Silver Nanoparticles Synthesized with Rumex hymenosepalus: A Strategy to Combat Early Mortality Syndrome (EMS) in a Cultivated White Shrimp

Early Mortality Syndrome (EMS) or Acute Hepatopancreatic Necrosis Syndrome (AHPNS) is a disease produced by gram-negative bacteria Vibrio parahaemolyticus (V. parahaemolyticus), which has caused declines in worldwide production of a white shrimp Litopenaeus vannamei (L. vannamei). In this work, we propose the implementation of silver nanoparticles (AgNPs) synthesized with Rumex hymenosepalus (Rh) extract as an alternative on V. parahaemolyticus control. AgNPs were characterized by UV-Vis spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and selected area electron diffraction (SAED). AgNP mean sizes by DLS were 80.82±1.16 nm and sizes between 2 and 10 nm by TEM, with a zeta potential of -47.72±1.05 mV. This study evaluated AgNPs and Rh antimicrobial capacity on V. parahaemolyticus at different concentrations; the minimum inhibitory concentration (MIC) found was 25 μg/mL for AgNPs and 220 μg/mL for Rh. Additionally, were carried out time-kill curves and reactive oxygen species (ROS) generation for 1 and 4 MIC. Both concentrations (MIC) were tested for toxicity on Artemia nauplii from Artemia franciscana (A. franciscana), because nauplii were used as biocarriers for AgNPs and Rh extract on L. vannamei. Once the shrimp were treated, they were challenged with Vibrio infection and it was found that those who were treated with both agents showed greater survival than the control. V. parahaemolyticus and postlarval samples were taken from the bioassay and fixed and prepared for TEM and SEM in order to search NPs in internal structure of bacteria and the hepatopancreatic area of shrimps; AgNPs were detected in both cases. AgNPs and Rh extract show antibacterial properties on the infected shrimp with V. parahaemolyticus. The action mechanisms are interaction with the bacterial membrane and ROS generation; these effects are produced by both agents.