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.

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.

In Italy, porcupines, badgers and red foxes share the same settlements.However, there is lack of information concerning their cohabitation.From 2012 to 2019, cohabitation by these three mammals was studied using camera-trapping and was found to occur only between porcupines and badgers, even in the presence of porcupettes.Cohabitation was associated with aggressive interaction between porcupines and badgers.Foxes were found to be scavengers of porcupine carcasses.Cohabitation among these semi-fossorial mammals and scavenging behaviour could play a role in disease transmission, including zoonotic diseases.

Free-ranging corvids-678 magpies (Pica pica) and 120 hooded crows (Corvus cornix) from nine protected areas of the Pisa province (central Italy)-were examined for Toxoplasma gondii and Trichinella pseudospiralis. The intracardiac blood clots from 651 magpies and 120 hooded crows were serologically examined for T. gondii. The DNA extracted from the hearts of seropositive birds was then used to perform a nested PCR for the amplification of the T. gondii B1 gene and for genotyping for SAG genetic markers. Breast muscle samples from 678 magpies and 91 hooded crows were tested by an artificial digestion method for Trichinella. Data were statistically analyzed. Forty-five (5.8%-41 magpies and four hooded crows) out of the 771 examined animals scored seropositive for T. gondii, with titers ranging from 1:25 to 1:100. T. gondii DNA was detected in 15 of the 45 positive birds and T. gondii genotypes II and III were identified. No positivity for T. pseudospiralis was found. No significant differences between the two species of corvids and among the different areas of origin were observed for seropositivity to T. gondii. This is the first extensive study on both T. gondii and T. pseudospiralis in magpies and hooded crows, as well as the first detection of T. gondii SAG genotypes in magpies.

The vegetarian diet of many herbivorous mammals is supplemented with proteins of animal origin, especially in young individuals and in breeding females, to provide key proteins necessary for both growth and breeding. Among porcupine species, only the Cape porcupine ( Hystrix africaeaustralis ) has been observed to consume carrion flesh. From June to August 2019, a pigeon carcass was placed together with corn in 7 study settlements and near 2 monitored capture-traps, in order to assess the carrion flesh feeding habits of the crested porcupine ( Hystrix cristata ) . Scavenging behaviour was recorded on four occasions. All the recorded individuals were adults and at least one was female. This demonstrates that the crested porcupine occasionally does eat flesh. Such evidence raises important questions concerning the relationship between feeding habits and the physiological needs of this herbivorous rodent.

The crested porcupine is a mainly nocturnal mammal that shows both moonlight avoidance and some diurnal motor activity, the latter as an extension of its night-time foraging behaviour. Furthermore, a peculiar daytime behaviour, described as "sunbathing", was reported as episodic in H. africaeaustralis. Between 2016 and 2019 a camera-trapping monitoring was performed within 10 porcupine settlements in order to detect the diurnal motor activity and to verify and describe the sunbathing behaviour in crested porcupine. Out of 1,003 trap days, a total of 148 events of diurnal motor activity were recorded. The diurnal motor activity occurred throughout the year mainly between December and June from 15:00 to 16:00, with no statistical difference between cubs, youngsters and adults. The sunbathing behaviour was detected for a total of 36 episodes recorded. Sunbathing was performed mainly by cubs. The sunbathing behaviour occurred only between April and June during the hottest hours of the day (11:00 to 12:00). Diurnal motor activity and sunbathing behaviour of porcupine are discussed in relation to food availability and porcupine physiology.

The crested porcupine ( Hystrix cristata L., 1758) is the largest rodent among the Italian fauna. It is nocturnal, monogamous, herbivorous and lives in burrows that could be self made as well as made by other mammals such as fox ( Vulpes vulpes ) and badger ( Meles meles ). It exhibits moonlight avoidance and male performs parental care. In Europe, distribution of the species is limited to Italy, although the presence of porcupine in Balkans is still debated. Outside Europe crested porcupine distribution includes parts of northwestern Africa from Morocco to Libya and Sub-Saharan Africa from Gambia to Tanzania.

We developed a new versatile strategy that allows the detection of several classes of RNases (i.e., targeting ss- or ds-RNA, DNA/RNA hetero-hybrid or junctions) with higher sensitivity than existing assays. Our two-step approach consists of a DNA-RNA-DNA chimeric Hairpin Probe (cHP) conjugated to magnetic microparticles and containing a DNAzyme sequence in its terminal region, and molecular beacons for fluorescence signal generation. In the first step, the digestion of the RNA portion of the cHP sequences in presence of RNases leads to the release of multiple copies of the DNAzyme in solution. Then, after magnetic washing, each DNAzyme molecule elicits the catalytic cleavage of numerous molecular beacons, providing a strong amplification of the overall sensitivity of the assay. We successfully applied our approach to detect very low concentrations of RNase A, E. coli RNase I, and RNase H. Furthermore, we analyzed the effect of two antibiotics (penicillin and streptomycin) on RNase H activity, demonstrating the applicability of our strategy for the screening of inhibitors. Finally, we exploited our system to detect RNase activity directly in crude biological samples (i.e., blood and saliva) and in cell culture medium, highlighting its suitability as cheap and sensitive tool for the detection of RNase levels.

The assessment of the risks exerted by nanoparticles is a key challenge for academic, industrial, and regulatory communities worldwide. Experimental evidence points towards significant toxicity for a range of nanoparticles both in vitro and in vivo. Worldwide efforts aim at uncovering the underlying mechanisms for this toxicity. Here, we show that the intracellular ion release elicited by the acidic conditions of the lysosomal cellular compartment--where particles are abundantly internalized--is responsible for the cascading events associated with nanoparticles-induced intracellular toxicity. We call this mechanism a "lysosome-enhanced Trojan horse effect" since, in the case of nanoparticles, the protective cellular machinery designed to degrade foreign objects is actually responsible for their toxicity. To test our hypothesis, we compare the toxicity of similar gold particles whose main difference is in the internalization pathways. We show that particles known to pass directly through cell membranes become more toxic when modified so as to be mostly internalized by endocytosis. Furthermore, using experiments with chelating and lysosomotropic agents, we found that the toxicity mechanism for different metal containing NPs (such as metallic, metal oxide, and semiconductor NPs) is mainly associated with the release of the corresponding toxic ions. Finally, we show that particles unable to release toxic ions (such as stably coated NPs, or diamond and silica NPs) are not harmful to intracellular environments.

Drosophila was the most important model organism used in the fields of medicine and biology over the last century. Recently, Drosophila was successfully used in several studies in the field of nanotoxicology. However, only a part of its potential has been exploited in this field until now. In fact, apart from macroscopic observations of the effect due to the interaction between nanomaterials and living organism (i. e. lifespan, fertility, phenotypic aberrations, etc.), Drosophila has the potential to be a very useful tool to deeply analyze the molecular pathways involved in response to the interactions at nano-bio level. The aim of this editorial is to encourage the use of Drosophila by the different research groups working in the fields of nanotoxicology and nanomedicine, in order to define the effects induced by nanomaterials at molecular level for their subsequent exploitation in the field of nanomedicine.

The continuous increasing of engineered nanomaterials (ENMs) in our environment, their combinatorial diversity, and the associated genotoxic risks, highlight the urgent need to better define the possible toxicological effects of ENMs. In this context, we present a new high-throughput screening (HTS) platform based on the cytokinesis-block micronucleus (CBMN) assay, lab-on-chip cell sorting, and automated image analysis. This HTS platform has been successfully applied to the evaluation of the cytotoxic and genotoxic effects of silver nanoparticles (AgNPs) and silica nanoparticles (SiO2NPs). In particular, our results demonstrate the high cyto- and genotoxicity induced by AgNPs and the biocompatibility of SiO2NPs, in primary human lymphocytes. Moreover, our data reveal that the toxic effects are also dependent on size, surface coating, and surface charge. Most importantly, our HTS platform shows that AgNP-induced genotoxicity is lymphocyte sub-type dependent and is particularly pronounced in CD2+ and CD4+ cells.

We demonstrate a general mechanism for the toxicity induced by metal-containing NPs, named “lysosome-enhanced Trojan horse effect”, which provides design rules to engineer safer NPs.

In this work, we propose a quantitative assessment of nanoparticles toxicity in vivo. We show a quantitative ranking of several types of nanoparticles (AuNPs, AgNPs, cadmium-based QDs, cadmium-free QDs, and iron oxide NPs, with different coating and/or surface chemistries), providing a categorization of their toxicity outcomes. This strategy may offer an innovative high-throughput screening tool of nanomaterials, of potential and broad interest to the nanoscience community.

Polydimethylsiloxane doped with colloidal CdSe/CdS nanocrystals is proposed as high-luminescent and biocompatible nanocomposite. A physical mixing procedure has been adopted to realize the optically active material and spectral and time-resolved optical analysis has been carried out in order to proof that nanocrystals quantum properties are preserved into the polymeric host matrix. Biocompatibility tests have shown no cadmium release in water ambient and no differences have been observed in cell viability cultured on pure PDMS substrates and on nanocomposite films. The obtained results let us envision possible applications of the proposed material as luminescent substrate for label-free cell imaging compatible with the most common optical microscopy techniques.

Bacterial adhesion onto abiotic surfaces is an important issue in biology and medicine since understanding the bases of such interaction represents a crucial aspect in the design of safe implant devices with intrinsic antibacterial characteristics. In this framework, we investigated the effects of nanostructured metal substrates on Escherichia coli adhesion and adaptation in order to understand the bio-molecular dynamics ruling the interactions at the interface. In particular, we show how highly controlled nanostructured gold substrates impact the bacterial behavior in terms of morphological changes and lead to modifications in the expression profile of several genes, which are crucially involved in the stress response and fimbrial synthesis. These results mainly demonstrate that E. coli cells are able to sense even slight changes in surface nanotopography and to actively respond by activating stress-related pathways. At the same time, our findings highlight the possibility of designing nanoengineered substrates able to trigger specific bio-molecular effects, thus opening the perspective of smartly tuning bacterial behavior by biomaterial design.

The interactions between biological systems and nanostructured materials are attracting great interest, due to the possibility to open up novel concepts for the design of smart nano-biomaterials that actively play a functional biological role. On the other hand, the assessment of the potential toxic effects arising from such interactions is gaining increasing attention, and a new field known as nanotoxicology is strongly emerging. In this frame, we investigated the response of human neurons to gold surfaces with different levels of nanoroughness, finding out that neurons are capable to sense and actively respond to these nanotopography features. These nanostructured substrates were also investigated to explore the impact of nanotopography on morphology and genomics of adherent bacteria. A multidisciplinary approach was exploited to characterize bacteria-nanostructured surface interactions, observing that type-1 fimbriae disappear in bacteria grown onto nanorough substrates. We also show how nanoparticles interact with biomolecules in culture media and in vitro and in vivo biological systems, by investigating the toxic effects of a wide range of nanomaterials (AuNPs, QDs, SiO2 NPs), demonstrating the key role of size, shape, and surface coating.

We show that water soluble InP/ZnS core/shell QDs are a safer alternative to CdSe/ZnS QDs for biological applications, by comparing their toxicity in vitro (cell culture) and in vivo (animal model Drosophila). By choosing QDs with comparable physical and chemical properties, we find that cellular uptake and localization are practically identical for these two nanomaterials. Toxicity of CdSe/ZnS QDs appears to be related to the release of poisonous Cd2+ ions and indeed we show that there is leaching of Cd2+ ions from the particle core despite the two-layer ZnS shell. Since an almost identical amount of In(III) ions is observed to leach from the core of InP/ZnS QDs, their very low toxicity as revealed in this study hints at a much lower intrinsic toxicity of indium compared to cadmium.

The expected potential benefits promised by nanotechnology in various fields have led to a rapid increase of the presence of engineered nanomaterials in a high number of commercial goods. This is generating increasing questions about possible risks for human health and environment, due to the lack of an in-depth assessment of the physical/chemical factors responsible for their toxic effects. In this work, we evaluated the toxicity of monodisperse citrate-capped gold nanoparticles (AuNPs) of different sizes (5, 15, 40, and 80 nm) in the model organism Drosophila melanogaster, upon ingestion. To properly evaluate and distinguish the possible dose-and/or size-dependent toxicity of the AuNPs, we performed a thorough assessment of their biological effects, using two different dose-metrics. In the first approach, we kept constant the total surface area of the differently sized AuNPs (Total Exposed Surface area approach, TES), while, in the second approach, we used the same number concentration of the four different sizes of AuNPs (Total Number of Nanoparticles approach, TNN). We observed a significant AuNPs-induced toxicity in vivo, namely a strong reduction of Drosophila lifespan and fertility performance, presence of DNA fragmentation, as well as a significant modification in the expression levels of genes involved in stress responses, DNA damage recognition and apoptosis pathway. Interestingly, we found that, within the investigated experimental conditions, the toxic effects in the exposed organisms were directly related to the concentration of the AuNPs administered, irrespective of their size.