Tilapia lake virus (TiLV) poses a major threat to global tilapia aquaculture and contributes to significant economic losses due to the absence of effective vaccines and treatments. Given the high mortality rates and severe pathological effects of TiLV on tilapia, alternative strategies, such as immunoglobulin-based therapies, are being considered for disease control. In this study, we developed specific immunoglobulin Y (IgY) antibodies against TiLV and evaluated their neutralization activity. Laying hens were immunized via intramuscular injections of recombinant TiLV segment 4 protein, and IgY antibodies were extracted and purified from their egg yolks using polyethylene glycol precipitation. Western blot analysis confirmed the specificity of the IgY, which demonstrated no cross-reactivity with nontarget proteins. Neutralization assays revealed a dose-dependent reduction in TiLV infectivity, which declined from 5.01 × 10⁶ TCID₅₀/mL to 5.01 × 10⁴–1.26 × 10⁵ TCID₅₀/mL, with the highest efficacy observed at a 1:2 dilution. Despite the variability in neutralization infectivity among the different hens, IgY effectively inhibited TiLV-induced cytopathic effects. Immunofluorescence assays further confirmed a significant reduction in the TiLV antigen levels in IgY-treated RHTiB cells. Our findings highlight IgY as a promising strategy for TiLV control and suggest its potential application in the prevention of emerging viruses.

Background and Aim: Immunoglobulin (Ig)Y, a specific type of Ig found in chicken eggs, has potential use in the diagnosis of human and animal diseases. This study assessed the feasibility of using spent laying hens to produce IgY. In addition, the effects of antigen injection on egg and antibody production in new and spent laying hens were compared. Materials and Methods: Hens were intramuscularly injected with three booster shots of antigens. IgY was extracted from egg yolks using polyethylene glycol 6000 precipitation followed by dialysis. Results: Spent laying hens (83 weeks) consistently showed lower egg production than new laying hens (27 weeks) throughout the study. Post-immunization, a further decline in egg production was observed in spent laying hens, and egg production stopped after the second antigen injection. Eggs from spent laying hens were less dense than eggs from new hens. Despite lower IgY levels in eggs from spent laying hens, the heavy-to-light chain ratio remained consistent in both groups. Notably, IgY from spent and new laying hens demonstrated effective hemagglutination against cat erythrocytes in the A blood group. Conclusion: This study demonstrated the potential of using spent laying hens to produce IgY, with significant implications for future research, immunotherapy, and diagnostic applications, despite the observed reduction in egg production compared with new laying hens. Keywords: antibody production, diagnosis, immunoglobulin Y antibody, immunological assays, spent laying hens.

Cholangiocarcinoma (CCA) is a heterogenous group of malignancies in the bile duct, which proliferates aggressively. CCA is highly prevalent in Northeastern Thailand wherein it is associated with liver fluke infection, or Opisthorchis viverrini (OV). Most patients are diagnosed in advanced stages, when the cancer has metastasized or severely progressed, thereby limiting treatment options. Several studies investigate the effect of traditional Thai medicinal plants that may be potential therapeutic options in combating CCA. Galangin is one such herbal flavonoid that has medicinal properties and exhibits anti-tumor properties in various cancers. In this study, we investigate the role of Galangin in inhibiting cell proliferation, invasion, and migration in OV-infected CCA cell lines. We discovered that Galangin reduced cell viability and colony formation by inducing apoptosis in CCA cell lines in a dose-dependent manner. Further, Galangin also effectively inhibited invasion and migration in OV-infected CCA cells by reduction of MMP2 and MMP9 enzymatic activity. Additionally, using proteomics, we identified proteins affected post-treatment with Galangin. Enrichment analysis revealed that several kinase pathways were affected by Galangin, and the signature corroborated with that of small molecule kinase inhibitors. Hence, we identified putative targets of Galangin using an in silico approach which highlighted c-Met as candidate target. Galangin effectively inhibited c-Met phosphorylation and subsequent signaling in in vitro CCA cells. In addition, Galangin was able to inhibit HGF, a mediator of c-Met signaling, by suppressing HGF-stimulated invasion, as well as migration and MMP9 activity. This shows that Galangin can be a useful anti-metastatic therapeutic strategy in a subtype of CCA patients.

The complete blood count (CBC) is a diagnostic test to analyze abnormalities of blood cells. Currently, automated hematology analyzers and artificial intelligence technology are being used with automated blood analyzers to ensure accuracy and reliability. This study aimed to evaluate the performance of artificial intelligence (AI) based automated blood cell analyzer, Awalife AI-100Vet Multifunctional Morphological Analyzer, in dog and cat blood samples by comparison with the CBC manual method. In dogs, PCV, hemoglobin, RBC, MCH, WBC, % Neutrophil, %Lymphocyte, %Monocyte, %Eosinophil and %Reticulocyte were all significantly correlated. While in cats, PCV, Hemoglobin RBC, WBC, % Neutrophil, % Lymphocyte, and % Eosinophil were all also significantly correlated. AUC values obtained by the Awalife AI-100Vet analyzer for Hematology testing in dogs and cats were 0.72 and 0.92 respectively. These findings suggest that the Awalife AI-100Vet analyzer demonstrated good accuracy using dog blood for hematology testing as well as excellent accuracy when using cat blood. The AI-based automated blood analyzer has the potential to analyze hematological data and is close to the reference method. However, there are still differences in some parameters. Further optimization of the AI algorithm, which will involve increasing the accuracy of identifying unusual cell shapes, improving stability against various samples, such as stains, and achieving good results when working with unique pathologies, should be carried out.