Dissections of Larval, Pupal and Adult Butterfly Brains for Immunostaining and Molecular Analysis

Butterflies possess impressive cognitive abilities, and investigations into the neural mechanisms underlying these abilities are increasingly being conducted. Exploring butterfly neurobiology may require the isolation of larval, pupal, and/or adult brains for further molecular and histological experiments. This procedure has been largely described in the fruit fly, but a detailed description of butterfly brain dissections is still lacking. Here, we provide a detailed written and video protocol for the removal of Bicyclus anynana adult, pupal, and larval brains. This species is gradually becoming a popular model because it uses a large set of sensory modalities, displays plastic and hormonally controlled courtship behaviour, and learns visual mate preference and olfactory preferences that can be passed on to its offspring. The extracted brain can be used for downstream analyses, such as immunostaining, DNA or RNA extraction, and the procedure can be easily adapted to other lepidopteran species and life stages.

Host preference of Thrips hawaiiensis for different ornamental plants

Thrips hawaiiensis is a common thrips pest that damages the flowers of various plants. The differing population sizes of T. hawaiiensis among host plants suggest its preference and performance vary among host plants. In this study, the host fitness of T. hawaiiensis for different flowers was assessed through field investigation. The behavioral responses of T. hawaiiensis to the color and volatiles of flowers eliciting different apparent fitness levels and their development and survival on the plants were also studied. Adults and larvae of T. hawaiiensis were found in the flowers of 21 species, which were classified into four fitness levels for this thrips species. T. hawaiiensis showed significantly different visual responses to the color and olfactory responses to the volatiles of four tested flowers (each representing one of the four fitness levels), with the rankings of visual preferences for Dianthus caryophyllus > Tulipa gesneriana > Hydrangea macrophylla > Rosa rugosa, and olfactory preferences for H. macrophylla ≥ T. gesneriana > D. caryophyllus > R. rugosa. Plant species had significant influences on the development and survival of T. hawaiiensis, with developmental times from egg to adult of 9.58 d, 9.92 d, 10.35 d and 10.75 d on H. macrophylla, T. gesneriana, D. caryophyllus and R. rugosa, respectively, and corresponding survival rates of 76.33%, 71.33%, 64.00% and 59.00%. In summary, this study shows that olfactory preferences were consistent with the field performance of T. hawaiiensis on the four flower plants tested. Further, fitness levels of host plant flowers are correlated with development rate and survivorship of T. hawaiiensis. Our study adds to the understanding of the mechanism of host selection by thrips and provides basic information to underpin the management of T. hawaiiensis on horticultural plants.

Something Stinks! Finding Ways to Manage Noxious Odours in the Operating Room and Other Clinical Settings A Randomized Controlled Trial

Objectives: The hospital can be saturated with noxious smells. Anecdotally, medical staff apply products to surgical masks to lessen the impact of these smells. This study aimed to determine the odour-masking ability of 4 inexpensive and convenient products. Methods: A randomized, single-blinded crossover study was conducted in Vancouver, Canada. Participants, 19 to 30 years old, were invited to participate. Participants with active allergies, upper respiratory tract infection, alteration to sense of smell, or failure of olfactory screen were excluded from the study. An experimental odour was used in lieu of a noxious surgical odour. After smelling the experimental odour without barriers, participants were re-exposed to the odour using 5 surgical masks in randomized order. Each mask was lined with a test product (cherry lip balm, tincture of benzoin, Mastisol, mint toothpaste, and control [plain mask]). Participants rated the effectiveness of products at masking the experimental odour from 0 to 100 (0 = completely ineffective, 100 = completely effective). Participants also rated the pleasantness of the products, recorded if the products made them feel unwell, and identified their preferred product overall. Results: Eighty participants were included in the study (33 male, 47 female), averaging 24.2 years of age. Mean odour-masking effectiveness for cherry lip balm was 66.5 (±24.6), tincture of benzoin: 62.6 (±25.0), Mastisol: 61.3 (±23.9), mint toothpaste: 57.5 (±27.4), and control: 21.9 (±21.8). All products performed better than the control ( P < .001), but there was no significant difference in performance between products. Cherry lip balm was the most preferred odour-masking product (29 participants), followed by mint toothpaste (22), Mastisol (14), tincture of benzoin (10), and control (5). Conclusions: All tested products demonstrated equivalent odour-masking abilities. If health care professionals choose to use an odour-masking product, they should consider their own olfactory preferences.

Chemosensory mechanisms of host seeking and infectivity in skin-penetrating nematodes

Approximately 800 million people worldwide are infected with one or more species of skin-penetrating nematodes. These parasites persist in the environment as developmentally arrested third-stage infective larvae (iL3s) that navigate toward host-emitted cues, contact host skin, and penetrate the skin. iL3s then reinitiate development inside the host in response to sensory cues, a process called activation. Here, we investigate how chemosensation drives host seeking and activation in skin-penetrating nematodes. We show that the olfactory preferences of iL3s are categorically different from those of free-living adults, which may restrict host seeking to iL3s. The human-parasitic threadwormStrongyloides stercoralisand hookwormAncylostoma ceylanicumhave highly dissimilar olfactory preferences, suggesting that these two species may use distinct strategies to target humans. CRISPR/Cas9-mediated mutagenesis of theS. stercoralis tax-4gene abolishes iL3 attraction to a host-emitted odorant and prevents activation. Our results suggest an important role for chemosensation in iL3 host seeking and infectivity and provide insight into the molecular mechanisms that underlie these processes.