In silico analysis of PirA- and PirB-like toxin genes of Vibrio spp., present in Asia and Costa Rica.

OBJECTIVE: Acute hepatopancreatic necrosis disease is an emerging infectious disease of Penaeus species. The causative agent is Vibrio species, which dispels binary toxin similar to pirA and pirB, which causes mortality in infected shrimp. The aim of this research was to investigate the evolutionary relationship of pirA and pirB homologous genes present in this Asia and Costa Rica in silico. MATERIALS AND METHODS: The sequences for in silico analysis were all retrieved from the Basic Local Alignment Search Tool Nucleotide (BLASTN) tool of the National Center for Biotechnology Center. For pirA, a total of 25 sequences submitted from different Asian countries and Costa Rica were retrieved for analysis. Meanwhile, for pirB, a total of 11 sequences submitted from five Asian countries were retrieved. Sequences were aligned using the CLUSTAL W alignment tool under Molecular Evolutionary Genetics Analysis (MEGA) 7 software. The evolutionary history was then estimated using the Unweighted Pair Group Method with Arithmetic mean (UPGMA) method, whereas the evolutionary distances were determined using the maximum composite likelihood model with 1,000 bootstrap replications. RESULTS AND DISCUSSION: The results show that, among 27 DNA sequences analyzed for pirA gene, three groups were generated, while for pirB, 13 DNA sequences yielded only one group. The analysis revealed low genetic variation among isolates for both pirA and pirB genes. CONCLUSION: This result suggests that the low frequency of polymorphism and geographic location cannot be attributed to the differences in V. parahaemolyticus isolates in Asian countries relative to Costa Rican isolates in pirA and pirB genes.

Comparative molecular characterization of Forkhead box protein 3 (FoxP3) gene of swamp-type (Bubalus carabanensis) and riverine-type (Bubalus bubalis) water buffaloes.

FoxP3 is a forkhead family member that plays an important role in the development and function of a type of CD4 + T cell called T regulatory cells. Molecular characterization of FoxP3 gene in swamp- and riverine-type water buffaloes was conducted to determine its homology and compare it to the FoxP3 gene of other animal species (cattle, goat, sheep, horse, pig, cat, and dog), determine its unique characteristics in water buffaloes, and provide a reference for future studies to analyze its immunological function. FoxP3 nucleotide sequence of swamp- and riverine-type water buffaloes was 99% identical, whereas its protein translation revealed 97% homology. FoxP3 of swamp- and riverine-type water buffaloes were compared to FoxP3 of other animal species and revealed a high degree of homology which suggests that they may have the same biological properties. This study is the first report that describes the genetic characteristic of FoxP3 gene in water buffalo.

Serological and molecular evaluation of Mycobacterium avium subspecies paratuberculosis (Johne's disease) infecting riverine-type water buffaloes (Bubalus bubalis) in the Philippines.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease and a possible cause of Crohn's disease in humans. A total of 70 blood and fecal samples were collected from water buffaloes in selected municipalities of Nueva Ecija for ELISA and qPCR assay. Results revealed presence of antibodies of MAP in 3 serum samples for ELISA. The qPCR assay was carried out using standard curve method targeting the MAP specific insertion element IS900. Results revealed that 10 of the samples were positive for MAP DNA in qPCR. ELISA was able to detect antibodies for MAP showing 2.48% infection rate among the 70 buffaloes tested using blood serum samples. On the other hand, qPCR was able to detect MAP using IS900 showed 14.28% infection rate among buffaloes tested using fecal samples. Nucleotide sequence of isolated MAP showed high homology (99-100%) among the reported MAP isolates in the GenBank.