Figure 1.
Specific IgG antisera to Africanized bee venom demonstrated by End Point Titration ELISA.
Figure 2.
Hemolysis and neutralization of Africanized honeybee (AHB) venom.
A, Minimum concentration of AHB venom in vitro able to cause 100% hemolysis was established in 312.5 µg/mL; B, Using this established venom concentration the least amount of antivenom needed to completely neutralize the hemolysis was 50-fold times higher than the venom protein content. Values represent average of three experiments.
Figure 3.
Cytotoxicity of AHB venom and neutralization by antivenom.
Cytotoxicity of HBV against CHO cells, highlighting the minimal amount able to cause 100% of cells death - 108.24 mg/mL (A); Neutralization of minimal cytotoxic dose by diluted antivenom - 1:58 (B). Values represent average of three experiments.
Figure 4.
Inhibition of hyaluronidase in vitro and myotoxic activity of AHB venom in vivo.
A, The activity of hyaluronidase is shown for the whole venom as well for the dialyzed venom using the same protein content. The inhibition of this enzyme by antivenom was completely achieved. Values represent an average of three experiments. B, The myotoxic activity was measured by creatine kinase release and expressed the amount of enzyme that will catalyze 1 μmol of substrate per minute under specified conditions. The specific antivenom was able to completely neutralize this activity in vivo. Values represent the average of three experiments. Statistical analysis – ANOVA. CK – Creatine kinase; AV – antivenom; AHBV – Africanized bee venom.
Figure 5.
Hemolysis and neutralization in vivo.
A, Determination of Hemolytic Dose 50% (HD50) collecting blood after 15 min of AHB venom injection in mice. B, Neutralization of hemolytic activity using 3-fold times HD50 (5.49 µg/g) and different dilutions of antivenom collecting blood after 15 min of AHB venom injection in mice. Higher values of absorbance at 540 nm indicate higher hemolytic activity.