Priming the immune system of Penaeid shrimp by bacterial HSP70 (DnaK).

This study was conducted to test the effect of DnaK on priming immune responses in Penaeid shrimp. Juvenile-specific pathogen-free (SPF) P. vannamei shrimp were injected with 0.05 µg recombinant DnaK. One hour post-DnaK priming, a non-lethal dose of Vibrio campbellii (10(5) CFU shrimp(-1)) was injected. Other treatments include only DnaK or V. campbellii injection or control with blank inocula. The haemolymph of three shrimp from each treatment was collected at 1.5, 6, 9 and 12 h post-DnaK priming (hpp). It was verified that injection with DnaK and V. campbellii challenge affected the transcription of 3 immune genes, transglutaminase-1 (TGase-1), prophenoloxidase-2 (proPO-2) and endogenous HSP70 (lvHSP70). In P. monodon, shrimp were first injected with DnaK at a dose of 10 µg shrimp(-1) and one hour later with 10(6) CFU of V. harveyi (BB120) shrimp(-1). Shrimp injected with DnaK showed a significant increase in proPO expression compared to the control (P < 0.05). Yet a double injection (DnaK and Vibrio) seemed to cause an antagonistic response at the level of expression, which was not equalled at the level of PO activity. Those results suggest that DnaK is able to modulate immune responses in P. vannamei and P. monodon.

Interference with the quorum sensing systems in a Vibrio harveyi strain alters the growth rate of gnotobiotically cultured rotifer Brachionus plicatilis.

AIMS: To evaluate the effect of Vibrio harveyi strains on the growth rate of the gnotobiotically cultured rotifer Brachionus plicatilis, and to establish whether quorum sensing is involved in the observed phenomena. METHODS AND RESULTS: Gnotobiotic B. plicatilis sensu strictu, obtained by hatching glutaraldehyde-treated amictic eggs, were used as test organisms. Challenge tests were performed with 11 V. harveyi strains and different quorum sensing mutants derived from the V. harveyi BB120 strain. Brominated furanone [(5Z)-4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone] as a quorum sensing inhibitor was tested in Brachionus challenge tests. Some V. harveyi strains, such as strain BB120, had a significantly negative effect on the Brachionus growth rate. In the challenge test with MM77, an isogenic strain of BB120 in which the two autoinducers (HAI-1 and AI-2) are both inactivated, no negative effect was observed. The effect of single mutants was the same as that observed in the BB120 strain. This indicates that both systems are responsible for the growth-retarding (GR) effect of the BB120 strain towards Brachionus. Moreover, the addition of an exogenous source of HAI-1 or AI-2 could restore the GR effect in the HAI-1 and AI-2 nonproducing mutant MM77. The addition of brominated furanone at a concentration of 2.5 mg l(-1) could neutralize the GR effect of some strains such as BB120 and VH-014. CONCLUSIONS: Two quorum sensing systems in V. harveyi strain BB120 (namely HAI-1 and AI-2-mediated) are necessary for its GR effect on B. plicatilis. With some other V. harveyi strains, however, growth inhibition towards Brachionus does not seem to be related to quorum sensing. SIGNIFICANCE AND IMPACT OF THE STUDY: Interference with the quorum sensing system might help to counteract the GR effect of some V. harveyi strains on Brachionus. However, further studies are needed to demonstrate the positive effect of halogenated furanone in nongnotobiotic Brachionus cultures and eventually, in other segments of the aquaculture industry.

Problems and solutions with the design and execution of an epidemiological study of white spot disease in black tiger shrimp (Penaeus monodon) in Vietnam.

White spot disease (WSD) is caused by white spot syndrome virus (WSSV) and is an acutely fatal pandemic disease of crustaceans. It has resulted in massive losses to the shrimp-farming industry in Asia and has now spread to the Americas. This paper reports the problems and solutions associated with the design and execution of a longitudinal epidemiological study of shrimp (Penaeus monodon) health on farms practising a crop rotation of rice and shrimp in the Mekong Delta of Vietnam. The pre-sampling phase of the project involved selecting an appropriate site and sampling variables, obtaining permission and establishing the necessary laboratory and logistic facilities. At the start of the sampling phase, 40 farmers were selected and 32 of these were visited and interviewed. This resulted in the enrolment of only 17 farmers. A further seven had to be enrolled to obtain the maximum number of farmers that could be sampled by the study team. Compliance was enhanced through meetings, regular visits by senior members of the project team and ensuring that visits were punctual and that all information was treated confidentially. The production cycle began in January 1998 and lasted for approximately 5 months. An attempt was made to collect 500 post larvae (PL) before each pond was stocked to assess the health of the batch and to test for the presence of WSSV by one-step PCR. After stocking, the wild crustaceans also were sampled from the pond for PCR analyses. Information was collected on the management practices and samples of water, pond bottom, feed and shrimp collected throughout the production cycle. Water quality variables with predictable diurnal variation were sampled in the morning and afternoon, twice a week. Two months after stocking, the first outbreak of WSD occurred; subsequently, 18 farms conducted a complete emergency harvest due to the actual or perceived presence of a WSD outbreak. Detectable mortalities were reported from 19 farms, and moribund shrimps were collected from four of these for PCR and histological analyses.