Bacteria and microalgae associations in periphytonmechanisms and biotechnological opportunities

Phototrophic and heterotrophic microorganisms coexist in complex and dynamic structures called periphyton. These structures shape the biogeochemistry and biodiversity of aquatic ecosystems. In particular, microalgaebacteria interactions are a prominent focus of study by microbial ecologists and can provide biotechnological opportunities for numerous applications (i.e. microalgal bloom control, aquaculture, biorefinery, and wastewater bioremediation). In this review, we analyze the species dynamics (i.e. periphyton formation and factors determining the prevalence of one species over another), coexisting communities, exchange of resources, and communication mechanisms of periphytic microalgae and bacteria. We extend periphyton mathematical modelling as a tool to comprehend complex interactions. This review is expected to boost the applicability of microalgaebacteria consortia, by drawing out knowledge from natural periphyton.This work was financially supported by: LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC); Portuguese funds through FCT—Fundação para a Ciência e a Tecnologia, I.P., under the Pluriannual Financing Programme for R&D Units for the Centre of Biological Engineering of University of Minho (ref. UIDB/BIO/04469/2020); LABBELS—Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/0029/2020; PRESAGE—Aquatic/0007/2020, funded by national funds through Foundation for Science and Technology FCT/MCTES (PIDDAC); Project Germirrad—POCI-01–0247-FEDER 072237 funded by FEDER funds through COMPETE2020–Programa Operacional Competitividade e Internacionalização (POCI) and by national funds (PIDDAC) through FCT/MCTESHealthyWaters (NORTE-01–0145-FEDER-000069), supported by Norte Portugal Re gional Operational Programme (NORTE 2020), under the PORTU GAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).info:eu-repo/semantics/publishedVersio

Sodium dodecyl sulfate allows the persistence and recovery of biofilms of Pseudomonas fluorescens formed under different hydrodynamic conditions

The effect of the anionic surfactant sodium dodecyl sulfate (SDS) on Pseudomonas fluorescens biofilms was investigated using flow cell reactors with stainless steel substrata, under turbulent (Re=5200) and laminar (Re=2000) flow. Steady-state biofilms were exposed to SDS in single doses (0.5, 1, 3 and 7 mM) and biofilm respiratory activity and mass measured at 0, 3, 7 and 12 h after the SDS application. The effect of SDS on biofilm mechanical stability was assessed using a rotating bioreactor. Whilst high concentrations (7 mM) of SDS promoted significant biofilm inactivation, it did not significantly reduce biofouling. Turbulent and laminar flow-generated biofilms had comparable susceptibility to SDS application. Following SDS exposure, biofilms rapidly recovered over the following 12 h, achieving higher respiratory activity values than before treatment. This phenomenon of posttreatment recovery was more pronounced for turbulent flow-generated biofilms, with an increase in SDS concentration. The mechanical stability of the biofilms increased with surfactant application, except for SDS concentrations near the critical micellar concentration, as measured by biofilm removal due to an increase in external shear stress forces. The data suggest that although SDS exerts antimicrobial action against P. fluorescens biofilms, even if only partial and reversible, it had only limited antifouling efficacy, increasing biofilm mechanical stability at low concentrations and allowing significant and rapid recovery of turbulent flow-generated biofilms.Fundação para a Ciência e a Tecnologia (FCT

Enhancement of Escherichia coli and Staphylococcus aureus antibiotic susceptibility using sesquiterpenoids

The present work examines the potential of sesquiterpenoids to sensitize Escherichia coli and Staphylococcus aureus, and modulate their susceptibility to the standard antibiotics ciprofloxacin, erythromycin, gentamicin and vancomycin. It was tested samples of three sesquiterpenoids: guaiazulene, nerolidol (racemic mixture of the cis and trans isomers) and germacrene D enriched natural extract. Experiments were conducted aiming to assess the antimicrobial effects of the antibiotic-sesquiterpenoid combination on bacterial growth inhibition, by the disc diffusion assay and the minimum inhibitory concentration (MIC) assessment, the bactericidal effects, the post-antibiotic effect (PAE) and the effect on membrane permeability. The data related with the antimicrobial activity evidenced, through the disc diffusion assay, an antibiotic S. aureus antimicrobial activity enhancement by sesquiterpenoids presence. The MIC value for E. coli decreased significantly by sesquiterpenoids combination with ciprofloxacin, erythromycin and gentamicin, and for S. aureus, with all four selected antibiotics. This combination also increased the PAE, with the exception of guaiazulene, which seemed to quench antibiotic antimicrobial action. A moderate correlation between antimicrobial action and impairment of cell membrane function was detected for germacrene D enriched extract, and nerolidol, as single treatments and in combination with antibiotic, while a poor correlation was obtained for guaiazulene. This study provides basis for the evaluation of sesquiterpenoids as alternative or possible synergistic compounds for current antimicrobial chemotherapeutics, showing the practical utility of natural derived products to increase the susceptibility of E. coli and S. aureus.The authors acknowledge the financial support provided by the Portuguese Foundation for Science and Technology (Project CHEMBIO-POCI/BIO/61872/2004, CIENCIA2007 - Manuel Simoes

Elucidating bacterial coaggregation through a physicochemical and imaging surface characterization

Bacterial coaggregation is a highly specific type of cell-cell interaction, well-documented among oral bacteria, and involves specific characteristics of the cell surface of the coaggregating strains. However, the understanding of the mechanisms promoting coaggregation in aquatic systems remains limited. This gap is critical to address, given the broad implications of coaggregation for multispecies biofilm formation, water quality, the performance of engineered systems, and diverse biotechnological applications. Therefore, this study aims to comprehensively characterize the cell surface of the coaggregating strain Delftia acidovorans 005P, isolated from drinking water, alongside a non-coaggregating strain, D. acidovorans 009P. By analyzing two strains of the same species, we aim to identify the factors contributing to the coaggregation ability of strain 005P. To achieve this, we employed a combination of physicochemical characterization, Fourier-transform infrared spectroscopy (FTIR), and advancing imaging techniques [transmission electron microscopy and cryo-electron tomography (cryo-ET)]. The coaggregating strain (005P) exhibited higher surface hydrophobicity, negative surface charge, and cell surface and co-adhesion energies than the non-coaggregating strain (009P). The chemical characterization of bacterial surfaces through FTIR revealed subtle differences, particularly in spectral regions linked to carbohydrates and phosphodiesters/amide III of proteins (860930 cm1 and 12121240 cm1, respectively). Cryo-ET highlighted significant differences in pili structures between the strains, such as variations in length, frequency, and arrangement. The pili in the 005P strain, identified as pili-like adhesins, serve as key mediators of coaggregation. By integrating physicochemical analyses and high-resolution imaging techniques, this study conclusively links the coaggregation ability of D. acidovorans 005P to its unique pili characteristics, emphasizing their crucial role in microbial coaggregation in aquatic environments.info:eu-repo/semantics/publishedVersio

Coaggregating Delftia acidovorans facilitates the metabolic activity of partner bacteria in drinking water biofilms

Although bacterial coaggregation has already been identified as a key mechanism in the multispecies biofilms development in diverse environments, little is known about this highly specific type of cell-cell interaction in aquatic systems. In this study, a strain of Delftia acidovorans isolated from drinking water (DW) and previously described with coaggregation ability, was studied for its role in biofilm development. Single and dual-species biofilms, of D. acidovorans and the DW representative bacteria Citrobacter freundii and Pseudomonas putida, were grown in 96-well microtiter plates and characterized in terms of metabolic activity. In addition, to prove that D. acidovorans can facilitate the metabolic activity of other bacteria, single biofilms were formed in the presence of D. acidovorans cell-free supernatant (CFS). The metabolic activity was higher when comparing single with dual-species biofilms. Furthermore, in the presence of CFS, it was evident an increase in the metabolic activity of C. freundii and P. putida biofilms. These results suggest the production of extracellular metabolites by D. acidovorans, that possibly act as public goods. That said, the presence of coaggregating species, namely D. acidovorans, in biofilms appears to create a functional cooperative microbial community, providing a favorable metabolic opportunity for partner bacteria.LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE), funded by national funds through FCT/MCTES (PIDDAC); UIDB/04469/2020 (CEB) and by LABBELS – Associate Laboratory in Biotechnology, Bioengineering and Microelectromechnaical Systems, LA/P/0029/2020; UIDB/04033/2020 (CITAB); Project Germirrad-POCI-01-0247-FEDER-072237, funded by FEDER funds through COMPETE2020–Programa Operacional Competitividade e Internacionalização (POCI) and by national funds (PIDDAC) through FCT/MCTES; project HealthyWaters (NORTE-01- 0145-FEDER-000069)- NORTE 2020/ERDF; and the FCT grant (2020.04773.BD).info:eu-repo/semantics/publishedVersio

Antibacterial activity and mode of action of selected glucosinolate hydrolysis products against bacterial pathogens

Abstract Plants contain numerous components that are important sources of new bioactive molecules with antimicrobial properties. Isothiocyanates (ITCs) are plant secondary metabolites found in cruciferous vegetables that are arising as promising antimicrobial agents in food industry. The aim of this study was to assess the antibacterial activity of two isothiocyanates (ITCs), allylisothiocyanate (AITC) and 2-phenylethylisothiocyanate (PEITC) against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Listeria monocytogenes. The antibacterial mode of action was also characterized by the assessment of different physiological indices: membrane integrity, intracellular potassium release, physicochemical surface properties and surface charge. The minimum inhibitory concentration (MIC) of AITC and PEITC was 100 μg/mL for all bacteria. The minimum bactericidal concentration (MBC) of the ITCs was at least 10 times higher than the MIC. Both AITC and PEITC changed the membrane properties of the bacteria decreasing their surface charge and compromising the integrity of the cytoplasmatic membrane with consequent potassium leakage and propidium iodide uptake. The surface hydrophobicity was also non-specifically altered (E. coli and L. monocytogenes become less hydrophilic; P. aeruginosa and S. aureus become more hydrophilic). This study shows that AITC and PEITC have strong antimicrobial potential against the bacteria tested, through the disruption of the bacterial cell membranes. Moreover, phytochemicals are highlighted as a valuable sustainable source of new bioactive products

Follicular dynamics in Serrana goats

Twenty-two Serrana goats were studied through two successive estrous cycles in order to characterize their follicular dynamics during the breeding season. The ovaries of the goats were scanned daily by realtime ultrasonography and all follicles ≥3mm were measured and classified. The data were classified by the number of follicular waves per goat to test the hypothesis that temporal and morphological differences between the last follicular wave of an ovary, irrespective of ovulation, will affect the selection of the next ovulatory wave. The mean interovulatory interval was 20.7±1.0 days (mean±S.D.). Three to five waves per estrous cycle were observed and 61.3% (19/31) of cycles had fourwaves. In estrous cycles with fourwaves, the day of onset of the first, second, third and fourth wave was 1.4±1.0, 6.9±1.4, 11.6±1.8 and 16.8±1.6, respectively. No differences (P > 0.05) were found between the day of onset of the first and second waves for estrous cycles with three, four or five waves. However, the day of onset of the third and fourth waves occurred later when the number of waves per estrous cycle increased (P < 0.001). The duration of the interwave interval (time between the day of onset of two consecutive waves) was longer when the second wave was ovulatory. The length of the growth phase (2.4±0.9 days) and size (5.9±0.7 mm) of the dominant follicle in the second wave were lower (P < 0.01) than for the first wave (3.3±1.2 days and 6.6±0.9 mm, respectively) and the fifth wave (4.1±1.2 days and 7.5±1.0 mm, respectively). Within pairs of ovaries, the onset of the last wave occurred later (P < 0.05) and was less variable in ovulatory ovaries (day 16.8±1.4, n = 20) than in anovulatory ovaries (day 15.1±3.7, n = 20). The length of the growing phase was longer (P < 0.001) in the last waves of ovulatory ovaries (3.1±0.9 days) than in the last waves of anovulatory ovaries (1.7±0.8 days). These results support the hypothesis that the day of onset of the ovulatory wave is related to or, at least, conditioned by the luteolysis and the decrease in plasma progesterone. In summary, the estrous cycle of Serrana goats is characterized by sequential follicular wave growth with a great variability in their onset and duration, with the exception of the ovulatory wave. The temporal and morphological differences observed in the last wave of estrous cycle provide strong evidence for the role of progesterone in their regulation

Seedless Cu electroplating on Co-W thin films in low pH electrolyte: early stages of formation

The use of Ta/TaN barrier bilayer systems in electronic applications has been ubiquitous over the last decade. Alternative materials such as Co-W or Ru-W alloys have gathered interest as possible replacements due to their conjugation of favourable electrical properties and barrier layer efficiency at reduced thicknesses while enabling seedless Cu electroplating. The microstructure, morphology, and electrical properties of Cu films directly electrodeposited onto Co-W or Ru-W are important to assess, concomitant with their ability to withstand the electroplating baths/conditions. This work investigates the effects of the current application method and pH value of the electroplating solution on the electrocrystallisation behaviour of Cu deposited onto a Co-W barrier layer. The film structure, morphology, and chemical composition were studied by X-ray diffraction, scanning electron microscopy and atomic force microscopy, as well as photoelectron spectroscopy. The results show that the electrolyte solution at pH 1.8 is incapable of creating a compact Cu film over the Co-W layer in either pulsed or direct-current modes. At higher pH, a continuous film is formed. A mechanism is proposed for the nucleation and growth of Cu on Co-W, where a balance between Cu nucleation, growth, and preferential Co dissolution dictates the substrate area coverage and compactness of the electrodeposited films.Portugal 2020 through European Regional Development Fund (FEDER) in the frame of Operational Competitiveness and Internationalisation Programme (POCI) and in the scope of the project USECoIN with grant number PTDC/CTM-CTM/31953/2017. This work was also supported by FCT, through IDMEC, under LAETA project UIDB/50022/202

The action of selected isothiocyanates on bacterial biofilm prevention and control

The activity of two selected isothiocyanates (ITCs), allylisothiocyanate (AITC) and 2-phenylethy-lisothiocyanate (PEITC) was evaluated on the prevention and control of biofilms formed by Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Listeria monocytogenes. In addition, the effect of ITCs was also tested on planktonic cell susceptibility, bacterial motility and adhesion. Biofilm prevention and control were tested using a microtiter plate assay and the effect of ITCs was assessed on biofilm mass and metabolic activity. The minimum bactericidal concentration for E. coli and P. aeruginosa was 1000 μg mL−1 (AITC) and >1000 μg mL−1 (PEITC), for S. aureus and L. monocytogenes was >1000 μg mL−1 (for both ITCs). AITC caused total inhibition of swimming (P. aeruginosa) and swarming (E. coli) motilities. PEITC caused total inhibition of swimming (E. coli, P. aeruginosa and L. monocytogenes) and swarming (E. coli and P. aeruginosa) motilities. Colony spreading of S. aureus was completely inhibited with PEITC. Adhesion assessed in terms of free energy was less favorable when bacteria were exposed to AITC for E. coli and P. aeruginosa and PEITC for P. aeruginosa. Both ITCs had preventive action on biofilm formation and showed a higher potential to reduce the mass of biofilms formed by the Gram-negative bacteria. AITC and PEITC promoted reductions in biofilm activity higher than 60% for all the biofilms tested. The overall study emphasizes the potential of ITCs as emergent products to inhibit bacterial motility and prevent/control biofilms of important human pathogenic bacteria.This work was supported by Operational Programme for Competitiveness Factors - COMPETE and by FCT - Portuguese Foundation for Science and Technology through Projects Bioresist - PTDC/EBB-EBI/105085/2008; Phytodisinfectants - PTDC/DTP-SAP/1078/2012 and the PhD grant awarded to Anabela Borges (SFRH/BD/63398/2009)