This study assessed the potential of five fluorochromes in marking she
lls of the abalone Haliotis rubra, using an immersion technique. Such
marks are required to 'time stamp' the shells and thus determine wheth
er shell layers are deposited regularly enough to be used to age abalo
ne. The stains used were: oxytetracycline and tetracycline at 300-1000
mg L(-1); calcein at 10-120 mg L(-1); alizarin red S at 10-60 mg L(-1
); and xylenol orange at 20-100 mg L(-1). Immersion times were 12, 24
and 48 h. Mortality rates were low in all treatments, and clearly disc
ernible marks were produced when abalone were immersed for 24 and 48 h
at high concentrations in all the stains. Three problems were encount
ered when tetracyclines were employed: (i) the solutions were acidic,
so the pH had to be adjusted with NaOH to prevent mortality; (ii) ther
e was excessive foaming of the solutions; and (iii) a natural fluoresc
ence in the shells closely resembled that of the tetracyclines. Proble
ms also arose in assessing alizarin red and xylenol orange because the
y have long emission wavelengths, so that simultaneously viewing natur
al layers on the sections is difficult, Calcein, although expensive, w
as the most effective, as at high concentrations it consistently produ
ced bright, extensive marks. The success of marking appeared to depend
on the growth rate of the abalone, as feeding before staining increas
ed the intensity of marks, and marking varied between batches of abalo
ne collected at different times.