Erythroid cells in the hemolymph of a bivalve Anadara kagoshimensis (Tokunaga, 1906) under hydrogen sulfide loading: Flow cytometry and light microscopy
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Abstract
The imbalance between organic matter oxidation and oxygen supply mediates the formation of time-stable redox zones in the water column. On the shelf, this typically occurs due to the absence of thorough vertical convection and the formation of localized decomposition zones. The functional mechanisms of the resistance of certain benthic organisms to such conditions are of particular interest. In this work, we study a bivalve Anadara kagoshimensis (Tokunaga, 1906) known for its tolerance to hydrogen sulfide contamination. Using flow cytometry and light microscopy, we examined the effect of hydrogen sulfide loading on morphofunctional characteristics of its erythroid cells under experimental conditions. The analysis was carried out on adult specimens with a shell height of 23–34 mm. The control group of molluscs was kept in an aquarium with an oxygen concentration of 7.0–8.2 mg O2·L−1 (normoxia). For the experimental group, oxygen content was first lowered to 0.1 mg O2·L−1 for 2 h (via nitrogen bubbling); then, Na2S was added to water to a final concentration of 6 mg S2−·L−1. Exposure to hydrogen sulfide revealed a significant increase in the volume of erythroid cells in A. kagoshimensis hemolymph (more than 40%, p < 0.01) accompanied by a substantial rise in fluorescence intensity of rhodamine 123 (R123) and 2′-7′-dichlorofluorescein-diacetate (DCF-DA) (2–3-fold, p < 0.01). This evidences for enhanced oxidative processes within cells and their possible lysis. The latter one may facilitate the release of hematin-containing granules, and hematin is capable of neutralizing sulfides. The observed response seems to be an adaptive one. A rise in values of side scatter and SYBR Green I fluorescence reflects an increase in abundance of granular inclusions within red blood cells under hydrogen sulfide loading and a gain in the functional activity of their nuclei.
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References
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