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Rodríguez Barahona, Julia Victoria [Sonstige Person, Familie und Körperschaft]

Lectin and immuno histochemical investigations on cellular alterations in chicken embryos following inoculation with Newcastle Disease Virus (NDV) of different virulence - [1. Aufl.]

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  • Medientyp: E-Book; Hochschulschrift
  • Titel: Lectin and immuno histochemical investigations on cellular alterations in chicken embryos following inoculation with Newcastle Disease Virus (NDV) of different virulence
  • Beteiligte: Rodríguez Barahona, Julia Victoria [Sonstige Person, Familie und Körperschaft]
  • Erschienen: Gießen: DVG Service, 2009
  • Ausgabe: 1. Aufl.
  • Umfang: Online-Ressource (173 S.= 4.652 kb, text); Ill., graph. Darst
  • Sprache: Englisch
  • Identifikator:
  • Schlagwörter: Hühnervögel > Embryo > Newcastle-Krankheit > Virulenz > Lectine > Immuncytochemie
  • Entstehung:
  • Hochschulschrift: Zugl.: Hannover, Tierärztl. Hochsch., Klinik für Geflügel, Diss., 2009
  • Anmerkungen:
  • Beschreibung: Lektin, Immunohistochemie, Newcastle-Krankheit-Virus. - Newcastle disease (ND) is a worldwide disease of poultry caused by a Paramyxovirus type 1, genus Avulavirus. The principal receptor of this virus is 2-deoxy-2,3-dehydro-N-acetyl-neuraminic acid. By means of histological techniques, including histochemistry, with special emphasis on lectin histochemistry and immunohistochemistry, tissue changes occurring in specific pathogen free (SPF) chicken embryos infected with NDV of different pathotypes (HB-1 and Komarov) were investigated in comparison to uninfected chicken embryos. For this purpose, our study was divided into two main parts: (1) Determination of the LD50 of the NDV pathotypes under study and (2) Histological studies in chicken embryos as mentioned above to determine the distribution of free sugar moiëties and presence of NDV in various organs of the chicken embryos. After euthanization, the embryos were fixed in Bouin’s solution, embedded in paraffin wax, and 5 μm sections were stained with four biotinylated lectins [Con A (sugar specifity: α-D-mannose/glucose), BSA-1 (sugar specifity: α-D-galactose), WGA (sugar specifity: β-D-N-acetylglucosamine) and MAA (sugar specifity: siaα-2,3-galactose)]. Immunohistochemical staining to detect the organ distribution of NDV, was done employing a polyclonal antibody against ND virus raised in rabbits. Different intensities of lectin staining were observed in the uninfected control group, indicating the presence of free sugar residues depending on the tissue type studied, incubation time and the lectin used. Apical parts of the epithelial cells of esophagus, proventriculus, trachea, lung and air sac exhibited strong to medium staining reactions with Con A, BSA-1 and WGA. These cells showed a significantly positive immunochemical reactions (Fisher's Exact Test <0,05) in both infected groups (NDV-Komarov and HB-1 pathotype) at day 11th compared to day 15th of incubation, when only 1 of the infected with NDV-Komarov pathotype and 1 infected with NDV-HB-1 pathotype were found NDV positive. Such coincidence was not observed when a rather weak MAA reaction and distinct immuno histochemical antivirus staining were found. The latter finding may be caused by the presence of 2-deoxy-2,6-dehydro-N-acetyl-neuraminic acid as a second receptor of NDV not recognized by MAA. At present, we can not explain the reaction difference in immuno histochemistry between day 11 and day 15 of incubation, but it may be speculated that this observation is attributable to resorption of possibly proteinaceous substances during the process of drinking amniotic fluid, both beginning about day 13 of incubation. Stronger lectin staining reactions were observed in infected organs of the chicken embryos infected at 11th; when compared to the uninfected controls. In these cases, significant differences were observed in the epithelium of the trachea with Con A, BSA-1, and the epithelium and goblet cells in the ventriculus with MAA when they were analyzed by Fisher´s Exact Test (<0,05). Further investigations are necessary to elucidate the role of the different sugar moiëties in virus infection, because they are very important structures in biological processes such as cell-cell interactions and pathogen entry into the host cell. In summary, the findings presented herein for the first time describe the (1) distribution of four different sugar moieties in various organs of the chicken embryo and (2) changes induced following infection with two different pathotypes of NDV. This may be the basis for further pathogenesis studies employing chicken embryos and their interaction with pathogens on the cellular level.
  • Zugangsstatus: Freier Zugang