Effect of freezing on the metabolic status of L3 larvae of Anisakis simplex s. s.

The fish-borne parasite, Anisakis simplex s. s., triggers a disease called anisakiasis, that is associated with a gastrointestinal infection. The Anisakis is also associated with allergic response which may lead to anaphylactic shock. The A. simplex s. s. L3 larvae may be freeze tolerant despite when the nematodes will be cooled rapidly to -20 degrees C according to the sanitary authorities of the USA and the EU. The aim of this work was to study the metabolic status of A. simplex s. s. L3 larvae when frozen in terms of viability, expression of genes involved in the nematodes' survival of freezing, as well content of carbohydrates which play a cryoprotective role in thermal stress and are the main source of energy. The levels of trehalose were significantly higher after slow freezing treatment (p<.0001), than the fast freezing (p<.002). The lower temperatures induce changes, especially in trehalose synthesis gene expression, genes responsible for oxidative metabolism, and chaperone proteins, but we cannot state clearly whether these changes occur during freezing, or because they are already prevalent during cold acclimation. The induction of mentioned genes seems to be a common trait of both cold- and dehydration tolerance.