The Euganean Thermal District (Italy) represents the oldest and largest thermal center in Europe, and its therapeutic mud is considered a unique product whose beneficial effects have been documented since Ancient Roman times. Mud properties depend on the heat and electrolytes of the thermal water, as well as on the bioactive molecules produced by its biotic component, mainly represented by cyanobacteria. The investigation of the healing effects of compounds produced by the Euganean cyanobacteria represents an important goal for scientific validation of Euganean mud therapies and for the discovering of new health beneficial biomolecules. In this work, we evaluated the therapeutic potential of exopolysaccharides (EPS) produced by Phormidium sp. ETS05, the most abundant cyanobacterium of the Euganean mud. Specifically, Phormidium EPS resulted in exerting anti-inflammatory and pro-resolution activities in chemical and injury-induced zebrafish inflammation models as demonstrated using specific transgenic zebrafish lines and morphometric and expression analyses. Moreover, in vivo and in vitro tests showed no toxicity at all for the EPS concentrations tested. The results suggest that these EPS, with their combined anti-inflammatory and pro-resolution activities, could be one of the most important therapeutic molecules present in the Euganean mud and confirm the potential of these treatments for chronic inflammatory disease recovery.

The Euganean Thermal District has been known since Roman times for the therapeutic properties of peloids, obtained from natural clays that have undergone a traditional maturation process. This leads to the growth of a green microbial biofilm with Cyanobacteria and the target species Phormidium sp. ETS-05 as fundamental components for their ability to synthetize anti-inflammatory molecules. Currently, in-depth studies on the microbiota colonizing Euganean peloids, as in general on peloids utilized worldwide, are missing. This is the first characterization of the microbial community of Euganean thermal muds, also investigating the effects of environmental factors on its composition. We analysed 53 muds from 29 sites (Spas) using a polyphasic approach, finding a stable microbiota peculiar to the area. Differences among mud samples mainly depended on two parameters: water temperature and shading of mud maturation plants. In the range 37–47 °C and in the case of irradiance attenuation due to the presence of protective roofs, a statistically significant higher mud Chl a content was detected. Moreover, in these conditions, a characteristic microbial and Cyanobacteria population composition dominated by Phormidium sp. ETS-05 was observed. We also obtained the complete genome sequence of this target species using a mixed sequencing approach based on Illumina and Nanopore sequencing.

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The nutraceutical supplementation of Spirulina (Arthrospira platensis) in dogs and cats has not yet been investigated. The aim of this study was to evaluate if the dietary supplementation of increasing amounts of Spirulina for 6 weeks is palatable to pets and to assess the owner’s perception of such supplementation. The owners of the 60 dogs and 30 cats that participated in this study were instructed to daily provide Spirulina tablets starting with a daily amount of 0.4 g, 0.8 g, and 1.2 g for cats as well as small dogs, medium dogs, and large dogs, respectively, and allowing a dose escalation of 2× and 3× every 2 weeks. The daily amount (g/kg BW) of Spirulina ranged from 0.08 to 0.25 for cats, from 0.06 to 0.19 for small-sized dogs, from 0.05 to 0.15 for medium-sized dogs, and from 0.04 to 0.12 for large-sized dogs. Each owner completed a questionnaire at the time of recruitment and the end of each 2-week period. No significant effect on the fecal score, defecation frequency, vomiting, scratching, lacrimation, general health status, and behavioral attitudes was detected by the owners’ reported evaluations. Most animals accepted Spirulina tablets either administrated alone or mixed with food in the bowl. Daily supplementation of Spirulina for 6 weeks in the amounts provided in this study is therefore palatable and well tolerated by dogs and cats.

Phormidium sp. ETS-05 is one of the target cyanobacteria species conferring anti-inflammatory properties to the therapeutic muds applied by spas of the Euganean Thermal District (Italy) to treat arthro-rheumatic pathologies. Beneficial mud is prepared by spas following a traditional method, called maturation, leading to the growth of a specific microbiota on natural raw clay irrigated by flowing thermal water at 37–47 °C for about two months. The effectiveness of the mud is related to heat, electrolytes and bioactive molecules synthesized by the microbiota. A clear role in the anti-inflammatory activity of muds has been demonstrated for the exopolysaccharides, EPS, produced by the entire microbiota and Phormidium sp. ETS-05. Considering the interest in this species, we assessed its optimal growth conditions to obtain the higher EPS production in relation to temperature, light spectra, and intensity and nitrogen availability. The production of biomass and pigments was also taken into account, as other high-value compounds can be obtained in parallel with EPS. We found that exposure to a temperature of 45 °C under white light at 100 µmol photons m−2 s−1 is optimal to reach the highest biomass (1.13 g L−1) and an average production of 75 mg gDW−1 phycocyanin and of 150 mg gDW−1 EPS for Phormidium sp. ETS-05 cultured in lab-scale photobioreactors for 9 days. Putative genes linked with EPS assembly and export have also been identified in its genome, some of which have been investigated for their expression levels, opening up the possibility of biotechnologically boost EPS production.