Calcein and manganese experiments for marking the shell of the common cockle (Cerastoderma edule): tidal rhythm validation of increments formation

Abstract

-This work focuses on investigating the potential of calcein and manganese as growth markers of the common cockle (Cerastoderma edule) in the bay of Somme (France). Recapture of shells, previously marked using a chemical marking and then bred in natural conditions, was performed in order to determine the shell growth patterns. Calcein marking has shown a fluorescent increment in shells after only 30 min immersion time at 150 mg L −1 , but also for shells immersed 3 h at 50 mg L −1 . Likewise, manganese shell marking was revealed under cathodoluminescence for shells immersed 1 h at 120 mg L −1 as well as for shells which spent 4 h at 90 mg L −1 . A numerical analysis performed on each marked cockles has revealed 23 micro-increments between the mark and the ventral edge of the valves, corresponding to the 23 tides that occurred during the 12 days at liberty post marking. The periodicity of increment formation is thus validated for a tidal frequency. The growth rates of C. edule, ranged from 11.67 to 19.94 μm d −1 , decreased significantly with increasing shell length. This preliminary study gives a clue to the understanding of cockle growth and could be used in shellfish production for cockle age monitoring, but also for chemical analysis to learn more about biomineralization process of this species.

Assessing the effect of calcein incorporation on physiological processes of benthic foraminifera

“…This was also reported for cockles (Mahé et al, 2010). We found that the sutures incorporated relatively more calcein and fluoresced brighter as compared to the rest of the chambers.…”

“…However, the susceptibility of organisms to calcein appears to vary among different organisms. No lethal effect was reported for gastropods and bivalves even after exposure to a calcein concentration as high as 150 mg/l (Moran, 2000;Riascos et al, 2007;Mahé et al, 2010).…”

Eficiencia de la tinción de bandas de crecimiento con calceina y azul de coomassie, para estimar la edad de jóvenes de Pinctada mazatlanica (Pterioida: Pteriidae)

“…Calcein used as a CaCO 3 -stainer was successful by immersion as a method for age validation producing a concise bright lime-green (Riascos et al, 2007), Donax hanleyanus (Herrmann et al, 2009), Cerastoderma edule (Mahé, Bellamy, Lartaud, & de Rafelis, 2010), Pinctada margaritifera (Linard et al, 2011), Loripes lacteus (van der Geest, van Gils, van der Meer, Olff, & Piersma, 2011) and other bivalves (Rowley & Mackinnon, 1995 Regarding the calcein concentration impact on marking, our individuals produced better marking (100%) at the lower (0.4g/L) than higher concentration (73% for 0.7g/L), as well as, a better quality mark at the lesser concentration (more perfect and good marks). The 27% of oysters at the greater concentration that showed no marking, actually showed green fluorescence over the whole shell thickness, as if the greater concentration affected the complete aragonite layer.…”

“…Herrmann (2009) found exposure time (3h vs 6h) to calcein to produce slightly more mortality for D. hanleyanus than the actual calcein concentration used (from 4 to 6% mortality), and thus recommended calcein staining as a non-lethal method. Mahé et al (2010) reported no mortality at 0.05 or 0.15g/L calcein concentration for (Cerastoderma edule). They observed clear marks at all concentrations and exposure times (submersion).…”

Use of calcein to estimate and validate age in juveniles of the winged pearl oysterPteria sterna

“…The existence of a sub-daily growth pattern has also been reported in the bivalves C. edule (House and Farrow 1968;Whyte 1975) and Chione cortezi (Goodwin et al 2001;Schöne et al 2002), although its relation with the physiology and periodicity of growth has not yet been established. In some cases, a subdaily growth rhythm has been associated with the presence of tides (Evans 1972(Evans , 1975Richardson et al 1979;Lonne and Gray 1988;Mahe et al 2010). Rodland et al (2006) suggested that these sub-daily growth patterns are controlled by internal rhythms or biological clocks, and that these mechanisms are in turn influenced by a complex interaction of physical (water temperature changes, daily light/dark cycles, tides, lunar cycle) and genetic cues (Beetjes and Williams 1985;Richardson 1988;Palmer 1995;Rodland et al 2006).…”