Aquatic animal species and organism relationship bacteriological culture techniques interpretation of biochemical identification tests and sets biochemical identification tables technical methods techniques for molecular identification of bacteria preparation of media for culture and identification. (Part contents.)

Bacteria from fish and other aquatic animals : a practical identification manual

Moribund shrimp affected by acute hepatopancreatic necrosis disease (AHPND) from farms in northwestern Mexico were sampled for bacteriological and histological analysis. Bacterial isolates were molecularly identified as Vibrio parahaemolyticus by the presence of the tlh gene. The tdh-negative, trh-negative, and tlh-positive V. parahaemolyticus strains were further characterized by repetitive extragenic palindromic element-PCR (rep-PCR), and primers AP1, AP2, AP3, and AP and an ems2 IQ2000 detection kit (GeneReach, Taiwan) were used in the diagnostic tests for AHPND. The V. parahaemolyticus strains were used in immersion challenges with shrimp, and farmed and challenged shrimp presented the same clinical and pathological symptoms: lethargy, empty gut, pale and aqueous hepatopancreas, and expanded chromatophores. Using histological analysis and bacterial density count, three stages of AHNPD (initial, acute, and terminal) were identified in the affected shrimp. The pathognomonic lesions indicating severe desquamation of tubular epithelial cells of the hepatopancreas were observed in both challenged and pond-infected shrimp. The results showed that different V. parahaemolyticus strains have different virulences; some of the less virulent strains do not induce 100% mortality, and mortality rates also rise more slowly than they do for the more virulent strains. The virulence of V. parahaemolyticus strains was dose dependent, where the threshold infective density was 10(4) CFU ml(-1); below that density, no mortality was observed. The AP3 primer set had the best sensitivity and specificity. Field and experimental results showed that the V. parahaemolyticus strain that causes AHPND acts as a primary pathogen for shrimp in Mexico compared with the V. parahaemolyticus strains reported to date.

Field and experimental evidence of Vibrio parahaemolyticus as the causative agent of acute hepatopancreatic necrosis disease of cultured shrimp (Litopenaeus vannamei) in Northwestern Mexico.

Expression of luminescence in the Penaeus monodon pathogen Vibrio harveyi is regulated by an intercellular quorum sensing mechanism involving the synthesis and detection of two signaling molecules, one of which is N-hydroxy butanoyl-L-homoserine lactone and the other of which is uncharacterized. Indirect evidence has suggested that virulence, associated with a toxic extracellular protein, and luminescence in V. harveyi are coregulated. In this study the effects of an acylated homoserine lactone antagonist produced by the marine alga Delisea pulchra on luminescence and toxin production in a virulent strain of V. harveyi were analyzed. Luminescence and toxin production were both inhibited by the signal antagonist at concentrations that had no impact on growth. Toxin production was found to be prematurely induced in V. harveyi cultures incubated in a 10% conditioned medium. Additionally, a significant reduction in the toxicity of concentrated supernatant extracts from V. harveyi cultures incubated in the presence of the signal antagonist, as measured by in vivo toxicity assays in mice and prawns, was observed. These results suggest that intercellular signaling antagonists have potential utility in the control of V. harveyi prawn infections.

Inhibition of Luminescence and Virulence in the Black Tiger Prawn (Penaeus monodon) Pathogen Vibrio harveyi by Intercellular Signal Antagonists

P.-C. LIU, K.-K. LEE AND S.-N. CHEN. 1996. The pathogenicity of six Vibrio harveyi strains in tiger prawn, Penaeus monodon, was studied, using both live bacteria and extracellular products (ECP). The organisms originally isolated from diseased penaeids were more virulent using both live bacteria and ECP (LD50, 4.87–8.65 times 104colony-forming units (cfu) and 1.20–1.51 μg protein g-1body weight) than the two reference strains originally isolated from either sea water (ATCC 25919; LD50, 3.18 times 106cfu and 2.70 μg protein g-1body weight) or diseased Talorchestia sp. (ATCC 14126, 0.418 times 106cfu and 2.34 μg protein g-1body weight). Each strain was reisolated from the haemolymph and the hepatopancreas of moribund prawns following each bacterial challenge. Both the live bacteria and the ECPs of the penaeid isolates exhibited stronger proteolytic (caseinase), phospholipase and haemolytic activities than those of the reference strains. These results indicate that there are differences between penaeid and non-penaeid isolates of V. harveyi in pathogenicity and reveal that proteases, phospholipases, haemolysins or exotoxins might play leading roles in the pathogenicity of V. harveyi in the tiger prawn, Penaeus monodon.

Pathogenicity of different isolates of Vibrio harveyi in tiger prawn, Penaeus monodon

http://www.vliz.be/imisdocs/publications/33359.pdf

Experimental infection models for shrimp vibriosis studies: a review.

Vibrio harveyi infection was found to occur in pond-reared black tiger prawn in Thailand. The diseased prawns with a hepatopancreatic infection of V. harveyi showed bacterial invasions and multiplication in the tubular lumens. This condition was followed by necrosis of hepatopancreatic cells and the thickened basal lamina, subsequent granulomatous encapsulation of the invaded tubules, and production of granulation tissue around granulomatous lesions. Heavy bacterial multiplication in the hepatopancreatic tubules caused systemic bacterial dissemination, which resulted in marked necrosis in the heart and lymphoid organ. On the other hand, two prawns with percuticular infection by V. harveyi had bacterial invasions in the subcuticular, spongy connective tissue in the telson, and systemic dissemination was followed by the occurrence of bacteriaphagocytizing hemocytes in the various tissues. Both types of isolates (chitinase positive and negative) were moderately pathogenic to prawn; intramuscular inj...

C. Limsuwan

Papers

Histopathology, Biochemistry, and Pathogenicity of Vibrio harveyi Infecting Black Tiger Prawn Penaeus monodon