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This study presents an effective technology for the olive processing industry to remediate olive washing water. A 14.5-L enclosed tubular photobioreactor was inoculated with a stable microalgal-bacterial consortium obtained by screening strains well adapted to olive washing water. The capacity of an enclosed tubular photobioreactor to remove toxic compounds was evaluated under photosynthesis conditions and without any external supply of oxygen. The results showed that the dominant green microalgae Scenedesmus obliquus, Chlorella vulgaris and the cyanobacteria Anabaena sp. and bacteria present in olive washing water (i.e. Pantoea agglomerans and Raoultella terrigena) formed a synergistic association that was resistant to toxic pollutants present in the effluent and during the initial biodegradation process, which resulted in the breakdown of the pollutant. Total phenolic compounds, COD, BOD, turbidity and colour removals of 90.3 ± 11.4, 80.7 ± 9.7, 97.8 ± 12.7, 82.9 ± 8.4 and 83.3 ± 10.4 %, respectively, were recorded in the photobioreactor at 3 days of hydraulic retention time. [ABSTRACT FROM AUTHOR]

Many studies on cold environments have been developed over the past two decades. High mountain freshwater presents high variability of nutrients and chemico-physical parameters, showing variations of pH, oxygen concentration, metals and temperature throughout the year. National Park of Sierra Nevada (Granada, Spain) (37°032 N 03°182 W), has almost 40 lakes that are reported to be both endemic and oligotrophic. However, very little information about their microbial diversity can be found in literature. In this work, a Response Surface Methodology (RSM) was used to find best nutritional conditions for the isolation of psychrophilic microalgae from La Caldera Lake. The results showed that best culture medium, was the Rodriguez-Lopez medium (RL); data were adjusted to a quadratic prediction model reporting a biomass concentration over 600 mg/L at 10 and 20°C. In this paper, the diversity of culturable freshwater microalgae in the La Caldera Lake was observed by PCR using specific primers for eukaryotic 18S rRNA genes. Samples were taken in early July and late Agust, 2011. In July presence of strains belonging to the Eustigmatophyceae, Bacillariophyceae, Trebouxiophyceae, Chlorophyceae and Scenedesmaceae families were found. In August, only microalgae from the Eustigmatophyceae, Trebouxiophyceae, Chlorophyceae and Scenedesmaceae families were found. An individual culture of each isolated strain was carried out. Microalgae S21 had phylogenetic similitude with Chlorophyceae, and showed best growth being biomass concentration in RL 393.73 mg/L and 128.52 mg/L at 20°C and 10°C, respectively. Moreover, specific growth rates (μmax), 0.25/h and 0.13/h at 20°C and 10°C, respectively, were detected for strain S21. [ABSTRACT FROM AUTHOR]

"La Caldera" is a high mountain lake located in Sierra Nevada, Spain. For its position and characteristics it is considered as an important "sensor" for global changes. Water samples were collected during the ice-free season in order to observe changes in the bacterial community structure. Experiments were carried out complementing culture and culture-independent methods. Strong variations of the bacterial communities were recorded along the sampling period in particular regarding number and phylogenetic relationships of the microorganisms detected. At the season beginning, bacterial counts were rather high but strains belonged only to β- and γ-Proteobacteria. By contrast, in the middle of the season, possibly due to external inputs, total number of bacteria decreased but other phylogenetic groups, such as α-proteobacteria, Actinobacteria and Firmicute appeared. On the whole, Proteobacteria were the dominant microbiota. Bacterial diversity and presence of most abundant species were analysed by PCR-TGGE. Sequencing of TGGE bands confirmed that prevalent Bacteria populations were evolutively close to Proteobacteria with predominance of α- and β-Proteobacteria. Analysis of diversity and functional organization suggested that "La Caldera" bacterial microbiota, typical of oligotrophic and/or extreme environments with low habitability, was characterized by a community organized in an adequate distribution of dominant and resilient species. Although rather specialized, the community presented a sufficient functionality and flexibility to react to changing and stressing conditions. However, along the ice-free season, the community evolved showing more specialized populations with less adaptation abilities. This is the first work complementing cultural and cultural-independent methods for the "La Caldera" lake. [ABSTRACT FROM AUTHOR]

Introduction: The marine environment is extremely complex and exerts strong evolutionary pressure often leading to the appearance of microbial strains with new metabolic competencies. Microorganisms in marine ecosystems are still largely unknown and should be explored and conserved for biodiversity preservation, possible ecosystem restoring, and other applications. Biodiversity conservation should become a basic ecological strategy of particular significance in relation to global change. In this context, the present research aimed at exploring the culturable mycobiota associated with the jellyfish Pelagia noctiluca, never studied before. In addition, the isolated strains were tested for potential application (antimicrobial activity and presence of genes related to the production of secondary metabolites)., Methods: Five jellyfishes were collected in the coastal area of Giglio Island and processed to isolate epizoic fungi. The strains were identified using a polyphasic approach (morphological, physiological, and molecular) and their salt preference was also investigated. The antifungal and antibacterial activity were tested for each strain with agar plug diffusion test. The presence of some key genes related to the main pathways for the production of secondary metabolites in fungi, polyketide synthases (PKSs), and non-ribosomal peptide synthase (NRPSs), was also assessed., Results: A total of 164 isolates were obtained; after the dereplication, 40 morphotypes, and 23 species were identified. The phylogenetic analyses suggested the presence of new taxa belonging to Pleosporales: two new genera and species, and a new species of Tamaricicola . The detected mycobiota showed a relatively high diversity, if compared to other epizoic fungal communities. All isolated strains were marine fungi as confirmed by their salt preference and marked euryhalinism. The genes related to the two main pathways for the production of secondary metabolites in fungi, PKSs and NRPSs, were identified in four and nine strains, respectively. The antimicrobial activity was revealed in 70% of the strains, including the new taxa. The abundance of bioactive strains may be related to the potential involvement of epizoic fungi in host defense strategies. Moreover, these strains could show a high potential for further biotechnological applications particularly in the case of new taxa. All strains are maintained in culture collections., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Pasqualetti, Braconcini, Barghini, Gorrasi, Schillaci, Ferraro, Della Sala, De Marino and Fenice.)

Nowadays, the discharge of wastewater is a global concern due to the damage caused to human and environmental health. Wastewater treatment has progressed to provide environmentally and economically sustainable technologies. The biological treatment of wastewater is one of the fundamental bases of this field, and the employment of new technologies based on granular biofilm systems is demonstrating success in tackling the environmental issues derived from the discharge of wastewater. The granular-conforming microorganisms must be evaluated as functional entities because their activities and functions for removing pollutants are interconnected with the surrounding microbiota. The deep knowledge of microbial communities allows for the improvement in system operation, as the proliferation of microorganisms in charge of metabolic roles could be modified by adjustments to operational conditions. This is why engineering must consider the intrinsic microbiological aspects of biological wastewater treatment systems to obtain the most effective performance. This review provides an extensive view of the microbial ecology of biological wastewater treatment technologies based on granular biofilms for mitigating water pollution.

In this study, 15 Lulworthiales strains isolated from the marine tunicate Halocynthia papillosa collected in the central Tyrrhenian Sea were characterized using a polyphasic approach (morpho-physiological, molecular, and phylogenetic analyses). Based on multi-locus phylogenetic inference and morphological characters, a new genus, Rambellisea , and two new species, R. halocynthiae and R. gigliensis (Lulworthiales), were proposed. Multi-locus phylogenetic analyses using the nuclear ribosomal regions of DNA (nrITS1-nr5.8S-nrITS2, nrLSU, and nrSSU) sequence data strongly supported the new taxa. Phylogenetic inference, estimated using Maximum Likelihood and Bayesian Inference, clearly indicates that Rambellisea gen. nov. forms a distinct clade within the order Lulworthiales. Moreover, the two new species were separated into distinct subclades, solidly supported by the analyses. This is the first report of Lulworthiales species isolated from animals.

Background: The Kuril-Kamchatka Trench (maximum depth 9604 m), located in the NW Pacific Ocean, is among the top seven deepest hadal trenches. The work aimed to investigate the unexplored abyssal-hadal prokaryotic communities of this fascinating, but underrated environment., Results: As for the bacterial communities, we found that Proteobacteria (56.1-74.5%), Bacteroidetes (6.5-19.1%), and Actinobacteria (0.9-16.1%) were the most represented bacterial phyla over all samples. Thaumarchaeota (52.9-91.1%) was the most abundant phylum in the archaeal communities. The archaeal diversity was highly represented by the ammonia-oxidizing Nitrosopumilus, and the potential hydrocarbon-degrading bacteria Acinetobacter, Zhongshania, and Colwellia were the main bacterial genera. The α-diversity analysis evidenced that both prokaryotic communities were characterized by low evenness, as indicated by the high Gini index values (> 0.9). The β-diversity analysis (Redundancy Analysis) indicated that, as expected, the depth significantly affected the structure of the prokaryotic communities. The co-occurrence network revealed seven prokaryotic groups that covaried across the abyssal-hadal zone of the Kuril-Kamchatka Trench. Among them, the main group included the most abundant archaeal and bacterial OTUs (Nitrosopumilus OTU A2 and OTU A1; Acinetobacter OTU B1), which were ubiquitous across the trench., Conclusions: This manuscript represents the first attempt to characterize the prokaryotic communities of the KKT abyssal-hadal zone. Our results reveal that the most abundant prokaryotes harbored by the abyssal-hadal zone of Kuril-Kamchatka Trench were chemolithotrophic archaea and heterotrophic bacteria, which did not show a distinctive pattern distribution according to depth. In particular, Acinetobacter, Zhongshania, and Colwellia (potential hydrocarbon degraders) were the main bacterial genera, and Nitrosopumilus (ammonia oxidizer) was the dominant representative of the archaeal diversity., (© 2023. The Author(s).)

Polyphenols and their intermediate metabolites are natural compounds that are spread worldwide. Polyphenols are antioxidant agents beneficial for human health, but exposure to some of these compounds can be harmful to humans and the environment. A number of industries produce and discharge polyphenols in water effluents. These emissions pose serious environmental issues, causing the pollution of surface or groundwater (which are used to provide drinking water) or harming wildlife in the receiving ecosystems. The treatment of high-polyphenol-content waters is mandatory for many industries. Nowadays, biotechnological approaches are gaining relevance for their low footprint, high efficiency, low cost, and versatility in pollutant removal. Biotreatments exploit the diversity of microbial metabolisms in relation to the different characteristics of the polluted water, modifying the design and the operational conditions of the technologies. Microbial metabolic features have been used for full or partial polyphenol degradation since several decades ago. Nowadays, the comprehensive use of biotreatments combined with physical-chemical treatments has enhanced the removal rates to provide safe and high-quality effluents. In this review, the evolution of the biotechnological processes for treating high-polyphenol-content water is described. A particular emphasis is given to providing a general concept, indicating which bioprocess might be adopted considering the water composition and the economic/environmental requirements. The use of effective technologies for environmental phenol removal could help in reducing/avoiding the detrimental effects of these chemicals. In addition, some of them could be employed for the recovery of beneficial ones.

Tomatoes are among the most consumed vegetables worldwide and represent a source of health-beneficial substances. Our study represents the first investigating the peel-associated epiphytic bacteria of red and purple (anthocyanin-rich) tomatoes subjected to organic and conventional farming systems. Proteobacteria was the dominant phylum (relative abundances 79-91%) in all experimental conditions. Enterobacteriaceae represented a large fraction (39.3-47.5%) of the communities, with Buttiauxella and Atlantibacter as the most represented genera. The core microbiota was composed of 59 operational taxonomic units (OTUs), including the majority of the most abundant ones. The occurrence of the most abundant OTUs differed among the experimental conditions. OTU 1 ( Buttiauxella ), OTU 2 ( Enterobacteriales ), and OTU 6 ( Bacillales ) were higher in red and purple tomatoes grown under organic farming. OTU 5 ( Acinetobacter ) had the highest abundance in red tomatoes subjected to organic farming. OTU 3 ( Atlantibacter ) was among the major OTUs in red tomatoes under both farming conditions. OTU 7 ( Clavibacter ) and OTU 8 ( Enterobacteriaceae ) had abundances ≥1% only in red tomatoes grown under conventional farming. PCA and clustering analysis highlighted a high similarity between the bacterial communities of red and purple tomatoes grown under organic farming. Furthermore, the bacterial communities of purple tomatoes grown under organic farming showed the lowest diversity and evenness. This work paves the way to understand the role of nutritional superior tomato genotypes, combined with organic farming, to modulate the presence of beneficial/harmful bacteria and supply healthier foods within a sustainable agriculture.

Pontimonas is currently described as a genus including only one species of slightly halophilic marine bacteria. Although some works revealed its presence in some hypersaline environments, the information on its habitat preference is still scant. This work investigated Pontimonas presence in selected ponds of the Saline di Tarquinia marine saltern and in the seawater intake area. The two-year metabarcoding survey documented its constant presence along the ponds establishing the salinity gradient and in a distinct basin with permanent hypersaline conditions (BSB). Pontimonas was higher in the ponds than in the sea, whereas it had similar abundances in the sea and in the BSB. Its representative OTUs showed significant trends according to different parameters. Along the salinity gradient, OTU1 abundance increased with decreasing water temperatures and increasing rainfalls, and it showed a maximum in January; OTU2 increased with increasing BOD 5 and it showed the highest abundances in the period August-October, and OTU 3194 increased at decreasing salinities. In BSB, a significant seasonal variation was shown by OTU 3194, which started increasing in spring to reach a maximum in summer. The results suggest that Pontimonas could easily settle in hypersaline habitats, having also broad euryhaline members and some possible extreme halophilic representatives.

The investigation for novel unique extremozymes is a valuable business for which the marine environment has been overlooked. The marine fungus Clonostachys rosea IG119 was tested for growth and chitinolytic enzyme production at different combinations of salinity and pH using response surface methodology. RSM modelling predicted best growth in-between pH 3.0 and 9.0 and at salinity of 0-40‱, and maximum enzyme activity (411.137 IU/L) at pH 6.4 and salinity 0‱; however, quite high production (>390 IU/L) was still predicted at pH 4.5-8.5. The highest growth and activity were obtained, respectively, at pH 4.0 and 8.0, in absence of salt. The crude enzyme was tested at different salinities (0-120‱) and pHs (2.0-13.0). The best activity was achieved at pH 4.0, but it was still high (in-between 3.0 and 12.0) at pH 2.0 and 13.0. Salinity did not affect the activity in all tested conditions. Overall, C. rosea IG119 was able to grow and produce chitinolytic enzymes under polyextremophilic conditions, and its crude enzyme solution showed more evident polyextremophilic features. The promising chitinolytic activity of IG119 and the peculiar characteristics of its chitinolytic enzymes could be suitable for several biotechnological applications (i.e., degradation of salty chitin-rich materials and biocontrol of spoiling organisms, possibly solving some relevant environmental issues).

The "Saline di Tarquinia" salterns have been scarcely investigated regarding their microbiological aspects. This work studied the structure and composition of their bacterial communities along the salinity gradient (from the nearby sea through different ponds). The communities showed increasing simplification of pond bacterial diversity along the gradient (particularly if compared to those of the sea). Among the 38 assigned phyla, the most represented were Proteobacteria , Actinobacteria and Bacteroidetes . Differently to other marine salterns, where at the highest salinities Bacteroidetes dominated, preponderance of Proteobacteria was observed. At the genus level the most abundant taxa were Pontimonas , Marivita , Spiribacter , Bordetella , GpVII and Lentibacter . The α-diversity analysis showed that the communities were highly uneven, and the Canonical Correspondence Analysis indicated that they were structured by various factors (sampling site, sampling year, salinity, and sampling month). Moreover, the taxa abundance variation in relation to these significant parameters were investigated by Generalized Linear Models. This work represents the first investigation of a marine saltern, carried out by a metabarcoding approach, which permitted a broad vision of the bacterial diversity, covering both a wide temporal span (two years with monthly sampling) and the entire salinity gradient (from the nearby sea up to the crystallisation ponds).

Background: Marine fungi are an important repository of bioactive molecules with great potential in different technological fields, the annual number of new compounds isolated from marine fungi is impressive and the general trend indicates that it is still on the rise. In this context, the antifungal and antimicrobial activity of the marine strain Mariannaea humicola IG100 was evaluated and two active terpenoids were isolated and characterized., Methods: Preliminary screening of activity of marine strain IG100 was carried out by agar plug diffusion methods against fungal (Penicillium griseofulvum TSF04) and bacterial (Bacillus pumilus KB66 and Escherichia coli JM109) strains. Subsequently, inhibition tests were done by using the cultural broth and the organic extract (ethyl acetate, EtOAc) by the agar well diffusion methods. The main active fractions were identified and tested for their antifungal activity against P. griseofulvum TSF04 in a 24 wells microplate at different concentrations (1000, 100, 10 and 1.0 µg/mL). Two active compounds were characterized and their relative MIC measured by the broth micro-dilution methods in a 96-well microplate against Aspergillus flavus IG133, P. griseofulvum TSF04, and Trichoderma pleuroticola IG137., Results: Marine strain IG100 presented significant antifungal activity associated with two active compounds, the terpenoids terperstacin 1 and 19-acetyl-4-hydroxydictyodiol 2. Their MIC values were measured for A. flavus (MIC of 7.9 µg/mL and 31.3 µg/mL for 1 and 2, respectively), P. griseofulvum (MIC of 25 µg/mL and 100 µg/mL for 1 and 2, respectively) and T. pleuroticola (MIC > 500 µg/mL and 125 µg/mL for 1 and 2, respectively). They showed a rather good fungistatic effect., Conclusions: In this study, the first marine strain of M. humicola (IG100) was investigated for the production of bioactive molecules. Strain IG100 produced significant amounts of two bioactive terpenoids, terperstacin 1 and 19-acetyl-4-hydroxydictyodiol 2. The two compounds showed significant antifungal activities against A. flavus IG133, T. pleuroticola IG137 and P. griseofulvum TSF04. Compound 2 was identified for the first time in fungi.

Vibrio species are ubiquitous in a number of different aquatic environments and promptly adapting to environmental changes due to high genome plasticity. The presence of these bacteria in marine salterns, in relation to a salinity gradient has been not investigated yet. Moreover, it is not clear if these hypersaline environments could represent a reservoir for Vibrio spp. This work investigated, through a metagenetic approach, the distribution of Vibrio (over 2 years) in different ponds along the salinity gradient within the 'Saline di Tarquinia' salterns, considering also the adjacent coastal waters and an isolated brine storage basin (BSB). Vibrio occurrence was higher in the sea than in the ponds and BSB, where it usually represented a rare taxon (abundance <1%). In the sea, it showed abundances in-between 1%-2.6% in 8 months out of 24. Four OTUs were assigned to the Vibrio genus; except for one that was more abundant in BSB, the others were much higher in the sea. Redundancy analysis (RDA) suggested a different distribution of the OTUs in relation to water temperature and salinity. Vibrio was found, even with low abundances, at the highest salinities also, suggesting the salterns as a possible reservoir for the bacterium., (© 2020 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Twenty-eight fungal strains have been isolated from different natural marine substrates and plate screened for their production of chitinolytic activity. The two apparent best producers, Trichoderma lixii IG127 and Clonostachys rosea IG119, were screened in shaken cultures in media containing 1% colloidal chitin, 1% yeast nitrogen base and 38‰ NaCl, for their ability to produce chitinolytic enzymes under halophilic conditions. In addition, they were tested for optimal growth conditions with respect to pH, salinity and temperature. The Trichoderma strain appeared to be a slight halotolerant fungus, while C. rosea IG119 clearly showed to be a halophilic marine fungus, its optimal growth conditions being very coherent for life in the marine environment (i.e., pH 8.0, salinity 38‰). Due to its high and relatively fast activity (258 U/L after 192 h of growth) accompanied by its halophilic behaviour (growth from 0 to 160‰ of salinity), C. rosea was selected for further studies. In view of possible industrial applications, its medium for chitinolytic enzyme production was optimized by Response Surface Methodology using 1% colloidal chitin and different concentrations of corn step liquor and yeast nitrogen base (0-0.5%). Time course of growth under optimized condition showed that maximum activity (394 U/L) was recorded after 120 h on medium containing Corn Steep Liquor 0.47% and Yeast Nitrogen Base 0.37%. Maximum of productivity (3.3 U/Lh) was recorded at the same incubation time. This was the first study that demonstrated high chitinolytic activity in a marine strain of C. rosea.

Background: Shewanella baltica KB30 was isolated from seawater collected in Kandalaksha Bay, White Sea (Russia). This strain is known for its ability to grow on a pool of different substrates, including carbohydrates, carboxylic and amino acids, and lipids. However, no data are available on its metabolic efficiency in relation to the use of different carbon sources typologies. This work represents the first attempt to characterize S. baltica by its heterotrophic kinetic performance., Results: Growth and substrate consumption, during the biodegradation of sodium acetate, glucose, tween 80 and peptone, were analyzed through a respirometric method. To find the model best fitting the experimental data and to obtain the kinetic parameters, the equations of Monod, Moser, Contois and Tessier were applied. The kinetic behavior of S. baltica was fitted to Monod model for sodium acetate and tween 80, while it was adjusted to Contois model for glucose and peptone. In this regard, peptone was consumed faster than the other substrates, as indicated by the highest values of substrate degradation rate, which exceeded 60 mg O 2  L -1  h -1 ., Conclusions: Proteolytic metabolism was favored than lipidic and glucidic metabolism, which could contribute much more to mineralization and recycling of proteins than lipids and carbohydrates.

MALDI-TOF Mass Spectrometry in association with the MALDI BioTyper 3.1 software has been evaluated for the identification and classification of 45 Arctic bacteria isolated from Kandalaksha Bay (White Sea, Russia). The high reliability of this method has been already demonstrated, in clinical microbiology, by a number of studies showing high attribution concordance with other credited analyses. Recently, it has been employed also in other branches of microbiology with controversial performance. The phyloproteomic results reported in this study were validated with those obtained by the "gold standard" 16S rDNA analysis. Concordance between the two methods was 100% at the genus level, while at the species level it was 48%. These percentages appeared to be quite high compared with other studies regarding environmental bacteria. However, the performance of MALDI BioTyper changed in relation to the taxonomical group analyzed, reflecting known identification problems related to certain genera. In our case, attribution concordance for Pseudomonas species was rather low (29%), confirming the problematic taxonomy of this genus, whereas that of strains from other genera was quite high (> 60%). Among the isolates tested in this study, two strains (Exiguobacterium oxidotolerans and Pseudomonas costantinii) were misidentified by MALDI BioTyper due to absence of reference spectra in the database. Accordingly, missing spectra were acquired for the database implementation.

Lecanicillium muscarium CCFEE 5003, isolated in Continental Antarctica, is a powerful producer of extracellular cold-tolerant enzymes. Chitin-hydrolyzing enzymes seems to be the principal extracellular catalytic activities of this psychrotolerant fungus. The production of chitinolytic activities is induced by chitin and other polysaccharides and is submitted to catabolite repression. The chitinolytic system of L. muscarium consists of a number of different proteins having various molecular weights and diverse biochemical characteristics, but their most significant trait is the marked cold-tolerance. L. muscarium and selected strains of the biocontrol agent of pathogenic fungi Trichoderma harzianum, have been compared for their ability to produce chitinolytic enzymes at different temperatures. At low temperatures the Antarctic strain was definitely much more efficient. Moreover, the fungus was able to exert a strong mycoparasitic action against various other fungi and oomycetes at low temperatures. The parasitic role of this organism appeared related to the production of cell wall degrading enzymes being the release of extracellular chitinolytic enzymes a key event in the mycoparasitic process. Due to the mentioned characteristics, L. muscarium could have an important role for potential applications such as the degradation of chitin-rich materials at low temperature and the biocontrol of pathogenic organisms in cold environments. For these reasons and in view of future industrial application, the production of chitinolytic enzymes by the Antarctic fungus has been up-scaled and optimised in bench-top bioreactor.