RESUMEN
Vibrio campbellii is a marine bacterium that is associated with luminous vibriosis, especially in the hatchery and nursery stages of penaeid shrimp cultivation worldwide, which has led to low survival rates of shrimp during aquaculture. Phage therapy has been reported as an alternative biocontrol agent which can reduce or replace the use of antibiotics and other chemicals. This study characterized a lytic V. campbellii bacteriophage, OPA17, originally isolated from bloody clams and investigated its biocontrol efficacy against V. campbellii infection in a model system, Artemia franciscana. Phage OPA17 lysed 83.89% of V. campbellii strains tested (n = 118) with clear plaque morphology. Some strains of Vibrio parahaemolyticus and Vibrio vulnificus were also infected by phage OPA17. Transmission electron microscopy and genetic features indicated that OPA17 belongs to the Siphoviridae family. The latent period and burst size of OPA17 were approximately 50 min and 123 PFU/cell, respectively. Moreover, it survived in artificial seawater throughout the 2-month study period and effectively destroyed Vibrio campbellii biofilms after 4 h of incubation. The addition of OPA17 significantly increased the survival of A. franciscana nauplii infected with V. campbellii. The genome sequence of OPA17 showed that it does not carry genes unsuitable for phage therapy. The phylogenetic tree analysis showed that OPA17 was closely related to the V. vulnificus lytic phage SSP002 (98.90% similarity), which was previously reported as a potential biocontrol agent. Accordingly, the results of this study provide valuable information regarding the potential biocontrol application of phage OPA17 against V. campbellii. IMPORTANCE V. campbellii is an emerging luminous pathogen associated with vibriosis, especially in marine shrimp hatcheries. Several strategies, including pond management and use of natural antimicrobials and probiotics, have been studied for control of this organism. Phage therapy is considered one of the effective biocontrol strategies against bacterial infections in aquaculture. However, there has been limited study of V. campbellii bacteriophages. In this study, V. campbellii-specific bacteriophage OPA17 was isolated, characterized, and investigated for its biocontrol efficacy against V. campbellii infection in an Artemia nauplii model. Phage OPA17 belongs to the Siphoviridae family and shares significant genome similarity to phage SSP002, a potential biocontrol agent against V. vulnificus infection in a murine model. However, the host range of OPA17 was broader than that of SSP002. Overall, we discuss the potential of OPA17 for phage therapy application in shrimp hatcheries.
RESUMEN
The number of patients with insulin-resistant diabetes has significantly increased. Thus, alternative insulin mimetics are required for such patients. Some evidences indicate that ribosomal protein L10a (RpL10a) is involved in the insulin pathway. In addition, we previously demonstrated that recombinant RpL10a from Fenneropenaeus merguiensis (Fm-RpL10a) could stimulate cell proliferation and trehalose metabolism in RpL10a-over-expressing flies by inducing insulin receptor (InR) expression and some insulin signaling mediators phosphorylation. In this study, we investigated the in silico binding between Fm-RpL10a and InR. The results indicated that Fm-RpL10a bound to InR at residues 635-640 and 697-702 of the FnIII2 domain. This binding was confirmed using a pull-down and immunofluorescence assay. Further analysis indicated that Fm-RpL10a could stimulate glucose utilisation by insulin-resistant cells (IRCs) and healthy cells. Additionally, Fm-RpL10a at a low concentration (1 µg/ml) altered some glucose metabolism-related genes expression in Fm-RpL10a treated IRCs. The qRT-PCR result revealed the up-regulation of Hk1, which encode key enzymes in glycolysis. Conversely, the expression of G6pc3, which participates in gluconeogenesis, was down-regulated. Overall, the results suggest that Fm-RpL10a can alleviate insulin resistance by stimulating insulin signaling via the FnIII2 domain of InR and activate glycolysis. Therefore, Fm-RpL10a may be a candidate insulin mimetic for the treatment of diabetes.
RESUMEN
Ribosome: machinery in control of messenger RNA's (mRNAs) and several ribosomal proteins are in the small and large subunit of the ribosome. Various aspects of ribosomal proteins have related to cell growth, cell cycle, and diseases. Ribosomal protein L10A (RpL10A) in shrimp and fruit fly has been demonstrated to play a role in oogenesis. Interestingly, deletion RpL10A gene (RpL10Ab-/-) germ line clone of the fruit fly showed a loss of follicle cells surrounding the egg chamber, but nurse cells appeared normal. This phenotype is reminiscent of insulin receptor mutants (InR-/-). In contrast, over-expression of RpL10A in the eyes of the fruit flies resulted in abnormal ommatidia with a loss of red pigment in the center of the eyes. In this study, the abnormal rearrangements of nuclei and lack of cell membranes in those eyes were demonstrated. Furthermore, the expression of InR gene and the InR protein were extensively increased as determined by real-time PCR and immunohistochemistry, respectively. In addition, some insulin signaling mediators were also detected. The Akt and FOXO proteins were highly phosphorylated in the RpL10A over-expressed mutant. The results revealed that RpL10A induced over-expression of the insulin receptor and consequently activated in insulin signaling pathway which affects cell proliferation and we suggest RpL10A stimulates cell proliferation via the insulin signaling pathway.
