552
administered i.p. (RPS = 92.3 %) and orally (RPS = 61.2 %) (Sun et al. 2009 ). Pang
et al. ( 2010 ) used OmpN mixed with FCA to vaccinate via i.p. (100 μl/fi sh), estuary
cod ( Epinephelus coioides ), that survived challenge after 28-days with a virulent
culture (RPS = 60 and 70 % depending on the nature of the challenge strain).
Meanwhile, Zhang et al. ( 2011a ) cloned, sequenced and characterised two OMP
genes, OmpK and GADPH, and expressed the recombinant proteins in the prokaryotic expression vector pET-30a(+), which were purifi ed and used to vaccinate intraperitoneally (100 μg/fi sh) large yellow croaker ( Pseudosciaena crocea ) with booster
doses after 3-weeks. The outcome was that r-OmpK and r-GADPH led to RPS values of 37.7 % and 40 %, respectively, after challenge. OmpK was expressed in yeast,
Pichia pastoris, and fed for 5 days to sea bass ( Lateolabrax japonicus ) on alginate
microspheres with a resultant production of antibodies, and protection against challenge (RPS = 61.5 %) (Mao et al. 2011 ). A denatured inactive cytotoxic 530 amino
acid recombinant secreted protease, Vhp1, was recovered from a pathogenic isolate
was determined to be an effective subunit vaccine (RPS = 70 %) with improved performance when expressed in Esch. coli as a live vaccine (RPS = 90 %) (Cheng et al.
2010 ). In a further study, Wang et al. ( 2011 ) used the purifi ed 35 kDa outer membrane protein OmpU (50 μg dose) and a DNA vaccine involving the insertion of the
ompU gene into pEGFP-N1 plasmid (10 μg) injected i.m. in turbot. Use of the purifi ed OmpU led to complete protection after challenge 5 weeks later (RPS = 100 %)
whereas lesser protection resulted with the DNA vaccine (RPS = 51.4 %). Two
potentially protective immunogens, DegQ and Vhp1, have been accommodated in
DNA vaccines and used to vaccine Japanese fl ounder with promising results for
pDV after challenge (RPS = 84.6 %) (Hu and Sun 2011 ). Hu et al. ( 2011 ) continued
the work by developing a recombinant product that expressed V. harveyi DegQ as a
soluble antigen that elicited signifi cant protection against both Edw. tarda and V.
harveyi in laboratory-based experiments with turbot when administered by i.p.
(RPS = 90.9 %) orally (RPS = 60.5 %) or immersion (RPS = 47.1 %) or a combination of oral plus immersion (RPS = 77.8 % after one month in a mock fi eld trial;
RPS = 81.8 % after two months). A serine protease has been reported as a protective
antigen, and enabling fi sh to resist challenge (Zhang et al. 2008 ).
The value of whole cell preparations was enhanced by entrapment within liposomes with the outcome being increased protection and a demonstrable stimulation
of immune parameters (Harikrishnan et al. 2012b ).
A bivalent vaccine (with Ph. damselae subsp. piscicida) based on formalised
cells and ECP administered to sole by immersion with booster or by i.p. injection
led to high levels of protection (RPS = ~88 %) for 4 months after which the benefi t
declined (Arijo et al. 2005 ). A combination vaccine with inactivated cells of V. alginolyticus , V. harveyi , V. vulnifi cus and infectious spleen and kidney necrosis virus
was evaluated in orange-spotted grouper ( Epinephelus coioides) with an overall
RPS of 80 % (Huang et al. 2012 ).
10 Vibrios
administered i.p. (RPS = 92.3 %) and orally (RPS = 61.2 %) (Sun et al. 2009 ). Pang
et al. ( 2010 ) used OmpN mixed with FCA to vaccinate via i.p. (100 μl/fi sh), estuary
cod ( Epinephelus coioides ), that survived challenge after 28-days with a virulent
culture (RPS = 60 and 70 % depending on the nature of the challenge strain).
Meanwhile, Zhang et al. ( 2011a ) cloned, sequenced and characterised two OMP
genes, OmpK and GADPH, and expressed the recombinant proteins in the prokaryotic expression vector pET-30a(+), which were purifi ed and used to vaccinate intraperitoneally (100 μg/fi sh) large yellow croaker ( Pseudosciaena crocea ) with booster
doses after 3-weeks. The outcome was that r-OmpK and r-GADPH led to RPS values of 37.7 % and 40 %, respectively, after challenge. OmpK was expressed in yeast,
Pichia pastoris, and fed for 5 days to sea bass ( Lateolabrax japonicus ) on alginate
microspheres with a resultant production of antibodies, and protection against challenge (RPS = 61.5 %) (Mao et al. 2011 ). A denatured inactive cytotoxic 530 amino
acid recombinant secreted protease, Vhp1, was recovered from a pathogenic isolate
was determined to be an effective subunit vaccine (RPS = 70 %) with improved performance when expressed in Esch. coli as a live vaccine (RPS = 90 %) (Cheng et al.
2010 ). In a further study, Wang et al. ( 2011 ) used the purifi ed 35 kDa outer membrane protein OmpU (50 μg dose) and a DNA vaccine involving the insertion of the
ompU gene into pEGFP-N1 plasmid (10 μg) injected i.m. in turbot. Use of the purifi ed OmpU led to complete protection after challenge 5 weeks later (RPS = 100 %)
whereas lesser protection resulted with the DNA vaccine (RPS = 51.4 %). Two
potentially protective immunogens, DegQ and Vhp1, have been accommodated in
DNA vaccines and used to vaccine Japanese fl ounder with promising results for
pDV after challenge (RPS = 84.6 %) (Hu and Sun 2011 ). Hu et al. ( 2011 ) continued
the work by developing a recombinant product that expressed V. harveyi DegQ as a
soluble antigen that elicited signifi cant protection against both Edw. tarda and V.
harveyi in laboratory-based experiments with turbot when administered by i.p.
(RPS = 90.9 %) orally (RPS = 60.5 %) or immersion (RPS = 47.1 %) or a combination of oral plus immersion (RPS = 77.8 % after one month in a mock fi eld trial;
RPS = 81.8 % after two months). A serine protease has been reported as a protective
antigen, and enabling fi sh to resist challenge (Zhang et al. 2008 ).
The value of whole cell preparations was enhanced by entrapment within liposomes with the outcome being increased protection and a demonstrable stimulation
of immune parameters (Harikrishnan et al. 2012b ).
A bivalent vaccine (with Ph. damselae subsp. piscicida) based on formalised
cells and ECP administered to sole by immersion with booster or by i.p. injection
led to high levels of protection (RPS = ~88 %) for 4 months after which the benefi t
declined (Arijo et al. 2005 ). A combination vaccine with inactivated cells of V. alginolyticus , V. harveyi , V. vulnifi cus and infectious spleen and kidney necrosis virus
was evaluated in orange-spotted grouper ( Epinephelus coioides) with an overall
RPS of 80 % (Huang et al. 2012 ).
10 Vibrios