Preparation and evaluation of multivalent infectious bronchitis vaccine from commercial vaccine strains

  • Hassan A. Hammadi Coll. of Vet. Med. / Univ. of Al-Qadisiyah
  • Abdul Ameer H. Zahid Coll. of Vet. Med. / Univ. of Baghdad


This study was designed to prepare and evaluate multivalent live attenuated (IBV) vaccine (lyophilized seed) from the commercial IBH120, 4\91 and MA5 strains in ALkindy Company for Drug and Vaccines Production, to reduce economic losses and increase the broad protection against infectious bronchitis disease in chickens. The viruses inoculated via chorioallantoic cavity route with 0.1 ml at 9-11 days embryonated eggs obtained from unvaccinated layers against IBV and were passaged for several times, then three equal doses of the strains were mixed, and the embryonated eggs were inoculated with 0.1 ml of the mixed vaccine. After 72 hours the allantoic fluids (Vaccine) were collected. The results of the infective dose of the new vaccine were 103.2 EID50 /Bird. This multivalent live attenuated IB vaccine was tested for (safety, purity, potency, and sterility test) according to the international protocol steps of vaccine production. Efficacy of multivalent vaccine was evaluated (depending on ELISA, HI, and challenge test) by using 160 one day old (Ross 308) chicks divided into four equal groups (40 chicks of each group) reared in well-isolated places. Group (A) was vaccinated with three doses (0.3 ml) of multivalent live attenuated IB vaccine, group (B) was vaccinated with two doses (0.2 ml), group (C) was vaccinated with one dose (0.1 ml) of the multivalent live attenuated IB vaccine via ocular route at the 8th day of age, and group (D) was left without vaccination as a control group. Blood was collected at (1, 8, 14, 21, 28, 35, 42) days for ELISA test. The mean titer of maternal immunity at day one of age was 4849.8±1161.8, decreased to 69.9±35.92 at the 8th day of age before vaccination. The serum of control group was seen avuncular from IB antibodies at the 14th day of age. The experiment showed significant differences (p<0.05) titers resulting from the use of different doses (0.3 ml, 0.2 ml, and 0.1 ml) at age of 14 days, it was 255.30±15.00, 736.60±94.00 and 545.50±17.00 respectively in comparison with zero in control group. The challenge test carried out for different groups with (IB- field isolated Variant 2 strain 105 EID50 / bird which was supplied from Veterinary Directorate) at 28 days old with 0.5 ml through ocular route. Ten birds from each group were taken randomly. According to the clinical signs, morbidity and mortality rate, gross lesion, and the antibody titer, the results carried out that protection rate in group B was higher than others groups, then group C and group A respectively, the test revealed significant differences (p<0.05) between the vaccinated groups, and also between vaccinated groups and the control group. This study has concluded the possibility of production IB multivalent live attenuated vaccine (broad protection vaccine) using different IB Vaccine strains in chickens.


1-Abro SH, Renstrom LH, Ullman K, Isaksson M, Zohari S, Jansson DS, Belak S, Baule C (2012) Emergence of novel strains of avian infectious bronchitis virus in Sweden. Veterinary Microbiology, 155(2-4): 237-46.
2-Ababneh M, Dalab AE, Alsaad S, Al-Zghoul M (2012) Presence of Infectious Bronchitis Virus Strain CK/CH/LDL/97I in the Middle East. Faculty of Vet. Med. Jordan University of Sci. and Technol., P.O. Box 3030, Irbid 22110, Jordan.
3-Worthington KJ, Currie RJ, Jones RC (2008) A reverse transcriptase-polymerase chain reaction survey of infectious bronchitis virus genotypes in Western Europe from 2002 to 2006. Avian Pathol 37(3), 247-57.
4-Seifi S, Asasi K, Mohammadi A (2009) A study of natural co-infection caused by avian influenza (H9 subtype) and infection bronchitis viruses in broiler chicken farms showing respiratory signs. Online J.Vet. Res, 13: 53-62.
5-Abbass HA (1987) A study of infectious bronchitis in chicken M.Sc. Thesis–College of Vet. Medicine Baghdad University – Iraq.
6-AL-Husairaji FF (2005) Isolation, identification and pathogenesis of infectious bronchitis virus in broilers in Northern area of Iraq. M.Sc. Thesis College of Veterinary Medicine, Mosul University
7-Khammas EJ (2008) Avian Influenza (H9N2) outbreak in Iraq. Iraqi J. Vet. Med., 32:(1)
8-Hussein MA (2012) Isolation and identification of infectious bronchitis virus and experimental infection in broilers. M.Sc. Thesis, Collage of Veterinary Medicine, Baghdad University – Iraq.
9-Zana H, Mahmood R, Sleman R, Aumaid UU (2010) Isolation and molecular characterization of Sul /01 /09 avian infectious bronchitis virus, indicate the emergence of anew genotype in the middle east. J. Vet. Mic. 10: 1016.
10-Office International des Epizooties (2008) Avian infectious bronchitis. In: "Manual of Standards for Diagnostic Tests and Vaccines" 5th Ed., Paris,: 1-18.
11-Allam S (1975) "Breeding and Management of Poultry" 2nd Ed. Cairo-Egypt. Pp: 32-38.
12-Toro H (1994) and Fernandez I: Avian infectious bronchitis: specific lachrymal IgA level and resistance against challenge. Zentralbl Veterinary med. (B), 41:467–472.
13-Rosner B (1995) Hypothesis testing: categorical data. In: Fundamentals of biostatistics. 4th ed. Belmont, CA: Duxbury Press, Wadsworth Publishing; 345-442.
14-Zahid AH, Kadhum LI, AK Al-Safy (2011) Interfering of infectious bronchitis vaccine (MA5 and 4/91) strains on the immune response of Newcastle and avian influenza vaccines in male layers, Iraqi J. Agrc. (Special Issue), Vol.16: No.2.
15-Terregino C, Toffan A, Beato MS, Nardi RD, Vascellari M, Meini A, Ortali G, Mancin M, Capua I (2008) Pathogenicity of a QX strain of infectious bronchitis virus in specific pathogen free and commercial broiler chickens and evaluation of protection induced by a vaccination program based on the Ma5 and 4191 serotypes. Avian Pathol., 37 (5): 487-493.
16-De Wit jj, H Van de Sande (2009) Efficay of combined vaccines at day of hatch against a D338 challenge in SPF and commercial chickens. Proceding of the 6th international Symposium on corona and Pneumoviruses and Complicating Pathogens, June 14-17, 2009, Rauischholzhausen, Germany,PP177-182.
17-Mase M, T Inoue, S Yamaguchi, T Imada, (2008) Existence of Avian IBV with a European- prevalent 4/91 genotype in Japan. Journal Vet. Med. Sci., 70: 1341-1344.
18-Cook J, Orbell S, Woods M, Huggins M (1999) Breadth of protection of the respiratory tract provided by different live-attenuated infectious bronchitis vaccines against challenge with infectious bronchitis viruses of heterologous serotypes. Avian Pathol., 28: 477–485.
19-Toro H, Fernandez I (1994) Avian infectious bronchitis: specific lachrymal IgA level and resistance against challenge. Zentralbl Veterinary med. (B), 41:467–472.