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Fig. 2. Long-term memory (LTM), operant conditioning in `standard' and H2S environments. (A) 18 naïve snails received operant conditioning training under standard conditions (i.e. two 45 min sessions separated by 1 h) with a memory test (MT) performed 24 h later. This cohort exhibited both learning and memory. Learning was shown as the second training session was significantly lower than the first, and memory was shown as the memory test (MT) was significantly lower than the first session but not significantly greater than the second training session (ANOVA F(17,2)=20.9304, P<0.01; sessions 2 and 3 are significantly different from session 1, P<0.01) (*signifies that a session is significantly different from session 1 but not from session 2). (B) A cohort of 23 snails that received the `more intense hypoxic challenge' also demonstrated learning and memory. That is, since the number of openings in session 2 was significantly less than in session 1 (ANOVA F(22,2)=40.6394, P<0.01) learning was demonstrated. Additionally, since the memory test session (MT) was significantly different from session 1 but not different from session 2 memory was shown (ANOVA F(22,2)=66.8919, P<0.01 and F(22,2)=2.5811, P=0.1224, respectively). (C) A separate cohort (N=23) of snails underwent the training and testing protocol in hypoxia + H2S (100 µmol l–1). These snails showed neither learning nor memory. That is, the data in session 2 were not significantly different than that in session 1 (i.e. no learning) and the memory test session (MT) was not significantly different from session 1 (i.e. criteria for memory not met; ANOVA F(22,2)=2.3095, P=0.1112; no significant difference between sessions).





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