Juana María Navarro Llorens

Extremophilic environments are rich reservoirs for discovering microorganisms with vast biotechnological potential. Among these, microalgae stand out for their pivotal role in sustainable wastewater treatment and nutrient recycling. This study introduces Coelastrella thermophile D14, a microalga isolated from a solar panel, identified through morphological studies and genomic sequencing. The genus Coelastrella has been characterized and classified as highly productive strains valuable for biofuel and bioproduct generation as well as for their ability to produce significant amounts of carotenoids. Experiments revealed the extraordinary resilience of this strain to prolonged desiccation and high-strength piggery wastewater. Notably, D14 cultivated in 10% pig effluent exhibited biostimulant properties, achieving a germination index 23% higher than the control on Lepidium sativum . In a groundbreaking development, we have successfully established an Agrobacterium -mediated transformation protocol for C. thermophila D14, optimizing key parameters for effective T-DNA transfer. This marks a pioneering achievement within the genus Coelastrella . These findings highlight the significant potential of D14 as a robust platform for future biotechnological applications, opening new opportunities for innovative solutions, especially in environmental protection and sustainable agriculture. Graphical Key points: • First microalga from solar panel biofilm: Coelastrella sp. D14 isolated and characterized. • Strain D14 tolerates prolonged desiccation and grows well in piggery wastewater. • Stable Agrobacterium-mediated transformation enables future metabolic engineering.

Honey processing is closely linked to water pollution due to the lack of a specific wastewater treatment. This study proposes a sustainable and innovative solution based on two sequential bioprocesses using a real effluent from an Argentine honey-exporting facility. In the initial stage, the honey wastewater was enriched with a non-Saccharomyces yeast (Candida ethanolica), isolated from the same effluent. Treatment with this yeast in a bioreactor nearly doubled the total sugar removal efficiency compared to the control (native flora). Subsequent clarification with diatomaceous earth reduced the optical density (91.6%) and COD (30.9%). In the second stage, secondary sewage effluent was added to the clarified effluent and inoculated with Chlorella vulgaris under different culture conditions. The best microalgae performance was observed under high light intensity and high inoculum concentration, achieving a fivefold increase in cell density, a specific growth rate of 0.752 d⁻¹, and a doubling time of 0.921 d. Although total sugar removal in this stage remained below 28%, cumulative COD removal reached 90% after nine days under both lighting conditions. This study presents the first integrated treatment approach for honey industry effluents using a native yeast–microalgae system, incorporating in situ effluent recycling and the potential for dual waste valorization.

Honey processing is closely linked to water pollution due to the lack of a specific wastewater treatment. This work presents a sustainable and innovative solution based on two sequential bioprocesses using a real effluent from an Argentine honey-exporting facility. In the first stage, the honey wastewater was enriched with a non-Saccharomyces yeast (Candida ethanolica), isolated and identified from the same effluent. Yeast treatment was carried out in a bioreactor, achieving nearly double the efficiency in total sugar removal compared to the control (native flora). Subsequent clarification with diatomaceous earth reduced optical density (91.6%) and COD (30.9%). In the second stage, secondary sewage effluent, was added to the clarified effluent and inoculated with Chlorella vulgaris under different culture conditions. The best microalgae performance was observed under high light intensity and high inoculum concentration, reaching a fivefold increase in cell density, a specific growth rate of 0.752 d⁻¹, and a doubling time of 0.921 d. Total sugar removal in this stage was under 28%, while cumulative COD removal reached 90% in nine days under both lighting conditions. The synergistic use of complementary microorganisms and different effluents enhances microalgal growth and supports to the overall sustainability of an integrated approach to managing all wastewater streams from honey production.

Microalga–bacteria consortia are increasingly recognized for their effectiveness in wastewater treatment, leveraging the metabolic synergy between microalgae and bacteria to enhance nutrient removal and overall treatment efficiency. These systems offer a sustainable approach to addressing pollutants such as nitrogen and phosphorus. However, their potential in removing specific contaminants like steroid hormones is less explored. In this study, a natural microbial consortium, A21, has been characterized and isolated from primary sewage treatment in Madrid and its potential for bioremediation of steroid hormone effluents has been evaluated. The A21 consortium includes Alphaproteobacteria genera Sphingopyxis and Pseudorhizobium and the Cyanobacterium Cyanobium. Sphingopyxis (31.78%) is known for biodegradation, while Pseudorhizobium (15.68%) exhibits detoxification abilities. Cyanobium (14.2%) may contribute to nutrient uptake and oxygen production. The effects of pH, nitrogen sources, and Sodium chloride concentrations on growth were evaluated. The optimal growth conditions were determined to be a pH range of 7 to 9, a salt concentration below 0.1 M, and the presence of a nitrogen source. The consortium also demonstrated effective growth across various types of wastewaters (primary, secondary, and tertiary treatment effluents). Additionally, A21 exhibited the ability to grow in the presence of steroids and transform them into other compounds, such as converting androstenedione (AD) into androsta-1,4-diene-3,17-dione (ADD) and β-estradiol into estrone.

Extremophile environments are an important source for finding microorganisms with a potential for biotechnological applications. Among these microorganisms, microalgae contribute to several sustainable processes such as wastewater treatments or nutrition. In this work it was characterized a microalga isolated from a solar panel. The morphological and phylogenetic analysis revealed that the isolate collected was a Coelastrella strain. Cultivation and stress experiments has shown that Coelastrella sp. D14 can resist a long period of desiccation and it can grow on cheap sources such as piggery wastewaters (PWW). This work reports that a Coelastrella strain displays biostimulant properties with a germination index of 123% on Lepidium sativum when D14 biomass grown at 10% piggery effluent was used. Altogether, these results suggest that this novel strain could be a good chassis for further biotechnological applications. Highlights: Coelastrella sp. D14, a xero-tolerant strain, has been isolated from a solar panel This strain can grow on piggery wastewater Coelastrella sp. D14 can promote germination of Lepidium sativum

Introduction: Microorganisms colonize a wide range of natural and artificial environments. Even though most of them are unculturable in laboratory conditions, some ecosystems are ideal niches for bioprospecting extremophiles with unique properties. Up today, there are few reports concerning microbial communities found on solar panels, a widespread, artificial, extreme habitat. Microorganisms found in this habitat belong to drought-, heat- and radiation-adapted genera, including fungi, bacteria, and cyanobacteria.Methods: Here we isolated and identified several cyanobacteria from a solar panel. Then, some strains isolated were characterizated for their resistance to desiccation, UV-C exposition, and their growth on a range of temperature, pH, NaCl concentration or diverse carbon and nitrogen sources. Finally, gene transfer to these isolates was evaluated using several SEVA plasmids with different replicons to assess their potential in biotechnological applications.Results and discussion: This study presents the first identification and characterization of cultivable extremophile cyanobacteria from a solar panel in Valencia, Spain. The isolates are members of the genera Chroococcidiopsis, Leptolyngbya, Myxacorys, and Oculatella all genera with species commonly isolated from deserts and arid regions. Four of the isolates were selected, all of them Chroococcidiopsis, and characterized. Our results showed that all Chroococcidiopsis isolates chosen were resistant up to a year of desiccation, viable after exposition to high doses of UV-C, and capable of being transformed. Our findings revealed that a solar panel is a useful ecological niche in searching for extremophilic cyanobacteria to further study the desiccation and UV-tolerance mechanisms. We conclude that these cyanobacteria can be modified and exploited as candidates for biotechnological purposes, including astrobiology applications.

The highly xerotolerant bacterium classified as Exiguobacterium sp. Helios isolated from a solar panel in Spain showed a close relationship to Exiguobacterium sibiricum 255-15 isolated from Siberian permafrost. Xerotolerance has not been previously described as a characteristic of the extremely diverse Exiguobacterium genus, but both strains Helios and 255-15 showed higher xerotolerance than that described in the reference xerotolerant model strain Deinococcus radiodurans. Significant changes observed in the cell morphology after their desiccation suggests that the structure of cellular surface plays an important role in xerotolerance. Apart from its remarkable resistance to desiccation, Exiguobacterium sp. Helios strain shows several polyextremophilic characteristics that make it a promising chassis for biotechnological applications. Exiguobacterium sp. Helios cells produce nanoparticles of selenium in the presence of selenite linked to its resistance mechanism. Using the Lactobacillus plasmid pRCR12 that harbors a cherry marker, we have developed a transformation protocol for Exiguobacterium sp. Helios strain, being the first time that a bacterium of Exiguobacterium genus has been genetically modified. The comparison of Exiguobacterium sp. Helios and E. sibiricum 255-15 genomes revealed several interesting similarities and differences. Both strains contain a complete set of competence-related DNA transformation genes, suggesting that they might have natural competence, and an incomplete set of genes involved in sporulation; moreover, these strains not produce spores, suggesting that these genes might be involved in xerotolerance.

The biochemistry and genetics of the bacterial steroid catabolism have been extensively studied during the last years and their findings have been essential to the development of biotechnological applications. For instance, metabolic engineering of the steroid-eater strains has allowed to obtain intermediaries of industrial value. However, there are still some drawbacks that must be overcome, such as the redundancy of the steroid catabolism genes in the genome and a better knowledge of its genetic regulation. KshABs and KstDs are key enzymes involved in the aerobic breakage of the steroid nucleus. Rhodococcus ruber Chol-4 contains three kshAs genes, a single kshB gene and three kstDs genes within its genome. In the present work, the growth of R. ruber ΔkshA strains was evaluated on different steroids substrates; the promoter regions of these genes were analyzed; and their expression was followed by qRT-PCR in both wild type and ksh mutants. Additionally, the transcription level of the kstDs genes was studied in the ksh mutants. The results show that KshA2B and KshA1B are involved in AD metabolism, while KshA3B and KshA1B contribute to the cholesterol metabolism in R. ruber. In the kshA single mutants, expression of the remaining kshA and kstD genes is re-organized to survive on the steroid substrate. These data give insight into the fine regulation of steroid genes when several isoforms are present.

Information about the paper titled "PANDEMIC: THE PHANTOM MENACE: LEARNING GENETIC ENGINEERING BY A GAME-BASED METHODOLOGY" at IATED Digital Library