The male annihilation technique (MAT) plays a crucial role in the pest management program of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae). However, a suitable method for real-time and accurate assessment of MAT’s control efficiency has not been established. Laboratory investigations found that motile sperms can be observed clearly under the microscope when the spermathecae dissected from mated females were torn, and no sperms were found in the spermathecae of virgin females. Furthermore, it was confirmed that sperms can be preserved in the spermathecae for more than 50 days once females have mated. Laboratory results also indicated that proportion of mated females decreased from 100% to 2% when the sex ratio (♀:♂) was increased from 1:1 to 100:1. Further observation revealed that there were no significant differences in the superficial area of the ovary or spermatheca between mated females and virgin females. Field investigations revealed that the proportion of mated females (PMF) could reach 81.2% in abandoned mango orchards, whereas the PMF was less than 36.4% in mango orchards where MAT was applied. This indicates that the PMF of the field population can be determined by examining the presence of sperms in the spermathecae. Therefore, we suggest that this method can be used to monitor the control efficiency when MAT is used in the field.

Genome-scale searches for insect olfactory genes are vulnerable to rapid receptor divergence, tandem duplication, fragmented gene models, circular labels, and evolutionary leakage between training and test data. Sequence foundation models trained on DNA or proteins may extend homology-based annotation, but available evidence does not support their use as autonomous annotators or direct predictors of sensory function. This critical review organizes the problem around annotation failures rather than model catalogues. We distinguish six inference levels, from candidate locus detection to organism-level function, and define task-specific high-confidence reference criteria. We also propose a benchmark using biological hard negatives, sequence-cluster and taxonomic holdouts, conventional baselines, calibration, abstention, and workload-aware retrieval metrics. Evidence integration is defined operationally as a pipeline that retains model scores alongside similarity, domains, topology, phylogeny, expression, and assay evidence, without promoting a weak signal to a stronger biological claim. A published ESM-2-supported analysis of remote insect chemoreceptor homology illustrates both the potential and the current evidence boundary. A convincing benefit will require an incremental gain over transparent baselines under frozen biologically realistic splits, with reliable uncertainty and a clear route from ambiguous predictions to review or experiment.

The male annihilation technique for Bactrocera dorsalis relies primarily on methyl eugenol (ME), which exclusively targets mature males and consequently misses the critical window for managing sexually immature males (SIM). To address this operational gap, we cross-validated male testicular development with ME responsiveness across a 1–12-day age gradient, improved a conventional sugar–vinegar–alcohol lure (SL), and evaluated the combinations of the optimized sugar–vinegar–alcohol lure (OSL) and ME using a 2 × 2 factorial field design. Laboratory assays revealed that ME capture rates and off-white-to-yellow testicular transitions were highly synchronized and strongly correlated; notably, males aged 1–4 days exhibited entirely off-white testes and negligible ME responsiveness, establishing testicular coloration as a robust physiological hallmark of immaturity. In field trials, OSL demonstrated stable attractancy across diurnal intervals, outperforming water controls but yielding captures statistically comparable to the conventional SL. Furthermore, incorporating ME into OSL significantly enhanced SIM captures; however, factorial analysis confirmed that their interaction was not statistically significant. Ultimately, these findings indicate that OSL and ME exert a significant, independent additive effect rather than a synergistic interaction on SIM attraction. This study provides a reliable physiological–behavioral diagnostic framework and a practical, low-cost direct-mixing strategy to extend the operational window of B. dorsalis management.