Masanobu Sakata

Elemental Characterization of Daphnia Resting Eggs by X-ray Analytical Microscopy

Abstract Resting eggs of Daphnia, a key crustacean zooplankton of freshwater food chains, can remain viable for more than a century. These eggs are able to withstand freezing and drying, and can survive the harsh environment of a predators digestive system. Until recently little was known about the chemical composition, microanatomy, and physical properties of the resting eggs. The current study utilized a physical technique, the X-ray analytical microscope, to identify and localize component elements of the Daphnia resting egg. The analysis demonstrated that phosphorus, sulfur, potassium, and calcium were detected as elemental components of the resting egg, and detection intensities of the four elements differed according to the position of the eggs. Phosphorus and calcium were mostly detected in regions of the eggshell that surrounded the two embryos. In addition, sulfur was distributed throughout the eggshell whereas potassium was localized to the areas that corresponded to where the embryos were encased. Through the use of X-ray analytical microscopy, the current study identifies elemental characteristics in relationship to the structure of the Daphnia resting eggs.

Crystalline calcium phosphate and magnetic mineral content of Daphnia resting eggs.

Abstract Daphnia is a key crustacean zooplankton of freshwater food chains. One factor that ensures successful propagation is the Daphnia resting eggs, which are able to retain structural integrity under extreme conditions. Until recently little was known about the chemical composition, microanatomy, and physical properties of the egg itself. The current study demonstrates that the resting eggs: (1) have shells that are made up of crystalline calcium phosphate and include a honeycombed structure, and (2) contain magnetic material having properties consistent with magnetite. These properties of the resting eggs may ensure Daphnia survival in harsh environments.

Estimation for diameter of superparamagnetic particles in Daphnia resting eggs

Ferromagnetic resonance (FMR) with an electron spin resonance (ESR) apparatus was investigated for super-paramagnetic particles within Daphnia resting eggs. High-field (HF) resonance lines near g=2 resulted from single superparamagnetic particles, were detected from ESR spectra of Daphnia resting eggs. The size of isolated superparamagnetic particles within Daphnia resting eggs was calculated to be approximately 13 nm in diameter by analysis of the temperature dependence of the HF line width. Small-angle X-ray scattering (SAXS) analysis of Daphnia resting eggs also showed that average size of superparamagnetic particles in diameter, equivalent to magnetite, was approximately 13 nm. The combination of FMR and SAXS measurement is very effective in estimating the size of superparamagnetic particles in biological organisms, with difficulties of preparing for samples for measurement by electron microscopy. However, Chlorella, with that Daphnia were raised, did not show FMR spectra, showing no magnetic particles within Daphnia resting eggs. Therefore, it suggested that superparamagnetic particles within Daphnia resting eggs, were mineralized in Daphnia as the result of biomineralization of Fe originated from Chlorella.