Vaccination of Quails with Bivalent Inactivated H5N1 AI Vaccine (Clades 2.1.3 and 2.3.2) at Laboratory Scale

Quails, Coturnix sp, are commercially bred for meat and egg production in order to support the needs for animal protein. Cases of H5N1 Avian Influenza still occur sporadically at quail farms. Vaccination become an option as a precaution against possible exposure to H5N1 AI virus. Thirty quails were vaccinated with bivalent inactivated H5N1 AI vaccine (clade 2.1.3 and 2.3.2) and 10 quails were used as control group. The quails were vaccinated with one dose (0.3 ml) per bird intramuscularly at the age of 23 days and booster was done at the age of 45 days. The response after a single vaccination showed that antibody titers were not optimal, but after the booster vaccination the antibody titers showed 4.2 log2 in average against the H5N1 AI antigen of clade 2.1.3 and 3.7 log2 against the antigen of clade 2.3.2. A challenge test with H5N1 influenza virus either with clade 2.1.3 or clade 2.3.2 indicated a 70% protection. Nevertheless, viral shedding was detected ≥7 days post-challenge. As conclusion, vaccination with inactivated bivalent vaccine H5N1 AI clades 2.1.3 and 2.3.2 induced antibody that were was not homogenous nor optimal

HOST-RESTRICTED RANGE OF H5N1 AVIAN INFLUENZA VIRUSES ASSOCIATED WITH CHARACTERS OF POLYMERASE COMPLEX OF PB2 AND PB1-F2 PROTEINS

Epidemiological studies on H5N1 avian influenza viruses indi-cated that the viruses do not transmit efficiently from human to human. Transmissibility of viruses among human population is very complex and polygenic. Studies on molecular determinants facilitating interspecies transmission of the viruses suggested that two polymerase complex proteins such as PB2 and PB1-F2 are important. PB2 is critical in determining the host specificity, whereas mutations in PB1-F2 increase the viral virulence. The study aimed to characterize the polymerase complex of PB2 and PB1-F2 proteins of H5N1 avian influenza viruses isolated from Indonesia. The DNA samples encoding the PB2 and PB1-F2 complex proteins of several H5N1 isolates were sequenced and analyzed. Pathogenicity of the viruses was studied in both avian and mammal models. The sequencing results showed that there was no mutation in both proteins of PB2 and PB1-F2 of the avian influenza virus isolates. The molecular character for host specificity was consistent with the animal experiment results. The H5N1 virus isolates were only infectious and pathogenic in chickens, but not in BALB/C mice as the mammal model. The study suggests that host range of H5N1 virus isolates of Indonesia is restricted to poultry and not transmisible to mammal model used in this study

Studi Efikasi Vaksin Bivalen AI Isolat Lokal Terhadap Beberapa Karakter Genetik Virus AI Subtipe H5N1

Studi vaksin inaktif bivalen AI isolat lokal subtipe H5N1 terhadap beberapa karakter genetik virus AI H5N1 padaayam layer dan broiler. Vaksin inaktif bivalen dari isolat lokal AI A/Ck/West Java/Smi-M6/2008 and A/Ck/westjava/Pwt-D10-39/2010. Ayam layer dan broiler komersial divaksinasi dengan vaksin inaktif bivalen AI isolat lokal,setelah 3 minggu vaksinasi ditantang dengan virus AI A/Ck/West Java/Smi-Part/2006, A/Ck/West Java/Subang-JAPFA-29/2007 and A /Ck/West Java/Smi-Rahm2/2011. Ayam layer vaksinasi mendapat perlindungan dari morbiditas,mortalitas dan penurunan ekskresi virus tantang dengan tingkat proteksi 90-100% sedangkan ayam layerkontrol mati dalam waktu 2-3 hari, sementara broiler yang divaksinasi tidak mendapatkan perlindungan dari morbiditasdan mortalitas setelah terinfeksi virus AI tantang. Hasil studi memperlihatkan vaksin inaktif bivalen AI isolatlokal subtipe H5N1 mampu memberikan perlindungan pada ayam layer dari infeksi beberapa karakter genetikvirus AI subtipe H5N1

Host-restricted Range of H5n1 Avian Influenza Viruses Associated with Characters of Polymerase Complex of Pb2 and Pb1-f2 Proteins

Epidemiological studies on H5N1 avian influenza viruses indi-cated that the viruses do not transmit efficiently from human to human. Transmissibility of viruses among human population is very complex and polygenic. Studies on molecular determinants facilitating interspecies transmission of the viruses suggested that two polymerase complex proteins such as PB2 and PB1-F2 are important. PB2 is critical in determining the host specificity, whereas mutations in PB1-F2 increase the viral virulence. The study aimed to characterize the polymerase complex of PB2 and PB1-F2 proteins of H5N1 avian influenza viruses isolated from Indonesia. The DNA samples encoding the PB2 and PB1-F2 complex proteins of several H5N1 isolates were sequenced and analyzed. Pathogenicity of the viruses was studied in both avian and mammal models. The sequencing results showed that there was no mutation in both proteins of PB2 and PB1-F2 of the avian influenza virus isolates. The molecular character for host specificity was consistent with the animal experiment results. The H5N1 virus isolates were only infectious and pathogenic in chickens, but not in BALB/C mice as the mammal model. The study suggests that host range of H5N1 virus isolates of Indonesia is restricted to poultry and not transmisible to mammal model used in this study

Sirkulasi Virus Avian Influenza Subtipe H5n1 Di Pasar Tradisional Di Jawa Timur Tahun 2012 [Circulation of the Avian Influenza Virus Subtype H5n1 at Traditional Markets of East Java in 2012]

Avian influenza virus subtype H5N1 outbreak has become endemic in Indonesia since 2003. The disease does not only cause immense economic losses but it also leads to significant fatality of human being. The existence of traditional markets including live bird trading is suspected to play important role in the spreading and evolution of the virus. The objective of this study was to identify the circulation of H5N1 virus at traditional markets of East Java in 2012 by using reverse transcriptase polymerase chain reaction (RT-PCR) test and virus isolation. As results, this study detected the presence of the H5N1 virus circulating in Gresik, Mojokerto, Lamongan and Surabaya in both of live birds and environmental samples. The successfulness of virus isolation indicated a potential transmission to other hosts, including to human. This study suggests that the improvement of the poultry trading system at traditional markets by implementing sanitation, hygiene and biosecurity is necessary to reduce the burden of virus contamination at the market environment

Enzyme-Linked Immunosorbent Assay for Detection of Infectious Bronchitis Antibody in Chickens Using Local Isolate of PTS III

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for screening of antibody to avian infectiousbronchitis (IBV). Antigen was prepared from whole virus of infectious bronchitis local isolate PTS-III serotype.Optimum dilution with minimum background for antigen concentration, rabbit anti-chicken conjugate and sera indeveloped ELISA were determined 0.4μg/well, 1:2000 and 1:100, respectively. Correlation optical densities (OD)were compared with a standard commercial ELISA (R2=0.933). The developed ELISA has a better sensitivity to hemagglutinationinhibition (HI) test. The developed local isolate ELISA can be used to detect antibody against infectiousbronchitis virus and it is suitable for sample screening at the diagnostic laboratories

Potensi Transmisi Virus Avian Influenza Dari Babi Dan Unggas Pada Peternakan Babi Di Wilayah Tangerang, Provinsi Banten [the Potential of Transmission of Aavian Influenza Virus From Pig and Bird at the Pig Farm in Tangerang District, Banten Province]

Pigs have an important role in the ecology of influenza virus since they are sensitive to influenza viruses from human and avian origin. Influenza A virus has a host specificity, although not absolute, so most of the AI virus circulating in various species is only limited to the species, but sometimes there are interactions between different AI virus species or strain. Farming systems that combine a variety of animal species together in the same or around the sites have an important role in the spread of disease and transmission between species. This study is aimed to investigate the cycle of AI virus in order to determine the potential occurrence of viral transmission among species pig and bird at the pig farm that also raising poultry. Influenza virus was identified by methods of RT-PCR and qRT-PCR. The results showed that the novel H1N1 pandemic virus was detected in one pig farm in Tangerang (Banten Province). The AI/H5 virus is also detected in the pig farm that also raises poultry or poultry/pig farmers and located adjacent each other. The AI virus / influence A is also detected in most of the pigs. Detection of AI viruses that infected in pig farm which kept birds or poultry farm around the pigs farm had potential of AI virus transmission from birds species to pig or vise versa. The pigs could serve as a mixing vessel, thus providing opportunities likelihood of reassortant viruses

Efikasi Vaksin Inaktif Bivalen Avian Influenza Virus Subtipe H5N1 (Clade 2.1.3. Dan Clade 2.3.2) Di Indonesia

Status of avian influenza virus subtype H5N1 in Indonesia until 2014 is still endemic in poultry and recorded, there were two types clade of circulating H5N1 namely clade 2.1.3 and the new introduction of lade 2.3.2 since the end of 2012. Both of the clade of avian influenza viruses subtype H5N1 (clade 2.1.3 and 2.3.2) caused the the AI vaccination program to control of AI in poultry needs to be evaluated. In this study, we developed a bivalent AI vaccine (which contains clade 2.1.3 and 2.3.2 viruses as a seed vaccine) that adapted with the circulation of AI viruses in the field. Result of the study showed that the bivalent vaccine which developed in this study has good efficacy that was challanged with both of AI clade AI and proven to reduce shedding / viral contamination to the environment. It is expected that the development of bivalent H5N1 vaccine will increase the effectiveness and efficacy of vaccination programs to control highly pathogenic avian influenza disease in Indonesia

Cross-Reaction of Duck and Chicken Sera Against Avian Influenza H5N1 Virus Clades 2.1.3 and 2.3.2 Antigens by Hemagglutination Inhibition Test

This study aims to determine the cross-reaction between the antigen of avian influenza (AI) H5N1 virus clades 2.1.3 and 2.3.2 in duck and chicken sera, which were vaccinated with inactivated AI H5N1 clade 2.1.3 vaccine against AI H5N1 clade 2.3.2 antigen and those vaccinated with inactivated AI H5N1 clade 2.3.2 vaccine against H5N1 clade 2.1.3 antigen. The sera tested were obtained from postvaccination and control (unvaccinated) chickens and ducks in the laboratory condition, and from AI H5N1 postvaccination ducks in the field condition. HI test was conducted by using AI H5N1 clades 2.1.3 and 2.3.2 antigens. The results of HI titer were analyzed by the geometric means and by ANOVA. The results show that cross-reactions in both chicken and duck sera after AI H5N1 clade 2.3.2 vaccination tested with AI H5N1 clade 2.1.3 antigen occurred with low antibody titers, whereas in chicken and duck sera postvaccination with avian influenza H5N1 virus clade 2.1.3 showed cross-reaction with high antibody titer against clade 2.3.2 antigen. The conclusion of this study, postvaccination sera of AI H5N1 clade 2.1.3 provide better cross-reaction compared to the postvaccination sera of AI H5N1 clade 2.3.2