Many organisms naturally defend themselves against microbial attachment and biofouling in the marine environment. In this study, we investigated microbial fouling on 2 molluscan egg capsules using scanning electron microscopy (SEM), two-photon laser scanning microscopy (TPLSM) with bacterial viability staining and bacterial attachment experiments with the biofilm-forming Pseudoalteromonas sp. S91 in flow chambers. Results indicated that early stage egg capsules of Dicathais orbita (Neogastropoda) are relatively free of surface microorganisms. Egg capsules during the trocophore stage had a regularly ridged microtexture, but as capsules matured, shedding of the outer wall was observed, followed by the extrusion of unidentified droplets, which then accumulated on the capsule surface in association with bacteria. By comparison, the egg capsules of Sepioteuthis australis (Cephalopoda) were found to have an irregular surface with many hills and valleys that accommodate colonization by a variety of microorganisms. At the later stages of development these squid egg capsules become heavily colonized by algal spores. Cross sections of egg capsules revealed that S. australis capsule walls were about 12 times thicker than D. orbita egg capsules. Staining the egg capsules with BacLight™ also revealed a significantly thicker biofilm, with more live and dead bacteria on S. australis capsules than on those of D. orbita (p < 0.05). Flow chamber experiments indicated that the surface of S. australis capsules provided a suitable substrate for colonization by Pseudoalteromonas sp. S91, whereas colonization was significantly less on D. orbita egg capsules after 24 and 72 h (p < 0.01). These experiments indicated that D. orbita egg capsules are better defended against fouling microbes than are the eggs of S. australis. D. orbita appears to use a combination of physical, mechanical and possibly chemical defense mechanisms to reduce fouling on their egg capsules.KEY WORDS: Biofilm · Egg capsules · Mollusc · Scanning electron microscopy · Two-photon laser scanning microscopy · Bacterial attachment
Resale or republication not permitted without written consent of the publisherAquat Microb Ecol 47: [275][276][277][278][279][280][281][282][283][284][285][286][287] 2007 an estimated density of 5 × 10 5 prokaryotic cells ml -1seawater (Whitman et al. 1998), sessile invertebrates and algae are exposed to a constant onslaught of potentially detrimental microbes. These include biofilm-forming bacteria along with single-cell diatoms that rapidly settle, attach and form colonies on any surface placed in the marine environment (Davis et al. 1989). The formation of a microbial biofilm promotes the attachment of algal spores, protozoa, barnacle cyprids and marine fungi, followed by the settlement of other marine invertebrate larvae and macroalgae (Maki 2000, Callow & Callow 2002. Heavy surface fouling could lead to the accumulation of toxic wastes, a reduction in oxygen and nutrient availability and increased drag, which can cause sessil...