Rapid heat-treatment attenuation of infectious bronchitis virus

Avian Pathol. 2010 Jun;39(3):227-33. doi: 10.1080/03079451003801516.

Abstract

In the present study we describe the rapid development of an attenuated live vaccine for GA08, a new serotype of infectious bronchitis virus, using a heat-treatment method. Incubation of the GA08 strain of IBV at 56 degrees C and passage in embryonated eggs was used as a method to fast track the attenuation process. The virus was incubated in a 56 degrees C water bath and aliquots were removed every 5 min for up to 1 h, and then each aliquot was inoculated into 10-day-old embryonated eggs. Virus with the longest incubation time that produced lesions in the embryos was harvested, again incubated at 56 degrees C as described and passaged in embryonated eggs. Attenuation of the virus, designated GA08/GA08HSp16/08, was verified in 1-day-old specific pathogen free chicks. A 10x dose of the vaccine was found to be safe for 2-week-old broiler chicks of commercial origin. The efficacy of the heat-treated attenuated virus was determined by vaccinating broiler chicks of commercial origin at 1 and 14 days of age intraocularly/intranasally. Vaccinated birds that were challenged with 10(4.5) median embryo infectious doses of pathogenic GA08 virus/bird at 28 days of age were protected from the disease, and challenge virus was only detected in the trachea of one of 21 birds by real-time reverse transcriptase-polymerase chain reaction at 5 days post challenge. The attenuation process took 10 weeks to complete, which is a substantially shorter time than required to attenuate infectious bronchitis virus by serial passage in embryonated eggs without heat treatment (38 weeks or more).

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Animals, Newborn / immunology
  • Animals, Newborn / virology
  • Base Sequence
  • Chick Embryo / immunology
  • Chickens / immunology
  • Chickens / virology
  • Coronavirus Infections / immunology*
  • Coronavirus Infections / veterinary
  • DNA Primers
  • Hot Temperature
  • Housing, Animal / standards
  • Infectious bronchitis virus / classification
  • Infectious bronchitis virus / genetics
  • Infectious bronchitis virus / immunology*
  • Phylogeny
  • Poultry Diseases / immunology*
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • Safety
  • Vaccines, Attenuated / immunology
  • Viral Vaccines / administration & dosage

Substances

  • DNA Primers
  • RNA, Viral
  • Vaccines, Attenuated
  • Viral Vaccines