Ecological and social factors influence interspecific pathogens occurrence among bees.

The interspecific transmission of pathogens can occur frequently in the environment. Among wild bees, the main spillover cases are caused by pathogens associated with Apis mellifera, whose colonies can act as reservoirs. Due to the limited availability of data in Italy, it is challenging to accurately assess the impact and implications of this phenomenon on the wild bee populations. In this study, a total of 3372 bees were sampled from 11 Italian regions within the BeeNet project, evaluating the prevalence and the abundance of the major honey bee pathogens (DWV, BQCV, ABPV, CBPV, KBV, Nosema ceranae, Ascosphaera apis, Crithidia mellificae, Lotmaria passim, Crithidia bombi). The 68.4% of samples were positive for at least one pathogen. DWV, BQCV, N. ceranae and CBPV showed the highest prevalence and abundance values, confirming them as the most prevalent pathogens spread in the environment. For these pathogens, Andrena, Bombus, Eucera and Seladonia showed the highest mean prevalence and abundance values. Generally, time trends showed a prevalence and abundance decrease from April to July. In order to predict the risk of infection among wild bees, statistical models were developed. A low influence of apiary density on pathogen occurrence was observed, while meteorological conditions and agricultural management showed a greater impact on pathogen persistence in the environment. Social and biological traits of wild bees also contributed to defining a higher risk of infection for bivoltine, communal, mining and oligolectic bees. Out of all the samples tested, 40.5% were co-infected with two or more pathogens. In some cases, individuals were simultaneously infected with up to five different pathogens. It is essential to increase knowledge about the transmission of pathogens among wild bees to understand dynamics, impact and effects on pollinator populations. Implementing concrete plans for the conservation of wild bee species is important to ensure the health of wild and human-managed bees within a One-Health perspective.

Exploring toxicological interactions in a changing sea: The case of the alkaloids caffeine and caulerpin.

The bisindolic alkaloid caulerpin (CAU) is a bioactive compound isolated from green algae of the genus Caulerpa that are highly invasive in the Mediterranean Sea. On the other side, the purine alkaloid caffeine (CAF) is one of the most globally consumed psychoactive substances and a widespread anthropogenic water pollutant. Both compounds display a large panel of biological properties and are well known to accumulate in the tissues of aquatic organisms and, in certain circumstances, co-occur in the human diet. On this premise, the present study aimed to investigate possible synergistic interactions between CAU and CAF by using the bivalve Mytilus galloprovincialis as a model organism. Mussels were exposed to CAF via medium while they were fed with food enriched with CAU. After treatments, biochemical analysis confirmed the toxic potential of CAF, with increased AChE activity and lipid peroxidation. Also, histopathological alterations were observed in the gills and digestive tubules. The NMR-based metabolomics analysis detected higher levels of free amino acids under CAF treatments. Conversely, the food administration of CAU did not affect the above toxicological biomarkers. In addition, we did not observe any cumulative effect between CAF and CAU toward increased cellular damage and neurotoxicity. On the other hand, a possible action of CAU in decreasing CAF toxicity could be hypothesized based on our results. This hypothesis is supported by the activity of CAU as an agonist of peroxisome proliferator-activated receptors (PPARs). PPARs mediate xenobiotic detoxification via cytochromes P450, which is involved in CAF metabolism. Overall, the results obtained not only rule out any cumulative adverse effects of CAF and CAU but also encourage further research to evaluate the possible use of CAU, a compound easily obtained through the valorization of biomass from invasive species, as a food additive to improve the clearance of xenobiotics.

Effects of Virgin Coconut Oil-Enriched Diet on Immune and Antioxidant Enzymatic Activity, Fat and Vitellogenin Contents in Newly Emerged and Forager Bees (Apis mellifera L.) Reared in Cages.

Searching for artificial diets positively affecting the survival, immune and antioxidant systems of honey bees is one of main challenges occurring in beekeeping. Among nutrients, lipids play a significant role in insect nutrition as structural components in cell membranes, energy sources and reserves, and are involved in many physiological processes. In this context, the aim of this work was to investigate the effect of 0.5% and 1% coconut oil-enriched diet administration on newly emerged and forager bees survival rate, feed intake, immune system, antioxidant system and both fat and vitellogenin content. In newly emerged bees, supplementation with 1% coconut oil determined a decrease in feed consumption, an increase in survival rate from the 3rd to 14th day of feeding, a short-term decrease in phenoloxidase activity, an increase in body fat and no differences in vitellogenin content. Conversely, supplementation with 0.5% coconut oil determined an increase in survival rate from the 3rd to 15th day of feeding and an increase in fat content in the long term (i.e., 20 days). Regarding the forager bee diet, enrichment with 0.5% and 1% coconut oil only determined an increase in fat content. Therefore, supplementation with coconut oil in honey bee diets at low percentages (0.5 and 1%) determines fat gain. Further investigations to evaluate the use of such supplement foods to prevent the fat loss of weak families during winter are desirable.

A food-web model as a tool for the ecosystem-level management of bivalves in an Atlantic coastal lagoon.

The Ria de Aveiro is an important coastal lagoon for wildlife in Portugal, where the production of bivalves reaches approximately 2700 tons annually. However, the illegal overfishing of bivalves is frequent in this lagoon, which causes critical changes in the ecosystem. In this study, using a developed food-web model (Ecopath model), the ecological carrying capacity (ECC) and maximum sustained yield (MSY) of the Manila clam, Ruditapes philippinarum were estimated, and the effects of further increases in clam biomass on other species were investigated. The results showed that 1) the current biomass and legal catch of R. philippinarum do not yet exceed the ECC (172.40 tons km-2) or the MSY (86.20 tons km-2 year-1) in Ria de Aveiro; 2) the harvested Manila clams of the MSY represent removing from the ecosystem âˆ¼ 581 tons carbon (C) and ∼83 tons nitrogen (N) annually, with substantial ecological and economic implications; and 3) a further increase in the biomass levels of this species may cause the ecotrophic efficiency of other groups to become unrealistic, potentially leading to decreases in ecosystem transfer efficiency, biodiversity and health. The results here are expected to guide the sustainable development and management of bivalve aquaculture in Ria de Aveiro and the protection of the local environment.

Nanoplastic exposure inhibits feeding and delays regeneration in a freshwater planarian.

The concentration of nanoplastics (NPs) is expected to increase in aquatic environments thus potentially threatening freshwater organisms through interactions with plastic particles that variously float, circulate in the water column or sink into the benthos. Studies into the mechanisms of any NP effects are still scarce, particularly with respect to the regenerative ability of biota for which there is no recognised model organism. The present study therefore aimed to investigate behavioural and regeneration responses of the freshwater planarian Girardia tigrina after 10 days exposed to along a gradient 0.01-10 mg/L of poly (styrene-co-methyl methacrylate) NPs (∼426 ± 175 nm). Exposure to NPs induced a significant reduction in planarian feeding rate even at low concentrations (LOEC of 0.01 mg/L), while head regeneration was delayed in a clear dose response way (LOEC of 0.1 mg/L for blastema length). Planaria locomotion assessed was not affected. Our results highlight the potential adverse effects of exposure to poly (styrene-co-methyl methacrylate) NPs and show that feeding behaviour and regeneration of a freshwater benthic organism can be indicators of the resulting toxicity. Planarians are becoming widely used model organisms in ecotoxicology and can help to address potential effects of plastic polymers on regeneration.

Porcupettes Management at Wildlife Rescue Centers and Liberation into the Wild: Implications for Post-Liberation Success.

The crested porcupine is a frequent host species in wildlife rescue centers and no guidelines for its management, liberation and post-liberation monitoring are yet available. Here, captive-grown porcupines' behavior and survival in the wild after liberation were investigated and described for the first time. Management strategies adopted at the centers could affect porcupine adaptation to the natural environment. The detention of porcupettes in single cages and fed only human-supplied food may not ensure ethological welfare, nor a suitable behavioral development compatible with wildlife. The liberation of captive-grown porcupines should also be carefully planned to promote and increase the possibility of post-liberation success.

Host-Parasitoid Relationship between Apis mellifera (Linnaeus, 1758) and Senotainia tricuspis (Meigen, 1838) (Diptera, Sarcophagidae): Fly Aggression Behavior and Infestation Rates of Senotainiosis.

Senotainia tricuspis (Meigen, 1838) is a Sarcophagid dipteran endoparasitoid of Apis mellifera L., and myiasis, caused by this fly, is reported in several European, North African and Middle Eastern countries. Nevertheless, very little knowledge concerning the aggression and parasitisation behavior of S. tricuspis toward A. mellifera is available in the scientific literature, and the temporal pattern of aggression remains unclear. The aim of this investigation was to describe the aggressive behavior of S. tricuspis and to provide data on pupation and adult emergence in order to identify further tools for the control of senotainiosis in beekeeping. Data were collected in an apiary in Pisa province (Tuscany, Italy), where observations of aggressive behavior were conducted indirectly by using a VHS camera and also directly by an observer. Four behavioral categories of the attack were described. A total of 55 aggressions, 21 beecatchers, 104 chases and 6 parasitization events were recorded with the camera. Slow-motion recording analyses of the parasitization episodes resulted in contact of at least 1/6 s between the parasitoid and the host. Through four days of direct observations, a total of 1633 aggression events were recorded. The daily temporal pattern of the number of aggressions showed two main peaks: one during the morning hours (i.e., from 10:00 to 11:00) and one in the afternoon (i.e., from 15:00 to 17:00). The morphometric data on the first-instars of S. tricuspis allowed us to hypothesize a penetration in the bee through its prothoracic spiracle as a modality of entrance in the host body. Third-instars successfully pupate when sinking in topsoil or clay soil, and adults emerge when left to a 4 °C overwintering period of six months. Furthermore, the high mortality rate of those larvae that did not sink and did not pupate successfully suggests that reaching a certain depth in the soil is a determining factor for larvae survival and that mulch and/or minimum soil tillage could prevent severe senotainiosis in apiaries.

An alien metabolite vs. a synthetic chemical hazard: An ecotoxicological comparison in the Mediterranean blue mussel.

Bioactive natural products from marine invasive species may dramatically impact native communities, while many synthetic pharmaceutical drugs are released into the marine environment and have long-lasting harmful effects on aquatic life. Sometimes, metabolites from alien species and synthetic compounds share similar mechanisms of action, suggesting comparable ecotoxicological impacts. This applies to the alkaloid caulerpin (CAU) from the green algae Caulerpa cylindracea, highly invasive in the Mediterranean Sea, and to the synthetic lipid-lowering drug fenofibrate (FFB), both acting as agonists of peroxisome proliferator-activated receptors (PPARs). Analogies with FFB, which is widely considered hazardous to the aquatic environment, have led to concerns about the ecotoxicological potential of CAU. The problem has implications for public health as CAU is well known to enter the food web accumulating in fish of commercial importance. Here, we compared the effects of FFB and CAU through biochemical and histopathological analysis on a relevant bioindicator molluscan species, the mussel Mytilus galloprovincialis. Under laboratory conditions, mussels were fed with food enriched with CAU or FFB. After treatment, biochemical markers were analyzed revealing metabolic capacity impairments, cellular damage, and changes in acetylcholinesterase activity in mussels fed with FFB-enriched food. NMR-based metabolomic studies also showed significant alterations in the metabolic profiles of FFB-treated mussels. In addition, dietary administration of FFB produced morphological alterations in the mussels' gills and digestive tubules. Obtained results confirm that FFB is harmful to aquatic life and that its release into the environment should be avoided. Conversely, dietary treatment with CAU did not produce any significant alterations in the mussels. Overall, our results pave the way for the possible valorization of the huge biomass from one of the world's worst invasive species to obtain CAU, a natural product of interest in drug discovery.

Bombus terrestris terrestris (Linnaeus, 1758) and hybrids with the endemic Bombus xanthopus spotted on Capraia Island (Tuscan Archipelago, Italy): some conservation management implications.

Expansion of wild and managed allochthonous species leads to potential negative consequences for the endemic wildlife, such as resource competition, pathogens spread, hybridization and native species replacements. On Capraia Island, the last sighting of Bombus terrestris terrestris dates back to 1917. All subsequent surveys carried out on the island only reported the presence of B. xanthopus and B. pascuorum melleofacies with B. t. terrestris apparently no longer existing in the area. In 2021 B. t. terrestris was again detected on the island raising two main hypotheses: (i) B. t. terrestris has always been present with a low population density, such as not to be detected in previous investigations, or (ii) its presence is the result of a more recent recolonization. The recolonization event may be promoted by either intentional or unintentional introduction or it may be the result of a natural migration from the mainland or surrounding islands. Hybridization between B. t. terrestris × B. xanthopus on Capraia Island has been also ascertained by the detection of hybrid queens, workers and males. These new finding provides insight on the distribution range of B. t. terrestris in the Tuscan Archipelago and raise concern on the conservation of the endemic B. xanthopus population.

Effect of Ageing in the Mating Behaviour Sequence of Osmia cornuta Latr. (Hymenoptera: Megachilidae).

Osmia cornuta Latr. is largely managed worldwide for the pollination of orchard crops, playing a key role in the maintenance of healthy ecosystems and ensuring economic and social benefits for human society. The management techniques of this pollinator include the possibility of delaying emergence from cocoons after diapause, allowing for the pollination of later-blooming fruit crops. In this study, the mating behaviour of bees emerging at the natural time (Right Emergence Insects) and of late-emerged bees (Aged Emergence Insects) was described in order to test if a delay in emergence could affect the mating sequence of O. cornuta. Markov analysis of the mating behaviour revealed the occurrence of antenna motion episodes that were repeated in a stereotyped manner at regular intervals during the mating sequence of both Right Emergence Insects and in Aged Emergence Insects. Pouncing, rhythmic and continuous emission of sound, motion of antennae, stretching of the abdomen, short and long copulations, scratching, inactivity, and self-grooming were identified as the stereotyped behavioural units of a behavioural sequence. The occurrence of short copulations, the frequency of which increased with the age of bees, could lead to a failure in the reproduction of the mason bee.

Ecotoxicological impacts of metals in single and co-exposure on mussels: Comparison of observable and predicted results.

Used in high-tech and everyday products, mercury (Hg), cobalt (Co), and nickel (Ni) are known to be persistent and potentially toxic elements that pose a serious threat to the most vulnerable ecosystems. Despite being on the Priority Hazardous Substances List, existing studies have only assessed the individual toxicity of Co, Ni and Hg in aquatic organisms, with a focus on the latter, ignoring potential synergistic effects that may occur in real-world contamination scenarios. The present study evaluated the responses of the mussel Mytilus galloprovincialis, recognized as a good bioindicator of pollution, after exposure to Hg (25 µg/L), Co (200 µg/L) and Ni (200 µg/L) individually, and to the mixture of the three metals at the same concentration. The exposure lasted 28 days at 17 ± 1 °C, after which metal accumulation and a set of biomarkers related to organisms' metabolic capacity and oxidative status were measured. The results showed that the mussels could accumulate metals in both single- and co-exposure conditions (bioconcentration factors between 115 and 808) and that exposure to metals induced the activation of antioxidant enzymes. Although Hg concentration in organisms in the mixture decreased significantly compared to single exposure (9.4 ± 0.8 vs 21 ± 0.7 mg/kg), the negative effects increased in the mixture of the three elements, resulting in depletion of energy reserves, activation of antioxidants and detoxification enzymes, and cellular damage, with a hormesis response pattern. This study underscores the importance of risk assessment studies that include the effects of the combination of pollutants and demonstrates the limitations of applying models to predict metal mixture toxicity, especially when a hormesis response is given by the organisms.

Combined effects of temperature rise and sodium lauryl sulfate in the Mediterranean mussel.

Personal care products (PCPs) are those compounds used daily (e.g., soaps, shampoos, deodorants, and toothpaste), explaining their frequent detection in aquatic systems. Still, scarce information is available on their effects on inhabiting wildlife. Among the most commonly used PCPs is the surfactant Sodium Lauryl Sulfate (SLS). The present study investigated the influence of temperature (CTL 17 ºC vs 22 ºC) on the effects of SLS (0 mg/L vs 4 mg/L) in the mussel species Mytilus galloprovincialis. Mussels' general health status was investigated, assessing their metabolic and oxidative stress responses. Higher biochemical alterations were observed in SLS-exposed mussels and warming enhanced the impacts, namely in terms of biotransformation capacity and loss of redox homeostasis, which may result in consequences to population maintenance, especially if under additional environmental stressors. These results confirm M. galloprovincialis as an excellent bioindicator of PCPs pollution, and the need to consider actual and predicted climate changes.

Effectiveness of Radiomic ZOT Features in the Automated Discrimination of Oncocytoma from Clear Cell Renal Cancer.

BACKGROUND: Benign renal tumors, such as renal oncocytoma (RO), can be erroneously diagnosed as malignant renal cell carcinomas (RCC), because of their similar imaging features. Computer-aided systems leveraging radiomic features can be used to better discriminate benign renal tumors from the malignant ones. The purpose of this work was to build a machine learning model to distinguish RO from clear cell RCC (ccRCC). METHOD: We collected CT images of 77 patients, with 30 cases of RO (39%) and 47 cases of ccRCC (61%). Radiomic features were extracted both from the tumor volumes identified by the clinicians and from the tumor's zone of transition (ZOT). We used a genetic algorithm to perform feature selection, identifying the most descriptive set of features for the tumor classification. We built a decision tree classifier to distinguish between ROs and ccRCCs. We proposed two versions of the pipeline: in the first one, the feature selection was performed before the splitting of the data, while in the second one, the feature selection was performed after, i.e., on the training data only. We evaluated the efficiency of the two pipelines in cancer classification. RESULTS: The ZOT features were found to be the most predictive by the genetic algorithm. The pipeline with the feature selection performed on the whole dataset obtained an average ROC AUC score of 0.87 ± 0.09. The second pipeline, in which the feature selection was performed on the training data only, obtained an average ROC AUC score of 0.62 ± 0.17. CONCLUSIONS: The obtained results confirm the efficiency of ZOT radiomic features in capturing the renal tumor characteristics. We showed that there is a significant difference in the performances of the two proposed pipelines, highlighting how some already published radiomic analyses could be too optimistic about the real generalization capabilities of the models.

Radiomic Features from Post-Operative 18F-FDG PET/CT and CT Imaging Associated with Locally Recurrent Rectal Cancer: Preliminary Findings.

Locally Recurrent Rectal Cancer (LRRC) remains a major clinical concern; it rapidly invades pelvic organs and nerve roots, causing severe symptoms. Curative-intent salvage therapy offers the only potential for cure but it has a higher chance of success when LRRC is diagnosed at an early stage. Imaging diagnosis of LRRC is very challenging due to fibrosis and inflammatory pelvic tissue, which can mislead even the most expert reader. This study exploited a radiomic analysis to enrich, through quantitative features, the characterization of tissue properties, thus favoring an accurate detection of LRRC by Computed Tomography (CT) and 18F-FDG-Positron Emission Tomography/CT (PET/CT). Of 563 eligible patients undergoing radical resection (R0) of primary RC, 57 patients with suspected LRRC were included, 33 of which were histologically confirmed. After manually segmenting suspected LRRC in CT and PET/CT, 144 Radiomic Features (RFs) were generated, and RFs were investigated for univariate significant discriminations (Wilcoxon rank-sum test, p < 0.050) of LRRC from NO LRRC. Five RFs in PET/CT (p < 0.017) and two in CT (p < 0.022) enabled, individually, a clear distinction of the groups, and one RF was shared by PET/CT and CT. As well as confirming the potential role of radiomics to advance LRRC diagnosis, the aforementioned shared RF describes LRRC as tissues having high local inhomogeneity due to the evolving tissue's properties.

The colors of Tuscan bee pollen: phytochemical profile and antioxidant activity.

Bee pollen's nutritional and beneficial health properties depend on the botanical origin and storage conditions. Palynological analysis determines the botanical composition of the multiflora and colour fractions. This study aimed to characterize the phytochemical profile and antioxidant activity of Tuscan bee pollen stored at freezing temperature for 2 years to verify the preservation of nutraceutical properties of the multiflora and colour fractions. Polyphenols, flavonoids content, antioxidant activity and volatile compounds profiles were measured. Non-terpene derivatives (acids and aldehydes) represented the main class of volatile compounds in most analysed samples. Among the colour fractions, coral showed significant differences in the antioxidant compounds. In the multiflora were also determined the soluble sugar content (128.33 mg/g of fresh weight) and mineral content, with the prevalence of K, organic N and Ca. The results suggest that the freezing storage of bee pollen for a long period can be still used as food.

Gray Wolf (Canis lupus italicus) and Red Fox (Vulpes vulpes) Parasite Survey in Anthropized and Natural Areas of Central Italy.

Gastrointestinal nematodes and protozoa and other parasite occurrences were evaluated in free-ranging wolf (Canis lupus italicus) and red fox (Vulpes vulpes) populations from natural and anthropized areas of Central Italy. Analyzed fecal samples were collected from 60 foxes and 40 wolves in the anthropized areas, and 41 foxes and 39 wolves in the natural areas. In foxes, hookworm infections (p < 0.0001) were more frequently recorded in the anthropized environment, while coccidia (p < 0.05) and Cryptosporidium spp. (p < 0.0001) were more frequent in the natural area. In wolves, a higher frequency of hookworms (p < 0.0001) was observed in natural areas, while coccidia were more common in the anthropized area (p < 0.05). Moreover, in the natural environment, trichuroid nematodes (p < 0.0001) were significantly more frequent in wolves than in foxes, while Cryptosporidium (p < 0.001) and Giardia duodenalis (p < 0.001) were more common in foxes. In the anthropic area, the occurrence of hookworms was found to be significantly higher in foxes (p < 0.0001), while trichuroid nematodes were more common in wolves (p < 0.0001). The obtained data are indicative of a different diffusion of specific parasite taxa in wolves and foxes living in the natural and/or anthropized environments examined herein.

Structured reporting of computed tomography in the polytrauma patient assessment: a Delphi consensus proposal.

OBJECTIVES: To develop a structured reporting (SR) template for whole-body CT examinations of polytrauma patients, based on the consensus of a panel of emergency radiology experts from the Italian Society of Medical and Interventional Radiology. METHODS: A multi-round Delphi method was used to quantify inter-panelist agreement for all SR sections. Internal consistency for each section and quality analysis in terms of average inter-item correlation were evaluated by means of the Cronbach's alpha (Cα) correlation coefficient. RESULTS: The final SR form included 118 items (6 in the "Patient Clinical Data" section, 4 in the "Clinical Evaluation" section, 9 in the "Imaging Protocol" section, and 99 in the "Report" section). The experts' overall mean score and sum of scores were 4.77 (range 1-5) and 257.56 (range 206-270) in the first Delphi round, and 4.96 (range 4-5) and 208.44 (range 200-210) in the second round, respectively. In the second Delphi round, the experts' overall mean score was higher than in the first round, and standard deviation was lower (3.11 in the second round vs 19.71 in the first round), reflecting a higher expert agreement in the second round. Moreover, Cα was higher in the second round than in the first round (0.97 vs 0.87). CONCLUSIONS: Our SR template for whole-body CT examinations of polytrauma patients is based on a strong agreement among panel experts in emergency radiology and could improve communication between radiologists and the trauma team.

Can temperature influence the impacts induced in Mytilus galloprovincialis by neodymium? Comparison between exposure and recovery periods.

Climate change-associated factors and pollutants, such as rare earth elements (REEs), have been identified as contributors to environmental changes. However, the toxicity resulting from the combination of these stressors has received little attention. Neodymium (Nd) is a REE that has been widely used, and this study aimed to evaluate the responses of Mytilus galloprovincialis to Nd exposure (10 µg/L), under actual (17 °C) and predicted warming conditions (21 °C), after fourteen days of exposure followed by fourteen days of recovery (without Nd), analyzing Nd accumulation, histopathological and biochemical alterations. The results showed that increased temperature and Nd exposure caused histopathological injuries in the gills. Contaminated mussels at 17 °C showed cellular damage, while at 21 °C, mussels were able to avoid cellular damage. After the recovery period, no improvements in gill's status were found and cellular damage was still present, highlighting the impacts caused by previous exposure to Nd.

Beyond Multiparametric MRI and towards Radiomics to Detect Prostate Cancer: A Machine Learning Model to Predict Clinically Significant Lesions.

The risk of misclassifying clinically significant prostate cancer (csPCa) by multiparametric magnetic resonance imaging is consistent, also using the updated PIRADS score and although different definitions of csPCa, patients with Gleason Grade group (GG) ≥ 3 have a significantly worse prognosis. This study aims to develop a machine learning model predicting csPCa (i.e., any GG ≥ 3 lesion at target biopsy) by mpMRI radiomic features and analyzing similarities between GG groups. One hundred and two patients with 117 PIRADS ≥ 3 lesions at mpMRI underwent target+systematic biopsy, providing histologic diagnosis of PCa, 61 GG < 3 and 56 GG ≥ 3. Features were generated locally from an apparent diffusion coefficient and selected, using the LASSO method and Wilcoxon rank-sum test (p < 0.001), to achieve only four features. After data augmentation, the features were exploited to train a support vector machine classifier, subsequently validated on a test set. To assess the results, Kruskal−Wallis and Wilcoxon rank-sum tests (p < 0.001) and receiver operating characteristic (ROC)-related metrics were used. GG1 and GG2 were equivalent (p = 0.26), whilst clear separations between either GG[1,2] and GG ≥ 3 exist (p < 10−6). On the test set, the area under the curve = 0.88 (95% CI, 0.68−0.94), with positive and negative predictive values being 84%. The features retain a histological interpretation. Our model hints at GG2 being much more similar to GG1 than GG ≥ 3.

Settlement of a stable wolf pack in a highly anthropic area of Pisan hills: Relationship with animal husbandry and hunting in a human-wolf coexistence perspective.

Anthropic areas play a pivot role for main wolf conservation challenges. Wolf presence in the higher Pisan hills has been well documented while wolf settlement in the lower Pisan hills is still uncertain. In this study, long-term information on wolf presence in a highly anthropic area of the lower Pisan hills was collected by using non-invasive monitoring techniques. Furthermore, both the relationship of this predator with human activity and the impact of hunting on wolf presence have been investigated. The results obtained indicate the presence of a stable and reproductive wolf pack composed by both Italian wolf and hybrids individuals in the municipalities of Crespina Lorenzana and Casciana Terme Lari. A high impact of wolf on livestock was recorded in this area since no prevention systems were adopted by farmers. Wolf appears not to have a negative impact on wild boar population. Similarly, wild boar drive hunting does not appear to affect the wolf pack presence in the area. Thereby wolf may play a key role as controller of wild population. Prevention strategies improvement becomes instrumental to promote wolf-human coexistence. Further investigation to monitor pack hybridization level and turnover and to assess the impact of packs on wild population and livestock in anthropic areas is desirable.