Efecto de la regulación de la cabecera del río Genil (Sierra Nevada, España) sobre la comunidad de macroinvertebrados acuáticos y la dieta larvaria de Rhyacophila nevada (Insecta : Trichoptera)

Se analiza el efecto que un embalse (Embalse de Canales), situado en la parte alta de un río de montaña (Río Genil, Sierra Nevada, Granada, España), tiene sobre la comunidad de macroinvertebrados, y en especial sobre la dieta de las larvas del tricóptero depredador Rhyacophila nevada Zamora-Muñoz, C. & Alba-Tercedor, 1992. Para ello se establecieron estaciones de muestreo aguas arriba y abajo del embalse, realizándose muestreos cuantitativos de macroinvertebrados con una periodicidad mensual. Los macroinvertebrados (MIB) se identificaron hasta el máximo nivel taxonómico posible (en la mayoría de los casos a nivel de especie). La mayor diversidad específica se encontró en los dípteros quironómidos. Se estimó también la biomasa por especie (PSLC) y los ciclos de vida de las especies de depredadores, efemerópteros y dípteros. El contenido de los tractos digestivos fue analizado y cuantificado. La diversidad de los MIB estuvo determinada por los cambios de flujo y los ciclos de vida de las presas. Rhyacophila nevada es el depredador más abundante, presenta un ciclo multivoltino, comportándose como un depredador que busca activamente sus presas que ingiere comenzando por la región caudal. El número de presas en R. nevada aumentó con su tamaño. Las diferencias significativas de la dieta entre las estaciones de muestreo son consecuencia de los cambios en la disponibilidad de recursos (presas) en el tiempo. El ancho de nicho trófico también mostró diferencias entre estaciones.The effect that a dam (the Canales dam), located in the upper part of a mountain river (Genil stream, Sierra Nevada, Spain), has on the macroinvertebrate community, is analysed, especially over the larval diet of the predator caddisfly Rhyacophila nevada Zamora-Muñoz, C. & Alba-Tercedor, 1992. To that end, sampling stations were established, upstream and downstream of the dam. Quantitative samplings of macroinvertebrates were realized every month. The macroinvertebrates were identified to the maximum taxonomical level (species level in the majority of cases). The highest specific diversity was found in the chironomidae Diptera. The biomass per species was estimated , as well as the life cycles of the predator's species, mayflies and dipterans. Gut contents were analysed and quantified. The diversity of the MIB was determined by flow shifts and prey life cycles. Rhyacophila nevada is the most abundant predator, with a multivoltine life cycle, and behaving as an active searcher of preys, ingesting them from the caudal region. The number of prey items consumed by R. nevada increased with its size. Significant differences on the diet between the sampling stations are a consequence of the shift in the resource availability of preys in time. The trophic niche breadth was different among sites

Cenozoic origins of the genus Calliarcys (Insecta, Ephemeroptera) revealed by Micro‑CT, with DNA barcode gap analysis of Leptophlebiinae and Habrophlebiinae

We are grateful to Michel Sartori (Lausanne, Switzerland) and José Ángel Martín del Arco (Salamanca, Spain) for donating the specimens of C. humilis to SMNS, BC CAS, and UŁ collections. We are also grateful to Christel and Hans-Werner Hoffeins (Hamburg, Germany), Mike Reich (BSPG, Munich, Germany), and Evgeny Perkovsky (Schmalhausen Institute of Zoology, Kyiv, Ukraine) for access to their collections of fossil mayflies from the Eocene Rovno amber. We would like to thank Kateřina Bláhová (IE, BC CAS) and Milan Pallmann (SMNS) for technical assistance with line drawings and the preparation of a set of macro photographs. Tomasz Mamos (UniLodz, Poland) is acknowledged for his help in the Bayesian reconstruction of phylogeny, and Łukasz Trębicki (UniLodz, Poland) for help in molecular laboratory. Comments from reviewers helped to improve the manuscript. JA-T thank the staff of Bruker SkyScan in Kontich (Belgium) for their effectiveness and fast support, for their constant improvements to the software, and for implementing the new options we requested. In this respect, we are especially indebted to Alexander Sasov (now at NeoScan, https:// neosc an. com), Stephan Boons, Xuan Liu, Phil Salmon, and Vladimir Kharitonov. We would like to thank the reviewers for their thoughtful comments and efforts towards improving our manuscript. LSID urn:lsid:zoobank. org:pub:C58BEE82-0EC6-4C59-A02D-1E5F796179B6Additional information Supplementary Information The online version contains supplementary material available at https:// doi. org/ 10. 1038/ s41598- 022- 18234-4.Funding Open access funding enabled and organized by the University of Łódź (Poland). RJG acknowledges the financial support of the Grant Agency of the Czech Republic (No. 21-05216S) and institutional support of the Institute of Entomology (Biology Centre of the Czech Academy of Sciences) RVO: 60077344. Acquisition of research equipment used in this study has been carried out within equipment subsidy granted by Alexander von Humboldt Foundation [Georg Forster Research Fellowship for Experienced Researchers] for RJG.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.Funding Open access funding enabled and organized by the University of Łódź (Poland)Grant Agency of the Czech Republic (No. 21-05216S)Institute of Entomology (Biology Centre of the Czech Academy of Sciences) RVO: 60077344Alexander von Humboldt Foundation [Georg Forster Research Fellowship for Experienced Researchers

Anatomical study of the female reproductive system and bacteriome of Diaphorina citri Kuwayama, (Insecta: Hemiptera, Liviidae) using micro-computed tomography

Huanglongbing (HLB) (citrus greening disease) is one of the most serious bacterial diseases of citrus. It is caused by (1) Candidatus Liberibacter africanus, transmitted by Trioza erytreae and (2) C.L. asiaticus and C.L. americanus, transmitted by Diaphorina citri. As part of a multidisciplinary project on D. citri (www.citrusgreening.org), we made a detailed study, using micro-computed tomography, of the female abdominal terminalia, reproductive system (ovaries, accessory glands, spermatheca, colleterial (= cement) gland, connecting ducts, and ovipositor) and bacteriome, which we present here. New terms and structures are introduced and described, particularly concerning the spermatheca, ovipositor and bacteriome. The quality of images and bacteriome reconstructions are comparable, or clearer, than those previously published using a synchrotron or fuorescence in situ hybridisation (FISH). This study: reviews knowledge of the female reproductive system and bacteriome organ in D. citri; represents the frst detailed morphological study of D. citri to use micro-CT; and extensively revises existing morphological information relevant to psylloids, hemipterans and insects in general. High quality images and supplementary videos represent a signifcant advance in knowledge of psylloid anatomy and are useful tools for future research and as educational aids.Kansas State University (KSU) S15192.01University of Granada, USDA-NIFA S15192.01 2014-70016-2302

Predicting River Macroinvertebrate Communities Distributional Shifts under Future Global Change Scenarios in the Spanish Mediterranean Area

Several studies on global change over the next century predict increases in mean air temperatures of between 1°C to 5°C that would affect not only water temperature but also river flow. Climate is the predominant environmental driver of thermal and flow regimes of freshwater ecosystems, determining survival, growth, metabolism, phenology and behaviour as well as biotic interactions of aquatic fauna. Thus, these changes would also have consequences for species phenology, their distribution range, and the composition and dynamics of communities. These effects are expected to be especially severe in the Mediterranean basin due its particular climate conditions, seriously threatening Southern European ecosystems. In addition, species with restricted distributions and narrow ecological requirements, such as those living in the headwaters of rivers, will be severely affected. The study area corresponds to the Spanish Mediterranean and Balearic Islands, delimited by the Köppen climate boundary. With the application of the MEDPACS (MEDiterranean Prediction And Classification System) predictive approach, the macroinvertebrate community was predicted for current conditions and compared with three posible scenarios of watertemperature increase and its associated water flow reductions. The results indicate that the aquatic macroinvertebrate communities will undergo a drastic impact, with reductions in taxa richness for each scenario in relation to simulated current conditions, accompanied by changes in the taxa distribution pattern. Accordingly, the distribution area of most of the taxa (65.96%) inhabiting the mid-high elevations would contract and rise in altitude. Thus, families containing a great number of generalist species will move upstream to colonize new zones with lower water temperatures. By contrast, more vulnerable taxa will undergo reductions in their distribution area.This work was funded by GUADALMED-II (REN2001-3438-C07-06/HID), a project of excellence from “Junta de Andalucía” (RNM-02654/FEDER), the Spanish “Ministerio de Ciencia e Innovación” (CGL2007-61856/BOS), projects and a collaboration agreement between the “Spanish Ministerio de Medio Ambiente, Medio Rural y Marino” and the University of Granada (21.812-0062/8511)

Micro-CT study of male genitalia and reproductive system of the Asian citrus psyllid, Diaphorina citri Kuwayama, 1908 (Insecta: Hemiptera, Liviidae)

The Asian citrus psyllid (ACP), Diaphorina citri, is a major vector of the bacteria Candidatus Liberibacter asiaticus and C.L. americanus, which cause Huanglongbing disease (HLB) (aka Citrus greening disease), considered the most serious bacterial disease of citrus trees. As part of a multidisciplinary project on psyllid biology (www.citrusgreening.org), the results presented here concern a detailed anatomical study of the male reproductive system (testes, seminal vesicles, accessory glands, sperm pump, connecting ducts, and aedeagus) using micro-computed tomography (micro-CT). The study summarizes current knowledge on psyllids male reproductive system and represents significant advances in the knowledge of ACP anatomy.This work was supported by USDA-NIFA Award 2014-70016-23028 ªDeveloping an Infrastructure and Product Test Pipeline to Deliver Novel Therapies for Citrus Greening Diseaseº, 2015-2020

Método específico para la evaluación medioambiental de los lagos de origen glaciar pirenaicos y su aplicación al lago de Sabocos

This study proposes a new method to asses the environmental state of the Pyrenean glacial lakes, based on the Water Framework Directive, the concept of ecological state and inspired by widely tested and used methodologies. Starting from a good ecological state as a reference term, a series of matrices are used to identify and characterize all anthropic impacts and pressures. Information regarding the most significant ones is then compared with the criteria of a panel of experts and finally the results are summarised in ICPA matrices of Impacts, Consequences, Proposals of Corrective Measures, and Applicability. This method has been tested in the glacial lake of Sabocos, finding that its ecological quality is lower than expected, based on the identification of some severe impacts. In order to subdue them, it has been proposed a plan of correcting measures and valued its applicability.En este trabajo se propone un nuevo método para la evaluación medioambiental de los lagos pirenaicos de origen glaciar a partir de la Directiva Marco del Agua, fundamentado en el concepto de estado ecológico e inspirado en metodologías ampliamente contrastadas. Partiendo de un estado ecológico de referencia se identifican y caracterizan todas las presiones e impactos antropogénicos mediante el empleo de una serie de matrices adaptadas. Los impactos más significativos son contrastados por un panel de expertos. Finalmente, los resultados se expresan mediante matrices ICPA de Impactos, Consecuencias, Propuestas de medidas correctoras y Aplicabilidad. Este método de evaluación medioambiental se ha aplicado al lago de Sabocos. Los resultados obtenidos muestran una calidad ecológica inferior a la esperada y que algunos de los impactos identificados son severos. Con el objetivo de mitigar tales afecciones, se han propuesto una serie de medidas correctoras y evaluado su aplicabilidad

Revealing the respiratory system of the coffee berry borer (Hypothenemus hampei; Coleoptera: Curculionidae: Scolytinae) using micro-computed tomography

The coffee berry borer (Hypothenemus hampei) is the most economically important insect pest of coffee globally. Micro-computed tomography (micro-CT) was used to reconstruct the respiratory system of this species for the first time; this is the smallest insect (ca. 2 mm long) for which this has been done to date. Anatomical details of the spiracles and tracheal tubes are described, images presented, and new terms introduced. The total volume and the relationship between tracheal lumen diameter, length and volume are also presented. The total length of the tracheal tubes are seventy times the length of the entire animal. Videos and a 3D model for use with mobile devices are included as supplementary information; these could be useful for future research and for teaching insect anatomy to students and the public in general.This paper benefitted from the sub-award agreement S15192.01 between Kansas State University (KSU) and the University of Granada, as part of a USDANIFA Award 2014-70016-23028 to S.J. Brown (KSU), “Developing an Infrastructure and Product Test Pipeline to Deliver Novel Therapies for Citrus Greening Disease” (2015–2020)

Micro-computed tomography and histology to explore internal morphology in decapod larvae

Traditionally, the internal morphology of crustacean larvae has been studied using destructive techniques such as dissection and microscopy. The present study combines advances in microcomputed tomography (micro-CT) and histology to study the internal morphology of decapod larvae, using the common spider crab (Maja brachydactyla Balss, 1922) as a model and resolving the individual limitations of these techniques. The synergy of micro-CT and histology allows the organs to be easily identified, revealing simultaneously the gross morphology (shape, size, and location) and histological organization (tissue arrangement and cell identification). Micro-CT shows mainly the exoskeleton, musculature, digestive and nervous systems, and secondarily the circulatory and respiratory systems, while histology distinguishes several cell types and confirms the organ identity. Micro-CT resolves a discrepancy in the literature regarding the nervous system of crab larvae. The major changes occur in the metamorphosis to the megalopa stage, specifically the formation of the gastric mill, the shortening of the abdominal nerve cord, the curving of the abdomen beneath the cephalothorax, and the development of functional pereiopods, pleopods, and lamellate gills. The combination of micro-CT and histology provides better results than either one alone.Financial support was provided by the Spanish Ministry of Economy and Competitiveness through the INIA project (grant number RTA2011-00004-00-00) to G.G. and a pre-doctoral fellowship to D.C. (FPI-INIA)