Combined effect of exposure to ammonia and hypoxia on the blue shrimp Litopenaeus stylirostris survival and physiological response in relation to molt stage

The effect of ambient ammonia, hypoxia and combination of both on survival and the physiological and immunological response of the blue shrimp Litopenaeus stylirostris in relation to molt stage was studied. Shrimp were submitted to 44.0-71.5 mg l1 total ammonia-N corresponding to 2.0 mg l1 unionized ammonia NH3-N and/or to 1.5 mg O2 l1 (4.3 kPa) for 24 hours. Survival was recorded and the molt stages of both dead and surviving shrimp determined. Only shrimp in intermolt and premolt stages were sampled for analysis of haemolymph. Haemolymph was assayed for osmoregulatory capacity (OC), magnesium ion (Mg2+), calcium ion (Ca2+), total proteins, oxyhaemocyanin, lactate, glucose and total haemocyte count (THC). Low mortalities were recorded for shrimp submitted independently to ammonia or hypoxia. Seventy five percent of dead shrimp were in early post molt (stage A) in ammonia treatment, while hypoxia affected mainly late premolt animals (stage D2). A synergic effect of ammonia and hypoxia combination (A + O2 treatment) on mortality was observed, affecting nearly exclusively shrimp in late premolt stage D2. Analysis of molt stage repartition at the end of the experiment suggests that ammonia treatment may have accelerated molting. The common physiological response of shrimp to the different treatments was characterized by a reduced OC and an increase in Ca2+. Increase in Mg2+ could not be validated by the statistical analysis, as well as glycaemia variations. Plasmatic lactate level increased and THC decreased in shrimp submitted to hypoxia and the combination of hypoxia and ammonia. Total proteins concentration was reduced in ammonia and A + O2 treatments. The effect was more pronounced in late premolt shrimp than in intermolt shrimp. Combination of ammonia and hypoxia led to a physiological response stronger than this observed for ammonia-alone and/or hypoxia-alone treatments, except for oxyhaemocyanin. The effects of each external factor (ammonia, hypoxia) and the combination of both, and internal one (molt stage) are discussed.