Biocleaning of starch glues from textiles by means of α-amylase-based treatments.

Glues based on starch are widely used for the consolidation of brittle fibres in historic and archaeological textiles. Ageing fabrics are affected by hydrolysis/oxidation and cross-linking of these glues, a decrease of glues' solubility, the formation of cracks, and discoloration. The hydrolytic action of enzymes on starch-based glues is promising, as molecular recognition offers great selectivity. However, a systematic assessment of the best methods for applying enzymatic formulations has not been explored yet. Here, α-amylase was applied either by pipetting a solution or combining with gellan gel (embedded in the gel or spread on the gel surface). The effectiveness of the different formulations on the removal of potato and wheat starch was evaluated by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and colorimetric measurements. Enzymes dispersed in gel showed weak diffusion at the surface, resulting in poor starch breakdown and removal. On the contrary, amylase applied by pipette and spread on gel resulted in high starch removal selectivity and efficiency, with neither swelling nor damage to the fibres. These results validate protocols for the assessment of the enzymatic activity on glue-consolidated fibres, identify best application methods and confirm the excellent properties of amylase dispersions for the conservation of historic and archaeological textiles.Key points• Application of α-amylase by pipette and combined with gellan gel to remove starch glues from wool.• Systematic assessment of the best application methods following a multi-analytical protocol.• Enzymes dispersed in gel exhibit poor diffusion at the surface, leading to weak starch removal.• Enzymes applied by pipette and spread on gel are efficient in starch cleaning, without damage to the fibres.

Immobilized Enzymes on Gold Nanoparticles: From Enhanced Stability to Cleaning of Heritage Textiles.

Enzyme-based treatments are used in heritage conservation for the effective removal of glues and other damaging organic layers from the surfaces of historic and artistic works. Despite their potential, however, the application of enzymatic treatments is currently limited because of their poor efficiency and low operational and environmental stability. We demonstrate the use of α-amylase immobilized on gold nanoparticles to improve the efficacy of enzymatic treatments enhancing both the reactivity and the stability of the formulations. Gold nanoparticles coated with α-amylase exhibit significant advantages compared to free enzymes. We report up to 5× greater resistance to environmental changes, up to 2× higher efficacy toward removal of starch-based glues from textiles and deeper penetration through the fibers, without causing damage or inducing salt precipitation. These results offer exciting prospects for the development of enzymatic formulations, both for heritage conservation and the wider application of enzymes, such as in medicine, the detergent industry, and green chemistry.

Stone/Coating Interaction and Durability of Si-Based Photocatalytic Nanocomposites Applied to Porous Lithotypes.

The use of hybrid nanocoatings for the protection of natural stones has received increasing attention over the last years. However, the interaction of these materials with stones and, in particular, its modification resulting from the blending of nanoparticles and matrices, are yet little explored. In this work, the interaction of two nanocomposite coatings (based on alkylalkoxysilane matrices and TiO2 nanoparticles in water and 2-propanol) with two different porous stones is examined in detail by comparing their absorption behaviour and protection performance with those of the respective TiO2-free matrices. It is shown that the protective effectiveness of both matrices is not negatively affected by the presence of TiO2, as the desired water barrier effect is retained, while a significant photocatalytic activity is achieved. The addition of titania leads to a partial aggregation of the water-based matrix and accordingly reduces the product penetration into stones. On the positive side, a chemical interaction between titania and this matrix is observed, probably resulting in a greater stability of nanoparticles inside the protective coating. Moreover, although an effect of TiO2 on the chemical stability of matrices is observed upon UV light exposure, the protective performance of coatings is substantially maintained after ageing, while the interaction between matrices and nanoparticles results in a good retention of the latter upon in-lab simulated rain wash-out.

Layered Nano-TiO2 Based Treatments for the Maintenance of Natural Stones in Historical Architecture.

Layered treatments of natural stones based on dispersions of experimental nano-TiO2 and commercial TEOS showing photocatalytic and self-cleaning properties were set up and tested. To enhance nano-TiO2 efficacy, a surface pre-treatment with tetraethyl orthosilicate was proposed to avoid the penetration of NPs into the crystalline porous substrates and to improve their adhesion to the stone. Two treatment applications (wet-on-wet and wet-on-dry) were compared, showing different results. A strong interaction Si-O-Ti was the key factor for the successful treatment, leaving the band gap and relevant properties of nano-TiO2 unaltered. The layered treatments were tested on a porous calcarenite (Noto stone) and a very compact marble (Carrara marble). The combined SiO2 -nano-TiO2 treatments can find application in suitable cases where a surface consolidation is needed, ensuring a depolluting and self-cleaning durable activity.

Photocatalytic Nanocomposites for the Protection of European Architectural Heritage.

In the field of stone protection, the introduction of inorganic nanoparticles, such as TiO2, ZnO, and Ag in polymeric blends can enhance the protective action of pristine treatments, as well as confer additional properties (photocatalytic, antifouling, and antibacterial). In the framework of the "Nano-Cathedral" European project, nanostructured photocatalytic protective treatments were formulated by using different TiO2 nanoparticles, solvents, and silane/siloxane systems in the blends. The results about the characterization and application of two promising nano-TiO2 based products applied on Apuan marble and Ajarte limestone are here reported, aiming at investigating the complex system "treatment/stone-substrate". The nanocomposites show better performances when compared to a commercial reference siloxane based protective treatment, resulting in different performances once applied on different carbonatic substrates, with very low and high open porosity, confirming the necessity of correlating precisely the characteristics of the stone material to those of the protective formulations. In particular, the TiO2 photocatalytic behavior is strictly linked to the amount of available nanoparticles and to the active surface area. The alkyl silane oligomers of the water-based formulation have a good penetration into the microstructure of Ajarte limestone, whereas the solvent-based and small size monomeric formulation shows better results for Apuan marble, granting a good coverage of the pores. The encouraging results obtained so far in lab will be confirmed by monitoring tests aiming at assessing the effectiveness of the treatments applied in pilot sites of historical Gothic Cathedrals.

A Photoluminescence Study of the Changes Induced in the Zinc White Pigment by Formation of Zinc Complexes.

It is known that oil paintings containing zinc white are subject to rapid degradation. This is caused by the interaction between the active groups of binder and the metal ions of the pigment, which gives rise to the formation of new zinc complexes (metal soaps). Ongoing studies on zinc white paints have been limited to the chemical mechanisms that lead to the formation of zinc complexes. On the contrary, little is known of the photo-physical changes induced in the zinc oxide crystal structure following this interaction. Time-resolved photoluminescence spectroscopy has been applied to follow modifications in the luminescent zinc white pigment when mixed with binder. Significant changes in trap state photoluminescence emissions have been detected: the enhancement of a blue emission combined with a change of the decay kinetic of the well-known green emission. Complementary data from molecular analysis of paints using Fourier transform infrared spectroscopy confirms the formation of zinc carboxylates and corroborates the mechanism for zinc complexes formation. We support the hypothesis that zinc ions migrate into binder creating novel vacancies, affecting the photoluminescence intensity and lifetime properties of zinc oxide. Here, we further demonstrate the advantages of a time-resolved photoluminescence approach for studying defects in semiconductor pigments.

Multiple drivers of decline in the global status of freshwater crayfish (Decapoda: Astacidea).

Rates of biodiversity loss are higher in freshwater ecosystems than in most terrestrial or marine ecosystems, making freshwater conservation a priority. However, prioritization methods are impeded by insufficient knowledge on the distribution and conservation status of freshwater taxa, particularly invertebrates. We evaluated the extinction risk of the world's 590 freshwater crayfish species using the IUCN Categories and Criteria and found 32% of all species are threatened with extinction. The level of extinction risk differed between families, with proportionally more threatened species in the Parastacidae and Astacidae than in the Cambaridae. Four described species were Extinct and 21% were assessed as Data Deficient. There was geographical variation in the dominant threats affecting the main centres of crayfish diversity. The majority of threatened US and Mexican species face threats associated with urban development, pollution, damming and water management. Conversely, the majority of Australian threatened species are affected by climate change, harvesting, agriculture and invasive species. Only a small proportion of crayfish are found within the boundaries of protected areas, suggesting that alternative means of long-term protection will be required. Our study highlights many of the significant challenges yet to come for freshwater biodiversity unless conservation planning shifts from a reactive to proactive approach.

Alien insects in Italy: comparing patterns from the regional to European level.

The introduction of species outside their native range contributes to the loss of biodiversity, alters the structure and functioning of ecosystems, and damages economy and human health. Insects are one of the taxa with the highest frequency of introduction due to their high diversity, biological properties, and close association with human activities. Here, the allodiversity of Italian entomofauna was analyzed, with a focus on Tuscany (Central Italy). A list of alien insects in Tuscany is included. The status of the alien entomofauna in Italy was updated. The number of alien insects amounts to 122 in Tuscany and 923 in Italy. An introduction rate of 98 species per decade was estimated in Italy. In Tuscany, alien insects belong to 10 orders, mostly Coleoptera (38%), Hemiptera (Sternorrhyncha and Auchenorrhyncha) (23%), and Hymenoptera (13%). They have been most often introduced through vegetable items (ornamental plants or crops). Most species come from the Nearctic region (26%) and are both phytophagous (63%) and amphigonic (80%). Differences and similarities in introduction patterns and in insect abundances across orders among regional, national, and European scales, also considering worldwide abundances, are discussed. Finally, a paucity of information regarding the negative impacts of many species, except for economic pests, phytosanitary threats, and vectors of disease, is underlined. A deeper understanding of the alien insects' ecological impact might help designate policies aimed at preventing further introductions and control the invasive populations of already established species.

Effects of climate change, invasive species, and disease on the distribution of native European crayfishes.

Climate change will require species to adapt to new conditions or follow preferred climates to higher latitudes or elevations, but many dispersal-limited freshwater species may be unable to move due to barriers imposed by watershed boundaries. In addition, invasive nonnative species may expand into new regions under future climate conditions and contribute to the decline of native species. We evaluated future distributions for the threatened European crayfish fauna in response to climate change, watershed boundaries, and the spread of invasive crayfishes, which transmit the crayfish plague, a lethal disease for native European crayfishes. We used climate projections from general circulation models and statistical models based on Mahalanobis distance to predict climate-suitable regions for native and invasive crayfishes in the middle and at the end of the 21st century. We identified these suitable regions as accessible or inaccessible on the basis of major watershed boundaries and present occurrences and evaluated potential future overlap with 3 invasive North American crayfishes. Climate-suitable areas decreased for native crayfishes by 19% to 72%, and the majority of future suitable areas for most of these species were inaccessible relative to native and current distributions. Overlap with invasive crayfish plague-transmitting species was predicted to increase. Some native crayfish species (e.g., noble crayfish [Astacus astacus]) had no future refugia that were unsuitable for the modeled nonnative species. Our results emphasize the importance of preventing additional introductions and spread of invasive crayfishes in Europe to minimize interactions between the multiple stressors of climate change and invasive species, while suggesting candidate regions for the debatable management option of assisted colonization.

Conceptual frameworks and methods for advancing invasion ecology.

Invasion ecology has much advanced since its early beginnings. Nevertheless, explanation, prediction, and management of biological invasions remain difficult. We argue that progress in invasion research can be accelerated by, first, pointing out difficulties this field is currently facing and, second, looking for measures to overcome them. We see basic and applied research in invasion ecology confronted with difficulties arising from (A) societal issues, e.g., disparate perceptions of invasive species; (B) the peculiarity of the invasion process, e.g., its complexity and context dependency; and (C) the scientific methodology, e.g., imprecise hypotheses. To overcome these difficulties, we propose three key measures: (1) a checklist for definitions to encourage explicit definitions; (2) implementation of a hierarchy of hypotheses (HoH), where general hypotheses branch into specific and precisely testable hypotheses; and (3) platforms for improved communication. These measures may significantly increase conceptual clarity and enhance communication, thus advancing invasion ecology.

Crustacean hyperglycemic hormone (cHH) as a modulator of aggression in crustacean decapods.

Biogenic amines, particularly serotonin, are recognised to play an important role in controlling the aggression of invertebrates, whereas the effect of neurohormones is still underexplored. The crustacean Hyperglycemic Hormone (cHH) is a multifunctional member of the eyestalk neuropeptide family. We expect that this neuropeptide influences aggression either directly, by controlling its expression, or indirectly, by mobilizing the energetic stores needed for the increased activity of an animal. Our study aims at testing such an influence and the possible reversion of hierarchies in the red swamp crayfish, Procambarus clarkii, as a model organism. Three types of pairs of similarly sized males were formed: (1) 'control pairs' (CP, n = 8): both individuals were injected with a phosphate saline solution (PBS); (2) 'reinforced pairs' (RP, n = 9): the alpha alone was injected with native cHH, and the beta with PBS; (3) 'inverted pairs' (IP, n = 9): the opposite of (2). We found that, independently of the crayfish's prior social experience, cHH injections induced (i) the expression of dominance behaviour, (ii) higher glycemic levels, and (iii) lower time spent motionless. In CP and RP, fight intensity decreased with the establishment of dominance. On the contrary, in IP, betas became increasingly likely to initiate and escalate fights and, consequently, increased their dominance till a temporary reversal of the hierarchy. Our results demonstrate, for the first time, that, similarly to serotonin, cHH enhances individual aggression, up to reverse, although transitorily, the hierarchical rank. New research perspectives are thus opened in our intriguing effort of understanding the role of cHH in the modulation of agonistic behaviour in crustaceans.

Revisiting social recognition systems in invertebrates.

Since the 1970s, the ability of some invertebrate species to recognize individual conspecifics has attracted increased scientific interest. However, there is still confusion in the literature, possibly due to the lack of unambiguous criteria for classifying social recognition in its different forms. Here, we synthesize the results of studies on invertebrates and provide a framework with the purpose of identifying research needs and directions for future investigations. Following in part Sherman et al.'s (Behavioural ecology: an evolutionary approach. Blackwell Science, Oxford, pp 69-96, 1997) definition of 'recognition systems' and Tibbetts and Dale's (Trends Ecol Evol 22:529-537, 2007) classification of 'individual recognition,' we first discuss different case studies that exemplify the categories of 'familiar recognition' and 'class-level recognition.' Then, through the analysis of the invertebrate literature, we illustrate eight key properties that characterize 'true individual recognition' systems. We are confident that the proposed framework will provide opportunities for exciting discoveries of the cognitive abilities in invertebrates.

Use of natural pyrethrum to control the red swamp crayfish Procambarus clarkii in a rural district of Italy.

BACKGROUND: The crayfish Procambarus clarkii inflicts severe ecological and economic damages in Europe. To develop an efficient method for its control, four experiments were carried out to assess the impact of natural pyrethrum (i.e. Pyblast) on crayfish: (1) the 24 h LC(100) and LC(50) were quantified on crayfish; (2) the breakdown time of the 24 h LC(100) was assessed using Daphnia magna as a bioindicator; the effects of 24 h LC(100) on crayfish were investigated by applying the biocide into burrows (3) and in a drainage channel (4). RESULTS: Pyblast concentrations of 0.05 and 0.02 mg L(-1) corresponded to 24 h LC(100) and LC(50) respectively. The concentration of 0.05 mg L(-1) broke down after 72 h, whereas 0.02 mg L(-1) did not cause any significant mortality in D. magna after 24 h. However, 0.05 mg L(-1) had no effect on crayfish when introduced into the burrows, but led to a mortality of 95% when applied in the water. CONCLUSION: Experimental evidence is provided for the efficacy of Pyblast to control invasive crayfish. Obviously, before its use on a large scale, further studies are needed to find a concentration that will achieve the target 100% mortality with the shortest recovery time of the environment.

I know my neighbour: individual recognition in Octopus vulgaris.

BACKGROUND: Little is known about individual recognition (IR) in octopuses, although they have been abundantly studied for their sophisticated behaviour and learning capacities. Indeed, the ability of octopuses to recognise conspecifics is suggested by a number of clues emerging from both laboratory studies (where they appear to form and maintain dominance hierarchies) and field observations (octopuses of neighbouring dens display little agonism between each other). To fill this gap in knowledge, we investigated the behaviour of 24 size-matched pairs of Octopus vulgaris in laboratory conditions. METHODOLOGY/PRINCIPAL FINDINGS: The experimental design was composed of 3 phases: Phase 1 (acclimatization): 12 "sight-allowed" (and 12 "isolated") pairs were maintained for 3 days in contiguous tanks separated by a transparent (and opaque) partition to allow (and block) the vision of the conspecific; Phase 2 (cohabitation): members of each pair (both sight-allowed and isolated) were transferred into an experimental tank and were allowed to interact for 15 min every day for 3 consecutive days; Phase 3 (test): each pair (both sight-allowed and isolated) was subject to a switch of an octopus to form pairs composed of either familiar ("sham switches") or unfamiliar conspecifics ("real switches"). Longer latencies (i.e. the time elapsed from the first interaction) and fewer physical contacts in the familiar pairs as opposed to the unfamiliar pairs were used as proxies for recognition. CONCLUSIONS: Octopuses appear able to recognise conspecifics and to remember the individual previously met for at least one day. To the best of our knowledge, this is the first experimental study showing the occurrence of a form of IR in cephalopods. Future studies should clarify whether this is a "true" IR.

Socioeconomic legacy yields an invasion debt.

Globalization and economic growth are widely recognized as important drivers of biological invasions. Consequently, there is an increasing need for governments to address the role of international trade in their strategies to prevent species introductions. However, many of the most problematic alien species are not recent arrivals but were introduced several decades ago. Hence, current patterns of alien-species richness may better reflect historical rather than contemporary human activities, a phenomenon which might be called "invasion debt." Here, we show that across 10 taxonomic groups (vascular plants, bryophytes, fungi, birds, mammals, reptiles, amphibians, fish, terrestrial insects, and aquatic invertebrates) in 28 European countries, current numbers of alien species established in the wild are indeed more closely related to indicators of socioeconomic activity from the year 1900 than to those from 2000, although the majority of species introductions occurred during the second half of the 20th century. The strength of the historical signal varies among taxonomic groups, with those possessing good capabilities for dispersal (birds, insects) more strongly associated with recent socioeconomic drivers. Nevertheless, our results suggest a considerable historical legacy for the majority of the taxa analyzed. The consequences of the current high levels of socioeconomic activity on the extent of biological invasions will thus probably not be completely realized until several decades into the future.

Disentangling the role of environmental and human pressures on biological invasions across Europe.

The accelerating rates of international trade, travel, and transport in the latter half of the twentieth century have led to the progressive mixing of biota from across the world and the number of species introduced to new regions continues to increase. The importance of biogeographic, climatic, economic, and demographic factors as drivers of this trend is increasingly being realized but as yet there is no consensus regarding their relative importance. Whereas little may be done to mitigate the effects of geography and climate on invasions, a wider range of options may exist to moderate the impacts of economic and demographic drivers. Here we use the most recent data available from Europe to partition between macroecological, economic, and demographic variables the variation in alien species richness of bryophytes, fungi, vascular plants, terrestrial insects, aquatic invertebrates, fish, amphibians, reptiles, birds, and mammals. Only national wealth and human population density were statistically significant predictors in the majority of models when analyzed jointly with climate, geography, and land cover. The economic and demographic variables reflect the intensity of human activities and integrate the effect of factors that directly determine the outcome of invasion such as propagule pressure, pathways of introduction, eutrophication, and the intensity of anthropogenic disturbance. The strong influence of economic and demographic variables on the levels of invasion by alien species demonstrates that future solutions to the problem of biological invasions at a national scale lie in mitigating the negative environmental consequences of human activities that generate wealth and by promoting more sustainable population growth.

Calibration of FI-ISK, an invasiveness screening tool for nonnative freshwater invertebrates.

The Freshwater Invertebrate Invasiveness Scoring Kit (FI-ISK) is proposed as a screening tool for identifying potentially invasive freshwater invertebrates. FI-ISK was adapted from the Fish Invasiveness Scoring Kit (FISK) of Copp, Garthwaite, and Gozlan, which is an adapted form of the Weed Risk Assessment (WRA) of Pheloung, Williams, and Halloy. Initial assessments using FI-ISK, which include confidence (certainty/uncertainty) rankings by the assessor to each response, were calibrated to determine the most appropriate score thresholds for classifying nonnative species into low-, medium-, and high-risk categories, using both the original medium-to-high risk threshold scores for the WRA (i.e., > or = 6) and for FISK (i.e., > or = 19). Patterns of the assessor's confidence, when making the responses during the FI-ISK assessments, were also examined. Using receiver operating characteristic (ROC) curves, FI-ISK was shown to distinguish accurately (and with statistical confidence) between potentially invasive and noninvasive species of nonnative crayfish (Decapoda: Astacidae, Cambaridae, Parastacidae), with the statistically appropriate threshold score for high-risk species scores being > or = 16. FI-ISK represents a useful and viable tool to aid decision- and policymakers in assessing and classifying freshwater invertebrates according to their potential invasiveness.

Sex identification in female crayfish is bimodal.

Sex identification has been studied in several species of crustacean decapods but only seldom was the role of multimodality investigated in a systematic fashion. Here, we analyse the effect of single/combined chemical and visual stimuli on the ability of the crayfish Procambarus clarkii to identify the sex of a conspecific during mating interactions. Our results show that crayfish respond to the offered stimuli depending on their sex. While males rely on olfaction alone for sex identification, females require the combination of olfaction and vision to do so. In the latter, chemical and visual stimuli act as non-redundant signal components that possibly enhance the female ability to discriminate potential mates in the crowded social context experienced during mating period. This is one of the few clear examples in invertebrates of non-redundancy in a bimodal communication system.

Behavioural indicators of pain in crustacean decapods.

Whether invertebrates are able or not to experience pain is a highly controversial issue. An operative way to solve such a controversy might be to investigate their responses to potentially noxious stimuli and to collect evidence of their behavioural complexities as proxies of cognitive capacities. The principle of argument-by-analogy can be then applied to these data: the behaviour displayed by invertebrates is compared with that of "higher" animals, its similarity denoting the former's capacity to have analogous experiences. Here, the author discusses some examples, extracted from the literature on crustacean decapods, that pinpoint their nature of "sentient" animals. This review, however, also shows that research is still scanty in the field. The studies that examine the potential links between stress responses and pain experience are few, and the several papers that help elucidate cognitive abilities in decapods have been limited to a few taxa and are not specifically directed to the question of "sentience". On the contrary, also in the light of the expected revision of the current EU legislation in the matter, more scientific efforts should be expended on exploring the issue of pain experience in invertebrates.