New Insights into the Mechanism of Action of PirAB from Vibrio Parahaemolyticus.

PirAB toxins secreted by Vibrio parahaemolyticus (Vp) harbor the pVA1 virulence plasmid, which causes acute hepatopancreatic necrosis disease (AHPND), an emerging disease in Penaeid shrimp that can cause 70-100% mortality and that has resulted in great economic losses since its first appearance. The cytotoxic effect of PirABVp on the epithelial cells of the shrimp hepatopancreas (Hp) has been extensively documented. New insights into the biological role of the PirBVp subunit show that it has lectin-like activity and recognizes mucin-like O-glycosidic structures in the shrimp Hp. The search for toxin receptors can lead to a better understanding of the infection mechanisms of the pathogen and the prevention of the host disease by blocking toxin-receptor interactions using a mimetic antagonist. There is also evidence that Vp AHPND changes the community structure of the microbiota in the surrounding water, resulting in a significant reduction of several bacterial taxa, especially Neptuniibacter spp. Considering these findings, the PirABvp toxin could exhibit a dual role of damaging the shrimp Hp while killing the surrounding bacteria.

Silver nanoparticles induce histopathological alterations in juvenile Penaeus vannamei.

The objective of this study was to evaluate the histopathological alterations in juvenile Penaeus vannamei caused by silver nanoparticles (AgNPs) for two types of experiments: at sublethal concentrations of 3.6 to 7.1 µg/µL of metallic silver (Ag) for a short period up to 72 h and for 2.6 to 7.9 µg of Ag/µL for the long period up to 264 h. The severity degree of the changes was evaluated and the histopathological index (Hi) was determined in both experiments using the necrosis (cellular dead) as an indicator. The pathological changes in the striated muscle, gills, antennal gland, circulatory system, heart, lymphoid organ, and connective tissue are described. The histopathological effects were similar for the two experiments without a direct relationship with the concentrations. In the short-term experiment, the values of Hi were higher (2.34 ± 0.41 at 48 hpi and 1.91 ± 0.39 at 72 hpi) compared with the long-term experiment (values between 0.57 ± 0.36 to 1.74 ± 0.57 at 264 hpi). The observed pathologies are similar to those caused by other metals, with the exception of the agglomerations of black particles in the gills, lymphoid organ, and muscle, which has not been previously reported. This work shows that silver nanoparticles cause damage to shrimp in sublethal concentrations.

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

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.

Histological effects of a combination of heavy metals on Pacific white shrimp Litopenaeus vannamei juveniles.

Exposure to different levels of a mixture in equitoxic concentrations of Cd, Cu, Fe, Hg, Mn, Pb and Zn, equivalent to between 5 and 0.5% the individual respective 96-h median lethal concentrations (0.05-0.005AF: application factors) caused dose- and time-dependent damages to the hepatopancreas, gills, epipodites and midgut tissues of Litopenaeus vannamei juveniles. After between 1 and 3-4 weeks, there was between 50 and 100% loss of the hepatopancreas R cells and of the tubules regular structure. Gill alterations were observed between 2 and 3 weeks with 0.05-0.025 and 0.005AF, respectively. Epipodites showed time- and dose-dependent increasing degrees of melanization, and hemocytic enteritis was observed with 0.025 and 0.01AF. Exposure to 0.05 and 0.025AF caused also 50 and 12% mortalities after 2 and 3 weeks respectively, showing that even at these low levels the mixture of these metals may have a mid-term lethal effect. For this reason, the assessment of risk and of safe levels of toxic substances added to any natural environment through human or natural sources, should not neglect the effects on biological systems caused by the interaction of minute amounts of toxicants, which would be harmless unless present in combination.