## 5 Discussion
### Standard Methods  
While standard methods for pollinator bioassays exist [32], such guidelines are still needed for SHBs. Topical bioassays have been used for evaluating the toxicity of potential control methods against adult SHBs [12; 33]. This allows the calculation of acute contact LD\({}_{508}\) for SHBs. To do this, a compound is administered directly to the exoskeleton of each SHB and then the SHB is monitored regularly to determine mortality. To calculate an oral LD\({}_{50}\) value, adult SHBs should be individually fed treated pollen and monitored regularly to measure mortality. A potential critique of our acute toxicity bioassay is the lack of certainty regarding oral or topical exposure of SHBs to the test compounds, though we argue that it is likely both. In our acute toxicity bioassay, groups of adult SHBs crawl throughout the pollen and feed on it, as seen when rearing SHBs with pollen [17]. As a result, adult SHB mortality may be caused by topical exposure, oral exposure, or a combination of both. Thus, we only calculate an LC\({}_{50}\) value using our assay. Nevertheless, our standard method is valuable in that it is field-realistic. The adult SHBs would be  
Figure 3: The 95% confidence intervals constructed from a generalized linear model with a negative binomial distribution for the average number of live small hive beetles (SHBs) aspirated from hives (**A**) and dead SHBs collected from traps (**B**) in each treatment. The averages for each treatment are depicted by a large dot with the confidence interval extending above and below it. Smaller dots represent the number of SHBs collected from the individual hives. Treatment means without overlapping confidence intervals are significantly different from one another.  
exposed topically and orally to treated pollen if beekeepers delivered SHB controls via pollen or other foodstuffs.  
A future SHB control agent should exhibit high toxicity to SHBs while maintaining relatively low toxicity to honey bees. While our proposed methods effectively assess the effectiveness of compounds against adult SHBs, they do not assess honey bee health. Thus, they cannot be used alone to determine the potential success of a future SHB control agent. These methods should be utilized alongside existing methodologies to assess bee health. These assessments include acute toxicity bioassays for bees [34], bee and brood scores taken at all stages of field trials [35], and residue analyses conducted on honey/pollen/wax samples [36]. The focus of our study was on SHBs specifically. Consequently, we opted to exclude piloting honey bee health assessments and advocate for using those that already exist [32; 34]. It is also possible that the method itself can impact the bees, given our reliance on the bounce method to catch and count SHBs. Nevertheless, this technique is essential for the accurate estimation of live beetle populations. The inclusion of negative controls can help one distinguish between adverse effects brought on by the bounce method and/or the compounds being tested.  
We list specific equipment to make the assay reproducible in Table 1. However, many specific items can be replaced with like items and still achieve the same desired outcome of assessing SHB control agents. For example, we choose to modify a CD case as traps for our field trial. The CD cases were easy to open and close for pollen placement, as well as facilitate the quantification of dead SHBs at the end of the trial. Other SHB trap designs could be used in place of the CD case and still effectively deliver treated pollen to adult SHBs. The most important aspect of the trap of choice is to ensure bees do not have access to the treated pollen. Finally, we identified thiamethoxam as an effective positive control for SHB acute toxicity bioassays.