ES2321787A1 - Procedure for obtaining the anti-edwardsiella tarda (edt) vaccine for the prevention of edwardsielosis rodaballo disease. (Machine-translation by Google Translate, not legally binding) - Google Patents

Abstract

Procedure for obtaining the anti-edwardsiella tarda vaccine (edt) for the prevention of edwardsielosis disease of turbot, cultivated in seawater produced by the microorganism edwardsiella tarda. The procedure is characterized by including in the vaccine the bacterial cells inactivated with formaldehyde of a strain of edwardsiella tarda isolated in galicia, strain deposited as cect 7215. (Machine-translation by Google Translate, not legally binding)

Description

Procedure for obtaining the Edwardsiella tard (EdT) vaccine for the prevention of Edwardsiellosis disease of turbot.

Edwardsielosis , produced by Edwardsiella takes was first diagnosed in turbot grown in 1994. However, from that date until 2006 no outbreaks of this disease have been described again in facilities dedicated to the cultivation of this species. In these last two years (2005-2006) this pathogen is isolated from diseased turbot in different geographical areas of Europe and today is considered an emerging pathogen in marine aquaculture (Castro et al., 2006: Journal of Fish Diseases , 29: 541-547). Edwardsiella strains of turbot isolates are very homogeneous both phenotypically and antigenically, and biotypes and serotypes cannot be established. However, by means of molecular methods, two different clonal lines have been detected within these isolates (Castro et al., 2006: op. Cited . Although chemotherapy can be effective in controlling this disease, effective vaccines need to be developed that allow us to prevent and control the mortalities produced by this pathogen (Castro et al., 2006: Aquatic Environment Microbiology Congress, Valencia).

Preparation

From pre-logs in the logarithmic phase of growth two liter flasks containing one liter are inoculated of tryptone soy broth culture medium (TSB: Tryptone Soy Broth: tryptone 17 gr / L; soybean peptone 3 gr / L; glucose 2.5 gr / L; bipotassium phosphate 2.5 gr / L; 10 g / L sodium chloride; pH 7.3 ± 0.2).

Incubation is performed at 25 ° C for 48 hours. When the crop reaches an optical density of 1 (A_ {550}: Absorbance_ {550}) (approximately 10 10 cells / mL), formalin is added to a final concentration of 0.35% to kill bacteria, and stays for three more hours in stirring, after which it is transferred at 4 ° C.

Sterility control is carried out sowing the vaccine preparation in TSA plates and tubes thioglycolate, incubating for 72 hours at 25 and 37 ° C, respectively.

The specificity control is carried out by slide agglutination, using cells as antigen Whole used in the manufacture of the vaccine.

The vaccine should be stored at 4 ° C until utilization.

Administration mode

This vaccine should be given by injection. intraperitoneal To do this, fish are inoculated with 0.1-0.2 mL / fish (depending on fish weight) of undiluted vaccine preparation.

The Edwardsiella strain used in this vaccine is RM 288.1, deposited in the Spanish Type Culture Collection as CECT 7215 isolated from the diseased turbot kidney in Galicia.

The efficacy of this vaccine was evaluated in turbot both laboratory and field scale. The power, expressed as Relative Survival Percentage (RSP) it has been over 80% after three months.

Claims (3)

1. Edwardsiella strain RM288.1 takes , isolated from an epizootic in Galicia (Spain), deposited in the Spanish Type Culture Collection (CECT) with accession number CECT 7215. 2. Procedure for obtaining the Edwardsiella tard (EdT) vaccine characterized by the growth of bacterial cells of strain RM288.1, CECT 7215, in liquid cultures of half-tripty soy broth (TSB), for its inactivation with formalin at a final concentration of 0.35% for 4 hours at room temperature and because the final concentration of bacterial cells in the bacterin is 10 10 cells / mL. 3. Anti- Edwardsiella tard (EdT) vaccine obtained by the method of claim 2, for the prevention of edwardsielosis disease in turbot grown in seawater, and to be administered by undiluted intraperitoneal injection.