Fowl adenovirus (FAdv) serotype 2 causes inclusion body hepatitis (IBH) disease which adversely affects the broiler industry in Thailand. We developed an indirect ELISA based on the recombinant hexon protein produced by E. coli. The recombinant hexon protein was tested with sera, in both infected and noninfected chickens. The recombinant hexon protein was standardized with an antigen concentration of 3.75 µg/ml and test sera. The intra- and inter-assays were repeatable. The cutoff value from TG-ROC curve analysis was 0.106. The specificity and sensitivity were 80 and 80%, respectively. The correlation coefficient (r) of absorbance values from this ELISA compared with the serum neutralization test was 0.76. This ELISA might be helpful for IBH diagnosis and surveillance.

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Background and Aim: Porcine epidemic diarrhea virus (PEDV) causes severe diarrhea in suckling piglets, leading to severe economic losses in the swine industry. Commercial vaccines have limited effectiveness against different genogroups of PEDV and the shedding of virus. The C-terminal of the S1 domain and the N-terminal of the S2 domain (S1-2) protein of the spike (S) protein have four neutralizing epitopes. However, research on the expression of the S1-2 segment of the S gene has been limited. In this study, we expressed a recombinant S1-2 protein of the S protein of the PEDV Thai isolate and characterized the immunological properties of the recombinant S1-2 protein. Materials and Methods: The S1-2 segment of the S gene of the PEDV Thai isolate (G2b) was amplified, cloned into the pBAD202/D-TOPO® vector (Invitrogen, Carlsbad, CA, USA), and expressed in Escherichia coli. The optimum concentration of arabinose and the optimum induction time for the expression of the recombinant S1-2 protein were determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The immunogenic reactivity of the recombinant S1-2 protein was determined using Western blot analysis with rabbit polyclonal antibodies against the SM98 strain of PEDV (G1a). Results: The recombinant S1-2 segment of the S gene of the PEDV Thai isolate protein was cloned and the recombinant S1-2 protein was successfully expressed. The optimum concentration of arabinose and the optimum induction time for the induction of the recombinant S1-2 protein were 0.2% and 8 h, respectively. The recombinant S1-2 protein reacted specifically with both rabbit anti-histidine polyclonal antibodies and rabbit anti-PEDV polyclonal antibodies. Conclusion: The recombinant S1-2 protein reacted with rabbit anti-PEDV polyclonal antibodies induced by the different PEDV genogroup. Therefore, the recombinant S1-2 protein may be a useful tool for the development of a diagnostic test for PEDV or for a vaccine against PEDV.

Snakeskin gourami (Trichogaster pectoralis) is a freshwater fish species that is being increasingly cultivated in Southeast Asia. The expansion of farms and intensive farming practices has led to the unexplained mortality of snakeskin gourami and tremendous economic losses in many farms. We investigated the unusual mortality of snakeskin gourami at 22 farms in Central Thailand. The moribund fish showed darkened skin, erratic swimming, exophthalmos, and haemorrhaging around the eyeballs, with cumulative mortality between 20% and 45%. Our necropsy findings revealed an enlarged liver and anterior kidney, splenomegaly, haemorrhage in most internal organs, pericarditis, and brain congestion. Histopathology revealed haemorrhaging and congestion of the blood vessels in the liver with infiltration of lymphocytes, enlarged blood vessels with mononuclear and lymphocyte infiltration in the meninges, and cerebral parenchyma were observed. Severe necrotising and suppurative pericarditis with myocardial infarction were found. Epidemiological studies and laboratory diagnosis revealed that Streptococcus agalactiae was predominantly isolated from the moribund fish. Laboratory investigations of the representative 33 isolates of S. agalactiae using mass spectrometry, multiplex polymerase chain reaction assay, pulse-gel electrophoresis, and serotyping suggested that all the isolates were S. agalactiae serotype VII, which is different from the serotype III isolated from diseased tilapia in Thailand. An experimental challenge using three representative isolates of S. agalactiae on snakeskin gourami caused clinical signs, gross lesions, and pathological changes, with high mortality exceeding 60%, which is similar to the mortality in most natural infections. Moreover, S. agalactiae was recovered from the spleen, kidneys, and liver of all the challenged fish. Taken together, this study provides important information that S. agalactiae serotype VII is virulent in snakeskin gourami and can potentially spread among these fish in fish farms. Appropriate preventive measures and the control of animal movements should thus be implemented.