The jelly coat of Aplysia depilans egg strings and the associated microorganisms

Egg masses of Aplysia depilans consist of long and intertwined strings containing numerous capsules with eggs. Light microscopy stains and transmission electron microscopy revealed four layers in the gelatinous sheath that encircled and aggregated the chain of egg capsules. The outermost layer has a fluffy structure. The second, third, and fourth layers consisted of reticulated matrices with different densities. The second and third layers were divided into 5‒6 strata each. The fourth and innermost layer of the gelatinous sheath has a higher density and no visible stratification. This layer glues the tightly packed capsules to one another and to the outer layers of the gelatinous sheath. The thin wall of the capsules is formed by a homogeneous and highly electron-dense material. Inside the capsules, the eggs or embryos were bathed in an electron-lucent aqueous medium. Bacteria and diatoms were the most abundant microorganisms on the surface of egg strings. Bacteria penetrate the gelatinous sheath and appear to be involved in the degradation of the upper strata, but were never found inside the egg capsules. Metagenomic analysis revealed a large taxonomic diversity of bacteria associated with egg masses of A. depilans . Although 15 phyla could be recognized, the families Flavobacteriaceae (Bacteroidota), Lentisphaeraceae (Lentisphaerota), and Rhodobacteraceae (Pseudomonadota) represented 67.9% ± 11.6% of the relative abundance in the microbiome of the egg string samples. The presence of genera capable of decomposing polysaccharides, such as Tenacibaculum and Cellulophaga , supports the idea that bacteria are responsible for the degradation of the gelatinous layers of the egg strings.