Vaccine efficacy of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Edwardsiella ictaluri against E. tarda in tilapia

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAP-DH), derived from the outer-membrane protein (OMP) fraction, has been used as a potential candidate for vaccine development. The gene-encoding 37 kDa GAPDH outer membrane protein (OMP) from Edwardsiella ictaluri was amplified using polymerase chain reaction (PCR) and was cloned and expressed in Escherichia coli BL21 (DE3). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting, and nucleotide and amino acid sequencing were used to analyze the expressed antigenic protein and gene encoding this protein. Comparative DNA and protein sequence analysis of GAPDH from E. ictaluri GAPDHs from several Gram-negative bacterial species within the Enterobacteriaceae family revealed that the GAPDHs within this group are highly conserved and share a sequence similarity of 75 100% with E. ictaluri GDPDH. Rabbit antiserum raised against the E. ictaluri recombinant GAPDH (rGAPDH) protein recognized purified GADPH, indicating that it has a strong immunogenicity. Tilapia fish were intraperitoneally immunized with formalin-killed E. ictaluri whole cells, and rGAPDH (30 µg fish 1 ) from E. ictaluri, both of which were emulsified in ISA 763A adjuvant. At 3 months after immunization, fish were challenged with the E. tarda strain to assess vaccine efficacy; the relative percent survival (RPS) values were found to exceed 71.4%. The specific mean antibody titer log 2 level of groups vaccinated with rGAPDH at 3 months was significantly higher than that of non-vaccinated fish (control group). Therefore, this recombinant protein can be considered a multi-purpose candidate vaccine against several pathogenic bacteria.Key words: channel catfish; Edwardsiella ictaluri; enteric septicemia of catfish; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) IntroductionEnteric septicemia of catfish (ESC), caused by the bacterial pathogen Edwardsiella ictaluri, was first identified in channel catfish (Ictalurus punctatus) by Hawke (1979). E. ictaluri has become the most lethal infectious disease of channel catfish and other ictalurids in the USA (Plumb, 1999). More than 50% of American catfish fry/fingerling losses are due to ESC (Williams and Lawrence, 2010). E. ictaluri was also isolated from cultured fish species in Indonesia, Vietnam, Japan, Turkey (Crumlish et al., 2002;Keskün et al., 2004;Nagai et al., 2008; Yuas et al., 2003) and China (Ye et al., 2009). ESC is currently controlled using antibiotics; however, extensive use of antimicrobial com pounds reduces their effectiveness by increasing the number of antibiotic-resistant bacterial strains, resulting in the need for higher dosages for effective treatment as well as higher residual amounts of drugs in the harvested fish Thompson and Adams, 2004). Vaccines that protect fish against bacterial diseases are an important alternative to antibiotics and are a safer strategy for disease prevention. Most vaccines for controlling ESC have been tested using Full Paper Vaccine efficacy of glyceraldehyde-3-phosphate dehydrogenase (G...

Synergism Between Bacterial GAPDH and OMVs: Disparate Mechanisms but Co-Operative Action

“…Bacterial GAPDH has already proven useful as a therapeutic target with the development of cross-protective GAPDH-based vaccines against Gram-negative and -positive bacteria for agri- and aqua-culture (Li et al, 2011 ; Vanden Bergh et al, 2013 ; Velineni and Timoney, 2013 ; Trung Cao et al, 2014 ). The GAPDH inhibitor pentalenolactone (Cane and Sohng, 1994 ) is known to act as an antibiotic due to its disruption of bacterial glycolysis, but it is also potent against mammalian homologs and is not used in the clinic.…”

State-of-the-Art Vaccine Research for Aquaculture Use: The Case of Three Economically Relevant Fish Species

“…This field has progressed significantly in the last decades: many vaccine types were developed (i.e., attenuated, inactivated, subunit, recombinant, and DNA) [17], and all proved at least partially effective against some pathogens. Many of the discussed studies produced encouraging results, achieving very high survival rates and specific antibody titers in challenge trials, even though it must be remembered that the correlates between antibody quantity/functional characteristics and induced immunity may be poor [106], as found in several studies [20,24,45,49,52,53,58]: a vaccine efficacy should never be solely investigated by means of serological assays.…”

“…Two promising vaccine candidates were developed against Edwardsiella tarda. Cao et al [49] used a recombinant GAPDH protein derived from the outer membrane protein fraction of the congener E. ictaluri as a potential candidate for vaccine development and compared it to a whole-cell formalin-inactivated formulation. Both recombinant and inactivated formulations were emulsified with Montanide TM ISA 763A VG and IP administered to 102 g BW specimens.…”

Comparative immunohistological study on using capsaicin, piperine, and okadaic acid for the transepithelial passage of the inactivated viral and bacterial vaccines in fish

“…Many research was done on injectable E. tarda killed vaccines, including ISA 763Aadjuvantedformalin-killed E. ictaluri whole cells exerted protection against virulent E. tarda in tilapia (Cao et al, 2014), also a killed, but metabolically active (KBMA) E. tarda vaccine resulted in protection in olive flounder (Choi, & Kim, 2015). Subunit vaccines such as recombinant rFimA (Wang, et al, 2013), recombinant vaccine rGAPDH (Liang et al, 2012) and recombinant FlgD protein (Zhang et al, 2012)induced protection in vaccinated fish, also DNA vaccine encoding molecular chaperone GroEL induced 60% survivability (Liu, et al, 2016).…”