Detection and characterization of Deformed Wing Virus (DWV) in apiaries with stationary and migratory management in the province of Entre Ríos, Argentina.

In Argentina, migratory activity in search of floral diversity has become a common approach to maximizing honey production. The Entre Ríos province possesses a floral diversity that allows beekeepers to perform migratory or stationary management. Beyond the impact caused by transhumance, migratory colonies in this province start and end the season in monoculture areas. To study the effect of these practices on viral infection, we assayed for the presence, abundance and genetic characterization of the Deformed Wing Virus (DWV) in honey bees from apiaries with both types of management. In migratory apiaries, DWV was detectable in 86.2% of the colonies at the beginning of the season (September 2018), and 66% at the end of the season (March 2019). On the other hand, DWV was detected in 44.11% and 53.12% of stationary samples, at the beginning and the end of the season, respectively. Sequence analysis from migratory and stationary colonies revealed that all samples belonged to DWV-A type. The highest viral loads were detected in migratory samples collected in September. Higher DWV presence and abundance were associated with migratory management and the sampling time. Based on our findings we propose that the benefit of migration to wild flowering areas can be dissipated when the bee colonies end the season with monoculture.

Morphological variation of the brachial plexus in four phyllostomid bat species (Chiroptera, Phyllostomidae).

Despite the remarkable morphological modifications that occurred in the thoracic limbs of bats, information about the brachial plexus in this group is still scarce. The present study aimed to describe the origin, structure, and distribution of these peripheral nerves in four Phyllostomidae species. Both antimeres of six Artibeus lituratus, five Desmodus rotundus, seven Glossophaga soricina, and five Phyllostomus hastatus-all adult males from the Adriano Lúcio Peracchi Collection (UFRRJ)-were dissected. After complete exposure of the structure, we found that the brachial plexus of D. rotundus and P. hastatus is formed by the same roots (C5-T1), whereas the fourth cervical spinal nerve and the second thoracic spinal nerve are present in G. soricina (C4-T1) and A. lituratus (C5-T2), respectively. There was intraspecific variation and asymmetry in the origin of the structure and the combinations of nerve segments forming terminal branches. The distribution to the target muscles and patagium, however, was not subject to significant variation in our sample. Data presented here support the presence of two prevailing conditions in distribution of nerves to the bat muscles, and the innervation of the membranes seems to be explained by embryogenesis. Although the brachial plexus in phyllostomid bats is similar to that of other terrestrial Laurasiatheria, aspects identified in these bats, apparently unique to Chiroptera, may be related to anatomical changes in the thoracic limbs functionally linked to flight.

Polyethylene microplastics and substrate availability can affect emergence responses of the freshwater insect Chironomus sancticaroli.

Pollution caused by polyethylene microplastics (MP) has been reported for aquatic environments worldwide. However, despite recurrent research for several aquatic organisms, the effects of MP on the emergence stage of freshwater insects from tropical environments are little known. This study is the first to assess the emergence of the Brazilian native species Chironomus sancticaroli Strixino & Strixino, 1981 when exposed to primary polyethylene microplastics (size 40-48 µm). We performed two exposure scenarios, with a substrate (standard assays) and without substrate (as a stressful experience), and recorded emergence responses. The MP did not affect the species' emergence rate, but these rates were statistically different for the standard and stressful exposure scenarios. In bioassays without substrate, the high concentrations of MP caused anticipation of the insect's emergence (5-6 days). On the other hand, female emergence time was longer than males in standard bioassays. The substrate absence caused a slight increase in the left female wing's length and the potential female fecundity. These findings suggest that the polyethylene microplastics and substrate availability can affect the emergence dynamics of the tropical insect C. sancticaroli.

Description of three new species of biting midge of the genus emAtrichopogon/em Kieffer (Diptera: Ceratopogonidae) from Brazilian Amazon.

Three new species of Atrichopogon with spotted wings of the family Ceratopogonidae (Diptera: Culicomorpha) from the Amazonas State of Brazil are described and illustrated. Male and female specimens of Atrichopogon janseni sp. nov., A. riopardensis sp. nov., and A. sergioluzi sp. nov. were associated by wing pigmentation patterns: with two darker spots, one over r-m and the other one in cell r3, posterior to the apex of R3. Male A. janseni sp. nov. have tergite 9 that is 2.5 × as broad as long, not extending to the apex of gonocoxite, and sternite 9 stout, sub-trapezoidal, with distal margin with a row of stout spines; gonostylus of A. janseni sp. nov. is spatulate, with distinct middle notch. Atrichopogon riopardensis sp. nov. is separated from other species with similar wing patterns and forked gonostylus, with gonostylus forked near midlength, with longitudinal furrow, inner portion short and fingernail-like, outer portion elongate and apically curved directed mesally; outer portion is 4 × longer than inner, with one long seta in basal 1/3. Contrastingly, A. sergioluzi sp. nov. has a small fork at the apex of gonostylus, and cercus broad basally with rounded tip. Females of the three new species are very similar (as are other females in this group), and it is probably not possible to distinguish them using the morphological features described. With the addition of the three new species described here, there are now known 15 Neotropical species of Atrichopogon with pigmented wings.

Structures related to pheromone storage in alar androconia and the female abdominal scent gland of Heliconius erato phyllis, Heliconius ethilla narcaea, and Heliconius besckei (Lepidoptera: Nymphalidae: Heliconiinae).

We describe the morphology of alar androconia and the female abdominal scent gland of Heliconius erato phyllis, Heliconius ethilla narcaea, and Heliconius besckei. Androconial scales of Heliconius, which are arranged in overlapping wing bands, release pheromones during courtship, probably through vibratory movements of male wings over the female to induce her to mate. An antiaphrodisiac is produced by glands located in the valves of the male and is transferred during copulation to the yellow dorsal abdominal sac present in the virgin female, causing this sac to emit a scent that reduces the attractiveness of the female for courtship with other males. Stereomicroscopy, SEM, and TEM analyses were conducted to describe the morphology of the internal and external scales and the external abdominal scent sac. The findings revealed different sizes of external androconial scales and an internal group of porous structural vesicles that are probably related to the preservation of internal space, reception and storage of secretions, and elimination of volatiles when the male is actively involved in courtship. Translucent projections on the female abdominal scent sac create open reservoirs for the reception, storage, and emission of antiaphrodisiac volatiles along with stink clubs. Male valve denticles vary in form and probably attach securely to the female sac during mating, thus ensuring secretion transfer. These features are discussed in the context of a comparative analysis.

Assessing the Use of Wing Morphometrics to Identify Fall Armyworm (Lepidoptera: Noctuidae) Host Strains in Field Collections.

The fall armyworm (Spodoptera frugiperda) (J. E. Smith) (Lepidoptera: Noctuidae), a major agricultural pest in the Western Hemisphere, has recently become established in Africa and Asia. This highly polyphagous species has potential to economically harm multiple crops. Contributing to this host range are two fall armyworm populations historically called 'host strains' that differ in host specificity. Understanding behaviors of the two strains is crucial to effective management of this pest. A major difficulty in such studies is that strains have long been considered morphologically indistinguishable, with molecular markers the only reliable means of identification. However, studies of fall armyworm in Colombia reported strain differences in wing morphology sufficiently large to potentially provide a more economical alternative method to determine strain. This study tested whether a similar phenotypic difference was present in Florida populations using geometric morphometric analysis of 15 anatomical landmarks on forewings of 182 specimens from three habitats associated with different host plants. Principle component and linear discriminant analyses identified significant differences in wing size and shape in comparison of strains from different habitats, but not between strains within the same habitat. Data indicate that apparent strain distinctions in wing phenotype are most likely a secondary consequence of differences in developmental growth patterns on different host plants combined with strain-biased host choice. Furthermore, Florida specimens showed much larger phenotypic overlap than observed for strains from Colombia. Together these findings suggest that wing morphology is probably not a reliable indicator of strain identity in field populations where different host plants are available.

Wing morphometrics of Aedes (Ochlerotatus) albifasciatus (Macquart, 1838) (Diptera: Culicidae) from different climatic regions of Argentina.

BACKGROUND: Gene flow restrictions between populations of Aedes albifasciatus, the vector of Western equine encephalitis and Dirophilaria immitis, have been described in the central region of Argentina. Genetic and eco-physiological variations usually result in local forms reflecting the climatic regions. Mosquito wings and their different parts have ecological functions in flight and communication. Therefore, wing shape could be considered an aspect of sexual dimorphism, and its eco-physiological responses can be expressed as morphological changes induced by the environment. METHODS: To compare the geographical and sexual variations with respect to wing shape and size in two Ae. albifasciatus populations from contrasting climates of Argentina (temperate: Buenos Aires, and the arid steppe of Patagonia: Sarmiento), the wings of adults reared in thermal trays at different constant temperatures (10-29 °C) were analyzed. RESULTS: The wing size of Ae. albifasciatus showed inverse linear relationships with the rearing thermal condition and higher slope for Buenos Aires. In the cool range (10-17 °C), geographical size variations responded to the converse Bergmann's rule, where Buenos Aires individuals were larger than those from Sarmiento. Sexual shape dimorphism occurred in both populations while geographical variation in shape was observed in both sexes. CONCLUSIONS: Buenos Aires individuals showed greater response sensitivity with respect to the size-temperature relation than those from Sarmiento. The converse Bergmann's rule in size variation could be due to a higher development rate in Sarmiento to produce more cohorts in the limited favorable season. The shape could be more relevant with respect to the size in the study of population structures due to the size being more liable to vary due to changes in the environment. The geographical variations with respect to morphology could be favored by the isolation between populations and adaptations to the environmental conditions. Our results demonstrate that the shape and size of wing provide useful phenotypic information for studies related to sexual and environmental adaptations.

Extracellular activity of NBAD-synthase is responsible for colouration of brown spots in Ceratitis capitata wings.

After the emergence of the Ceratitis capitata imago, the pale and folded wings are expanded and sclerotized to acquire the definitive form and to stabilize the cuticle. The wings of this fly show a specific pattern of brownish and black spots. Black spots are pigmented by melanin, whereas there was scarce information about the development of the brownish spots. N-beta-alanydopamine (NBAD) is the main tanning precursor in C. capitata body cuticle, and we hypothesized that it may be responsible for the colouration of the brownish spots. We determined the topology and timing of NBAD synthesis and deposition to attain the species-specific colouration pattern. We demonstrated that during the first hours the colour of the brownish spots was principally determined by the tanning of the hairs. Haemolymph circulation through the veins is required to tan the wings. We confirmed that soon after wing spreading, most of the wing epidermal cells disappeared. Thus, the tanning of the brown spots was accomplished when the wing lamina was devoid of cells. NBAD synthase (NBAD-S; Ebony protein in D. melanogaster) activity in wings was detected in pharate adults and lasted several days after the emergence, even after the end of the tanning process. This observation is in contrast to epidermal NBAD-S activity in the body, where it was nearly undetectable 48 h post emergence. Our results indicate that NBAD-S was exported and deposited into the extracellular matrix of the brown spot areas before cell death and that tanning occurs through gradual export of NBAD precursors (dopamine and b-alanine) from veins.

Genotoxicity and Antigenotoxicity Assessments of the Flavonoid Vitexin by the Drosophila melanogaster Somatic Mutation and Recombination Test.

Vitexin is a C-glycosylated flavone found in various medicinal plants with several proven biological properties such as anti-inflammatory, antispasmodic, antimicrobial, cytotoxic, and antioxidant activities. Considering that the human population consumes many foods that contain this flavonoid, the objective of this study was to investigate the genotoxic and the antigenotoxic potentials of vitexin by the SMART (Somatic Mutation and Recombination Test) in Drosophila melanogaster wings, in vivo. To evaluate the genotoxic activity, larvae obtained from standard (ST) and high bioactivation (HB) crosses were chronically treated with different concentrations of vitexin (0.15, 0.3, and 0.6 mM). For the evaluation of antigenotoxic activity, the same vitexin concentrations were associated with two damage inducing agents: doxorubicin (0.2 mM) and benzo[a]pyrene (1.0 mM). The results obtained for genotoxic activity showed that vitexin did not induce a statistically significant increment in the frequency of mutant spots, when compared to control. The results for the evaluation of antigenotoxicity indicated that the flavonoid statistically reduced the frequency of mutant spots, when compared to those treated with only the damage inducing agents. Thus, this article presents results that have demonstrated the antigenotoxic activity of vitexin, which could be applied in new studies for the development of drugs with chemoprotective effects.

Effects of chemical stimulation of the lateral wings of the dorsal raphe nucleus on panic-like defensive behaviors and Fos protein expression in rats.

The lateral wings subnucleus of the dorsal raphe nucleus (lwDR) has been implicated in the modulation of panic-like behaviors, such as escape. Infusion of non- excitotoxic doses of the excitatory amino acid kainic acid into this subnucleus promptly evokes a vigorous escape response. In addition, rats exposed to panic-inducing situations show an increase in Fos protein expression in neurons within the lwDR. In the present study, we first investigated whether key structures associated with the mediation of escape behavior are recruited after chemical stimulation of the lwDR with kainic acid. We next investigated whether the infusion of the GABAA receptor antagonist bicuculline into the lwDR also evoked escape responses measured both in a circular arena and in the rat elevated T-maze. The effects of bicuculline in the circular arena were compared to those caused by the infusion of this antagonist into the ventrolateral periaqueductal gray (vlPAG), an area in close vicinity to the lwDR. The results showed that kainic acid infusion into the lwDR increased Fos protein immunostaining in brain structures deeply involved in panic-like defensive behaviors, such as the periaqueductal gray and hypothalamus, but not the amygdala. As observed with kainic acid, bicuculline evoked a pronounced escape response in the circular arena when microinjected in the lwDR, but not in the vlPAG. The escape-promoting effect of bicuculline in the lwDR was also evidenced in the elevated T-maze. These findings strength the view that dysfunction in mechanisms controlling escape in the lwDR is critically implicated in the pathophysiology of panic disorder.

Tergal and pleural structures contribute to the formation of ectopic prothoracic wings in cockroaches.

Wings were a fundamental morphological innovation for the adaptive radiation of insects, the most diversified group among all animals. Pterygote insects have two pairs of wings, the mesothoracic (T2) forewings and the metathoracic (T3) hindwings, whereas the prothorax (T1) is wingless. Using RNA interference approaches, we have found that the gene Sex combs reduced (Scr) determines the wingless identity of T1 in the cockroach Blattella germanica. Interference of Scr triggers the formation of ectopic wing structures in T1, which are formed from the expansion of the latero-posterior region of the pronotum, along with a contribution of the epimeron, a pleurite of T1. These data support the theory of a dual origin for insect wings, from pronotal (tergal origin theory) and pleural (pleural origin theory) structures and genes.

Triatoma maculata, the Vector of Trypanosoma cruzi, in Venezuela. Phenotypic and Genotypic Variability as Potential Indicator of Vector Displacement into the Domestic Habitat.

Triatoma maculata is a wild vector of Trypanosoma cruzi, the causative agent of Chagas disease; its incursion in the domestic habitat is scant. In order to establish the possible domestic habitat of T. maculata, we evaluated wing variability and polymorphism of genotypic markers in subpopulations of T. maculata that live in different habitats in Venezuela. As markers, we used the mtCyt b gene, previously apply to evaluate population genetic structure in triatomine species, and the ß-tubulin gene region, a marker employed to study genetic variability in Leishmania subgenera. Adults of T. maculata were captured in the period 2012-2013 at domestic, peridomestic (PD), and wild areas of towns in the Venezuelan states of Anzoátegui, Bolívar, Portuguesa, Monagas, Nueva Esparta, and Sucre. The phenotypic analysis was conducted through the determination of the isometric size and conformation of the left wing of each insect (492 individuals), using the MorphoJ program. Results reveal that insects of the domestic habitat showed significant reductions in wing size and variations in anatomical characteristics associated with flying, in relation to the PD and wild habitats. The largest variability was found in Anzoátegui and Monagas. The genotypic variability was assessed by in silico sequence comparison of the molecular markers and PCR-RFLP assays, demonstrating a marked polymorphism for the markers in insects of the domestic habitat in comparison with the other habitats. The highest polymorphism was found for the ß-tubulin marker with enzymes BamHI and KpnI. Additionally, the infection rate by T. cruzi was higher in Monagas and Sucre (26.8 and 37.0%, respectively), while in domestic habitats the infestation rate was highest in Anzoátegui (22.3%). Results suggest domestic habitat colonization by T. maculata that in epidemiological terms, coupled with the presence in this habitat of nymphs of the vector, represents a high risk of transmission of Chagas disease.