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Sulfide is a natural and widely distributed toxicant. It can be commonly found on the interface between water and sediment in the
aquatic environment. The Pacific white shrimp Litopenaeus vannamei starts life in the benthic zone soon after the mysis stage,
an early stage of post larvae. Therefore, L. vannamei is inevitably affected by exposure to sulfide released from pond sediment. The
toxicity and poisoning mechanisms of sulfide were studied in Litopenaeus vannamei from the perspective of energy metabolism,
metabolomics, gut health and microbiota. The lethal concentrations of sulfide in L. vannamei (LC50) at 24 h, 48 h, 72 h, and 96 h were
determined. Sulfide at a concentration of 0, 1/10 (425.5 �¼g/L), and 1/5 (851 �¼g/L) of the LC50 at 96 h was used to test the responses
of L. vannamei for 21 days. The results of the study showed that chronic sub-lethal sulfide exposure could lead to the dysfunction
of mitochondrial respiration, as evidenced by the change of cytochrome C oxidase activity, disturbed protein synthesis, enhanced
gluconeogenesis, and increased substrate consumption for ATP synthesis. The accelerated tricarboxylic acid cycles could provide
extra energy for dealing with sulfide stress. Chronic sulfide exposure could adversely affect the health status of shrimp and therefore
lower L. vannamei survival. Dose-dependent relationships were found in exposed to sulfide. Also chronic exposure to
sub-lethal sulfide could lead to damage of the gut structure, stimulate the response of the inflammatory and immune systems, and
shape the microbiota structure in the gut of L. vannamei. The microbiota structure includes three aspects. First, the abundance of
pathogenic bacteria increased significantly with the increasing concentration of sulfide. Next, the abundance of some anti-stress
bacteria decreased. Last, adaptation of sulfide-stimulated bacteria was commonly found and down-regulated.