Basri, C.

Hermans, P.G.

Muljono, A.T.

Nell, A.J.

Nugroho, E.

Purnawarman, T.

Soejoedono, R.D.

English

Name not available

Antibody responses to avian influeza vaccination in broiler chickens in Indonesia

Wageningen Yield 

Annex 5.4 
 
ANTIBODY RESPONSES TO AVIAN INFLUENZA VACCINATION  
IN BROILER CHICKENS IN INDONESIA 
 
Erianto Nugroho1, R.D. Soejodono2, T. Purnawarman 2, C. Basri1,2, P. Hermans3, A.T. Muljono1, 
A.J. Nell 4 
1
 Center for Indonesian Veterinary Analytical Studies (CIVAS) 
Jl. RSAU No. 4, Atang Sandjaja, Bogor. 16310. West Java Indonesia 
2
 Department of Infectious Diseases and Veterinary Public Health  
Faculty of Veterinary Medicine, Bogor Agricultural University 
Jl Agatis. Kampus IPB Dramaga Bogor. 16680. West Java.  Indonesia. 
3 Central Veterinary Institute, part of Wageningen UR, 
 P.O. Box 65, 8200 Lelystad, The Netherlands 
4
 Centre for Development Innovation, Wageningen University and Research Centre 
P.O. Box 88, 6700 AB Wageningen, The Netherlands 
 
Introduction 
Avian Influenza (AI) is a respiratory disease of poultry caused by type A influenza viruses from 
the Orthomyxoviridae family. In Indonesia, avian influenza cases in humans were first detected in 
2005 with 20 cases and 13 fatalities. The number of cases and fatalities peaked in 2006 with 55 
cases and 45 fatalities. To control this disease, one of the policies implemented by the 
Indonesian government is vaccination of poultry in high risk areas (targeted vaccination). The 
World Animal Health Organization (OIE) has recommended vaccination as a way to control AI, 
although acknowledges that this program alone will not succeed without the support of other 
control measures, such as biosecurity implementation, surveillance, and management of poultry 
trade. There is evidence that AI vaccination reduces virus shedding (van der Goot et al, 2005, 
Poetri et al, 2009) which in turn would reduce virus spread and the risk of human exposure. 
However, there has been some concern regarding the inconsistency of field protection of poultry 
after vaccination, possibly related to vaccine quality, vaccine strain or inadequate administration 
(Swayne, 2008). This study aims to measure the development of AI antibody titers after 
vaccination of broiler chickens at different ages during the production cycle. 
 
Material and Methods 
The study was conducted for two months, from September to November 2008, in West Bogor 
subdistrict. A total of 1500 Cobb broiler day-old chicks were divided at random into 5 groups of 
300 chicks. Group 1 was vaccinated against AI on day 1 (1T), group 2 on day 7 (2T), group 3 on 
day 10 (3T), and group 4 on day 14 (4T). Group 5 (5T) was a control group and was not 
vaccinated against AI. The AI vaccine used in this study was locally produced H5N1 killed oil 
emulsion vaccine. Serum samples were collected from 20 randomly selected chickens on days 1, 
7, 14, 21, 28, 35, 42 and 49 and were tested for AI H5 antibodies with the Haemagglutination 
Inhibition (HI) test. Tracheal and cloacal swabs were collected from 10 chickens of each group on 
the last day of the experiment (day 49) and were tested using an AI H5 Reverse Transcription 
Polymerase Chain Reaction (rt-PCR) test. Data was analyzed using ANOVA (analysis of 
variance) tests and Duncan tests (Duncan multiple range test) with a critical probability of 0.05.  
 
Result and Discussion 
The highest antibody titers for all groups were found on day 1 (range 22.8 – 23.4), indicating 
relatively high levels of maternal immunity. Antibody titers reached their lowest levels (zero) on 
day 21 for the vaccinated groups and on day 28 for the control group. Mean titers increased to 
reach peak levels at day 42 for all groups except the control group after which titers started to 
decline in all four vaccinated groups. Highest mean antibody titers were found in the group 
vaccinated on day 10 (22.4) but these mean titers were not significantly different from those in the 
group vaccinated on day 7 (21.5) or those in the group vaccinated on day 14 (21.6). From the 100 
swab samples that were tested with the rt-PCR on day 49, none were positive for H5N1. In this 
study we assumed a titer of 25 or higher to be required to protect an individual bird against a 
challenge of H5N1 virus (Kumar et al 2007) with at least 80% of birds needing to acquire this titer 
to obtain herd immunity (Tiensin et al 2007). Even at peak antibody levels on day 42, only a 
maximum of 30% of serum samples had titers of 25 or higher, which was far below levels required 
for herd immunity. Possible reasons for low antibody titers in vaccinated birds are poor vaccine 
quality, unsuitable vaccination schedules, improper vaccine administration, or impaired immune-
competence. According to Vui et al., (2002) poor vaccine quality is a common problem in 
developing countries and could be the result of poor manufacturing standards, lack of storage 
facilities (cold chain), and use of expired batches. Because the manufacturing standards of the 
vaccine and the quality of the storage facilities at the manufacturer and distributor were outside of 
our control, it cannot be excluded that this had an effect on the results of this experiment. 
Impaired immune-competence can be a result of immunosuppressive diseases, 
immunosuppressive substances in the feed such as mycotoxins or a poor innate immune 
response of the host. Serological results demonstrated antibody titers for Infectious Bursal 
Disease (IBD) which were higher than would be expected from vaccination alone in all 
experimental groups. No clinical signs of IBD were observed during the course of this experiment 
but to what extent a concurrent sub-clinical IBD infection had impaired AI antibody production is 
unknown. Feed was not tested for mycotoxins but the overall performance of the birds (i.e growth, 
morbidity, mortality), did not suggest that these were present at significant levels. An alternative 
explanation of the poor titer development in this experiment could be the innate immune system 
of the host. Broiler chickens have been genetically programmed towards high performance (fast 
growth, high feed efficiency). There is some evidence that this genetic selection has adversely 
affected some of the innate immune responses of broilers (Kirschermann et al, 2006)  and it could 
well be that it also has had a negative effect on the capacity of the modern broiler to produce 
antibodies. However, Ka Oud et al., (2008) reported titers of 25.2 and 26.2 after  vaccination of 
broiler chickens with an inactivated H5N1  vaccine given on day 7 and day 10 respectively, 
indicating that at least some broiler chicken strains are able to mount sufficient immune 
responses after vaccination. Differences in experimental set-up, vaccine manufacturer, vaccine 
dose or broiler strain might well be possible explanations for these differences in study results 
and it highlights the need for further studies. 
 
Conclusion 
One-time vaccination of broiler chickens on day 1, 7, 10 or 14 with an inactivated H5N1 vaccine 
did not result in mean levels of antibody titers which are considered to be protective. Negative 
results from the PCR tests and absence of clinical signs in the unvaccinated control group 
indicate there was no AI virus circulation at the study site which could have possibly affected the 
results of the experiment. More studies are needed to determine the optimum AI vaccination 
protocol for broilers in Indonesia. Ideally, these should be combined with challenge tests in order 
to obtain a more accurate assessment of the afforded protection against AI.  
 
References 
Ka Oud, H.A., M.A. Zakia, M.M. Kamel, 2008. Evaluation of Immune Response in AI Vaccinated 
Broiler: Effect of Biosecurity Fault on Immune Response. Int. J. Poult. Sci. 7(4):390-396. 
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An inactivated H5N2 vaccine reduces transmission of highly pathogenic H5N1 avian influenza virus among native chickens.

Association of serologic and protective responses of avian Influenza vaccines in chickens. Avian Dis.

Quantification of the effect of vaccination on transmission of avian influenza (H7N7) in chickens.

Transmission of the Highly Pathogenic Avian Influenza Virus H5N1 within flocks during the 2004 epidemic in Thailand.

Antibody responses to avian influeza vaccination in broiler chickens in Indonesia

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