Fertile Hen's Egg Research Articles

Electron Microscopy of the Pathology and Chemotherapy of Legionella pneumophila Infection in the Chick Embryo

The ultrastructural pathology induced in organs of chick embryos previously infected with 100 times the yolk sac LD50 of Legionella pneumophila was examined by electron microscopy. Subtle pathological changes were observed three days post-inoculation while more extensive cellular and sub-cellular necrosis occurred at four days post-inoculation. Selected antimicrobial agents were administered three days after infection and evaluated for their ability to reduce or reverse ultrastructural lesions. In this study, ciprofloxacin was the only antibiotic that reversed tissue damage induced in response to infection with L. pneumophila while doxycycline, erythromycin, and rifampicin were also effective, but to a lesser degree. These results demonstrated the superior antimicrobial activity of ciprofloxacin and further illustrated the efficacy of the fertile hen's egg as a useful in vivo assay system for the evaluation at the ultrastructural level of putative chemotherapeutic agents for the treatment of Legionnaires' disease.

Pathogenesis and Chemotherapy of Experimental Legionella pneumophila Infection in the Chick Embryo

The pathogenicity of Legionella pneumophila, serogroup 1, strain Nottingham N7, was assessed in terms of LD50 data and the ability of the organism to induce pathological lesions in the fertile hen's egg. Histopathological examination of embryo organs after inoculation with 1, 10, 100 and 1000 times the yolk sac LD50 revealed a disseminated infection. Systemic spread of the organism resulted in widespread necrosis and evidence of consolidation together with generation of copious amounts of oedema fluid. These were particularly severe in the liver, heart, spleen and kidney. The infection elicited a massive inflammatory response typified by infiltration with polymorphonuclear leucocytes and lymphocytes. Selected antimicrobial chemotherapeutic agents were investigated in protection studies for their capacity to ameliorate or control disease processes in this test system. Of those examined ciprofloxacin was most effective in reducing the incidence of lesions in these tissues and for prolonging embryo viability. Rifampicin, and to a lesser degree, erythromycin and doxycycline, also showed antimicrobial activity in these in vivo trials. These results indicate that the fertile hen's egg may be a useful alternative to other animal systems for the in vivo testing of clinically putative antimicrobial agents in the treatment of Legionnaires' disease.

Penetration studies and residue determinations of 2,4-dichlorophenoxyacetic acid butyl ester in fertile hen eggs and chicks hatched from treated eggs.

Fertilized hen eggs were topically applied with 2,4-Dichlorophenoxyacetic acid butyl ester (3.1 mg/egg) before starting incubation. During the incubation time 20% of the compound applied on eggs was lost by volatilization. At different incubation times (0, 1, 5, 10 and 15 days) the wash-off technique was applied to fertile hen eggs and residue analyses of 2,4-D ester were determined and quantified by gas chromatography with an electron capture detector. We found that 2,4-D ester began to be detectable at the embryo from the 5th incubation day and the amount of the compound progressively increased during chick embryonic development. Furthermore chicks hatched from treated eggs showed the presence of the compound in all studied tissues and the highest levels were found in organs such as brain and kidney.

Evidence for embryotoxicity of gossypol in mice and chicks with no evidence of mutagenic activity in the ames test

Effects of gossypol treatment were studied in pregnant mice and chick embryos. Pregnant Balb C mice were treated orally with 60 or 120 mg/kg of gossypol acetic acid on days 6–13 of pregnancy and killed on day 18. The uteri were removed, the number of resorptions and late fetal deaths were recorded, and the fetuses were weighed and assessed for malformations. Fertilized hen eggs were injected with 0.25 mg gossypol/egg at 24, 48, 72, or 96 h of incubation. The embryos were examined at day 9 of incubation. The mutagenic potential of gossypol was determined by the Ames test. Treatment of mice with gossypol produced significant adverse effects on the dam and offspring including decreased pregnancy weight gain of the dam and growth retardation of the offspring. There were increased resorptions and late fetal deaths in mice and high mortality in chick embryos. Exencephalic fetuses were observed in one of four litters exposed to the higher dose of gossypol, micromelia was observed in one of 26 chick embryos treated at 24 h, and gastroschisis was observed in one of 21 chick embryos treated at 72 h. No malformations were observed in the controls. The number of revertants per plate in the gossypol treatment group (100 or 500 mg gossypol/plate) did not differ significantly from that of control. This study provides evidence that gossypol has embryotoxic and possibly teratogenic activity in mouse and chick embryos but no mutagenic activity according to the Ames assay.

Aeration of the shell membranes of avian eggs

The inner and outer shell membranes (ISM, OSM) of fertile hen's eggs become aerated during the first ten days of incubation. The volume of gas in the compound membrane increased from 0 to 65% by day 11 when practically all of the space between the fibers is gas filled. The amount of gas depends on a balance between the capillary tension produced by fluid menisci in the interstices between fibers and the colloid osmotic tension of the albumen. Capillary tension, measured directly with a pressure plate apparatus, and colloid osmotic pressure, measured with a new technique, are in virtual equilibrium during aeration. These tensions increase from 0.15 bar at laying to 2.25 bar on day 11 because of evaporation from the membranes and active removal of water from the albumen by the embryo. Water leaving the membranes by evaporation is only partly replaced from the albumen. Thus the water contents of the membranes and albumen decrease (ISM: 70 to 56%, OSM: 71 to 52%, albumen: 86 to 58%). As the membranes dry, gas enters the larger pores first. Physiological estimates of effective radius show smaller ( x = 1.3 μ m ), bimodally distributed pores in the ISM and larger ( x = 3.4 μ m , unimodally distributed pores in the OSM.

Identification of Legionella pneumophila antigens and antibodies by immunoferritin electron microscopy.

Material from 18 lungs positive for Legionella pneumophila and 21 strains of legionella grown on bacteriological media or in the yolk sac of fertile hen's eggs were examined by direct negative stain and immunoferritin electronmicroscopy. The 18 lung samples and all preparations of cultivated organisms showed the presence of bacteria with the typical morphology of L. pneumophila. By immunoferritin electronmicroscopy, with specific antisera, it was possible to serogroup the organisms. The immunoferritin method in reverse made it possible to detect and titrate antibody in sera from patients. Direct and immunoferritin electronmicroscopy were as sensitive as immunofluorescent antibody tests for detecting the antigens and antibodies of Legionnaires' disease. Additional advantages of the ultrastructural technique are discussed.

A Technique For Illuminating Early Chick Embryos In Ovo With A Glass Rod As A Light Guide

A glass rod mounted on a microscope condenser acts as a light guide through the yolk of a fertile hen's egg. Two apertures must first be made in the shell: the one below covered with plastic film which seals around the rod; the one above covered with a glass coverslip to allow microscopic observation of the embryo.

Gas exchange of the fertile hen's egg: Components of resistance

The resistance to diffusion of respiratory gases falls by 50% from day 10 to day 18 in the incubating fertile hen's egg (Temple ande Metcalf, 1970). We have calculated the change in the components of this resistance with respect to incubation age. The egg shell resistance remains unchanged. The chorioallantoic membrane (including capillary endothelium) resistance decreases dramatically from day 10 to day 14 and then remains relatively unchanged. The resistance offered by the blood is unchanged from day 10 to day 12, it decreases by day 14 due to an increase in oxygen capacity and continues to decline at days 16 and 18 because of the combined effects of a continued increase in oxygen capacity and an increase in capillary volume.

The effects of chemical irritants and tobacco smoke condensate on the chorioallantoic membrane of the fertile hen's egg

The effects of various materials on the chorioallantoic membrane of the 10-day incubated fertile hen's egg have been studied. A detailed sequential histological study of the effects of croton oil and tobacco smoke condensate showed that both substances caused local haemorrhage and necrosis, followed by hyperplasia. The degree of thickening was found to be both dose-and time-dependent. From these observations a test procedure for comparing the irritant activity of tobacco smoke condensates has been developed; this procedure involves measuring the maximum thickness of the membranes 72 h after application of the condensates.

Ouabain-induced growth retardation and death in chick embryos.

SummaryOuabain injected into the air space of fertile hen eggs on the seventh day of incubation produced retardation of growth of the embryos incubated over a further period of five days. An injected close of 35 μg of ouabain per embryo caused 24% mortality of the eggs as against 9% in the saline‐injected controls. The survivors showed marked retardation of growth as ouabain‐treated embryos had a significantly lower weight than the saline‐injected controls. On doubling the dose of ouabain, the mortality rate increased to 57% and the growth retardation in the survivors was even more pronounced. Apart from growth inhibition, no gross structural abnormalities were noticed. The activity of the enzyme Na+‐K+ activated adenosine triphosphatase was depressed in the erythrocytes of the ouabain‐treated chicks.

High School Students Teach Science in Elementary Grades

I t is common for grade schools in small, primarily rural school districts to offer curricula relatively barren of meaningful science activities. This deficiency is a result of lack of funds, space, and equipment; of inadequate preparation of teachers in the sciences; and of the continuing influence of traditional, textbook-centered elementary programs. It is also common in many high school science classes, regardless of the size of the district, for a number of students to work far below their potential, primarily because they find the work irrelevant and boring. In the school district where I teach, most of the students are bussed in from the countryside to one senior-junior high and to four grade schools, two of which are located within a few minutes' walking distance of the secondary-school complex. This district has long suffered from the inadequacies I have mentioned. However, the recent initiative of two high school science teachers and a budding cooperation between administrators and teachers in all the schools have helped to alleviate these conditions and have significantly enriched the entire science program. No extra cost has been incurred in this endeavor, nor have extensive program changes been necessary. It has been the use of a rich and untapped resource Fig. 1. Jane Kessler explains to second-graders a sterile technique for opening and maintaining fertile hen eggs. This technique has allowed students to observe a single developing embryo for as long as eight days.

A survey of the occurrence of Vibrio parahaemolyticus and V. alginolyticus on mussels and oysters and in estuarine waters in the Netherlands.

Summary: In a survey on the occurrence of Vibrio parahaemolyticus and V. alginolyticus on mussels and oysters and in estuarine waters in The Netherlands it appeared necessary to use at least 2 different enrichment procedures as well as isolation methods in parallel to obtain reliable results. A consistent identification of V. parahaemolyticus on biochemical grounds remains a difficult task; further research in this area is definitely required. The ligated intestinal segment technique gave inconsistent results; injection of organisms into the yolk sac of fertilized hen eggs seems to be a more reliable diagnostic procedure. Of 288 samples of mussels examined 2·4% were V. parahaemolyticus positive, 80 samples of oysters were all negative and amongst 64 water samples 4·7% were positive. V. alginolyticus was isolated in similar percentages of samples of mussels and water, but oysters were positive to an extent of 6·8%.

The isolation of influenza viruses.

Of 129 strains of the Hong Kong variant of influenza A2 virus isolated from 176 respiratory infections, 105 (81.4%) were isolated in tissue culture, the remainder being detected only by the inoculation of fertile hen's eggs. Organ cultures of chick embryo trachea were at least as sensitive as monkey kidney tissue culture for the isolation of this virus but organ cultures of human embryonic ciliated epithelium were much less efficient. The reverse was true for strains of influenza B virus studied in 1970 and 1971 when eggs and chick embryo tracheal organ culture were of little value and organ cultures of human embryonic ciliated epithelium and monkey kidney tissue cultures were the systems of choice.

Growth of Newcastle disease virus in fertile eggs of the Japanese quail ( Coturnix coturnix japonica)

Five strains of Newcastle disease virus (NDV) were inoculated into the allantoic cavities of six-day fertile quail eggs and nine-day fertile hen eggs. The mean embryo infective dose (EID 50) was higher and the mean embryo death time (EDT 50) was shorter in quail eggs than in hen eggs. The F strain killed all the quail embryos, but hardly any chick embryos. The HA and EID 50 titres of allantoic fluids were higher from hen eggs than from quail eggs. It is considered that fertile quail eggs have some advantages over hen eggs for use in the virological laboratory.

The effect of injection of chlorinated hydrocarbon pesticides on hatchability of eggs

Dieldrin, aldrin, endrin, DDT, heptachlor, toxaphene, endosulfan, lindane, and strobane were injected via a corn oil carrier into fertile hen eggs as a test of their toxicity. Dieldrin, 5 or 10 mg, injection into the albumin of fertile eggs prior to incubation caused decreases in hatchability of 30 and 45%, respectively. In addition, chicks from the 10-mg treatment exhibited tremors and died within 24 hours, while the chicks given 5 mg all died within 1 week of hatching. Injection of 0.2 mg of endrin into fertile eggs after 7 days of incubation decreased hatchability to 40%. Higher levels were very toxic and resulted in as low as 1.8% hatchability. Similarly, 3.0 mg of DDT per egg resulted in 10% lower hatchability. Lindane, at 2.0 mg and strobane at 6.0 mg/egg caused no decrease in hatchability. Aldrin, at 2.0 mg per egg and toxaphene and endosulfan, at 1.5 mg/egg resulted in no decrease in hatchability. The 1.5 mg injection of heptachlor resulted in 12% reduction in hatchability. All tested chicks appeared normal except those given dieldrin.

Production and viability of eggs from hens treated with paraquat.

IN the communication from Dunachie and Fletcher1 it is shown that when paraquat is injected into fertile hen's eggs to give a final concentration greater than approximately 0.1 p.p.m. the subsequent rate of hatching was reduced. Because hens may eat food containing residues of paraquat from previous sprayings I made the following experiment to determine whether the amount of paraquat likely to be present would cause similar effects.

INFECTION OF THE ONE-DAY OLD FERTILE HEN'S EGG WITH RABIES VIRUS. VIII. BETA-PROPIOLACTONE INACTIVATED VACCINE PRODUCED BY INOCULATION OF ONE-DAY EGGS WITH NISHIGAHARA-O30-M (NOPM) STRAIN.

Nishigahara-O30M (NOPM) strain of rabies virus, which had been produced by passing the rabbit-fixed Nishigahara strain 30 times consecutively in 1-day eggs and then transferring it to the mouse brain passage, was used to prepare the 1-day egg vaccine employing beta-propiolactone inactivation. The vaccines produced exhibited satisfactory potencies in mouse protection tests, Habel indices being consistently higher than 1,000. Preservation of the vaccine in an ice box for 4 months did not reduce the potency. A standard method of vaccine preparation suited for mass-production was also worked out. A pool of 3 lots of this vaccine was given to 30 volunteers in 3 or 4 doses of 0.1 ml. by the intradermal route with intervals of 3 to 7 days, and all the vaccinees responded to the vaccination with fairly good rises of neutralizing antibodies occurring before the end of the second week, except one person who showed only a very weak response. Little systemic reactions were observed, and local reactions appeared as erythema and induration lasting for a few days, which nevertheless became less intense upon continuation of the vaccination. It was estimated therefore that the NOPM-1-day egg vaccine might be able to replace the brain type vaccine currently used for human prophylaxis.

Infection of the one-day old fertile hen's egg with rabies virus. VI. Strain differences in the egg adaptability.

Infection of the one-day old fertile hen's egg with rabies virus. VI. Strain differences in the egg adaptability.

Infection of the one-day old fertile hen's egg with rabies virus. VII. Comparison of viral yields obtained by inoculation of eggs of different ages with different strains and production of vaccine.

Nishigahara-O30M (NOPM) strain of rabies virus, which had been produced by passing the rabbit-fixed Nishigahara strain 30 times consecutively in 1-day eggs and then transferring it to the mouse brain passage, was used to prepare the 1-day egg vaccine employing beta-propiolactone inactivation. The vaccines produced exhibited satisfactory potencies in mouse protection tests, Habel indices being consistently higher than 1,000. Preservation of the vaccine in an ice box for 4 months did not reduce the potency. A standard method of vaccine preparation suited for mass-production was also worked out. A pool of 3 lots of this vaccine was given to 30 volunteers in 3 or 4 doses of 0.1 ml. by the intradermal route with intervals of 3 to 7 days, and all the vaccinees responded to the vaccination with fairly good rises of neutralizing antibodies occurring before the end of the second week, except one person who showed only a very weak response. Little systemic reactions were observed, and local reactions appeared as erythema and induration lasting for a few days, which nevertheless became less intense upon continuation of the vaccination. It was estimated therefore that the NOPM-1-day egg vaccine might be able to replace the brain type vaccine currently used for human prophylaxis.

Recent advances in the study of measles virus.

Editorials1 February 1959RECENT ADVANCES IN THE STUDY OF MEASLES VIRUSF. R. MCCRUMB JR., M.D.F. R. MCCRUMB JR., M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-50-2-522 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptMeasles was first transmitted artificially from man to man in 1758 by Home,1who exposed skin incisions of nonimmune volunteers to blood from patients ill with the disease. In 1905 Hektoen2injected fresh blood from patients with measles subcutaneously into two individuals who developed measles 11 and 13 days later, respectively. Susceptibility of species of the genusMacacawas established by the experiments of Goldberger and Anderson in 1911,3when they demonstrated that more than 50% of monkeys inoculated with blood and nasopharyngeal washings from patients with measles developed fever, catarrhal symptoms and exanthem. Transmission of this disease through six...1 Home F: Medical facts and experiments, Gentleman's Magazine and Historical Chronicle 28: 285, 1758. Google Scholar2 Hektoen L: Experimental measles, J. Infect. Dis. 2: 238-255, 1905. CrossrefGoogle Scholar3 GoldbergerAnderson JJE: An experimental demonstration of the presence of the virus of measles in the mixed buccal and nasal secretions, J. A. M. A. 57: 476-478, 1911a. CrossrefGoogle Scholar4 BlakeTrask FGJD: Studies on measles. I. Susceptibility of monkeys to the virus of measles, J. Exper. Med. 33: 385-412, 1921a. CrossrefMedlineGoogle Scholar5 Plotz H: Culture "in vitro" du virus de la rougeole, Bull. Acad. de méd., Paris 119: 589-601, 1938. Google Scholar6 RakeShaffer GMF: Propagation of the agent of measles in the fertile hen's egg, Nature, London 144: 672-673, 1939. CrossrefGoogle Scholar7 EndersPeebles JFTC: Propagation in tissue cultures of cytopathogenic agents from patients with measles, Proc. Soc. Exper. Biol. and Med. 86: 277-286, 1954. CrossrefMedlineGoogle Scholar8 CohenGordonRappMacaulayBuckley SMIFJCSM: Fluorescent antibody and complement-fixation tests of agents isolated in tissue culture from measles patients, Proc. Soc. Exper. Biol. and Med. 90: 118-122, 1955. CrossrefMedlineGoogle Scholar9 RuckleRogers GKD: Studies with measles virus. II. Isolation of virus and immunologic studies in persons who have had the natural disease, J. Immunol. 78: 341-355, 1957. MedlineGoogle Scholar10 GirardiWarrenGoldmanJeffries AJJCB: Growth and CF antigenicity of measles virus in cells deriving from human heart, Proc. Soc. Exper. Biol. and Med. 98: 18-22, 1958. CrossrefMedlineGoogle Scholar11 DekkingMcCarthy FK: Propagation of measles virus in human carcinoma cells, Proc. Soc. Exper. Biol. and Med. 93: 1-2, 1956. CrossrefMedlineGoogle Scholar12 BlackReissigMelnick FMJL: Propagation of measles virus in a strain of human epidermoid cancer cells (Hep-2), Proc. Soc. Exper. Biol. and Med. 93: 107-108, 1956. CrossrefMedlineGoogle Scholar13 FrankelWest JWMK: Cultivation of measles virus in a stable line of human amnion cells, Proc. Soc. Exper. Biol. and Med. 97: 741-742, 1958. CrossrefMedlineGoogle Scholar14 FrankelBurnsteinWest JWTMK: Propagation of measles virus in tissue cultures of dog kidney cells (abstract), Federation Proc. 17: 551, 1958. Google Scholar15 EndersPeeblesMcCarthyMilovanovicMitusHolloway JFTCKMAA: Measles virus: a summary of experiments concerned with isolation, properties and behavior, Am. J. Pub. Health 47: 275-282, 1957. CrossrefGoogle Scholar16 KatzMilovanovicEnders SLMJF: Propagation of measles virus in cultures of chick embryo cells, Proc. Soc. Exper. Biol. and Med. 97: 23-29, 1958. CrossrefMedlineGoogle Scholar17 KatzMedearisEnders SLDNJF: Experiences with a live attenuated measles virus (abstract), Am. J. Dis. Child. 96: 183-184, 1958. Google Scholar This content is PDF only. To continue reading please click on the PDF icon. Author, Article, and Disclosure InformationAuthors: F. R. MCCRUMBJR., M.D.Affiliations: PreviousarticleNextarticle Advertisement FiguresReferencesRelatedDetails Metrics 1 February 1959Volume 50, Issue 2Page: 522-526KeywordsBloodFeversMeasles ePublished: 1 December 2008 Issue Published: 1 February 1959 PDF downloadLoading ...