With an accelerating negative impact of anthropogenic actions on natural ecosystems, non-invasive biodiversity assessments are becoming increasingly crucial. As a consequence, the interest in the ...application of environmental DNA (eDNA) survey techniques has increased. The use of eDNA extracted from faeces from generalist predators, have recently been described as "biodiversity capsules" and suggested as a complementary tool for improving current biodiversity assessments. In this study, using faecal samples from two generalist omnivore species, the Eurasian badger and the red fox, we evaluated the applicability of eDNA metabarcoding in determining dietary composition, compared to macroscopic diet identification techniques. Subsequently, we used the dietary information obtained to assess its contribution to biodiversity assessments. Compared to classic macroscopic techniques, we found that eDNA metabarcoding detected more taxa, at higher taxonomic resolution, and proved to be an important technique to verify the species identification of the predator from field collected faeces. Furthermore, we showed how dietary analyses complemented field observations in describing biodiversity by identifying consumed flora and fauna that went unnoticed during field observations. While diet analysis approaches could not substitute field observations entirely, we suggest that their integration with other methods might overcome intrinsic limitations of single techniques in future biodiversity surveys.
Tropical forests shelter an unparalleled biological diversity. The relative influence of environmental selection (i.e., abiotic conditions, biotic interactions) and stochastic–distance‐dependent ...neutral processes (i.e., demography, dispersal) in shaping communities has been extensively studied for various organisms, but has rarely been explored across a large range of body sizes, in particular in soil environments. We built a detailed census of the whole soil biota in a 12‐ha tropical forest plot using soil DNA metabarcoding. We show that the distribution of 19 taxonomic groups (ranging from microbes to mesofauna) is primarily stochastic, suggesting that neutral processes are prominent drivers of the assembly of these communities at this scale. We also identify aluminium, topography and plant species identity as weak, yet significant drivers of soil richness and community composition of bacteria, protists and to a lesser extent fungi. Finally, we show that body size, which determines the scale at which an organism perceives its environment, predicted the community assembly across taxonomic groups, with soil mesofauna assemblages being more stochastic than microbial ones. These results suggest that the relative contribution of neutral processes and environmental selection to community assembly directly depends on body size. Body size is hence an important determinant of community assembly rules at the scale of the ecological community in tropical soils and should be accounted for in spatial models of tropical soil food webs.
see also the Perspective by Dumbrell
Continued advancements in environmental DNA (eDNA) research have made it possible to access intraspecific variation from eDNA samples, opening new opportunities to expand non‐invasive genetic studies ...of wildlife populations. However, the use of eDNA samples for individual genotyping, as typically performed in non‐invasive genetics, still remains elusive. We present successful individual genotyping of eDNA obtained from snow tracks of three large carnivores: brown bear (Ursus arctos), European lynx (Lynx lynx) and wolf (Canis lupus). DNA was extracted using a protocol for isolating water eDNA and genotyped using amplicon sequencing of short tandem repeats (STR), and for brown bear a sex marker, on a high‐throughput sequencing platform. Individual genotypes were obtained for all species, but genotyping performance differed among samples and species. The proportion of samples genotyped to individuals was higher for brown bear (5/7) and wolf (7/10) than for lynx (4/9), and locus genotyping success was greater for brown bear (0.88). The sex marker was typed in six out of seven brown bear samples. Results for three species show that reliable individual genotyping, including sex identification, is now possible from eDNA in snow tracks, underlining its vast potential to complement the non‐invasive genetic methods used for wildlife. To fully leverage the application of snow track eDNA, improved understanding of the ideal species‐ and site‐specific sampling conditions, as well as laboratory methods promoting genotyping success, is needed. This will also inform efforts to retrieve and type nuclear DNA from other eDNA samples, thereby advancing eDNA‐based individual and population‐level studies.
The conservation of wildlife requires management based on quantitative evidence, and especially for large carnivores, unraveling cause-specific mortalities and understanding their impact on ...population dynamics is crucial. Acquiring this knowledge is challenging because it is difficult to obtain robust long-term data sets on endangered populations and, usually, data are collected through diverse sampling strategies. Integrated population models (IPMs) offer a way to integrate data generated through different processes. However, IPMs are female-based models that cannot account for mate availability, and this feature limits their applicability to monogamous species only. We extended classical IPMs to a two-sex framework that allows investigation of population dynamics and quantification of cause-specific mortality rates in nonmonogamous species. We illustrated our approach by simultaneously modeling different types of data from a reintroduced, unhunted brown bear (Ursus arctos) population living in an area with a dense human population. In a population mainly driven by adult survival, we estimated that on average 11% of cubs and 61% of adults died from human-related causes. Although the population is currently not at risk, adult survival and thus population dynamics are driven by anthropogenic mortality. Given the recent increase of human-bear conflicts in the area, removal of individuals for management purposes and through poaching may increase, reversing the positive population growth rate. Our approach can be generalized to other species affected by cause-specific mortality and will be useful to inform conservation decisions for other nonmonogamous species, such as most large carnivores, for which data are scarce and diverse and thus data integration is highly desirable. La conservación de la vida silvestre requiere de manejo basado en evidencia cuantitativa. Para los carnívoros, en especial, es crucial cuantificar las tasas de mortalidad por causas específicas y entender su impacto sobre las dinámicas poblacionales. Adquirir este conocimiento es un reto, ya que es complicado obtener bases de datos a largo plazo de poblaciones amenazadas y, usualmente, los datos provienen de distintas estrategias de muestreo. Los modelos de población integrados (MPIs) ofrecen una forma de integrar los datos generados por medio de procesos diferentes. Sin embargo, los MPIs son modelos basados en hembras que no tienen en cuenta de la disponibilidad de pareja y esta característica limita su aplicabilidad solamente a las especies monógamas. Extendimos los MPIs clásicos a un marco de trabajo de dos sexos que permite la investigación de las dinámicas poblacionales y la cuantificación de las tasas de mortalidad por causas específicas en especies no monógamas. Ilustramos nuestra estrategia modelando simultáneamente diferentes tipos de datos de una población de osos (Ursus arctos), reintroducida y no sujeta a caza, que vive en un área con una población humana densa. En una población impulsada principalmente por la supervivencia de adultos, estimamos que en promedio el 11% de los cachorros y el 61% de los adultos murieron por causas relacionadas con humanos. Aunque actualmente la población no se encuentra en riesgo, la supervivencia adulta y, por lo tanto, la dinámica poblacional están dirigidas por la mortalidad antropogénica. Debido al incremento reciente de los conflictos entre humanos y osos en la zona, la extracción de individuos por razones de manejo o por caza furtiva puede incrementar, lo que invertiría la tasa positiva de crecimiento poblacional. Nuestra aproximación analítica puede generalizarse a otras especies afectadas por distintas fuentes de mortalidad y será útil para informar las decisiones de conservación en otras especies no monógamas, como la mayoría de los grandes carnívoros, para las cuales los datos son escasos y diversos, y por lo tanto es deseable la integración de los sus datos.
Environmental DNA (eDNA) metabarcoding is increasingly used to study the present and past biodiversity. eDNA analyses often rely on amplification of very small quantities or degraded DNA. To avoid ...missing detection of taxa that are actually present (false negatives), multiple extractions and amplifications of the same samples are often performed. However, the level of replication needed for reliable estimates of the presence/absence patterns remains an unaddressed topic. Furthermore, degraded DNA and PCR/sequencing errors might produce false positives. We used simulations and empirical data to evaluate the level of replication required for accurate detection of targeted taxa in different contexts and to assess the performance of methods used to reduce the risk of false detections. Furthermore, we evaluated whether statistical approaches developed to estimate occupancy in the presence of observational errors can successfully estimate true prevalence, detection probability and false‐positive rates. Replications reduced the rate of false negatives; the optimal level of replication was strongly dependent on the detection probability of taxa. Occupancy models successfully estimated true prevalence, detection probability and false‐positive rates, but their performance increased with the number of replicates. At least eight PCR replicates should be performed if detection probability is not high, such as in ancient DNA studies. Multiple DNA extractions from the same sample yielded consistent results; in some cases, collecting multiple samples from the same locality allowed detecting more species. The optimal level of replication for accurate species detection strongly varies among studies and could be explicitly estimated to improve the reliability of results.
Cancer spheroids are in vitro 3D models that became crucial in nanomaterials science thanks to the possibility of performing high throughput screening of nanoparticles and combined nanoparticle-drug ...therapies on in vitro models. However, most of the current spheroid analysis methods involve manual steps. This is a time-consuming process and is extremely liable to the variability of individual operators. For this reason, rapid, user-friendly, ready-to-use, high-throughput image analysis software is necessary. In this work, we report the INSIDIA 2.0 macro, which offers researchers high-throughput and high content quantitative analysis of in vitro 3D cancer cell spheroids and allows advanced parametrization of the expanding and invading cancer cellular mass. INSIDIA has been implemented to provide in-depth morphologic analysis and has been used for the analysis of the effect of graphene quantum dots photothermal therapy on glioblastoma (U87) and pancreatic cancer (PANC-1) spheroids. Thanks to INSIDIA 2.0 analysis, two types of effects have been observed: In U87 spheroids, death is accompanied by a decrease in area of the entire spheroid, with a decrease in entropy due to the generation of a high uniform density spheroid core. On the other hand, PANC-1 spheroids' death caused by nanoparticle photothermal disruption is accompanied with an overall increase in area and entropy due to the progressive loss of integrity and increase in variability of spheroid texture. We have summarized these effects in a quantitative parameter of spheroid disruption demonstrating that INSIDIA 2.0 multiparametric analysis can be used to quantify cell death in a non-invasive, fast, and high-throughput fashion.
Urinary incontinence (UI) is a very common condition, negatively affecting social, occupational, domestic, and psychophysical wellbeing. In particular, a peculiar and detrimental effect of UI has ...been described concerning sexual function. However, the impact of UI on quality of life is not fully understood yet, and further investigation into this issue is warranted. With this narrative review, we aimed to report the current evidence from recent literature regarding the quality of life and psychological wellbeing in patients with urinary incontinence, with a special focus on sexual function and its evolution after UI treatment. There is strong evidence that urinary incontinence-in its different forms, including stress urinary incontinence, urge urinary incontinence, mixed urinary incontinence, and coital urinary incontinence-negatively affects female sexual function. Treatments aimed to cure urinary incontinence-including pelvic floor muscles training, medications, and surgery-seem to improve quality of life by recovering, at least in part, sexual function. In conclusion, there is a substantial association between involuntary urinary loss and sex life quality. However, few studies are available and more evidence is needed before consistent conclusions can be made.
: A consensus regarding the optimal sonographic technique for measuring vaginal wall thickness (VWT) is still absent in the literature. This study aims to validate a new method for measuring VWT ...using a biplanar transvaginal ultrasound probe and assess both its intra-operator and inter-operator reproducibility.
: This prospective study included patients with genitourinary syndrome of menopause-related symptoms. Women were scanned using a BK Medical Flex Focus 400 with the 65 × 5.5 mm linear longitudinal transducer of an endovaginal biplanar probe (BK Medical probe 8848, BK Ultrasound, Peabody, MA, USA). Vaginal wall thickness (VWT) measurements were acquired from the anterior and posterior vaginal wall at three levels.
: An inter-observer analysis revealed good consistency between operators at every anatomical site, and the intra-class coefficient ranged from 0.931 to 0.987, indicating high reliability. An intra-observer analysis demonstrated robust consistency in vaginal wall thickness measurements, with an intra-class coefficient exceeding 0.9 for all anatomical sites.
: The measurement of vaginal wall thickness performed by transvaginal biplanar ultrasound was easy and demonstrated good intra- and inter-operator reliability.
: Uterosacral ligaments (USLs) suspension is a well-studied, safe, and long-lasting technique for central compartment correction. Preliminary clinical experiences showed encouraging data for this ...technique, also for post-hysterectomy vaginal vault prolapse surgical treatment. However, up-to-date evidence for post-hysterectomy vaginal vault prolapse repair through high uterosacral ligaments suspension is limited. Consequently, with this study, we aimed to assess the efficiency, complications frequency, and functional results of native-tissue repair through USLs in vaginal vault prolapse.
: This was a retrospective study. Women with symptomatic vaginal vault prolapse (≥stage 2) who underwent surgery with transvaginal native-tissue repair by high uterosacral ligaments were included. Patient characteristics, preoperative assessment, operative data, postoperative follow-up visits, and re-interventions were collected from the hospital's record files. High uterosacral ligament suspension was performed according to the technique previously described by Shull. A transverse apical colpotomy at the level of the post-hysterectomy scar was performed in order to enter the peritoneal cavity. USLs were identified and transfixed from ventral to dorsal with three absorbable sutures. Sutures were then passed through the vaginal apex and tightened to close the transverse colpotomy and suspend the vaginal cuff. At the end of the surgical time, a diagnostic cystoscopy was performed in order to evaluate ureteral bilateral patency. Using the POP-Q classification system, we considered an objective recurrence as the descensus of at least one compartment ≥ II stage, or the need for a subsequent surgery for POP. The complaint of bulging symptoms was considered the item to define a subjective recurrence. We employed PGI-I scores to assess patients' satisfaction.
: Forty-seven consecutive patients corresponding to the given period were analyzed. No intraoperative complications were observed. We observed one postoperative hematoma that required surgical evacuation. Thirty-three patients completed a minimum of one-year follow-up (mean follow-up 21.7 ± 14.6 months). Objective cure rate was observed in 25 patients (75.8%). No patients required reintervention. The most frequent site of recurrence was the anterior compartment (21.2%), while apical compartment prolapse relapsed only in 6% of patients. An improvement in all POP-Q parameters was recorded except TVL which resulted in a mean 0.5 cm shorter. Subjective recurrence was referred by 4 (12.1%) patients. The mean satisfaction assessed by PGI-I score was 1.6 ± 0.8.
: This analysis demonstrated that native-tissue repair through high USL suspension is an effective and safe procedure for the treatment of post-hysterectomy vaginal vault prolapse. Objective, subjective, functional, and quality of life outcomes were satisfactory, with minimal complications.
PDF
