely resembling Guarnieri bodies are demonstrated in all the lesions occurring in the various other organs. 5. It was not possible to demonstrate conclusively the presence of Paschen bodies in the lesions of the internal organs by the Morosow method usually used for the demonstration of these bodies in the membranal lesion.