Although vaccination of poultry for control of highly pathogenic avian influenza virus (HPAIV) H5N1 has been practiced during the last decade in several countries, its effectiveness under field conditions remains largely unquantified. Effective HPAI vaccination is however essential in preventing incursions, silent infections and generation of new H5N1 antigenic variants. The objective of this study was to asses the level and duration of vaccine induced immunity in commercial layers in Indonesia. Titres of H5N1 haemagglutination inhibition (HI) antibodies were followed in individual birds from sixteen flocks, age 18–68 week old (wo). The study revealed that H5N1 vaccination had highly variable outcome, including vaccination failures, and was largely ineffective in providing long lasting protective immunity. Flocks were vaccinated with seven different vaccines, administer at various times that could be grouped into three regimes: In regime A, flocks (n = 8) were vaccinated two or three times before 19 wo; in regime B (n = 2), two times before and once after 19 wo; and in regime C (n = 6) three to four times before and two to three times after 19 wo. HI titres in regime C birds were significantly higher during the entire observation period in comparison to titres of regime A or B birds, which also differed significantly from each other. The HI titres of individual birds in each flock differed significantly from birds in other flocks, indicating that the effectiveness of field vaccination was highly variable and farm related. Protective HI titres of >4log2, were present in the majority of flocks at 18 wo, declined thereafter at variable rate and only two regime C flocks had protective HI titres at 68 wo. Laboratory challenge with HPAIV H5N1 of birds from regime A and C flocks confirmed that protective immunity differed significantly between flocks vaccinated by these two regimes. The study revealed that effectiveness of the currently applied H5N1 vaccination could be improved and measures to achieve this are discussed.

Background and AimInfectious coryza (IC) or snot is an infectious upper respiratory disease affecting chickens and birds, including quails, and it is caused by Avibacterium paragallinarum. The symptoms of IC are facial swelling, malodorous nasal discharge, and lacrimation. This study aimed to isolate, identify, and serotype the A. paragallinarum of snot in quails and to determine the sensitivity and resistance to several antibiotics.Materials and MethodsNine quails from Yogyakarta, Indonesia with typical snot disease symptoms were used in this study. The nasal swab was obtained and directly streaked onto a chocolate agar plate and blood agar plate (BAP), then incubated in 5% CO2 at 37°C for 24-48 h. Staphylococcus spp. was cross-streaked onto the BAP to show the satellite growth. The observation of the morphology of the suspected colony, Gram staining, and biochemical tests (catalase test, oxidase test, urease test, peptone test, and carbohydrate fermentation such as maltose, mannitol, lactose, and sorbitol) are done to identify the species of bacteria. This research also detects the serovar of A. paragallinarum using hemagglutination inhibition test.The antibiotic sensitivity tests were also performed using several antibiotics against five A. paragallinarum isolates that were cultured on Mueller–Hinton Agar and added with antibiotic discs, then incubated in 5% CO2 at 37°C for 24–48 h.ResultsFive isolates out of nine suspected isolates (55.5%) were A. paragallinarum. The growth of isolates from quails did not depend on the nicotinamide adenine dinucleotide (NAD) (NAD-independent). Sensitivity test was done using the five identified A. paragallinarum isolates, results showed that they were 100% sensitive to amoxicillin (AMC) and ampicillin (AMP); 100% resistant toward amikacin (AK), erythromycin (E), gentamycin (CN), and tetracycline (TE); 80% resistant toward kanamycin (K) and trimethoprim (W); 60% resistant toward chloramphenicol (C); and 20% toward enrofloxacin (ENR). The antibiotics that have an intermediate sensitivity (in between sensitive and resistant) were ENR and K, 80% and 20%, respectively. Three out of five A. paragallinarum isolates were identified as serovar B of A. paragallinarum using HI test.ConclusionFive out of nine isolates (55.5%) from quails with typical IC disease symptoms identified as A. paragallinarum and sensitive toward AMC and AMP. Three out of five A. paragallinarum isolates were identified as serovar B.

To help guide surveillance and control of highly pathogenic avian influenza subtype H5N1 (H5N1-HPAI), the Food and Agriculture Organization of the United Nations in 2004 devised a poultry farm classification system based on a combination of production and biosecurity practices. Four "Sectors" were defined, and this scheme has been widely adopted within Indonesia to guide national surveillance and control strategies. Nevertheless, little detailed research into the robustness of this classification system has been conducted, particularly as it relates to independent, small to medium-sized commercial poultry farms (Sector 3). Through an analysis of questionnaire data collected as part of a survey of layer farms in western and central Java, all of which were classified as Sector 3 by local veterinarians, we provide benchmark data on what defines this sector. A multivariate analysis of the dataset, using hierarchical cluster analysis, identified three groupings of the farms, which were defined by a combination of production-and biosecurity-related variables, particularly those related to farm size and (the lack of) washing and disinfection practices. Nevertheless, the relationship between production-related variables and positive biosecurity practices was poor, and larger farms did not have an overall higher total biosecurity score than small or medium-sized ones. Further research is required to define the properties of poultry farms in Indonesia that are most closely related to effective biosecurity and the prevention of H5N1-HPAI.

We report the complete genome sequences of 11 virulent Newcastle disease viruses. The isolates were obtained from vaccinated broiler and layer chickens in three different provinces of Indonesia in 2013 and 2014. Phylogenetic analysis revealed that all isolates belong to subgenotype VII.2 in the class II cluster.

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Kasus penyakit Infectious Bursal Disease (IBD) dewasa ini masih sering ditemukan pada peternakan ayam komersial baik layer maupun broiler di Indonesia. Diagnosis penyakit IBD sejauh ini mengandalkan lesi patologik spesifik dan kultur in ovo dengan mengamati lesi makroskopis embrio, serta diidentifikasi dengan uji agar gel presipitasi (AGP). Penelitian ini bertujuan menerapkan diagnosis dengan teknik reverse transcriptase polymerase chain reaction (RT-PCR) dari sampel Bursa Fabrisius (BF) sebagai konfirmasi pada kasus terdiagnosa IBD. Deteksi serologis virus IBD dengan uji AGP dengan sumber antigen chorioallantoic membrane (CAM) dan embrionya, untuk melihat potensinya sebagai sumber antigen uji AGP. Sampel BursaFabrisius sebanyak 5 yang diperoleh pada kasus terdiagnosa IBD, dikoleksi dari peternakan ayam komersial di Yogyakarta. Konfirmasi diagnosis dilakukan dengan metode RT-PCR. Sampel positip uji RT-PCR yang mengamplifikasi fragmen gen VP2. Isolasi virus IBD yang dilakukan kultur in ovo pada telur ayam berembrio (TAB) antibodi negatif terhadap virus IBD, berumur 11 hari. Desposisi materi inokulasi dilakukan pada (CAM), diinkubasi selama lima hari. Panen virus dilakukan dengan mengkoleksi membran korioalantois dan embrio, selanjutnya diamati lesi makroskopis yang timbul akibat infeksi virus IBD. Membran korioalantois dan embrioselanjutnya digerus dan diproses sebagai suspensi antigen yang digunakan dalam uji AGP. Hasil uji RT-PCR terhadap lima sampel Bursa Fabrisius yang dikoleksi dari peternakan ayam terdiagnosa penyakit IBD, tiga sampel menunjukkan hasil positif teramplifikasi fragmen gen VP-2 virus IBD dengan produk amplifikasi sebesar 440 bp, sedangkan dua sampel sisanya menunjukkan hasil negatif. Uji AGP dengan sumber antigen CAM menunjukkan hasil positip 2 dari 3 sampel yang diuji, sedangkan sumber antigen embrio menunjukkanhasil negatif. Berdasarkan data yang diperoleh dalam penelitian ini dapat disimpulkan bahwa uji RT-PCR dapat digunakan dalam mendeteksi virus IBD dari sampel BF terdiagnosa IBD. Uji AGP dengan sumber antigen CAM menunjukkan hasil lebih baik dari pada embrionya.

Avian influenza virus is a poultry viral disease, which causes high economic losses. Various efforts have been made to control the disease. One effort required fast and accurate screening diagnostic test. This study aimed to determine the potential of a rapid test kit, namely AIV/H5 Anigen Rapid Test for the detection of AI virus types A and subtype H5 in the fields. Some tests were carried out, e.g. the potential test, crossreaction test, sensitivity and specificity test. The potential test was done to evaluate detection limits of the kit, by having the test of a serial dilution of AI virus positive control. Crossreaction test was done to detect antigens other than AI virus H5N1, e.g. IB virus of Massachusetts strain, IBV strain 491, Newcastle Disease virus, and Escherichia coli. Sensitivity and specificity test were applied to the field samples which clinically and laboratory were confirmed as H5N1 positive. To confirm the result of rapid test was then being done by reverse transcriptase polymerase chain reaction. Based on these results it can be concluded that Anigen Kit AIV/H5 Ag Rapid Test can detect antigencontaining samples having AI virus HA titer up to 2 6 of type A virus, and up to 2 5 for subtype H5 virus. Anigen Kit AIV/H5 Ag Rapid Test showed no crossreactions with Infectious Bronchitis virus, Newcastle Disease virus, and Escherichia coli. Anigen A Rapid Test Kit AIV Ag showed a sensitivity of 50% and specificity of 100%, while Anigen Ag Rapid Test Kit AIV/H5 showed a sensitivity of 25% and specificity of 100%.

Background: Pyocyanin (PCN), a highly pathogenic pigment produced by Pseudomonas aeruginosa, induces caspase 3-dependent human B cell (Raji cells) death. The aim of the present study, therefore, was to assess whether antigen-specific IgY antibodies may be protective on PCN-induced Raji cell death. Methods: Chickens were subcutaneously immunized with Freund's complete adjuvant containing PCN, and then given two boosted immunizations.  Anti-PCN IgY antibodies were purified from egg yolk and detected using an agar gel precipitation test (AGPT) and ELISA. Protective effects of antigen-specific IgY on Raji cells were tested using a cell viability assay. Results: AGPT results showed the formation of strong immune complex precipitates, whilst ELISA further confirmed the presence of IgY antibodies specific to PCN at significant concentration. Further experiments showed that anti-PCN IgY antibodies significantly increased PCN-treated Raji cell viability in a dose-dependent fashion (p<0.05). Conclusions: The results of the present study suggest that anti-PCN IgY antibodies may be protective on PCN-induced Raji cell death.