Charles Rangga Tabbu

Isolation, identification, and serotyping of Avibacterium paragallinarum from quails in Indonesia with typical infectious coryza disease symptoms

Background and Aim Infectious 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 Methods Nine 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. Results Five 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. Conclusion Five 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.

Use of M2e ELISAs for longitudinal surveillance of commercial poultry in Indonesia vaccinated against highly pathogenic avian influenza

In countries where highly pathogenic avian influenza virus (HPAIV) H5N1 is endemic and controlled by vaccination, post-vaccination serological monitoring is essential to differentiate vaccinated poultry from those that are infected. The objectives of this study were to validate two experimental ELISAs that detect antibodies raised against the M2e protein of avian influenza virus that can be used for DIVA purposes. Results from the sM2e and tM2e ELISAs were compared with other conventional tests for the detection of H5N1influenza virus (virus isolation and RT-PCR) using samples collected from 16 commercial flocks in Indonesia. These comprised vaccinated layers aged between 18 and 68 weeks old that were sampled at ten-weekly intervals. A small number of sera were positive in sM2e and tM2e ELISA, 14 (0.6%) and 17 (0.7%) respectively, with low OD420 (0.1-0.3), but only 4 sera were positive in both tests. At the flock level, the incidence of M2e positive sera was low (4%), well below previously established minimum of 40% for an HPAIV H5N1-infected flock. Conventional M and H5 gene RT-PCRs indicated that none of 16 flocks were infected at any time during the study. No virus was isolated from any of the 480 pooled swab samples, except from one, for which the combined data analysis suggest to be the result of a laboratory cross-contamination. Clinical disease, mortalities or reduction in production performance, indicative of field H5N1 challenge, were not observed either in any of the flocks. Birds from two surveyed flocks, challenged in the laboratory with an Indonesian HPAIV H5N1 developed M2e antibodies in 50% and 55% of surviving birds with OD420 in the range of 0.35-1.47 in tM2e ELISA, confirming the validity of the criteria established for use of M2e ELISA for DIVA purposes. Overall these results showed that the tM2e ELISA could be a useful monitoring tool to ascertain freedom from H5N1 infections in vaccinated commercial poultry.

Isolasi dan Identifiicasi Serologis Virus Avian Influenza Dari Sampel Unggas Yang Diperoleh di D.I. Yogyakarta dan Jawa Tengah = Isolation and Serological Identification of Avian Influenza Virus From Poultry Sample ...

Avian Influenza (AI) merupakan penyakit penting pada unggas, karena dapat menyebabkan kerugian ekonomi secara signifikan, dengan tingkat morbiditas dan mortalitas penyakit sangat tinggi. L,ebih dan itu potensi penularan penyakit AI dari hewan ke manusia, memberikan dampak ekonomi tersendiri. Beberapa kasus yang diduga sebagai AI banyak mewabah di beberapa daerah di Indonesia. Penyakit tersebut cukup membingungkan peternak dan sangat dikacaukan dengan penyakit Newcastle (ND) karena kedua penyakit mempunyai kemiripan karakter dan gejala klinis. Penelitian ini bertujuan untuk mengkonfirmasi apakah wabah penyakit tersebut disebabkan oleh virus AI atau virus ND. Sampel isolasi diambil dari paru atau trakhea, kemudian diproses lebih lanjut untuk diisolasi, dipropagasi secara in ovo menggunakan telur ayam berembrio umur 9 sampai 12 hari, spesific pathogen free atau telur yang setidaknya bebas antibodi terhadap virus AI. Teknik isolasi menurut standar prosedur Office International des Epizooties (01E) dan kemungkinan adanya pertumbuhan virus diuji terhadap kemampuan mengaglutinasi sel darah merah ayam atau hemaglutinasi (HA). Uji HA positif, mengindikasikan ada pertumbuhan virus ND atau virus AL Kedua jenis virus tersebut dapat dibedakan dengan uji hemaglutinasi inhibisi (HI) menggunakan serum anti dari masing-masing virus yang diuji. Berdasarkan hasil penelitian ini dapat diambil suatu kesimpulan bahwa beberapa sampel unggas, yaitu: ayam petelur, ayam broiler, ayam kampung, dan burung puyuh, yang di peroleh dari beberapa daerah di D.I. Yogyakarta dan Jawa Tengah dan secara klinis menunjukkan gejala tersifat maupun tidak tersifat AI, secara serologis dapat dikonfirmasi sebagai virus avian influenza sub-tipe 145Ni.