A Multiplex PCR Method for Simultaneous Detection of Infectious Laryngotracheitis Virus and Ornithobacterium rhinotracheale.

To date, many fluorescence- and gel-based multiplex polymerase chain reaction (PCR) assays have been developed for the simultaneous detection of multiple infectious agents of respiratory disease in poultry. However, PCR assays are not available for other important emerging respiratory bacteria, such as Ornithobacterium rhinotracheale (ORT). We aimed to fill this gap by establishing a new duplex PCR method for the simultaneous detection of infectious laryngotracheitis virus (ILTV) and ORT. Multiplex primer design software was used to select the compatible multiplex primer pairs. It was determined that an annealing temperature of 65 °C and an initial concentration of 2.5 pmol/µL for each primer set were the most suitable conditions for multiplex PCR. The assay was confirmed to be specific, as it only detected the target pathogens, even in the presence of six non-target agents. The limit of detection was up to 103 copies/µL of template DNA for both ILTV and ORT. In the screening of 304 field samples, 23, 88, and 44 were positive for both ILTV and ORT, solely for ILTV, and solely ORT, respectively.

Dynamic Models of Within-Herd Transmission and Recommendation for Vaccination Coverage Requirement in the Case of African Swine Fever in Vietnam.

African swine fever (ASF) is a highly contagious disease that is caused by the ASF virus (ASFV) with a high fatality rate in domestic pigs resulting in a high socio-economic impact. The pig business in Vietnam was recently affected by ASF for the first time. This study thus aimed to develop a disease dynamic model to explain how ASFV spreads in Vietnamese pig populations and suggest a protective vaccine coverage level required to prevent future outbreaks. The outbreak data were collected from ten private small-scale farms within the first wave of ASF outbreaks in Vietnam. Three methods were used to estimate the basic reproduction number (R0), including the exponential growth method, maximum likelihood method, and attack rate method. The average R0 values were estimated at 1.49 (95%CI: 1.05-2.21), 1.58 (95%CI: 0.92-2.56), and 1.46 (95%CI: 1.38-1.57), respectively. Based on the worst-case scenario, all pigs in a herd would be infected and removed within 50 days. We suggest vaccinating at least 80% of pigs on each farm once a commercially approved ASF vaccine is available. However, an improvement in biosecurity levels in small-scale farms is still greatly encouraged to prevent the introduction of the virus.

Serological and Molecular Characterization of Avian Metapneumovirus in Chickens in Northern Vietnam.

Avian Metapneumovirus (aMPV) is a causative agent of respiratory disease complex in turkeys and chickens that has recently been detected in Vietnam. Due to its novelty, this study was conducted to elucidate the distribution of aMPV in several provinces in northern Vietnam. By the application of Enzyme-Linked Immunosorbent Assay (ELISA) and nested Reverse Transcription-Polymerase Chain Reaction (RT-PCR), this study demonstrated the circulation of aMPV in 12 out of 14 cities/provinces with positive rates of 37.6% and 17.2%, respectively. All nested RT-PCR positive samples were aMPV subgroup B. By pairing the detection results with age groups, it was observed that aMPV infections occurred in chickens of all ages. Additionally, by genetic characterization, aMPV strains were demonstrated to not be attenuated vaccine viruses and to belong to at least two genetic clades. Overall, the obtained results provided insights into the prevalence of aMPV and indicated a greater complexity of respiratory diseases in chickens in Vietnam.