Anti-IgY monoclonal antibody, clone 27D8

Outbreaks of mass mortality among cultured zoea of M. rosenbergii occurred in different hatcheries of Gaoyou country of China in April 2017. Of the isolates obtained from the diseased zoea, XL3-1 was pathogenic to the zoea and postlarva of M. rosenbergii in the virulence test, with a LD50 value 3.2 x 10(6) CFU/mL and 3.5 x 10(6) CFU/mL, respectively. XL3-1 was identified as Enterobacter cloacae by biochemical characteristics and molecular tools. Detection of virulence-associated genes by PCR indicated that XL3-1 was positive for irp2, fhuA, sodB, sltA, flaD and OmpX. According to the antibiotic resistant assay, the isolate was sensitive to quinolones, while they were resistant to cephalosporins and penicillins. This study also investigated the protection of specific chicken egg yolk antibodies (IgY) in E. doacae-infected M. rosenbergii. The highest titer against E. cloacae of the obtained specific IgY by ELISA was 1: 2(13). M. rosenbergii treated with specific egg yolk powder could achieve the survival rate of 67% after E. cloacae infection, while M. rosenbergii treated with non-specific egg yolk powder or no egg yolk powder could achieve survival rates of 20% and 17%, respectively. As well, the bacterial burden in hepatopancreas was significantly lower in M. rosenbergii treated with specific egg yolk powder than those treated with non-specific egg yolk powder. This study clarified the virulence characteristics and antibiotic resistance of E. cloacae, and provided a strategy to control the disease by using specific IgY.

The present study investigates the enhancement of immunogenicity and protection efficacy of Salmonella Typhimurium ghosts surface-displayed with FliC against chicken salmonellosis. The membrane-anchored FliC is a potential TLR-5 agonist, delivers an essential adjuvant effect for the ghost vaccine candidate. The present ghost plasmid pJHL184 construct carries a convergent dual promoter system that has the temperature-dependent induction of the phage lysis gene E and the target antigen FliC at the same time. Under permissible conditions of temperatures, less than 30 degrees C at the presence of 20 mM c-arabinose effectively suppresses expression of the lysis gene. Once the temperature is up-lifted to 42 degrees C without arabinose, cause the generation of ST ghosts expelling the cytoplasmic content. The addition of FliC adjuvant significantly enhanced the IgY response, cell-mediated immune responses, regulatory cytokine induction and subsequently enhanced protection against Salmonella challenge. Further, intramuscular immunization with ST ghosts displaying FliC induced particularly high CD8(+) response demarcating its proficiency to elicit Type I immune responses. Further, ST ghosts displaying FliC caused an increase in both CD4(+) and CD8(+) response compared to the PBS control suggesting its capability to engage both cell-mediated and humoral immune responses essential for the elimination of Salmonella. Upon the virulent challenge, we could observe a significant reduction in challenged bacterial load on spleen, liver and cecum tissues in the ST ghosts surface-displaying FliC adjuvant. Our study suggests the biological incorporation of FliC on ST ghosts enhances vaccine immunogenic potency and acts as a safe and effective prevention strategy against chicken salmonellosis.