Global warming scenarios for the Eastern Adriatic Sea indicate a higher risk of invasiveness of non-native marine organisms relative to current climate conditions.

Globally, marine bioinvasions threaten marine ecosystem structure and function, with the Mediterranean Sea being one of the most affected regions. Such invasions are expected to increase due to climate change. We conducted a risk screening of marine organisms (37 fishes, 38 invertebrates, and 9 plants), both extant and 'horizon' (i.e., not present in the area but likely to enter it). Based on expert knowledge for the Eastern Adriatic Sea coasts of Slovenia, Croatia, and Montenegro, screenings were conducted under both current and predicted climate conditions indicating with an increase in sea surface temperature and salinity of the Adriatic Sea together with changes in precipitation regime. Our aims were to: (1) identify non-native extant and horizon marine species that may pose threats to native biodiversity and (2) evaluate the risk of invasiveness of the selected species under current and predicted climate conditions. Of the 84 species screened, there was an increase in those ranked as 'high risk' from 33 (39.3%) under current climate conditions and to 47 (56.0%) under global warming scenarios. For those ranked as 'very high' risk, the increase was from 6 (7.1%) to 21 (25.0%). Amongst the screened species, the already established high-risk species Pacific oyster Magallana gigas and Atlantic blue crab Callinectes sapidus represent a threat to ecosystem services. Given the under-representation of marine species in the current European Union List, the species we have ranked as high to very high risk should be included. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-023-00196-9.

pH-Dependent Behavior of Novel 5-FU Delivery System in Environmental Conditions Comparable to the Gastro-Intestinal Tract.

A biogenic carrier for 5-fluorouracil (5-FU) loading and subsequent tableting as a new drug formulation for slow release has been proposed using the biomineral from blue crab carapace. Due to its highly ordered 3D porous nanoarchitecture, the biogenic carbonate carrier could achieve increased effectiveness in colorectal cancer cure provided that the formulation would successfully pass through the gastric acid conditions. Following the recently proven viability of the concept by demonstrating the slow release of the drug from the carrier using the highly sensitive SERS technique, here we investigated the 5-FU release from the composite tablet drug in pH conditions replicating the gastric environment. The released drug from the tablet was studied in solutions with three relevant pH values, pH 2, pH 3, and pH 4. The 5-FU SERS spectral signature for each pH value was used to build calibration curves for quantitative SERS analysis. The results suggested a similarly slow-releasing pattern in acid pH environments to that in neutral conditions. Although biogenic calcite dissolution was expected in acid conditions, the X-ray diffraction and Raman spectroscopy showed preservation of calcite mineral along with the monohydrocalcite during acid solution exposure for two hours. The total released amount in a time course of seven hours, however, was lower in acidic pH solutions, with a maximum fraction of ~40% of the total amount of loaded drug, for pH 2, as opposed to ~80% for neutral values. Nonetheless, these results clearly prove that the novel composite drug retains its slow-releasing character in environmental conditions compatible with the gastrointestinal pH and that it is a viable and biocompatible alternative for oral delivery of anticancer drug to reach the lower gastro-intestinal tract.

Novel flexible sensing technology for nondestructive detection on live fish health/quality during waterless and low-temperature transportation.

Fish health/quality issues are increasingly attracting attention during waterless and low-temperature transportation. Nondestructive detection has become a great need for an effective method to improve fish health/quality. Currently, emerging Internet of Things, novel flexible electronics and data fusion technology have received great interest for nondestructive detection on live fish health/quality. This paper analysized nondestructive detection mechanisms using novel flexible sensing technology to achieve high-precision sensing of key parameters, and machine learning based data fusion modeling to achieve live fish health/quality nondestructive evaluation during waterless and low-temperature transportation. Recent studies on novel flexible electrochemical and physiological biosensors development and application for solving key ambient and physiological parameter sensing were summarized. The ML based data fusion modeling framework and application for live fish health/quality nondestructive evaluation was also highlighted. The future perspective is also proposed to provide promising solutions for accurate sensing of multi-parameter and real applications of live fish health/quality nondestructive detection during waterless and low-temperature transportation.

Impact of COVID-19 on Croatian mariculture: Findings from the first national surveys.

An overview of the negative effects of the COVID-19 pandemic on the mariculture industry of the republic of Croatia is provided. An initial online survey was circulated early after the onset of the pandemic and a follow-up field survey was performed a year into the pandemic. The surveyed companies varied in size (micro to medium enterprises), location (north, central and southern coast) and cultured organism (European flat oyster, Mediterranean mussel, European sea bass, Gilthead sea bream and/or Bluefin tuna) and were asked questions on the subject of economic and job losses, aquaculture supply chain processes and implemented or proposed measures for mitigation of negative effects. Results from the online survey showed higher economic loss than job loss, but companies reported increased job loss in the period leading to the field survey. Most companies reported reductions in sales and avenues of procurement, which, in addition to direct stressors, indirectly affected business processes. Micro enterprises fared well due to their part-time nature, low capital investments and running costs, while small to medium enterprises were under the most pressure. Large enterprises were barely affected as they had secure local and/or international distribution chains and dominated the market. Producers most affected were those that relied on the HoReCa market for product placement and/or had difficulty coping with existing stressors. Bivalve producers generally experienced a higher drop in sales than finfish farms and companies with specialized production were not able to adapt to market changes to the degree that more versatile businesses seemed capable of.

Comparative screening the life-time composition and crystallinity variation in gilthead seabream otoliths Sparus aurata from different marine environments.

Differences in crystallinity, structure and composition variation along the growing direction in gilthead seabream, Sparus aurata otoliths that inhabited different environments were determined to evaluate the correlation of spectroscopic and chemical data with the lifetime development and movement pattern. The Raman spectroscopy signal provided the characteristic bands whose Full Width at Half Maximum (FWHM) were used to track the signal variability. The FWHM showed an initial increase in the core area, followed by a decrease depicting two minima coinciding growth rings. The crystal discontinuity linked to annual rings was confirmed. The FWHM pattern followed cycle in the individual's activity. However, no significant correlation with FWHM and environmental factors although the slope of the FWHM variation distinguished aquaculture and costal groups from open sea and transitional, estuarine waters. Raman data were further correlated with morphological and elemental composition obtained via SEM-EDX and by LA-ICP-MS. SEM clearly confirmed CRM findings. Finally, multiparameter analysis of Ba/Ca concentrations obtained by LA-ICP-MS indicated the separation of groups associated with aquaculture and transitional waters due lowest variability in the elemental composition. Other groups are more variable possibly due to the water oligotrophic character and greater variability in prey availability in each environment. Results of the present study showed the additional potential of Raman spectroscopy as a complementary tool for inference of migration or origin of fish based on otolith composition and structure like other well-established technique.

Rapid and Application-Tailored Assessment Tool for Biogenic Powders from Crustacean Shell Waste: Fourier Transform-Infrared Spectroscopy Complemented with X-ray Diffraction, Scanning Electron Microscopy, and Nuclear Magnetic Resonance Spectroscopy.

Due to their chemical composition, richness in calcium carbonate, chitin, proteins, and pigments, and nanoporous structure, crustacean shell waste shows great potential for a wide variety of applications. Large quantities of waste shells are produced annually, meaning that they can be considered a renewable source of ecofriendly biogenic materials, which can be turned into value-added byproducts. In this paper, an IR-based technique is developed to differentiate various biogenic powders originated from crude or food-processed crustacean shells. The validity of the method is supported by cross-checking with XRD, NMR, and SEM-EDX analyses. Our goal was to determine changes in properties of waste crab shells after the two most common treatments, deproteinization and milling. We discovered that deproteinization with NaOH could be tracked from the IR absorbance intensity ratio of the υ(CH2,3) and υasym(CO3 2-) bands while milling time less influenced this ratio but induced changes in powder particle size distribution and morphology. The relative organic/inorganic ratio was different for different colored shells. Unexpectedly, waste shells stored for an average of 6 months or more were found to contain hydrated calcium carbonate (monohydrocalcite), which was absent in equivalent fresh shell composition. Deproteinization caused changes in mechanical properties of shells, making them more brittle, which resulted in a larger fraction of fine particles after powdering.

Novel Drug Carrier: 5-Fluorouracil Formulation in Nanoporous Biogenic Mg-calcite from Blue Crab Shells-Proof of Concept.

The ever-growing demand for novel, cheaper, and more effective drugs has put nanomedicine and targeted drug delivery to the forefront of scientific innovation. Owing to its porous three-dimensional (3D)-nanostructure and properties, the biogenic calcite from wasted blue crab shells is employed in the present work as a new drug carrier for 5-fluorouracil (5-FU), a drug widely used in cancer therapy. The drug solution has been loaded in the porous nanoarchitecture of the powdered biogenic material and further pelleted in tablets with a 5-FU concentration of 1.748 mg/g. Their structural and morphological properties were characterized using Raman, X-ray diffraction, and scanning electron microscopy. Confocal micro-Raman spectra of tablet surface showed a typical signal of biogenic carbonate with preserved carotenoids and carotenoproteins found in the native waste shell, while the drug Raman signal was absent, indicating its adsorption in the intricate nanoporous biogenic carrier. The slow release of the drug from the newly formulated tablet was investigated by tracking the surface-enhanced Raman scattering (SERS) signal of the tablet solution in a series of time-dependent experiments. The SERS signal quantification is achieved using the well-known SERS spectral fingerprint of 5-fluorouracil aqueous solution adsorbed on Ag nanoparticles. The proof of concept is demonstrated by quantifying the slow release of the drug through the characteristic SERS band intensity of 5-FU in a time course of 26 h. This proof of concept boosted further investigations concerning the released drug identity in simulated solutions that mimic the pH of the upper- and lower gastrointestinal tract, as well as the multiple possibilities to control porosity and composition during powdering and treatment of biogenic material, to achieve the most convenient formulation for relevant biomedical drug delivery. Nonetheless, the present results showed great promise for innovative reusing waste biogenic 3D-nanomaterials of aquatic origin as advantageous drug carriers for slow release purposes, in line with the concept of blue bioeconomy.

Wasted Biomaterials from Crustaceans as a Compliant Natural Product Regarding Microbiological, Antibacterial Properties and Heavy Metal Content for Reuse in Blue Bioeconomy: A Preliminary Study.

The compliance of crab shells traditionally used as a complex natural product for agricultural soil amendment with modern biofertilizers' quality and safety requirements was investigated. Shells waste from the Blue crab, Callinectes sapidus and the Green crab, Carcinus aestuarii were tested for macronutrients, heavy metals, bacteria content, and antimicrobial properties. Such information is crucial for further utilization of the biogenic powders for any composite formulation in added-value by-products. The calcium carbonate-rich hard tissue yield was 52.13% ± 0.015 (mean ± S.D.) and 64.71% ± 0.144 from the blue and green crabs, respectively. The contents of Pb, Ni, Zn, Cr (VI), and Cu were several orders of magnitude below the prescribed limit by EU biofertilizer legislation, with Fe, Mn (not prescribed), and As being the most abundant. The content of As and Cd from the material considered here was within limits. The shells contain no colony-forming units of Salmonella spp. and compliant levels of Escherichia coli; moreover, the shell micro-powder showed dose-dependent growth inhibition of Pseudomonas aeruginosa and Staphylococcus aureus. In summary, the waste crab shells present a complex natural product as plant biofertilizer following the circular economy concepts.

A global-scale screening of non-native aquatic organisms to identify potentially invasive species under current and future climate conditions.

The threat posed by invasive non-native species worldwide requires a global approach to identify which introduced species are likely to pose an elevated risk of impact to native species and ecosystems. To inform policy, stakeholders and management decisions on global threats to aquatic ecosystems, 195 assessors representing 120 risk assessment areas across all six inhabited continents screened 819 non-native species from 15 groups of aquatic organisms (freshwater, brackish, marine plants and animals) using the Aquatic Species Invasiveness Screening Kit. This multi-lingual decision-support tool for the risk screening of aquatic organisms provides assessors with risk scores for a species under current and future climate change conditions that, following a statistically based calibration, permits the accurate classification of species into high-, medium- and low-risk categories under current and predicted climate conditions. The 1730 screenings undertaken encompassed wide geographical areas (regions, political entities, parts thereof, water bodies, river basins, lake drainage basins, and marine regions), which permitted thresholds to be identified for almost all aquatic organismal groups screened as well as for tropical, temperate and continental climate classes, and for tropical and temperate marine ecoregions. In total, 33 species were identified as posing a 'very high risk' of being or becoming invasive, and the scores of several of these species under current climate increased under future climate conditions, primarily due to their wide thermal tolerances. The risk thresholds determined for taxonomic groups and climate zones provide a basis against which area-specific or climate-based calibrated thresholds may be interpreted. In turn, the risk rankings help decision-makers identify which species require an immediate 'rapid' management action (e.g. eradication, control) to avoid or mitigate adverse impacts, which require a full risk assessment, and which are to be restricted or banned with regard to importation and/or sale as ornamental or aquarium/fishery enhancement.

Promoting hidden natural design templates in wasted shells of the mantis shrimp into valuable biogenic composite.

The mineralized cuticle of the mantis shrimps Squilla Mantis which serve as natural hammers, spears and armors, have attracted research attention from various fields due to its amazing mechanical properties which were studied from evolutionary and ecological points of view. Here we aimed to valorize the astonishing mantis shrimp shell waste resulted from fishery and seafood industry as valuable biogenic composite derived from nature, potentially re-usable for novel, smart materials or added-value by-products, aspect which was not deeply considered before. Employing multi-laser Raman spectroscopy and imaging, supported by x-ray diffraction and high-resolution electron microscopy, we discover that the peripheral segments anatomically known as claws and telson, featured completely different composition and morphology, suggesting different applicability. The claw presents a bulk Mg-CaCO3 structure reinforced with fluorapatite coating, while the carotenoid-rich telson presents a porous and anisotropic structure of an amorphous mixture of CaCO3 and CaPO4 in gradient deposition on the chitin-protein scaffold. Resonance Raman spectroscopy showed concentrated pools of astaxanthin carotenoid within the bright red spots visible on telson, Based on our findings, we discuss this material's potential for selective applicability, as a natural source of phosphate-carbonate minerals, antioxidants, biofertilizer, pollutant adsorbent, valuable material for regenerative medicine or even as a cell culture substrate. Knowledge-based approach on this bio-template is the basis for smart recycling of such fishery waste for sustainable development, by opening channels for blue bioeconomy avenue.

Color-specific porosity in double pigmented natural 3d-nanoarchitectures of blue crab shell.

3D-engineered nano-architectures with various functionalities are still difficult to obtain and translate for real-world applications. However, such nanomaterials are naturally abundant and yet wasted, but could trigger huge interest for blue bioeconomy, provided that our understanding of their ultrastructure-function is achieved. To date, the Bouligand pattern in crustaceans shell structure is believed to be unique. Here we demonstrated that in blue crab Callinectes sapidus, the 3D-nanoarchitecture is color-specific, while the blue and red-orange pigments interplay in different nano-sized channels and pores. Thinnest pores of about 20 nm are found in blue shell. Additionally, the blue pigment co-existence in specific Bouligand structure is proved for the green crab Carcinus aestuarii, although the crab does not appear blue. The pigments interplay, simultaneously detected by Raman spectroscopy in color-specific native cuticles, overturns our understanding in crustaceans coloration and may trigger the selective use of particular colored natural nanoarchitectures for broaden area of applications.

Fishers and groupers (Epinephelus marginatus and E. morio) in the coast of Brazil: integrating information for conservation.

BACKGROUND: Groupers are a vulnerable but economically important group of fish, especially for small-scale fisheries. We investigated catches and local ecological knowledge (LEK) of diet, habitat, and past fishing experiences. METHODS: Landings, prices, interviews, and restaurants demand for two species, Epinephelus marginatus (dusky grouper) and Epinephelus morio (red grouper), were registered. RESULTS: We visited 74 markets and 79 sites on the coast of Brazil in 2017-2018, and we interviewed 71 fishers: Bahia (NE), Rio de Janeiro and São Paulo (SE), and Santa Catarina (S). The landings sampled of dusky grouper (2016-2017) in Rio de Janeiro were: n = 222, size 38-109 cm, weight 1-24 kg, average 3.84 kg; in São Paulo, São Sebastião were: n = 47, size 39-106 cm, weight 2-8 kg, average of 2.77 kg; and at Santos: n = 80, 26-120 cm, weight 0.36-15 kg, average 2.72 kg. Red grouper was observed in markets in the northeastern Brazil. We did not observe Epinephelus marginatus from Bahia northward; a maximum size of 200 cm was reported south of the Bahia, besides Rio de Janeiro and São Paulo coasts, 20 years ago (or longer) by 12 fishers. Local knowledge of fishers was important for grouper data of habitat and diet; the reproduction period was identified by fishers as September to March. CONCLUSIONS: Groupers can be considered as a cultural and ecological keystone species. We suggest protective measures: 1) fishing zoning, 2) islands (MPAs) with the surveillance of fishers, 3) late Spring and early Summer as key periods for management (grouper reproduction), 4) studies on grouper larvae, 5) mapping of fishing spots, 6) studies on local knowledge. Collaboration with small-scale fishers and local knowledge could contribute to low-conflict management measures. In that regard, integrative models of management from Latin America, by using local knowledge and citizen science, could produce successful grouper management for Brazilian data-poor fisheries, a contrasting reality to the Mediterranean areas. Finally, the distribution of E. marginatus in Brazil leave us with questions: a) Have dusky groupers disappeared from Bahia because of a decline in the population? b) Was it uncommon in Northeast Brazil? c) Did changes in water temperatures forced a movement southward?

Lipophilic marine biotoxins SERS sensing in solutions and in mussel tissue.

To detect and recognise three structurally related marine biotoxins responsible for the diarrheic shellfish poisoning (DSP) symptom, namely okadaic acid (OA), dinophysistoxin-1 (DTX-1) and dinophysistoxin-2 (DTX-2) respectively, as well as the structurally different yessotoxin (YTX), we developed a novel surface-enhanced micro-Raman scattering (micro-SERS) approach to investigate for the first time their micro-SERS signalling in solution and jointly analysed them in conjunction with the normal and toxic mussel tissue. YTX provided the main SERS feature surprisingly similar to DTX-1 and DTX-2, suggesting similar molecular adsorption mechanism with respect to the AgNPs. A fingerprint SERS band at 1017 cm-1 characteristic for the C-CH3 stretching in DTX-1 and DTX-2 and absent in OA SERS signal, allowed direct SERS discrimination of DTX-1,2 from OA. In acid form or as dissolved potassium salt, OA showed reproducible SERS feature for 0.81 µM to 84.6 nM concentrations respectively, while its ammonium salt slightly changed the overall SERS signature. The inherently strong fluorescence of the shellfish tissue, which hampers Raman spectroscopy analysis, further increases when toxins are present in tissue. Through SERS, tissue fluorescence is partially quenched. Artificially intoxicated mussel tissue with DSP toxins and incubated with AgNPs allowed direct SERS evidence of the toxin presence, opening a novel avenue for the in situ shellfish tracking and warning via micro-SERS. Natural toxic tissue containing 57.91 µg kg-1 YTX (LC-MS confirmed) was micro-SERS assessed to validate the new algorithm for toxins detection. We showed that a portable Raman system was able to reproduce the lab-based SERS results, being suitable for in situ raw seafood screening. The new approach provides an attractive, faster, effective and low-cost alternative for seafood screening, with economic, touristic and sustainable impact in aquaculture, fisheries, seafood industry and consumer trust.

Citrus fruits freshness assessment using Raman spectroscopy.

The freshness of citrus fruits commonly available in the market was non-destructively assessed by Raman spectroscopy. Intact clementine, mandarin and tangerine species were characterised concerning their carotenoids skin Raman signalling in a time course from the moment they were acquired as fresh stock, supplying the market, to the physical degradation, when they were no longer attractive to consumers. The freshness was found to strongly correlate to the peel Raman signal collected from the same area of the intact fruits in a time course of a maximum of 20days. We have shown that the intensity of the carotenoid Raman signal is indeed a good indicator of fruit freshness and introduced a Raman coefficient of freshness (CFresh), whose time course is linearly decreasing, with different slope for different citrus groups. Additionally, we demonstrated that the freshness assessment could be achieved using a portable Raman instrument. The results could have a strong impact for consumer satisfaction and the food industry.

Lampetra soljani, a new brook lamprey from the southern Adriatic Sea basin (Petromyzontiformes: Petromyzontidae).

Lampetra soljani, new species, is described from the lower Neretva River in the Adriatic Sea basin. Previously it was identified as L. zanandreai. Based on morphological similarity and mitochondrial genetic data, it is related to L. lanceolata and L. ninae from the Black Sea basin. Lampetra soljani is distinguished from all other species of Lampetra by having a marmorate flank pattern in live, fully grown ammocoetes, and in some adults (vs. plain colour pattern). Lampetra soljani is distinguished from other Lampetra species by having three velar tentacles, 54-57 trunk myomeres between the last branchial opening and the anus, no posterial teeth and a bicuspid middle endolateral tooth. Lampetra soljani is also well distinguished by COI barcode data from its congeners. The new species is widespread in the Neretva River drainage and lampreys from Lake Skadar basin are likely to belong to this species also.

Amnesic shellfish poisoning biotoxin detection in seawater using pure or amino-functionalized Ag nanoparticles and SERS.

Domoic acid (DA) biotoxin responsible for the amnesic shellfish poisoning (ASP) has been unambiguously detected in seawater in a broad range of concentration, with both pure and amino-functionalized Ag nanoparticles employed for surface enhanced Raman scattering (SERS). To achieve this, a comprehensive SERS study on DA dissolved in distilled water has been conducted. SERS of DA dissolved in seawater in concentrations ranging from 3.3 × 10(-4) to 3.3 × 10(-8) mol l(-1) exhibited specific signal, completely different to those of the corresponding DA aqueous solutions, due to the seawater interference in the overall SERS effect. In order to assess the capability of the technique as a cheaper alternative for rapid and unambiguous detection of the DA biotoxin in seawater, three detection schemes have been proposed. DA was detectable at 0.33 nmoll(-1) concentration (0.33) dissolved in distilled water and 0.033 nmol l(-1) (0.033 ppb) in seawater respectively, much lower than the admitted level by the current regulation. A solvent specific interaction of DA with the NPs was concluded, since DA aqueous solution added to Ag nanoparticles provided different SERS signal compared to that of DA directly dissolved in seawater. Employing amino-functionalized Ag nanoparticles with 4-aminothiophenol as SERS tag, SERS signal of DA on amino-AgNPs revealed significant specificity associated with the aromatic primary amine interaction of the SERS tag with DA, thus allowing DA detection in seawater at 4.16 × 10(-4) mol l(-1) concentration, much higher than in the case of pure NPs. To highlight the findings, a brief literature review to date on the DA biotoxin detection was also provided.

Occurrence of Philometra lateolabracis (Nematoda: Philometridae) in the gonads of marine perciform fishes in the Mediterranean region.

Gravid females of the nematode Philometra lateolabracis (Yamaguti, 1935), a parasite of gonads of marine perciform fishes, were found in wild and cultured dusky grouper Epinephelus marginatus (Lowe) from waters near the Balear Islands (Spain, Mediterranean Sea) and Sicily (Italy, Thyrrenean Sea), and in the greater amberjack Seriola dumerili (Risso) in Croatia (south-eastern Adriatic Sea). In wild E. marginatus in Spain, the overall prevalence was 21% and the intensity of infection 1 nematode per fish. The nematodes are briefly described and illustrated. The species Sanguinofilaria jordanoi López-Neyra, 1951, described from the ovary of Epinephelus gigas Brünich from Morocco, is synonymized with P. lateolabracis. This is the first documented record of P. lateolabracis from fishes of the Mediterranean region and its finding in S. dumerili represents a new host record. The possible importance of this pathogenic parasite for cultures of marine perciform fishes in the region is stressed.