# Impacts of Diverse Natural Products on Honey Bee Viral Loads and Health
## 3 Results  
### Chrysin, Curcumin, Hesperidin, Vanillin, and Tocopherol  
For these compounds, there were no significant differences in DWV compared to the control for any treatment (DF = 6, \(p\) = 0.6336), (Figure 1). The only significant difference in VDV levels when the bees were fed each of these treatments was seen when bees were fed hesperidin (mean control \(=-5.302\)\(+\)\(-\)\(2.6289\) (SE); mean hesperidin \(=-14.230\)\(+\)\(2.6289\) (SE), \(n\) = 8, \(p\) = 0.0208), where a \(\sim\)9-cycle difference indicates a 500-fold decrease in VDV when bees are fed hesperidin (Figure 1). There were no significant differences in the virus LSV compared to the control for any treatment (DF = 6, \(p\) = 0.2751). Although all treatments had generally higher levels of trypanosomes when compared to the controls, there was no significant difference for any of the treatments. Finally, there were no significant differences in _Nosema ceranae_ compared to the control for any treatment (DF = 6, \(p\) = 0.6101).  
Figure 1: One-way analysis of viral/bacterial loads and vitellogenin by treatment. Normalized Cq values of two honey bee viruses, DWV and VDV, the honey bee gut bacterial symbiont, _Gilliamella_, and transcripts for the storage protein vitellogenin. Treatments were supplemented at 100 ppm in sugar syrup. The diamond tips represent the 95% confidence intervals for the ANOVA, with the mean designated with the middle horizontal line across each diamond. Comparison circles shown the means, where circles that overlap are not significantly different, whereas circles that do not overlap are significantly different.  
_Gillianella_ was lower in all treatments tested, some significantly so (ANOVA DF = 6, \(p\) = 0.0210), such as tocopherol with an \(\sim\)8 cycle, \(2^{7}-2^{8}\)-fold difference compared to the control (mean control = \(-1.6958\) + 1.7447 (SE); mean octopherol = \(-9.8808\) + 1.7623 (SE), \(n\) = 8, \(p\) = 0.0019). Vanillin showed a \(\sim\)6-cycle difference (mean control = \(-1.6958\) + 1.7447 (SE); mean vanillin = \(-6.2737\) + 1.76447 (SE), \(n\) = 8, \(p\) = 0.0706), curcumin a \(\sim\)7-cycle difference (mean control = \(-1.6958\) + 1.7447 (SE); mean curcumin = \(-7.3573\) + 1.7447 (SE), \(n\) = 8, \(p\) = 0.0268), hesperidin a \(\sim\)5-cycle difference (mean control = \(-1.6958\) + 1.7447(SE); mean hesperidin = \(-5.7193\) + 1.7447 (SE), \(n\) = 8, \(p\) = 0.1104), and chrysin a \(\sim\)3 cycle difference (mean control = \(-1.6958\) + 1.7447 (SE); mean chrysin = \(-2.8945\) + 1.7447 (SE), \(n\) = 8), with lower levels of _Gillianella_ detected, but not significantly different (\(p\) = 0.629) (Figure 1).  
There are significant differences in the eater expression between some of the treatments and the control (DF = 5, \(p\) = 0.0002). Curcumin and octopherol led to lower expression levels of the immune effector eater compared to the control (mean control = \(-13.938\) + 1.3716 (SE); mean curcumin = \(-19.284\) + 1.3716 (SE), \(n\) = 8, \(p\) = 0.0086 and mean control = \(-13.938\) + 1.3716 (SE); mean octopherol = \(-17.306\) + 1.3716 (SE), \(n\) = 8, \(p\) = 0.0899). Curcumin showed a significant \(\sim\)6-cycle reduction in Cq values, reflecting a 40-fold reduction in expression with this supplement (Figure 2). Chrysin, hesperidin, and vanillin remained consistent with the expression levels of the control group. With the exception of hesperidin (mean control = 2.6212 + 1.9925 (SE); mean hesperidin = \(-3.1320\) + 1.9925 (SE), \(n\) = 8, \(p\) = 0.0475), which had a \(\sim\)6 cycle difference, reflecting 64 times more expression than in the control, there were no significant differences in the expression of hypenotaein in any other treatments tested when compared to the control (Figure 2). There were no differences in the expression of vittllogenin in any treatment when compared to the control (DF = 5, \(p\) = 0.9428) (data not shown).