Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish

Khushiramani, Rekha; Girisha, S.K.; Karunasagar, Indrani; Karunasagar, Iddya

doi:10.1016/j.pep.2006.07.021

Cited by 56 publications

(35 citation statements)

References 15 publications

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“…Fang et al (2000) showed significant protection against 2 isolates of A. hydro phila in blue gourami Trichogaster trichopterus (75 and 87.5% relative percent survival, RPS) immunized with a recombinant 43 kDa OMP. The 37 kDa OMP of A. hydrophila was immunogenic in rohu carp (Khushiramani et al 2007). Among the different types of vaccine treatment in the present study, the C. auratus group treated with OMP and BF with adjuvant showed higher immunogenic performance including survival and decreased specific bacterial load.…”

Section: Discussionmentioning

confidence: 54%

Vaccination strategies to protect goldfish Carassius auratus against Aeromonas hydrophila infection

Viji¹,

Kumari²,

Velmurugan³

et al. 2013

Dis. Aquat. Org.

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Ornamental goldfish Carassius auratus were treated with whole cell (WC), extracellular product (ECP), outer membrane protein (OMP) and biofilm (BF) vaccines developed from the virulent Aeromonas hydrophila (AHV1; GenBank HQ331525.1) with and without the immunoadjuvant Asparagus racemosus. On various days post-vaccination (dpv), the treated fish were challenged with virulent A. hydrophila. These fish were monitored for survival, growth, specific bacterial reduction, and biochemical, haematological and immunological parameters. C. auratus attained 100% mortality within 7 d in non-vaccinated groups, whereas the vaccines helped to significantly (p ≤ 0.001) increase survival after 25 and 50 dpv. The vaccines with immunoadjuvant (ECP 2 , OMP 2 and BF 2 treatments) helped to reduce the Aeromonas load after the challenge, and serum albumin, globulin and protein levels were significantly (p < 0.01) improved in the OMP 2 -and BF 2 -treated groups. Haemoglobin and red blood cell counts were also significantly improved (p < 0.05) in the vaccinated groups compared to the control group. Additionally, haemagglutination occurred at the 1:12 dilution level in the vaccine plus immunoadjuvant-treated groups. Supplementing the vaccines with immunoadjuvant helped to improve phagocytosis to 54.07%, serum bactericidal activity to 14.6% and the albumin:globulin ratio to 7.6% in BF 2 after 50 dpv. Its positive effect significantly (p < 0.05) increased in vaccinated groups compared to controls. Based on the results, especially with the OMP and BF vaccines, the immunoadjuvant A. racemosus helped to improve the efficiency of the vaccines. This approach will aid in the development of more efficient vaccines against bacterial infections affecting the aquaculture industry. KEY WORDS:Ornamental fish · Biofilm · Outer membrane protein · OMP · Herbal immunoadjuvants Resale or republication not permitted without written consent of the publisher

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Section: Discussionmentioning

confidence: 54%

Vaccination strategies to protect goldfish Carassius auratus against Aeromonas hydrophila infection

Viji¹,

Kumari²,

Velmurugan³

et al. 2013

Dis. Aquat. Org.

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“…Some of the important attributes of a candidate molecule for vaccine development are that it should be highly conserved among various members of the same genus, should be expressed on the surface of pathogens so that antigen-presenting cells can easily recognize them, and should be immunogenic (Khushiramani et al 2007). Vaccination with OmpK and OmpW has been shown to be effective against Vibrio infections in fish (Qian et al 2007, Li et al 2008.…”

Section: Discussionmentioning

confidence: 99%

“…Antiserum against the isolated OmpU was prepared according to the method of Khushiramani et al (2007). This antiserum was used for analysis of antibody levels.…”

Section: Methodsmentioning

confidence: 99%

“…Control of vibriosis through antibiotic treatment is not always successful and could be hazardous for the environment since resistant bacteria may be selected and the resistance may be transferred to aquatic animal or human pathogens. Therefore, the development of vaccines to protect aquatic animals against vibriosis is important (Kumar et al 2007, Khushiramani et al 2007, Li et al 2008. Some virulence factors may contribute to the overall virulence of V. alginolyticus.…”

Section: Introductionmentioning

confidence: 99%

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Immune response in Lutjanus erythropterus induced by the major outer membrane protein (OmpU) of Vibrio alginolyticus

Cai¹,

Yao²,

Lu³

et al. 2010

Dis. Aquat. Org.

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The outer membrane proteins (OMPs) of the marine aquatic animal pathogen Vibrio alginolyticus play an important role in the virulence of the bacterium and are potential candidates for vaccine development. In this study, the major 35.6 kDa OMP of V. alginolyticus was isolated by gel excision from the crude OMP fraction from V. alginolyticus. The sequence of the first 27 amino acid residues from the N-terminal end of the protein is ATV YKD GGT ELL VGG RVE FRG DFI GSD, which has high homology with OmpU proteins from other Vibrio spp. (92%). Lutjanus erythropterus were vaccinated with OmpU, and immunogenicity was confirmed by subsequent western blotting. Enzyme-linked immunosorbent assay (ELISA) analysis demonstrated that OmpU produced an observable antibody response in all sera of the vaccinated fish. L. erythropterus vaccinated with OmpU produced specific antibodies, and were highly resistant to infection with virulent V. alginolyticus. These results indicate that OmpU is an effective vaccine candidate against V. alginolyticus for L. erythropterus. KEY WORDS: Vibrio alginolyticus · Outer membrane protein · OmpU · Immunogenicity · VaccineResale or republication not permitted without written consent of the publisher

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“…However, little is known about the virulence factors of E. ictaluri responsible for antibody effectiveness (Panangala et al, 2009b). Recent studies have revealed that OMPs of pathogenic bacteria may be effective vaccine candidates for protection against bacterial infections in fish (Kawai et al, 2004;Khushiramani et al, 2007;Li et al, 2010;Mao et al, 2007;Zhang et al, 2007). Notably, OMPs with epitopes exposed on the cell surface, such as fimbriae and porins, are easily recognized as foreign substances by the host immune system (Kawai et al, 2004).…”

Section: Vaccine Efficacy Of Glyceraldehyde-3-phosphate Dehydrogenasementioning

confidence: 99%

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

Cao¹,

Tsai²,

Yang

et al. 2014

J. Gen. Appl. Microbiol.

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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...

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Cloning and expression of an outer membrane protein ompTS of Aeromonas hydrophila and study of immunogenicity in fish

Cited by 56 publications

References 15 publications

Vaccination strategies to protect goldfish Carassius auratus against Aeromonas hydrophila infection

Vaccination strategies to protect goldfish Carassius auratus against Aeromonas hydrophila infection

Immune response in Lutjanus erythropterus induced by the major outer membrane protein (OmpU) of Vibrio alginolyticus

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

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