In the traditional fishing sector of Ecuador, integrating a gender perspective enhances sustainability and addresses climate change by recognizing the vital roles and knowledge of women in fisheries management. The objective of this study was to validate the content, comprehension, and construction of the designed questionnaire. Additionally, it is intended to identify perceptions of sustainability and climate change amongst women and men in fisheries, and compare their views on the dimensions of sustainability and climate change. The research implemented a questionnaire with a metrically satisfactory scale, providing information on how men and women perceive sustainability and climate change in the artisanal fishing sector. The data were collected by designing an ad hoc questionnaire denominated “Gender perception as regards sustainability and climate change” following a process in which the validity of the content (experts > 0.5823; Content Validity Ratio (CVR) > 6.1; Content Validity Index (CVI) > 0.852), decompression (pilot study, N = 30) and construct (KMO = 0.71) were analyzed. The results of the internal consistency reliability analysis indicated that the Likert scale had a high index of reliability, as evidenced by Cronbach’s Alpha coefficient of 0.70. This analysis provided evidence of the structure and validity of the scale, resulting in a reliable, integral, and short tool with which to measure perceptions of sustainability and climate change. The questionnaire was administered to 1464 people employed in the traditional fishing industry (17.14% women) in 40 fishing community ports near the coastal profile of Ecuador (Esmeraldas and Manabí). Most of those surveyed stated that the current traditional fishing quotas are not very sustainable in the long term (47.10%, n = 690) and that climate change may significantly negatively impact catch rates (50.30%, n = 736). Finally, the Wilcoxon test of tests confirmed that women had significantly more positive attitudes than men towards sustainability (3.03 ± 0.69) and climate change (3.35 ± 0.64).