Chemical signals act as the main reproductive barrier between sister and mimetic Heliconius butterflies.

Colour pattern is the main trait that drives mate recognition between Heliconius species that are phylogenetically close. However, when this cue is compromised such as in cases of mimetic, sympatric and closely related species, alternative mating signals must evolve to ensure reproductive isolation and species integrity. The closely related species Heliconius melpomene malleti and H. timareta florencia occur in the same geographical region, and despite being co-mimics, they display strong reproductive isolation. In order to test which cues differ between species, and potentially contribute to reproductive isolation, we quantified differences in the wing phenotype and the male chemical profile. As expected, the wing colour pattern was indistinguishable between the two species, while the chemical profile of the androconial and genital males' extracts showed marked differences. We then conducted behavioural experiments to study the importance of these signals in mate recognition by females. In agreement with our previous results, we found that chemical blends and not wing colour pattern drive the preference of females for conspecific males. Also, experiments with hybrid males and females suggested an important genetic component for both chemical production and preference. Altogether, these results suggest that chemicals are the major reproductive barrier opposing gene flow between these two sister and co-mimic species.

Characterization and Comparative Analysis of Voids in Class II Composite Resin Restorations by Optical Coherence Tomography.

PURPOSE: This study aimed to characterize and analyze the number of voids and the percentage of void volume within and between the layers of class II composite restorations made using the bulk fill technique or the incremental technique by optical coherence tomography (OCT). METHODS AND MATERIALS: Class II cavities (4×4×2 mm) were prepared in 48 human third molars (n=24 restorations per group, two class II cavities per tooth). Teeth were divided into four groups and restored as follows: group 1 (FOB), bulk filled in a single increment using Filtek One Bulk Fill (3M Oral Care); group 2 (FXT), incrementally filled using four oblique layers of Filtek Z350 XT (3M Oral Care); group 3 (FBF+FXT), bulk filled in a single increment using Filtek Bulk Fill Flowable Restorative (3M Oral Care) covered with two oblique layers of Filtek Z350 XT (3M Oral Care), and group 4 (FF+FXT), incrementally filled using Filtek Z350 XT Flow (3M Oral Care) covered with two oblique layers of Filtek Z350 XT (3M Oral Care). After the restorative procedure, specimens were immersed into distilled water and stored in a hot-air oven at 37°C. Forty-eight hours later, thermal cycling was conducted (5000 cycles, 5°C to 55°C). Afterward, OCT was used to detect the existence of voids and to calculate the number of voids and percentage of voids volume within each restoration. Data were submitted to chi-square and Kruskal-Wallis tests (α=0.05). Comparisons were made using the Dunn method. RESULTS: Voids were detected in all groups, ranging from 0.000002 (FBF+FXT and FF+FXT) to 0.32 mm3 (FBF+FXT). FF + FXT presented voids in all of the restorations and had a significantly higher number of voids per restoration when compared to the other groups (p<0.05), but restorations with the presence of voids were significantly higher only when compared to FXT (p<0.05). FBF + FXT presented a significantly higher percentage of voids volume than that of FXT (p<0.05). When comparing restorations made using high-viscosity resin-based composites (FOB and FXT), no significant differences regarding number of voids or percentage of voids volume were detected (p≥0.05). CONCLUSIONS: The use of flowable resin-based composites can result in an increased number of voids and percentage of voids volume in restorations, and this appears to be more related to voids present inside the syringe of the material than to the use of incremental or bulk fill restorative techniques.