Molecular characterization and immunohistochemical localization of tilapia piscidin 3 in response to Aeromonas hydrophila infection in Nile tilapia
| Autor: | Alkhateib Y. Gaafar, Dalia M. Mabrouk, Mohamad M. Aboelenin, Mohamed S. Hassanane, Heba A.M. Abd El-Kader, Mahmoud A. Mahmoud, Wagdy K. B. Khalil, Karima F. Mahrous, Abdelgayed M. Younes |
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| Rok vydání: | 2020 |
| Předmět: |
food.ingredient
Microbial Sensitivity Tests Biology 010402 general chemistry 01 natural sciences Biochemistry Fish Diseases Nile tilapia Minimum inhibitory concentration food Structural Biology Drug Discovery Gene expression Animals Molecular Biology Pharmacology chemistry.chemical_classification 010405 organic chemistry Aquaculture of tilapia Organic Chemistry Tilapia Cichlids General Medicine Antimicrobial biology.organism_classification Immunohistochemistry Molecular biology Aeromonas hydrophila Anti-Bacterial Agents 0104 chemical sciences Amino acid chemistry Molecular Medicine Gram-Negative Bacterial Infections Antimicrobial Cationic Peptides |
| Zdroj: | Journal of Peptide Science. 26 |
| ISSN: | 1099-1387 1075-2617 |
| DOI: | 10.1002/psc.3280 |
| Popis: | The antimicrobial activity of tilapia piscidin 3 (TP3) was determined in vitro against a locally isolated Aeromonas hydrophila. A 388 bp fragment was amplified from the TP3 cDNA and sequenced. The coding sequence (CDS) of TP3 was estimated to be 231 bp codes for 76 amino acids long and stop codon. In silico analysis was performed to detect both the signal peptide and the prodomain cleavage sites to follow the amino acids number 22 and 70, respectively. Based on this, a peptide 23 amino acids long with a remarkably high computed antimicrobial probability was synthesized and used in the subsequent experiments. The antimicrobial activity of TP3 was determined with minimum inhibitory concentration (MIC) and minim um bactericidal concentration (MBC) methods. TP3 exhibited relatively weak antimicrobial activities against the tested bacteria. A challenge experiment was then performed in Nile tilapia with low and high doses of A. hydrophila, followed by timely recognition; after 3, 6, 24 h, and 7 days of the specific TP3 gene expression, immunohistochemical localization was also performed. Histopathological examination revealed provoked inflammatory responses and congestion in the same organs of TP3 expression. Immunohistochemical localization showed that A. hydrophila induced tilapia fish to express TP3 after 24 h within the gills, intestine, hepatopancreas, spleen, and posterior kidney. In quantitative real time (RT)-polymerase chain reaction analysis, the high dose showed higher mRNA expression levels than the low dose, and its expression levels increased in the A. hydrophila-infected fish. It was therefore concluded that TP3 plays an essential role in fish immunity. |
| Databáze: | OpenAIRE |
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