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Abstract A complete three-dimensional reconstruction of the internal damage (oviposition holes, entry and exit galleries, cavities caused by fungal infection) of three destructive pests of olive fruit was obtained using micro-computed tomography. In the case of the olive fruit fly (Bactrocera oleae), complete reconstruction of the galleries was achieved. The galleries were colour-coded according to the size of the internal lumens produced by larval instars. In the case of the olive moth (Prays oleae), we confirmed that the larvae only consume olive stones, leaving pulp tissue intact. This study revealed the evolutionary defensive adaptation developed by larvae, creating entrance/exit gallery in the form of a zigzag with alternating angles to avoid the action of possible parasitoids. In the case of olive fruit rot, caused by fungal infection transmitted by the midge (Lasioptera berlesiana), microtomography revealed the infection cavity, which was delimited by a protective layer of tissue produced by the plant to isolate the infection zone, which contained fungal hyphae and reproductive organs of the fungus. Two ovoid cavities were observed below a single external orifice in the concave necrotic depression. These results were interpreted as successive ovipositions of B. oleae, followed by the parasitoid L. berlesiana. High-resolution 3D rendered images are included as well as supplementary videos that could be useful tools for future research and teaching aids.

Abstract With micro-computed tomography techniques, using the single-distance phase-retrieval algorithm phase contrast, we reconstructed enhanced rendered images of soft tissues of Aedes aeqypti fourth instar larvae after Bti treatment. In contrast to previous publications based on conventional microscopy, either optical or electron microscopy, which were limited to partial studies, mostly in the form of histological sections, here we show for the first time the effects of Bti on the complete internal anatomy of an insect. Using 3D rendered images it was possible to study the effect of the bacterium in tissues and organs, not only in sections but also as a whole. We compared the anatomy of healthy larvae with the changes undergone in larvae after being exposed to Bti (for 30 min, 1 h and 6 h) and observed the progressive damage that Bti produce. Damage to the midgut epithelia was confirmed, with progressive swelling of the enterocytes, thickening epithelia, increase of the vacuolar spaces and finally cell lysis, producing openings in the midgut walls. Simultaneously, the larvae altered their motility, making it difficult for them to rise to the surface and position the respiratory siphon properly to break surface tension and breathe. Internally, osmotic shock phenomena were observed, resulting in a deformation of the cross-section shape, producing the appearance of a wide internal space between the cuticle and the internal structures and a progressive collapse of the tracheal trunks. Taken together, these results indicate the death of the larvae, not by starvation as a consequence of the destruction of the epithelia of the digestive tract as previously stated, but due to a synergic catastrophic multifactor process in addition to asphyxia due to a lack of adequate gas exchange.

Abstract Mayflies (Ephemeroptera) are among the oldest pterygote insects, with the earliest fossils dating back to the Late Carboniferous. Within mayflies, Leptophlebiidae are a highly diverse and widespread group, with approximately 140 genera and 640 species. Whereas taxonomy, systematics, and phylogeny of extant Leptophlebiidae are in the focus of extensive studies, little is known about leptophlebiid fossil taxa. Because fossil remains of Ephemeroptera in sedimentary rocks are relatively rare, inclusions of mayflies in amber are a unique source of information on their evolution and diversity in the past. Leptophlebiidae found in Cenozoic resins mostly belong to the subfamilies Leptophlebiinae (in Eocene Baltic amber) and Atalophlebiinae (in Miocene Dominican and Mexican ambers). In the present contribution, we confirm the first finding of the genus Calliarcys from Eocene Baltic amber by using Micro-CT, which allowed confirming its generic placement by visualizing diagnostic key characters otherwise hidden by a cloud of turbidity. Additionally, we present first molecular data on the extant species Calliarcys humilis Eaton, 1881 from the Iberian Peninsula and the barcode gap analysis for Leptophlebiinae and Habrophlebiinae.

Abstract Background The decapods are a major group of crustaceans that includes shrimps, prawns, crayfishes, lobsters, and crabs. Several studies focused on the study of the digestive system of the decapods, constituted by the oesophagus, stomach, midgut tract, midgut gland, and hindgut. Nevertheless, in the midgut tract there are associated a set of organs called “midgut caeca”, which are among the most controversial and less studied digestive organs of this group. This work used the common spider crab Maja brachydactyla Balss, 1922 as a model to resolve the origin, development, and potential role of the midgut caeca. Such organs were studied in the larvae (zoea I, zoea II, megalopa), first juveniles, and adult phases, being employed traditional and modern techniques: dissection, micro-computed tomography (Micro-CT), and light and electron microscopical analyses (TEM and SEM). Results The common spider crab has a pair of anterior midgut caeca and a single posterior caecum that originate from the endoderm germ layer: they develop from the midgut tract, and their epithelium is composed by secretory cells while lacking a cuticle lining. The midgut caeca are small buds in the newly hatched larvae, enlarge linearly during the larval development, and then continue growing until became elongated and coiled blind-tubules in adults. The adult midgut caeca are internally folded to increase their inner surface. The electron microscopy observations showed that the midgut caeca are highly active organs with important macroapocrine and microapocrine secretory activity. Our results suggest that the role of the caeca might be related to the digestive enzyme secretion. The secretory activity should increase as the animal grows in size. Conclusion The present study resolves the embryonic origin of the midgut caeca (endoderm derived organs), development (general lengthening starting from small buds), and role (active secretory organs). The secretory activity of the midgut caeca should be incorporated in the current models of the digestive physiology in different decapod taxa.

Hace ya varias décadas que el impacto mediático de la aparición de un gran agujero en la capa ozono, a nivel de la Antártida, fue un punto de inflexión por el cual se tuvo conciencia de lo que la actividad humana estaba produciendo a nivel planetario, y se tomaron medidas prohibiendo los clorofluorocarbonos (CFC) responsables de esa degradación. Lo que ha tenido un efecto muy positivo en la capa protectora de ozono que rodea la tierra. Pero los datos científicos vienen dando señales de que nuestro planeta está experimentando importantes cambios que se manifiestan, entre otros, por un incremento muy rápido de las temperaturas, con los efectos colaterales que ello implica.....