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Abstract Accurate identification and typing of microbes are crucial steps in gaining an awareness of the biological heterogeneity and reliability of microbial material within any proprietary or public collection. Paenibacillus polymyxa is a bacterial species of great agricultural and industrial importance due to its plant growth-promoting activities and production of several relevant secondary metabolites. In recent years, matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) has been widely used as an alternative rapid tool for identifying, typing, and differentiating closely related strains. In this study, we investigated the diversity of three P. polymyxa strains. The mass spectra of ATCC 842T, DSM 292, and DSM 365 were obtained, analysed, and compared to select discriminant peaks using ClinProTools software and generate classification models. MALDI-TOF MS analysis showed inconsistent results in identifying DSM 292 and DSM 365 as belonging to P. polimixa species, and comparative analysis of mass spectra revealed the presence of highly discriminatory biomarkers among the three strains. 16S rRNA sequencing and Average Nucleotide Identity (ANI) confirmed the discrepancies found in the proteomic analysis. The case study presented here suggests the enormous potential of the proteomic-based approach, combined with statistical tools, to predict and explore differences between closely related strains in large microbial datasets.

Abstract Karst hydrosystems represent one of the largest global drinking water resources, but they are extremely vulnerable to pollution. Climate change, high population density, intensive industrial, and agricultural activities are the principal causes of deterioration, both in terms of quality and quantity, of these resources. Samples from 172 natural karst springs were collected in the whole territory of Greece. To identify any geogenic contamination and/or anthropogenic pollution, analyses of their chemical compositions, in terms of major ions and trace elements, were performed and compared to the EU limits for drinking water. Based on chloride content, the collected karst springs were divided into two groups: low-chloride ( 100 mg L−1). An additional group of springs with calcium-sulfate composition was recognised. Nitrate concentrations were always below the EU limit (50 mg L−1), although some springs presented elevated concentrations. High contents in terms of trace elements, such as B, Sr, As, and Pb, sometimes exceeding the limits, were rarely found. The Greek karst waters can still be considered a good quality resource both for human consumption and for agriculture. The main issues derive from seawater intrusion in the aquifers along the coasts. Moreover, the main anthropogenic pollutant is nitrate, found in higher concentrations mostly in the same coastal areas where human activities are concentrated. Finally, high levels of potentially harmful trace elements (e.g. As, Se) are very limited and of natural origin (geothermal activity, ore deposits, etc.).

Background Chemotherapy drugs are widely used to treat cancer, but their active compounds represent a danger for workers who could be exposed to them. However, they aren’t yet included in directive CE No. 1272/2008 and the European Biosafety Network has only recommended a limit value of 100 pg/cm 2 for surface contamination. Thus, it is crucial to assess surface contaminations in healthcare environments. Currently, the technique of choice is surface wipe test combined with liquid chromatography tandem mass spectrometry to achieve high sensibility. Material and Methods A campaign involving Careggi University Hospital (Florence, Italy) was performed from January 2020 to December 2021, collecting 1449 wipe samples between administration units, preparation unit, and personnel gloves. From the obtained data, the 90th percentile was calculated for 30 antiblastic drugs and proposed as surface exposure levels (SELs); while from data concerning personnel glove contamination, weekly contamination was estimated. Results In the 2-year period only 417 wipe samples were found positive (28.8%), the majority of which regard samples coming from administration unit bathrooms. The proposed SELs are almost all

Mucocele is a common lesion localized in the oral cavity; it originates from the lesion of a salivary duct and the consequent accumulation of mucin in soft tissues. It is a common lesion of young patients and is frequently associated with areas subject to traumas. Surgical treatment is needed especially if it reaches considerable dimensions, as it can cause discomfort in the patient and tends to increase in size if subjected to further traumatism. This scoping review aims to investigate which treatments are used for this type of lesion, which are the most suitable to prevent recurrence, and if laser-assisted surgery is considered more efficient than other methods. An electronic search was performed within the PubMed (MEDLINE) and Scopus databases. Articles published in the years 2010 to 2023 were selected. Techniques like marsupialization and laser-assisted excision effectively treat mucoceles, with no clear prevalence between them. Laser surgery offers speed, precision, and minimal post-operation discomfort, but randomized trials are needed for conclusive comparisons.

Desquamative gingivitis is a clinical condition with a chronic course, not specific to a particular disease, characterized by intense erythema, scaling, vesicles, and/or blisters that may involve both the marginal free gingiva (MG) and the neighboring adherent gingiva (AG). This scoping review aimed to investigate whether there is a correlation between oral hygiene and gingival lesions induced by autoimmune diseases of the oral cavity and whether periodontal disease can negatively influence a clinical picture of desquamative gingivitis due to an immune disorder of the oral cavity. Case series studies and randomized controlled trials were considered for this scoping review; studies that did not comply with the inclusion criteria were excluded. A total of seven studies were selected for this review. The PRISMA-ScR (preferred reporting items for scoping reviews) consensus has been followed. Based on the included studies, it is possible to state that improvement in disease and patient-reported outcomes may be the result of appropriate oral hygiene education when patients are found to have autoimmune diseases with gingival manifestations.

Plain language summary Antibodies are proteins produced by the immune system that are released into the bloodstream and help fight infections. To understand how the immune system responds to COVID-19 vaccination in individuals who have had a previous SARS-CoV-2 infection, we compared the types and levels of antibodies produced after first and second doses of the vaccine with those of individuals who had never been infected. We found that one dose of vaccine, even several months after the infection, was sufficient to boost a very efficient response by eliciting specific types of antibodies, some of which were and some that were not able to neutralize the virus. We also observed an unusual antibody profile in almost half of individuals who had not been infected. These findings may help better understand the immune response to COVID-19 vaccines.

La littérature sur l’« interdisciplinarité » est imposante. Toutefois, peu de travaux fournissent des taxonomies descriptives des différentes figures du travail scientifique au croisement entre savoirs différents. Ceux qui le font adoptent soit un point de vue normatif et internaliste relativement à une discipline donnée, soit le point de vue de la sociologie de l’identité. En outre, le concept de « zone de transaction » est si utilisé en sciences sociales et au-delà qu’il semble pouvoir recouvrir la plupart des échanges « interdisciplinaires », mais se révèle en fait insuffisant. À partir d’un terrain auprès d’équipes de recherche en sciences de la complexité, cet article se propose de fournir une taxonomie descriptive du travail scientifique « interdisciplinaire » selon cinq types (transfrontaliers, ambassadeurs, polyglottes, binationaux et traducteurs) illustrés par des extraits d’entretiens et d’archives.

The concentrations of trace elements in atmospheric bulk depositions (wet plus dry) were investigated from two highly industrialised areas of Sicily (southern Italy) from June 2018 to July 2019, in order to recognise the main natural and anthropogenic sources. A side objective of this study was to improve the common sampling procedures and analytical methods used for monitoring trace elements in atmospheric deposition. The trace element VWM (Volume-Weighted Mean) concentrations ranged from less than 0.01 μg L−1 for trace elements such as Cs, Tl, and U, up to 24 μg L−1 for minor elements (Al, Zn, Sr), in the filtered aliquot, while they reached concentrations up to 144 μg L−1 for the same elements, in the unfiltered aliquot. Therefore, significant differences in concentrations between these two aliquots were found, particularly for Al, Fe, Ti, Zn, Cr, Pb, Se, Cs, and U. This implies that filtering operations may produce a consistent underestimation of concentrations of certain ‘constituents’ of the atmospheric deposition. Natural (marine spray, local and regional geogenic input, volcanic emanations) and anthropogenic sources (industrial emissions, auto vehicular traffic, and diffuse background pollution) which influence rainwater chemistry were identified. Enrichment factors (EFs), with respect to the upper crust composition, provided clear evidence of the different sources above mentioned: Ti, Fe, Al, Cs, Cr, Rb, and Co have low EFs (10), highlight the influence of marine and/or industrial sources. The study produced a novel dataset on the atmospheric deposition rate of several trace elements, which had never been studied in the investigated areas. Finally, a comparison of trace element deposition rates in the studied areas with the atmospheric deposition reported for 53 different sites, belonging to 20 different European nations, was made. The comparison showed that some elements, such as Al, V, Zn, and Mo had higher median deposition fluxes in the Sicilian sites than in European monitoring sites.

Abstract Duvalo “volcano” is a site of anomalous geogenic degassing close to Ohrid (North Macedonia) not related to volcanic activity, despite its name. CO2 flux measurements made with the accumulation chamber (321 sites over ∼50,000 m2) showed fluxes up to nearly 60,000 g m−2 d−1, sustaining a total output of ∼67 t d−1. Soil gas samples were taken at 50 cm depth from sites with high CO2 fluxes and analyzed for their chemical and isotope composition. The gas is mainly composed by CO2 (>90%) with significant concentrations of H2S (up to 0.55%) and CH4 (up to 0.32%). The isotope compositions of He (R/RA 0.10) and of CO2 (δ13C ∼ 0‰) exclude significant mantle contribution, while δ13C‐CH4 (∼−35‰) and δ2H‐CH4 (∼−170‰) suggest a thermogenic origin for CH4. The area is characterized by intense seismic activity and Duvalo corresponds to an active tectonic structure bordering the Ohrid graben. The production of H2S within the stratigraphic sequence may be explained by thermochemical reduction of sulfate. The uprising H2S is partially oxidized to sulfuric acid that, reacting with carbonate rocks, releases CO2. The tectonic structure of the area favors fluid circulation, sustaining H2S production and oxidation, CO2 production and allowing the escape of the gases to the atmosphere. In the end, Duvalo represents a tectonic‐related CO2 degassing area whose gases originate mostly, if not exclusively, in the shallowest part of the crust (1,000 t km−2 d−1).

Un certain nombre d’historiens et de sociologues ont interrogé la relation entre socialisation et architecture. Mais rares sont les auteurs ayant réfléchi au lien entre architecture et production des savoirs scientifiques. Dans cet article, nous présentons une piste de recherche pour approfondir le rapport entre architecture et production de savoirs dits interdisciplinaires. Nous proposons d’enquêter sur les discours, les stratégies et les éléments matériels que les chercheurs déploient pour mettre en place des collaborations scientifiques entre domaines différents. Les cas d’étude sont deux « instituts de la complexité » : le Santa Fe Institute (SFI) et l’Institut des systèmes complexes − Paris Île-de-France (ISC-PIF). Le matériau empirique se fonde sur des entretiens, des ethnographies et des archives.

The present article describes the epistemic project of complex systems sciences. The Santa Fe Institute founded in 1984 (New Mexico, United States) was the first theoretical research institute to focus on complex systems sciences as a field of research for studying the network of relationships within and between systems. Described by some as post-Laplacian, holist and antireductionist, the field of complex systems sciences relies heavily on a multidisciplinary approach to fundamental theoretical questions across the domains of physical and social science. Proponents such as renowned journalists and sociologists have declared the developing domain as the advent of a new revolutionary paradigm for all fields of knowledge. The specific aim of this text is to show that the epistemic project of complexity science is less pluralist, anti-positivist and antireductionist than some would claim. Further, I will examine why the epistemological and ontological framework of these sciences have remained both analytic and reductionist in physicalist, computational, mathematical and biological manners.

A detailed analysis of the elemental and molecular composition of Posidonia oceanica (PO) and Ascophyllum nodosum (AN) is presented. In particular, an in-depth study of the molecular identification via NMR spectroscopy of aqueous and organic extracts of PO and AN was carried out, exploiting 2D COSY and pseudo-2D DOSY data to aid in the assignment of peaks in complex 1D proton NMR spectra. Many metabolites were identified, such as carbohydrates, amino acids, organic acids, fatty acids, and polyphenols, with NMR complementing the characterization of the two species by standard elemental analysis, HPLC analysis, and colorimetric testing. For PO, different parts of the live plant (roots, rhizomes, and leaves) were analysed, as well as the residues of the dead plant which typically deposit along the coasts. The combination of the various studies made it possible to recognize bioactive compounds naturally present in the two plant species and, in particular, in the PO residues, opening the door for their possible recycling and use in, for example, fertilizer. Furthermore, NMR is proven to be a powerful tool for the metabolomic study of plant species as it allows for the direct identification of specific biomarkers as well as providing a molecular fingerprint of the plant variety.

Abstract We report on the results of an extensive geochemical survey of fluids released in the Vardar zone (central‐western Serbia), a mega‐suture zone at the boundary between Eurasia and Africa plates. Thirty‐one bubbling gas samples are investigated for their chemical and isotopic compositions (He, C, Ar) and cluster into three distinct groups (CO2‐dominated, N2‐dominated, and CH4‐dominated) based on the dominant gas species. The measured He isotope ratios range from 0.08 to 1.19 Ra (where Ra is the atmospheric ratio), and reveal for the first time the presence of a minor (

Background Paranasal sinuses act as bacterial reservoirs and contribute to transmitting bacteria to the lower airway of patients with cystic fibrosis (CF). Also, passage of bacteria from the oral cavity to the lungs may occur. Methods We evaluated the presence of Pseudomonas aeruginosa, Staphylococcus aureus, Stenotrophomonas maltophilia, Achromobacter xylosoxidans and Serratia marcescens in sputum and nasal lavage of 59 patients with CF, and also collected saliva and used toothbrushes from 38 of them. We assessed the clonal identity of the strains isolated from the different samples by pulsed-field gel electrophoresis. Results About 80% of the patients were positive for at least one of the bacterial species examined in nasal lavage and sputum. Among the subjects with positive sputum, 74% presented the same species in the nasal lavage and saliva, and 26% on their toothbrush. S. aureus was the most abundant species in all samples. Clonal identity (≥80% similarity) of the strains isolated among the different samples from each patient was confirmed in almost all cases. Longitudinal observation helped to identify five patients who were colonised in the lower airways after an initial period of nasal or oral colonisation. Conclusion Nasal and oral sites act as bacterial reservoirs, favouring the transmission of potentially pathogenic microorganisms to the lower airway. The lack of eradication from these sites might undermine the antibiotic therapy applied to treat the lung infection, allowing the persistence of the bacteria within the patient if colonisation of these sites is not assessed, and no specific therapy is performed.

Abstract Background and methods Wall paintings and architectural surfaces in outdoor environments are exposed to several physical, chemical and biological agents, hence they are often treated with different products to prevent or slow down their deterioration. Among the factors that have to be taken into account in the selection of the most suitable treatment for decorated surfaces, the aesthetic compatibility with the substrate is of great importance in the cultural heritage field; minimizing colour variation after treatment application is a crucial issue in particular for painted surfaces. In the framework of the European Project Nanomatch the color variation induced on wall painting mock-ups by the two innovative consolidants (calcium alkoxides) developed was evaluated using colorimetry in comparison with two traditional products. In this work these innovative consolidants have been also tested in combination with two commercial biocides and the results of colorimetric measurements discussed. Moreover, as the univariate approach didn’t allow to draw clear conclusions on the relation between the different sources of data variability, multivariate analysis was performed on colorimetric data. Results Principal Component Analysis and multi-way Parallel Factor Analysis (PARAFAC) were successfully applied to colorimetric data to investigate the short-term effects of the application of different consolidants on wall painting surfaces, making it possible to study at the same time the different sources of data variability, i.e. treatments, painting techniques, pigments. Finally, a ranking list of the treatments under study in terms of colour variation induced on the surface was established, in function of the painting technique and pigment, taking also in consideration the combination consolidant/biocide. In particular, given the true multi-way nature of the data, PARAFAC model turned out to be extremely useful in the study of the dependence of colour variation on pigments, a critical issue for painted surfaces, that was not clear using univariate approach. Conclusions Multivariate approach to colorimetric data and especially 3-way PARAFAC method resulted a powerful technique to evaluate in short-term the color compatibility of consolidants for wall paintings, improving data interpretation and visualization, and thus outperforming the univariate statistical analysis.

Forty-five gas samples have been collected from natural gas manifestations at the island of Kos—the majority of which are found underwater along the southern coast of the island. On land, two anomalous degassing areas have been recognized. These areas are mainly characterized by the lack of vegetation and after long dry periods by the presence of sulfate salt efflorescence. Carbon dioxide is the prevailing gas species (ranging from 88 to 99%), while minor amounts of N2 (up to 7.5%) and CH4 (up to 2.1%) are also present. Significant contents of H2 (up to 0.2%) and H2S (up to 0.3%) are found in the on-land manifestations. Only one of the underwater manifestations is generally rich in N2 (up to 98.9%) with CH4 concentrations of up to 11.7% and occasionally extremely low CO2 amounts (down to 0.09%). Isotope composition of He ranges from 0.85 to 6.71 R/RA, indicating a sometimes-strong mantle contribution; the highest values measured are found in the two highly degassing areas of Paradise beach and Volcania. C-isotope composition of CO2 ranges from -20.1 to 0.64‰ vs. V-PDB, with the majority of the values being concentrated at around -1‰ and therefore proposing a mixed mantle—limestone origin. Isotope composition of CH4 ranges from -21.5 to +2.8‰ vs. V-PDB for C and from -143 to +36‰ vs. V-SMOW for H, pointing to a geothermal origin with sometimes-evident secondary oxidation processes. The dataset presented in this work consists of sites that were repeatedly sampled in the last few years, with some of which being also sampled just before and immediately after the magnitude 6.6 earthquake that occurred on the 20th of July 2017 about 15 km ENE of the island of Kos. Changes in the degassing areas along with significant variations in the geochemical parameters of the released gases were observed both before and after the seismic event; however, no coherent model explaining those changes was obtained. CO2 flux measurements showed values of up to about 104 g×m−2×d−1 in both the areas of Volcania and Kokkino Nero, 5×104 g×m−2×d−1 at Paradise beach, and 8×105 g×m−2×d−1 at Therma spring. CO2 output estimations gave values of 24.6, 16.8, 12.7, and 20.6 t×d−1, respectively, for the above four areas. The total output of the island is 74.7 t×d−1 and is comparable to those of the other active volcanic/geothermal systems of Greece (Nisyros, Nea Kameni, Milos, Methana, and Sousaki).

Nisyros Island (Greece) is an active volcano hosting a high-enthalpy geothermal system. During June 2013, an extensive survey on Hg concentrations in different matrices (fumarolic fluids, atmosphere, soils, and plants) was carried out at the Lakki Plain, an intracaldera area affected by widespread soil and fumarolic degassing. Concentrations of gaseous elemental mercury (GEM), together with H2S and CO2, were simultaneously measured in both the fumarolic emissions and the atmosphere around them. At the same time, 130 samples of top soils and 31 samples of plants (Cistus creticus and salvifolius and Erica arborea and manipuliflora) were collected for Hg analysis. Mercury concentrations in fumarolic gases ranged from 10,500 to 46,300 ng/m3, while Hg concentrations in the air ranged from high background values in the Lakki Plain caldera (10-36 ng/m3) up to 7100 ng/m3 in the fumarolic areas. Outside the caldera, the concentrations were relatively low (2-5 ng/m3). The positive correlation with both CO2 and H2S in air highlighted the importance of hydrothermal gases as carrier for GEM. On the other hand, soil Hg concentrations (0.023-13.7 μg/g) showed no significant correlations with CO2 and H2S in the soil gases, whereas it showed a positive correlation with total S content and an inverse one with the soil pH, evidencing the complexity of the processes involving Hg carried by hydrothermal gases while passing through the soil. Total Hg concentrations in plant leaves (0.010-0.112 μg/g) had no direct correlation with soil Hg, with Cistus leaves containing higher values of Hg with respect to Erica. Even though GEM concentrations in the air within the caldera are sometimes orders of magnitude above the global background, they should not be considered dangerous to human health. Values exceeding the WHO guideline value of 1000 ng/m3 are very rare (

Raman spectroscopy, and handheld spectrometers in particular, are gaining increasing attention in food quality control as a fast, portable, non-destructive technique. Furthermore, this technology also allows for measuring the intact sample through the packaging and, with respect to near infrared spectroscopy, it is not affected by the water content of the samples. In this work, we evaluate the potential of the methodology to model, by multivariate data analysis, the authenticity of Parmigiano Reggiano cheese, which is one of the most well-known and appreciated hard cheeses worldwide, with protected denomination of origin (PDO). On the other hand, it is also highly subject to counterfeiting. In particular, it is critical to assess the authenticity of grated cheese, to which, under strictly specified conditions, the PDO is extended. To this aim, it would be highly valuable to develop an authenticity model based on a fast, non-destructive technique. In this work, we present preliminary results obtained by a handheld Raman spectrometer and class-modeling (Soft Independent Modeling of Class Analogy, SIMCA), which are extremely promising, showing sensitivity and specificity of 100% for the test set. Moreover, another salient issue, namely the percentage of rind in grated cheese, was addressed by developing a multivariate calibration model based on Raman spectra. It was possible to obtain a prediction error around 5%, with 18% being the maximum content allowed by the production protocol.

In order to reduce the mismatch effect caused by non-uniform shadows in PV arrays, reconfigurable interconnections approaches have been recently proposed in the literature. These systems usually require the knowledge of the solar radiation affecting every solar module. The aim of this work is to evaluate the effectiveness of three irradiance estimation approaches in order to define which can be well suited for reconfigurable PV arrays. It is presented a real-time solar irradiance estimation device (IrradEst), implementing the three different estimation methods. The proposed system is based on mathematical models of PV modules enabling to estimate irradiation values by sensing a combination of temperature, voltage and current of a PV module. Experimental results showed generally good agreement between the estimated irradiances and the measurements performed by a standard pyranometer taken as reference. Finally one of the three methods was selected as possible solution for a reconfigurable PV system.

This work presents a preliminary study on the implementation of a new system for power output maximization of photovoltaic generators under non-homogeneous conditions. The study evaluates the performance of an efficient switching matrix and the relevant automatic reconfiguration control algorithms. The switching matrix is installed between the PV generator and the inverter, allowing a large number of possible module configurations. PV generator, switching matrix and the intelligent controller have been simulated in Simulink. The proposed reconfiguration system improved the energy extracted by the PV generator under non-uniform solar irradiation conditions. Short calculation times of the proposed control algorithms allow its use in real time applications even where a higher number of PV modules is required.

The interview is based on some thesis of Semir Zeki’s last book, Splendours and Miseries of the Brain. The founder of the International Institute of Neuroaesthetics (London 2001) wrote that a very different kind of knowledge can be acquired by the brain in the experience of art. It seems that he recognizes there can be a kind of “intelligence” in perception, while he writes about microcosciences of perception being located in the same areas where the information coming from the outside is elaborated and perceived. He explains the distinction between inherited “concepts” and acquired synthetic “concepts” of the visual brain. The theory of acquired concepts could also help in explaining the problem of taste: it seems that what we like in our lives is determined by the result (the synthesis) of those probabilities coming from our acquired synthetic brain concepts. Thus there wouldn’t be any free will in our aesthetical and love choices.

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The frequency of erythromycin resistance in Streptococcus pneumoniae isolates from human specimens, in different Piemonte Region hospitals between 2005-2007, has been studied. Erythromycin-susceptible isolates were 137/198 and erythromycin-resistant 61/198.Among resistant S. pneumoniae isolates 26/61 belonged to constitutive resistance phenotype, 26/61 to M phenotype and 9/61 to inducible resistance phenotype.

The amphibians of the pond complex “Stagni di Belangero” in the Po Plain, Asti Province, have been studied. The species living in the pond are Bufo bufo, Bufo viridis, Hyla intermedia, Pelobates fuscus, Rana dalmatina, Rana synklepton esculenta, Rana cf. kurtmuelleri, Triturus carnifex and Triturus vulgaris. Species composition, migration period of P. fuscus, and biometric data are provided. Relevant importance has been given to P. fuscus, since we reconfirm its presence in one of the ponds 13 years after its first finding in the area. This toad is also one of the most threatened species of amphibians in Europe and needs particular attention in order to be protected adequately.

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Copyright of Physis is the property of Casa Editrice Leo S. Olschki SRL and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The article presents information on the herpetological study in Sicily, Italy on the Xenopus laevis, African clawed frogs first seen on the island in the early 2000's. The research focused on the consequences of the introduction of the frogs to the native Sicilian amphibians, including distribution, predation, competition, and other possible negative effects. Included are photographs of the animal and the research conducted in a pond in Sicily.

The article focuses on the Pleurodeles waltl, a ribbed newt found in the Iberian Peninsula of Spain and Portugal. It belongs to the subfamily Pleurodelinae Tshudi of the Salamandridae family. The habitat, distribution and habits of the ribbed newt are discussed. The Pleurodeles waltl has a rounded snout and a wide and flattened head. It is classified as Near Threatened in the International Union for Conservation of Nature (IUCN) Red Data List of amphibians and reptiles in Spain.