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.
The state of Sinaloa in Mexico is an industrialized agricultural region with a documented pesticide usage of 700 t year; which at least 17 of the pesticides are classified as moderately to highly toxic. Pollutants in the water column of rivers and drains are of great concern because the water flows into coastal lagoons and nearshore waters and thereby affects aquatic organisms. This study was done in four municipalities in the state of Sinaloa that produce food intensively. To investigate the link between pollution in the lagoons and their proximity to agricultural sites, water was sampled in three coastal lagoons and in the rivers and drains that flow into them. Seawater from the Gulf of California, 10 km from the coast, was also analyzed. Concentrations of nutrients, organochlorines, and organophosphorus pesticides were determined. Nutrient determination showed an unhealthy environment with N/P ratios of <16, thus favoring nitrogen-fixing cyanobacteria. The organochlorine pesticides showed a clear accumulation in the coastal lagoons from the drains and rivers, with ΣHCH showing the highest concentrations. In the southern part of the region studied, pollution of the coastal lagoon of Pabellones could be traced mainly to the drains from the agricultural sites. Accumulation of OC pesticides was also observed in the Gulf of California. Tests for 22 organophosphates revealed only five (diazinon, disulfoton, methyl parathion, chlorpyrifos, and mevinphos); diazinon was detected at all the sites, although methyl parathion was present at some sites at concentrations one order of magnitude higher than diazinon.
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