Identification and purification of a local isolate of infectious laryngotracheitis virus

Local infectious laryngotracheitis [ILT] virus strain was isolated on ECE for ten passages. The isolated virus produced characteristics pock lesions on chorio-allantoic membranes of the embryonated eggs, which increased in size and number when the number of passage increases. Moreover, the isolated virus was further propagated on CER and Vero cells where the virus gave high titers only when propagated on Vero cells. The virus was purified by sucrose gradient ultra centrifugation where a white band was collected at the end of the centrifugation cycle. The isolated ILT virus was identified by means of Electron Microscopy, Dot-ELISA and serum neutralization

Preparation of cell-culture inactivated infectious laryngotracheitis virus vaccine adjuvanted with nigella sativa oil

Isolated and identified infectious laryngotracheitis virus was inactivated by [BEI] and tested for its sterility, safety and potency then the inactivated ILT virus was mixed by addition of oil adjuvant, which consists of Nigella sativa and paraffin oil [PANISA oil] in the ratio of 1: 1. Two vaccines were made, the first where the aqueous phase to the oil phase was 1: 4 while the second vaccine where the ratio was 1: 2. Quality control tests were made such as drop test, safety and sterility test. The emulsion was oil in water, safe and free from bacterial, fungal and Mycoplasma contamination. Three experiments were made; the first where the vaccines were giving to chicken and the antibody pattern during 16 weeks were demonstrated using ELISA. The second experiment where comparison between the inactivated vaccines and the live attenuated vaccine was done. The third experiment included three successive challenges with four weeks intervals. Vaccines prepared adjuvanted with Nigella sativa oil produced higher humeral response than live attenuated vaccine

Preparation and evaluation of newcastle disease virus inactivated vaccines adjuvanated with crude or fractionized Nigella sative oil

Different types of inactivated oil emulsion Newcastle disease vaccines were prepared using different extractions of the Nigella Sativa oil. The physical properties of emulsions were earned out and included emulsion type, emulsion stability and emulsion viscosity. The vaccinated chicks were bled at one-week intervals post-vaccination over six weeks and the collected sera were tested by the HI test. After that, they were challenged 21-days and 42-days post-vaccination by the intramascular inoculation with VVNDV. From this study we can conclude that the non-specific immunostimulant effect of Nigella Saliva oil is acquired when it is used as a crude oil and this improved its ability as a good adjuvant for viral vaccines

Preparation and evaluation of an inactivated Infectious Bursal Disease [IBD] virus vaccines from recent Egyptian virus isolate adjuvated with Nigella sativa oil

Inactivated IBD virus vaccines were prepared from a recent Egyptian isolate and adjuvated with Nigella sativa. The first passages of propagated viruses in SPF-embryonated chicken eggs [ECE], Vero and chicken embryo fibroblast [CEF] cell cultures were inactivated with binary ethyleneimine [BEI] and supplemented with Nigella sativa oil, as adjuvant. The prepared vaccines proved to be highly immunogenic and elicited high titers of neutralizing antibodies [17-20 log2] at weekly interval till 7 months post-vaccination [PV] and high values of lymphocyte blastogenesis [0.598 versus control 0.06]. Besides, they were able to protect vaccinated chickens [100% protection] when challenged 21 days PV. The superior potential effect of these vaccines, when compared with imported one, may be due to the use of recent local IBDV isolate and Nigella sativa oil for its nonspecific immune stimulation effect. In addition, the keeping quality of prepared vaccines proved to be sterile, safe, stable and potent when preserved at 4C for 6 months [the end of the experiment] as they produced 100% and 80% protection after 3 and 6 months of preservation, respectively

Preparation and laboratory evaluation of live attenuated Infectious Bursal Disease [IBD] virus vaccine from recent Egyptian isolate propagated on chicken embryo fibroblast [CEF] cell culture.

Very virulent IBDV was propagated for 3 passages in SPF-ECE followed by 3 passages in young susceptible chicks then for 3 passages in SPF-ECE then for further 12 passages in SPF-ECE. The first passage of last propagation was further propagated for 3 passages on Vero cells and for 60 passages in CEF-cell culture. The passage 40 of propagated IBDV proved to be safe and highly immunogenic by virus neutralization test. It still elicited high neutralizing antibody titers [18 log2] for a stationary phase about 7 months [the end of the experiment] and high values of lymphocyte blastogenesis [0.560 versus control 0.03]. Vaccinated chicks well protected against challenge with highly virulent IBDV after 3 weeks post-vaccination [PV]. The prepared vaccine has superior potential immunogenic effect than commercial live mild and intermediate plus [hot] vaccines. The vaccine is effective even when preserved for 8 months [the end of experiment] at -20C

Comparison between solid phase ELISA, dot ELISA and agar gel precipitation test for detection of bursal disease viral antigen in bursal homogenates

Forty-four bursal samples were collected from naturally infected and normal broiler chicks. The samples were homogenized and the homogenates were tested by agar gel precipitation test [AGPT], solid- phase ELISA [by indirect double antibodies s and wich and single antibody ELISA systems] and dot-ELISA for detection of infectious bursal disease virus [IBDv] antigen. The reacted positive samples showed 86.3, 88.6 and 88.6 percentage in AGPT, solid-phase ELISA and dot-ELISA, respectively. The findings suggested that the Dot-ELISA is faster, more economic and easily applicable procedure than the other two techniques. The indirect double antibody s and wich technique is more sensitive than the indirect single antibody ELISA system

Infectious bursal disease virus infection among Egyptian poultry flocks 1- Detection and isolation of the virus

A total of 46 suspected bursae have been collected from different governorates in Egypt. These samples represented Dakahlia, Ismailia, Fayoum, Sharkia, Giza and Kaliobia [Banha] 12, 10, 10, 4, 5, 5, respectively. These bursae have been firstly tested by the AGPT, using specific chicken anti-IBDV serum, for the presence of specific Gumboro viral antigen [s] before being subjected to viral isolation. Gumboro virus was isolated from every infected bursa that was giving positive reaction in the AGPT, using SPF-9 days old embryonated chicken eggs [ECE], chicken embryo fibroblast [CEF] and QT-35 cells