In recent years, the lipopolysaccharide (LPS)-induced immune-endocrine interactions and its implications have been demonstrated in amphibians. Moreover, when considering immune challenges, the immune-endocrine alterations can vary with time of injection (day or night). In this study, we explored the LPS effects on the immune response (plasma bacterial killing ability – BKA, phagocytosis of blood cells – PP, and neutrophil: lymphocyte ratio – NLR) and endocrine mediators (corticosterone – CORT and melatonin – MEL plasma levels) in Rhinella icterica toads injected at day (10h) and night (22h). Our results showed LPS-induced increases in CORT, NLR, and PP when compared with saline-treated individuals. For NLR, the increase happened at both times (day and night) in the LPS-treated toads. Interestingly, for CORT levels, the response was more pronounced during the night, while for the PP, the effect was more evident during the day. However, no changes were observed in BKA and plasma MEL levels. Overall, our preliminary results demonstrated the LPS-injection promoting inflammatory response in R. icterica toads (higher PP and NLR), followed by activation of the hypothalamic-pituitary-interrenal axis (higher CORT levels) at both times of stimulation. Additionally, when the toads received the immune challenge (day or night) can differently favor the endocrine and immune mediators. It is also worth highlighting the relevance of investigating distinct immune components (cellular and protein) for a better understanding of the LPS-induced immunomodulation.