Thioredoxin fused CHH1 protein as antigen for polyclonal antisera: Application to regulate glycemia in Penaeus monodon

Crustacean hyperglycemic hormone (CHH) family neuropeptides have been in research limelight for the past two decades due to their importance in the regulation of glycemia, moulting and gonad development in crustaceans. Under natural conditions, low levels of CHH neuropeptide and the structural similarity of the three CHH family neuropeptides limit their purification directly from the animal. In this study, we isolated the mature region of the CHH1 gene, constructed the recombinant translation expression vector (pET32a+ - PmCHH1) and produced thioredoxin fused protein in E scherichia coli (BL21 (DE3) pLysS). The translation expression vector construct (pET32a+ - PmCHH1) was successfully built for production of thioredoxin fused mature CHH1 protein (mf-PmCHH1-29.47 kDa). Mf-PmCHH1 produced a hyperglycemic effect similar to that of the eyestalk extract when experimentally injected into adult eyestalk ablated Penaeus monodon. The polyclonal antiserum (anti-mf-PmCHH1) was developed in mice against the purified thioredoxin fused mf-PmCHH1 protein. A hypoglycemic effect was induced in adult P. monodon by the polyclonal antiserum which was raised against thioredoxin fused CHH1 protein. Immunolocalization of CHH1 producing neurosecretory cells in the eyestalk of P. monodon was a practical result obtained with the polyclonal antiserum anti-mf-PmCHH1. Therefore, mf-PmCHH1 and its antiserum (anti-mf-PmCHH1) are added to the list of tools to better understand the endocrine mechanisms regulating glycemia and reproduction in P. Monodon

Evaluation of Direct Rapid Immunohistochemical test (DRIT) of Canis lupus familiaris hippocampal touch impression smears using a monospecific polyclonal antibody for rabies virus detection

@#<p style="text-align: justify;"><strong>BACKGROUND:</strong> Rabies is an important zoonotic disease that needs to be eradicated worldwide. It is still prevalent in the Philippines, thus development of a relatively affordable but still accurate and rapid post-mortem detection test for the virus is desired, especially in regional laboratories.<br /><strong>METHODS:</strong>The study evaluated the Direct Rapid Immunohistochemical Testing (DRIT) of hippocampal touch impressions of suspected rabid Canis lupus familiaris using monospecific N protein polyclonal antibody developed by the Research Institute for Tropical Medicine (RITM). One hundred sixty (160) acetone-fixed hippocampal touch impressions were subjected DRIT.<br /><strong>RESULTS:</strong> One hundred thirteen (70.6%) out of 160 samples tested positive for rabies viral antigen (RVA) and 47 (29.4%) out of 160 samples tested negative for RVA. No false positive and false negative results were obtained. The results agree with the gold standard, dFAT.<br /><strong>CONCLUSION:</strong> DRIT was able to detect low to high concentrations of RVA in the hippocampal touch impressions based on the grading distribution. DRIT had 100% sensitivity, specificity and over-all accuracy using monospecific polyclonal antibodies, which suggests its use as a more affordable alternative to the gold standard dFAT.</p>