Additional file 1: of First serine protease inhibitor isolated from Rhinella schneideri poison

Poison inhibitory assays. (DOCX 725 kb

Additional file 1: of Tityus serrulatus envenoming in non-obese diabetic mice: a risk factor for severity

Representative comparison of eyeball of Ts envenomed and non-envenomed mice. Mice were injected with Ts venom (1 mg/kg). (A) Envenomed NOD mice with glucose basal level ≥ 200 mg/dL, retinopathy indication. (B) Envenomed BALB/c mice, typical ptosis. (C) BALB/c mice control (non-envenomed), normal eyeball. (PPTX 81 kb

Additional file 1: of Peptidomic investigation of Neoponera villosa venom by high-resolution mass spectrometry: seasonal and nesting habitat variations

Peptide assignment of N. villosa venom extracted during winter and summer identified by nano-LC-ESI-MS/MS. (DOCX 13 kb

Additional file 2: of Peptidomic investigation of Neoponera villosa venom by high-resolution mass spectrometry: seasonal and nesting habitat variations

Peptide assignment of N. villosa venom extracted from arboreal and ground-dwelling ants identified by nano-LC-ESI-MS/MS. (DOCX 13 kb

Molecular and electrophysiological characteristics of the neurotoxins used in the study.

Molecular and electrophysiological characteristics of the neurotoxins used in the study.</p

Nociception assays with Ts5 response neutralized.

(A) Nociceptive behavior was evaluated in BALB/c male mice (n = 6) using 0.5 μg/animal paw of TsV as positive control animals. In the tested group of animals, TsV (1 μg/animal paw) was pre-incubated 1 h at 37° C with 2 eqM of purified IgGs anti-Ts5 (considering the % of Ts5 from the whole venom). The same rate was used in the positive control group, using 0.5 μg of Ts5/animal paw pre-incubated 1 h at 37° C with 2 eqM of IgGs anti-Ts5. Animals were observed for 35 min to record the time the animals spent either licking or lifting/shaking the injected paw. (B) Area under curve (AUC) over any individual parameter in the evaluation of nociceptive response with Ts5 neutralized. Data was analyzed using one-way ANOVA and Tukey’s post hoc test. * p < 0.05, when compared to the control group. Graphs were generated using GraphPad Prism Version 9.3.1.</p

<i>T</i>. <i>serrulatus</i> venom and toxins: Mass and molarity used for nociceptive assays.

T. serrulatus venom and toxins: Mass and molarity used for nociceptive assays.</p

Nociceptive assays.

(A) Nociceptive behavior was evaluated in C57BL/6 male mice (n = 6) using 0.5 μg paw injections of TsV and isolated toxins (Ts1, Ts5, Ts8, Ts19 frag II). Animals were observed for 35 min to record the time the animals spent either licking or lifting/shaking the injected paw. (B) Area under curve (AUC) over any individual parameter in the evaluation of nociceptive response. Data was analyzed using one-way ANOVA and Tukey’s post hoc test. * p p (C) Mice nociceptive behavior following paw injection with different quantities of Ts5 (0.1–4 μg). Graphs were generated using GraphPad Prism Version 9.3.1.</p

Ts5 recognition by scorpion antivenom and predicted epitopes.

(A) TsV + NI (-): TsV + non-immune horse serum; TsV + AS: TsV + scorpion antivenom (Butantan Institute); Ts5 + NI (-): Ts5 + non-immune horse serum; Ts5 + AS: Ts5 + scorpion antivenom (Butantan Institute); C (+): wells coated with non-immune horse serum. Rabbit anti-horse-HRP antibodies were used for the ELISA detection. The ELISA assay was performed in duplicate and results are plotted as mean ± SD. Data was analyzed using one-way ANOVA and Tukey’s post hoc test. Graph was generated using GraphPad Prism Version 9.3.1. *p (B) Ts5 primary sequence (P46115) [32] with the B-cell epitopes predicted by ABCpred Server tool highlighted and their scores. (C) Ts5 molecular model from AlphaFold Protein Structure Database [38, 39], modified by PyMol tool [40], with front and back view. Colors (blue, red, orange, and magenta) are not related to the alpha-helix and beta-pleated sheets, they represent the four B-cell epitopes predicted in different types of 3D representation: surface (up) and ribbon (down).</p

Nociception video.

(WMV)</p