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DNA Extraction Optimisation for Minute Land Snails of Vertigo Müller, 1773 (Gastropoda: Vertiginidae): A Comparative Evaluation of Six Methods, Including a Non-Destructive Shell-Preserving Protocol.

No systematic comparison of DNA extraction strategies exists for minute Vertiginidae (shell height <&#x2009;3&#x2009;mm), a group posing a dual analytical challenge: extremely low tissue input and co-purified PCR-inhibitory mucus. For legally protected species, an additional requirement to preserve the shell voucher further constrains available protocols. Using Vertigo antivertigo as the model species, we compared six approaches applied to specimens preserved in 96% ethanol (n&#x2009;=&#x2009;10 per method): two HotSHOT alkaline-lysis protocols (destructive and non-destructive shell-preserving variants), a modified CTAB protocol supplemented with PVP-40 and DTT, and three commercial silica-column kits (GeneJET Genomic, DNeasy Blood & Tissue, QIAamp DNA Micro). DNA yields were quantified by QuantiFluor fluorometry, and PCR performance was subsequently assessed across four loci (COI barcode, COI mini-barcode, ITS1, ITS2). DNeasy Blood & Tissue produced the highest fluorometric concentrations; QIAamp DNA Micro and CTAB&#x2009;+&#x2009;PVP-40 gave intermediate values. The shell-preserving HotSHOT variant yielded lower concentrations but improved A260/230 ratios. BSA and trehalose supplementation increased PCR success in inhibition-prone HotSHOT extracts from 70% to 100%. ITS1 Sanger sequencing of three Vertigo species listed in Annex II of the EU Habitats Directive, all extracted with the shell-preserving protocol, confirmed species-level identification (99.8%-100% BLASTn identity; mean Phred Q&#x2009;>&#x2009;51). The shell-preserving non-destructive HotSHOT protocol yields sequenceable DNA from protected Vertiginidae while retaining the morphological voucher, making it the preferred option for conservation-genetic monitoring. The practical decision framework documented here-integrating voucher preservation, amplification robustness and per-sample cost-has broad applicability to other minute terrestrial gastropods processed in large-scale biodiversity surveys.

Habitats Directive

Beauty bias in butterfly research and conservation.

Conservation biases have been documented since the first emergence of the concept of biodiversity in the 1980s,1,2,3 showing a systematic disproportion in the allocation of research and conservation efforts among taxa.4,5,6,7,8,9,10,11 One factor underlying this disproportion, gaining prominence in recent literature, is species' perceived beauty, shaped by human visual preferences.12,13,14,15,16,17 Here, we integrate a large-scale survey of the perceived beauty of European butterflies yielding >21,000 survey completions from >100 countries into a time-explicit network linking species' beauty, public attention, research and conservation efforts, and the EU regulatory framework. We found that species beauty is consistently associated with public attention, research, and conservation efforts in a temporally structured pattern compatible with a cumulative beauty bias. Research effort and public attention concentrate on widespread and visually attractive species, whereas species included in the legal conservation framework, particularly the Convention on the Conservation of European Wildlife and Natural Habitats (hereafter, Bern Convention, BC, 1979)18 and the EU Habitats Directive (hereafter, HD, 1992)19 are disproportionately represented by visually appealing and historically protected taxa. Because these frameworks guide funding and management actions, early associations between species beauty and BC/HD inclusion have contributed to long-lasting institutional patterns in butterfly research and conservation. By contrast, European IUCN Red Lists20,21 do not overrepresent beautiful species and identify more inconspicuous taxa as threatened. This mismatch reveals a tension between scientific assessments of extinction risk and historically embedded conservation priorities. Our findings suggest that recognizing beauty bias is vital for aligning conservation with actual ecological urgency. VIDEO ABSTRACT.

Animals

Eye regionalization and spectral tuning of retinal pigments in insects.

The spatial and spectral properties of an eye can often be directly linked to the behaviour and habitat of the animal. In a honey bee (Apis mellifera) society, the drones use the well-developed dorsal part of the eye to detect the queen against the sky during her nuptial flight. Recently it has become clear that the dorsal area of the drone's eye serves its task by cleverly combining a number of optical mechanisms, thus achieving a high spatial acuity as well as a high sensitivity precisely in the wavelength range of interest--the ultraviolet to blue range. Since the various optical specializations in the drone eye have now been recognized, they can be traced in the eyes of other species: thus, the drone eye serves as a model to give a better understanding of the relationship between structure and function of compound eyes in particular, but also of visual systems in general.

Animals

[Orientation of the catfish in uniform and nonuniform electric fields].

Experiments with conditioning revealed that catfish showed compass orientation with precision about 20 degrees using a direction of homogeneous horizontally directed weak electric field with current density of 1.5 . 10(-11) A/mm2 in water with a specific resistance of 15 Ohm-m. Similar fields occur in their habitat. The catfish used the current density gradient for directed movements in the unhomogenous field. The lowest current density gradient in which the catfish showed the orientation behaviour proved to be 3.6 x 10(-12) A/mm2 per 1-mm distance. The possible mechanism of electro-orientation and its functional significance, are discussed.

Animals

Conservation genetics of whales and dolphins.

Whales and dolphins (cetaceans) are found in all the world's oceans and in some of the major rivers, yet little is known about the distribution and behaviour of many species. At the same time, cetaceans are under threat from a variety of pressures including direct and indirect takes, pollution, and competition for habitat and prey. To ensure their long-term survival it will be necessary to preserve genetic diversity through the identification and protection of differentiated populations, the assessment of variation within local populations, and through a better understanding of reproductive and dispersal behaviour. The application of molecular genetic techniques is helping to provide answers to some of these previously intractable questions. Early results suggest few consistent patterns. Obvious geographic boundaries correlate to genetic distance in some species, and not in others. Furthermore, morphological variation within species can be fairly extensive without correlating to genetic distance, or relatively minor between morphotypes that are as genetically distinct as some species. These examples emphasize the need for further study.

Animals

The influence of selective logging on primates and some other animals in East Kalimantan.

A brief survey was conducted in East Kalimantan, Indonesia, to determine the influence of selective logging operations on primate populations. Data were also collected on some birds and squirrels. Selective logging that causes only moderate habitat disturbance does not seem to result in a significant decrease in the densities of several primate species. Whereas some species adjust readily to moderately disturbed habitats, other species may have difficulty in maintaining a breeding population in logged areas. A review of the literature plus our own data suggest that rain forest birds and squirrels seem to be the most disrupted by selective logging of their habitats. While some of the other mammals are directly harmed by selective logging, others are endangered only by the human encroachment that often follows. Suggestions regarding enlightened tropical forestry practices, additional studies of the influence of selective logging, and the establishment of forest preservation areas are also discussed.

Animals

Adaptive Evolution for Freshwater Adaptation in Coilia nasus by Directional Selection on Osmoregulation Genes.

The molecular mechanisms underlying the adaptation to freshwater habitats in fish of marine origin remain unclear. Grenadier anchovies, such as Coilia nasus, originate from marine environments and include both anadromous and freshwater-resident conspecifics, making them ideal for studying adaptive evolution from marine to freshwater habitats. We conducted a comparative population genomic and transcriptome analysis of two distinct C. nasus lineages, one anadromous and the other freshwater-resident, collected from mainstream and estuarine regions of the Yangtze River, China. By genome-wide genotyping of the anadromous and the freshwater-resident populations, we observed significant divergence in osmoregulation, energy metabolism, and immune response pathways associated with ecological adaptation and energy expenditure for migration. Some ion transport genes such as CAMK1, ATP1&#x3b1;3, KCNJ1 and SLC30A2 were identified that may contribute to freshwater adaptation. Notably, numerous mineralocorticoid signalling genes (e.g., NR3C2, SGK1, ATP1&#x3b1;3, KCNJ1) exhibit dynamic change between the anadromous and freshwater populations, suggesting an important role for the hormone cortisol in regulating salinity acclimation in euryhaline fish. Among these genes, the ion channel ATP1&#x3b1;3 experienced adaptive amino acid substitutions (Val317Ile and Thr329Ser), which appear to be evolutionary hotspots across migratory species based on ortholog comparisons. These variants may facilitate sodium/potassium transport and highlight salinity tolerance as a key driver of divergence in anadromous fish transitioning to freshwater. These results enhance our understanding of the genetic basis underlying freshwater adaptation for an anadromous fish across osmotic boundaries.

Animals

Distribution and environmental synchronization of the marine insect, Halobates robustus, in the Galapagos islands.

The following three aspects of the biology of the marine insect Halobates robustus were studied, during a two week observation period, at several sites in the Galapagos Islands: distribution, aggregation behaviour and rhythmicity of locomotory activity. H. robustus occurred in highest numbers on the water surface at shores fringed with mangroves. The aggregations of H. robustus varied according to their location and density. Copulating adults formed dense, floating aggregations, which tended to be close to rocks or mangroves. Late instar nymphs were less aggregated and, in lagoons (where there was some shelter from direct tidal forces), were furthest from the shore. In two types of habitat (mangrove-fringed, sandy shores and rocky shores) the aggregations of H. robustus showed a pronounced ability to maintain a floating station in relation to the surrounding environment, irrespective of tidal movements (in one case at 34 m from the nearest fixed objects). Evidence of the ability of the aggregations to maintain station on the water surface was also obtained by comparing the movements of H. robustus with those of floating polystyrene particles, which move passively with wind and tide. Laboratory observations and experiments indicated no clear periodicity in locomotory movements throughout a 24 h period. However, the frequency of encounters between individuals showed two daily peaks, post-dawn and pre-dusk, with fewer encounters during the day and only occasional encounters during the night. By shifting the light-dark cycle it was demonstrated that the daily bimodal rhythm of encounters is triggered by dawn and, since it is not maintained in constant light or dark, an 'hour-glass' mechanism is suggested. The contributions of single adults, of copulating pairs and of immature stages to the overall pattern of activity were also determined. Immature stages did not affect the overall rate of encounters significantly and the interactions between single and copulating pairs of adults appear to have been responsible for the bimodal pattern.

Animals

Isolation of Legionella pneumophila from nonepidemic-related aquatic habitats.

Continuous centrifugation of large volumes of water from natural southeastern lakes allowed quantitative detection of Legionella pneumophila by direct immunofluorescent staining. Positive samples were injected intraperitoneally into guinea pigs, and the L. pneumophila were isolated and identified by their morphological, cultural, physiological, and serological characteristics.

Animals

A controlled environmental culture system for the production of erythropoietin.

A controlled environmental culture system in which consistent growth of fetal liver cells typical of the tissue of origin and capable of producing erythropoietin in culture is described. This system attempts to duplicate the major components of the natural habitat of the mammalian cell allowing for meaningful studies to be conducted on the regulation and control of erythropoietin biosynthesis. The direct effects of changes in pO2, pCO2, pH and various cell nutrients on cells producing erythropoietin is now possible.

Animals

Growth of Cryptococcus neoformans in UV-irradiated excreta of pigeons.

UV irradiation of pigeon droppings resulted in an increased concentration of some inhibitors (peroxides) of growth of Cryptococcus neofarmans. This may be, in addition to the direct germicidal action of sunshine, another cause of the rare occurrence of this fungus in pigeon droppings on unsheltered sites in natural habitats.

Ammonia

Metagenomic insights into ecological risk of antibiotic resistome and mobilome in riverine plastisphere under impact of urbanization.

Microplastics (MPs) are of increasing concern due to their role as reservoirs for antibiotic resistance genes (ARGs) and pathogens. To date, few studies have explored the influence of anthropogenic activities on ARGs and mobile genetic elements (MGEs) within various riverine MPs, in comparison to their natural counterparts. Here an in-situ incubation was conducted along heavily anthropogenically-impacted Houxi River to characterize the geographical pattern of antibiotic resistome, mobilome and pathogens inhabiting MPs- and leaf-biofilms. The metagenomics result showed a clear urbanization-driven profile in the distribution of ARGs, MGEs and pathogens, with their abundances sharply increasing 4.77 to 19.90 times from sparsely to densely populated regions. The significant correlation between human fecal marker crAssphage and ARG (R2&#xa0;=&#xa0;0.67, P=0.003) indicated the influence of anthropogenic activity on ARG proliferation in plastisphere and natural leaf surfaces. And mantel tests and random forest analysis revealed the impact of 17 socio-environmental factors, e.g., population density, antibiotic concentrations, and pore volume of materials, on the dissemination of ARGs. Partial least squares-path modeling further unveiled that intensifying human activities not only directly boosted ARGs abundance but also exerted a comparable indirect impact on ARGs propagation. Furthermore, the polyvinylchloride plastisphere created a pathogen-friendly habitat, harboring higher abundances of ARGs and MGEs, while polylactic acid are not likely to serve as vectors for pathogens in river, with a lower resistome risk score than that in leaf-biofilms. This study highlights the diverse ecological risks associated with the dissemination of ARGs and pathogens in varied MPs, offering insights for the policymaking of usage and control of plastics within urbanization.

Urbanization

[Serologic survey on human leptospirosis in Argentina].

This paper presents the results obtained with the sera of persons having direct or indirect contact with ethiological agents of leptospirosis. Exposure could have ocurred during the performance of their duties, by contact with contaminated habitats and/or animals. One thousand thirty eight sera from persons living in different regions of Argentina were received at the laboratory and studied by the microscopic agglutination tecnique. Of these, 164 were positive (15.7%) to serveral serotypes. One hundred thirty eight sera of the 857 males studied (16.1%) were positive, and 16 out of 173 female sera (15%) were positive. Males between 31 and 40 years of age had the highest proportion of positives (21.7%); however, among females the group between 21 and 30 years of age had the highest rate (14.2%). Most of the sera reacted to L. brastislava (15%), followed by L. icterohaemorrhagiae (17.7%) and L. butembo (14%), with nine other serotypes at lower rates.

Adolescent

Landscape heterogeneity, forest structure, and mammalian host diversity shape tick density and prevalence of the causative agent of Lyme borreliosis.

Ticks, particularly Ixodes ricinus, and the associated Lyme borreliosis risk, represent key concerns within the One Health framework, prompting extensive research in this field. However, comprehensive studies that jointly consider landscape characteristics, local forest structure and management, climate, and host community composition-alongside direct measures of tick density and infection status with Borrelia spp., the bacterial agents causing Lyme borreliosis, are scarce. In this study, we test the hypothesis that habitat diversity exerts a dilution effect, primarily by supporting greater diversity of mammal hosts. Therefore, we examined I. ricinus tick density and Borrelia spp. prevalence in relation to a comprehensive set of habitat and host-related variables. Ticks were collected using the flagging method and mammal hosts were monitored using an innovative camera-trapping approach across 25 forest plots along a land-use gradient within the Schw&#xe4;bische Alb exploratory in Germany. Both tick density and Borrelia spp. prevalence are influenced by a complex combination of habitat factors across different spatial scales, as well as the mammal host community composition. Overall, our results provide novel support to the dilution effect hypothesis, suggesting that greater habitat and host diversity contribute to a reduced Lyme borreliosis risk in this region.

Animals

Wear striations on the incisors of ceropithecid monkeys as an index of diet and habitat preference.

Wear striations on the incisiors of Old World monkeys were examined in order to determine associations between the distributions of striations, diet and habitat preference. Significant differences exist between the Cercopithecinae and the Colobinae in respect to the orientation of incisor wear striations. In the Colobinae striations are oriented in a predominantly mesiodistal direction. In the Cercopithecinae straitions usually have a labiolingual orientation. Comparisons of terrestrial and arboreal genera indicate that significant differences exist between the two groups in respect to the density of striations on the occlusal wear facets of maxillary central incisiors. Arboreal and terrestrial monkeys also differ in the frequency of individual incisor wear facets completely devoid of wear striations.

Animals

Reproduction on the Rocks: Life History of a Freshwater Macrobioeroding Bivalve.

Macrobioerosion, the excavation and removal of consolidated mineral substrates by macrofauna, is well established in marine systems, where macrobioeroders drive carbonate cycling, sediment production and habitat formation. In freshwater ecosystems, however, it has been documented in only a small number of invertebrate taxa and remains a poorly resolved ecological process. Among these, the teredinid shipworm Lithoredo abatanica represents a remarkable departure from the wood-boring ecology of its family, having evolved to excavate and ingest limestone in fresh water. Despite this remarkable ecological transition, its reproductive biology and life history remain unknown. Here, we investigate the reproductive mode and life history strategy of this species using population size structure, in&#xa0;situ observations of siphonal morphology, and sperm morphometrics. We show that L.&#x2009;abatanica reaches exceptional dimensions, with measured body lengths exceeding 100&#x2009;mm, one intact empty burrow exceeding 200&#x2009;mm and in-water observations indicating burrows possibly exceeding 500&#x2009;mm, establishing it as the largest known freshwater macrobioeroder. Its large size and dense aggregations indicate considerable capacity for local rock breakdown and habitat modification within the Abatan River system (Bohol, Philippines). Recently settled juveniles (<&#x2009;10&#x2009;mm) alongside reproductively mature individuals indicate ongoing recruitment. The morphology of the siphons and calcareous tube appears to preclude direct sperm transfer via pseudocopulation, while sperm morphometrics are consistent with external fertilisation. Together, these findings indicate that L.&#x2009;abatanica reproduces via broadcast spawning with external fertilisation and likely possesses a planktotrophic larval phase. This raises a fundamental question: how does a broadcast-spawning species with planktotrophic larvae maintain populations up to 15&#x2009;km upstream in a flowing freshwater river subject to persistent downstream advection? By resolving the life history of the largest known freshwater macrobioeroder, this study provides critical insight into the persistence, dispersal and ecological role of a globally unique riverine ecosystem engineer.

broadcast spawning

Native edaphoclimatic regions shape soil communities of crop wild progenitors.

Unveiling the soil biological communities ecologically associated with crop wild progenitors (CWPs) in their habitats of origin is essential for advancing productive and sustainable agriculture. A field survey was conducted to investigate the edaphoclimatic conditions and soil bacterial, fungal, protist, and invertebrate communities of 125 populations of direct progenitors of major crops for world agriculture. The wild populations clustered into four ecoregions shaped by two edaphoclimatic dimensions: one summarizing variations in soil sand contents and nutrients concentrations, and the other featuring changes in aridity, soil pH, and carbon storage potential. We identified a common soil core community across CWPs that varied significantly along deserts to tropical seasonal forests and savannas. The assembly of the soil core community was driven by varying environmental preferences amongst soil biodiversity kingdoms, reflecting potential shifts in their functional profiles. The tropical ecoregion exhibited higher proportion of acidophilic bacteria, fungal, and protist parasites, whilst desert ecosystems harboured greater abundances of saprophytic fungi and heterotrophic protists. Moreover, CWPs displayed unique microhabitats that incorporate variability into the soil community assembly. Our work reveals the biogeography of soil communities associated with CWPs, the first step towards the development of microbial rewilding initiatives.

centres of origin

Converting standard air decompression tables for no-stop diving from altitude or habitat.

Using the phase equilibration theory of Hills (1966), as modified by Hennessy and Hempleman (1977), it is possible to predict formulas for converting standard air decompression tables for no-stop diving at altitude or from a normoxic habitat, breathing air. For diving following equilibration at altitude, the Royal Navy, Royal Naval Physiological Laboratory, and Haldane-type rules appear to be too conservative, with the opposite result for diving after excursion to altitude. Predictions in the latter case are in fair agreement with the Swiss (Boni, Schibli, Nussberger, and Bühlmann 1976) no-stop altitude tables. In the case of habitats, close agreement is found between the Hamilton, Kenyon, Freitag, and Schreiner (1973) normoxic tables for no-stop downward excursions and indefinite dive upward-excursions on air. In the case of flying directly after no-stop diving, the US Navy rule of using repetitive group D appears to be conservative for dives less than 50 fsw, and possibly unsafe for dives over 50 fsw. It is concluded that for no-stop diving a single tissue and single safe ascent pressure formula are all that is necessary to generate equivalent air dives. This enforces the hypothesis that it is the volume of gas released on ascent that governs marginal type I bends, and that in a no-stop ascent, all excess dissolved gas is released in the worst case.

Aerospace Medicine