“…Since the study by Corpuz (1981), it has been known that fertilization success can be enhanced using an artificial maturation treatment consisting of an alkaline seawater bath, generally consisting of NaOH solutions at pH 9.0-9.5 for several hours (Aquino De Souza et al, 2009;Peŕez et al, 2016;Guallart et al, 2020b;Cañizares et al, 2021;Castejoń et al, 2021), or based on NH 4 OH solutions at pH 8.5-9.0 lasting no more than 10 min (Wanninger et al, 1999;Gould et al, 2001;Hodgson et al, 2007;Ribeiro, 2008;Peŕez et al, 2016;Seabra et al, 2019;Nakano et al, 2020). The alkaline treatment promoted chorion removal and the acquisition of a spherical shape and significantly reduced the oocyte diameter to 141 ± 6 µm in P. aspera (Yuen's test, pvalue = 0) and 146 ± 6 µm in P. candei (Yuen's test, p-value = 0) (Figures 5Q, S; assay 5) when using NaOH at pH 8.4-9.0 for 3 h on these species (Cañizares et al, 2021;Castejoń et al, 2021;Castejoń et al, 2022). The duration of the alkaline bath should be controlled to avoid the degradation of the oocytes (Figure 5R) and to limit the ratio of abnormal development (Figures 9J, K) (Cañizares et al, 2021;Castejoń et al, 2021).…”