Environmental control strategies are commonly practised to limit biodeterioration issues threatening indoor cultural heritage objects, while they are still poorly exploited for the conservation of outdoor stone heritage surfaces, including rock art. In this study, we evaluated the environmental factors driving the diversity and abundance of lithobiontic communities in the Rock Engravings National Park of Naquane (UNESCO WHS n. 94, Italy). The survey considered 23 rocks that had been cleaned in the last three (3YC) or twelve (12YC) years or more than 40 years ago (NRC). A cyanobacteria-dominated biofilm and lichens (37 taxa) were the most widespread and abundant lithobiontic components, prevailing on 3YC–12YC and NRC rocks, respectively. On the latter, a turnover of xerophytic and meso-hygrophytic lichen communities was observed. On 3YC–12YC rocks lichen colonization, if present, was limited to nitrophytic species, including common epiphytes from surrounding trees and a small number of meso-hygrophytic species, with a prevalence of asexual reproductive strategies. Multivariate analyses including environmental parameters (canonical correspondence analyses) indicated that tree cover and the presence of bare or vegetated ground upstream of the rocks, probably prolonging wetness and providing nutrients by water transport, are the factors mostly related to the microbial and lichen recolonization of 3YC–12YC surfaces. On this basis, an experiment on preventive conservation was conducted, consisting of a new cleaning of a strongly recolonized 3YC surface combined with the building of a small wall to protect part of the rock from prolonged water fluxes. The fluorimetric and colorimetric monitoring of the rock surface, carried out 40 months after this new cleaning intervention, displayed recolonization on the unprotected area only, indicating the potential of preventive conservation strategies in outdoor environments.

The research described in this Research Communication addresses the hypothesis that intramammary infections with Streptococcus uberis (S. uberis) are associated with biofilm formation, which limits antibiotic efficacy. This retrospective study investigated biofilm expression and antimicrobial resistance (AMR) patterns of 172 S. uberis infections. Isolates were recovered from milk samples of subclinical, clinical, and intramammary infection cases on 30 commercial dairy herds. We determined the presence and intensity of biofilm expression of S. uberis isolates in vitro in three somatic cell count categories to recognise their AMR patterns. An automated minimum inhibitory concentration system with a commercially available panel of 23 antimicrobial agents evaluated AMR, while biofilm determination was conducted using a microplate method. The study established that all the S. uberis isolates assessed expressed biofilm with the following varying degrees of intensities: 30 (17.8%) strong, 59 (34.9%) medium and 80 (47.3%) weak biofilms. The newly registered UBAC mastitis vaccine containing biofilm adhesion components may, therefore, be a viable option for proactive mastitis management under field conditions. No differences were identified between biofilm intensity and the three somatic cell count groups. Most S. uberis isolates indicated a high-level sensitivity to the antimicrobial agents tested. Resistances were present in 8.7, 8.1 and 7.0% cases to rifampin, minocycline and tetracycline, respectively. Multidrug resistance was observed in 6.4%, emphasising AMR to antibiotics used in human medicine only. The low overall resistance suggests that farmers adhere to the prudent use of antimicrobials in the dairy industry.

The starting objective of this research communication was to determine the prevalence of subclinical mastitis in buffalo in Turkey. We also seeked to isolate and identify staphylococci, determine their antimicrobial susceptibilities and biofilm-forming abilities as well as investigating the presence of biofilm-related genes and microbial surface components recognizing adhesive matrix molecules. A total of 107 (66.9%) staphylococci (28 S. aureus and 79 coagulase-negative staphylococci, CoNS) were isolated from 160 mastitic milk samples collected from 200 lactating water buffalos. The staphylococci were especially resistant to beta-lactams except for cefoxitin but were less resistant to the other antimicrobials that were tested. Based on the Congo red agar method, 92.9% of the S. aureus and 70.9% of the CoNS isolates were positive for biofilm-forming ability, while all S. aureus and 97.5% of CoNS isolates were positive by a microtiter plate analysis. The presence of icaA and icaD genes was not always correlated with biofilm synthesis, and even in the absence of these genes, the isolates were able to synthesize biofilm.

With the increase of crewed space missions and the rise of space microbiology, the research of microbes grown under microgravity environment has been attracting more attention. The research scope in space microbiology has been extended beyond pathogens directly related to spaceflight. Y. pestis, the causative agent of plague, is also of interest to researchers. After being cultivated for 40 consecutive passages in either simulated microgravity (SMG) or normal gravity (NG) conditions, the Y. pestis strain 201 cultures were analysed regarding their phenotypic features. By using crystal violet staining assays, increased biofilm amount was detected in Y. pestis grown under SMG condition. Besides that, the damage degrees of Hela cell caused by SMG-grown Y. pestis were found diminished in comparison to those under NG condition. Consistent with this observation, the death course was delayed in mice infected with SMG-grown Y. pestis, suggesting that microgravity condition can contribute the attenuated virulence. RNA-seq-based transcriptomics analysis showed that a total of 218 genes were differentially regulated, of which 91 upregulated and 127 downregulated. We found that dozens of virulence-associated genes were downregulated, which partially explained the reduced virulence of Y. pestis under SMG condition. Our study demonstrated that long-term exposure to SMG influences the pathogenesis and biofilm formation ability of Y. pestis, which provides a novel avenue to study the mechanism of physiology and virulence of this pathogen. Microgravity enhanced the ability of biofilm formation and reduced the virulence and cytotoxicity of Y. pestis. Many virulence-associated genes of Y. pestis were differentially regulated in response to the stimulated microgravity. However, there is no molecular evidence to explain the enhanced biofilm formation ability, which requires further research. Taken together, the phenotype changes of Y. pestis under SMG conditions can provide us a new research direction of its potential pathogenesis.

In clinical environments, orthopedic implants are associated with a risk of infection during implantation. However, the growth paths of bacteria on metal, which is nontransparent, are difficult to observe. In this study, we visualized the DH5-alpha Escherichia coli bacterial growth path on the surface of magnesium by using scanning electron microscope (SEM) images and constructed a convolutional neural network-based artificial intelligence (AI) system to identify metal surfaces, bacteria, and its generated products to grade the growth stage of the bacteria implanted on the magnesium. The detection result of the E. coli growth stage by the AI system was close to that manually marked by experts, and it may greatly accelerate the investigation of the bacterial growth process in various types of metallic material.

Biofilm formation is a central feature to guarantee staphylococcal persistence in hosts and is associated with several diseases that are difficult to treat. In this research paper, biofilm formation and antimicrobial susceptibility were investigated in staphylococcal strains belonging to several species. These strains were isolated from the milk of cows with subclinical mastitis and most of them were coagulase-negative, with the prevalence of Staphylococcus chromogenes. High genetic diversity was observed among the strains by pulsed field gel electrophoresis. Antimicrobial resistance was assessed by disk diffusion and more than 50% of the strains were resistant to ampicillin and penicillin G, with multi-resistance profiles (13.6%) also being observed. Most strains (65.9%) formed biofilms when cultivated in BHI supplemented with 1% glucose. Most strains (72.7%) carried the intercellular adhesion gene (icaA), while less than half (36.3%) carried the biofilm-associated protein gene (bap). Concentrations of up to 10xMIC of erythromycin and tetracycline were not sufficient to suppress cell viability in preformed biofilms. Our results revealed that a genetically diverse group of biofilm-forming Staphylococcus species can be involved in subclinical mastitis. Since high antimicrobial concentrations cannot eradicate biofilm cells in vitro, their use in dairy animals may be ineffective in controlling infections, while supporting selection of resistant microorganisms. These data reinforce the need for alternative therapies aiming at disrupting biofilms for effective disease control.

Finely laminated (cm–μm scale) metalliferous precipitates are widespread in the surficial environment, especially around mineral deposits and reflect biogeochemical processes that can pervade near-surface environments on a larger scale. Examples in this paper involve precipitates of the transition metals Fe, Cu and Mn with minor Co, Ni, V and Zn; the metalloids As and Sb; and authigenic Au. Mobility and re-precipitation are driven primarily by geochemical disequilibrium, especially with respect to pH and redox states, that arises from complex interactions between biological processes, geological processes, and variations in the surrounding environment. Different degrees of chemical disequilibrium arise on small spatial scales on time scales of days to millennia. Interactions between biota, waters and rocks in these small near-surface settings affect the biogeochemical environments. Sulfur- and iron-oxidising bacteria are common biogeochemical agents associated with sulfide-bearing lithologies, but localised reductive environments can also develop, leading to gradients in pH and redox state and differential metal mobility. In general, there is commonly a spatial separation of Fe-rich precipitates from those with Cu and Mn, and other transition metals also follow Cu and Mn rather than Fe. Metalloids As and Sb have a strong affinity for Fe under oxidising conditions, but not under more reducing conditions. However, complex biogeochemical parageneses of laminated metalliferous deposits preclude prediction of finer formation details. The textures, mineral species, and metal associations within these deposits are likely to be encountered in all facets of mineral deposit development: initial exploration activity of near-surface locations, mining of shallow portions of orebodies, especially supergene zones, and downstream environmental management with respect to discharging metalliferous waters.

Scedosporium and Lomentospora species are ubiquitous saprophytic filamentous fungi that emerged as human pathogens with impressive multidrug-resistance profile. The ability to form biofilm over several biotic and abiotic surfaces is one of the characteristics that contributes to their resistance patterns against almost all currently available antifungals. Herein, we have demonstrated that Scedosporium apiospermum, Scedosporium minutisporum, Scedosporium aurantiacum and Lomentospora prolificans were able to form biofilm, in similar amounts, when conidial cells were incubated in a polystyrene substrate containing Sabouraud medium supplemented or not with different concentrations (2%, 5% and 10%) of glucose, fructose, sucrose and lactose. Likewise, the glucose supplementation of culture media primarily composed of amino acids (SCFM, synthetic cystic fibrosis medium) and salts (YNB, yeast nitrogen base) did not modulate the biofilm formation of Scedosporium/Lomentospora species. Collectively, the present data reinforce the ability of these opportunistic fungi to colonize and to build biofilm structures under different environmental conditions.

The circular economy aspects of PET (polyethylene terephthalate) waste conversion into value-added products are discussed concerning different governmental policies and industrial protocol for plastic degradation.

The use of microbial enzymes such as PET hydrolase is discussed regarding PET (polyethylene terephthalate) degradation.

The primary purpose of this perspective is a critical analysis of the correlation of the current state-of-the-art rising circular economy platform enacted across the world with close looping of PET (polyethylene terephthalate)-based plastic polymer disposal and sustainable recycling and upcycling technology. The goal of the upcycling process is to get the low-cost value-added monomer than those obtained from the hydrocarbon industry from the sustainability prospect. A summary of the circular bio-economic opportunities has also been described. Next, how the PET hydrolase enzyme degrades the PET plastic is discussed. It is followed by an additional overview of the effect of the mutant enzyme for converting 90% of plastics into the terephthalate monomer. A site-directed mutagenesis procedure obtains this particular mutant enzyme. The diversity of different microbial organism for producing PET hydrolase enzyme is finally discussed with a suggested outlook of the circular economy goal from the viewpoint of plastic degradation objectives soon.

Liquid phase analysis dominates the field of blood diagnostics and requires drawing blood volumes of several ml for each test. To achieve acceptable accuracy, each single liquid blood test requires ∼7 mL per blood sample, and repeated blood tests are often needed. Frequent testing ca result in Hospital Acquired Anemia for infants, chronically ill, and critically ill patients. Blood testing methods that can be utilized with small amounts of blood are a critical need to save lives. Theranos claimed to have developed novel methods requiring only a few nL of blood. However, Theranos’ techniques led to errors that exceeded beyond the medically acceptable threshold of 10%. This work investigates solid state blood analysis using low volumes of several µL. The most common blood tests used as first line for diagnostics and monitoring patients’ status, always include blood electrolytes, iron, and in some cases, heavy metals.

The present work investigates the formation of rapidly solidified Homogeneous Thin Solid Films (HTSFs) formed from blood drops, in order to make them suitable for solid state analysis in vacuo and in air. The solidification of ∼5 micro-liter (µL)-sized blood droplets into HTSFs is studied with two goals: achieve reproducible HTSFs optimized for producing accurate analysis, and successfully measure the potential accuracy of measurements made on HTSFs for blood electrolytes Na, K, Mg, Ca, and Cl and heavy metals such as Fe.

The blood volumes selected for this work are in the µL range, one thousandth volumes drawn for current liquid phase analysis. Balanced Saline Solution (BSS) is used as an initial liquid for testing solidification uniformity and a potential calibration material. Next, canine and human blood are studied on two types of HemaDropTM coatings for solidification: super-hydrophilic and hyper-hydrophilic. HTSF formation from BSS and blood drops are compared on both coated and uncoated surfaces.

Three solid state analytical methods are investigated in parallel to probe composition at different depths and test each for reproducibility and accuracy: Ion Beam Analysis (IBA), X-ray Fluorescence (XRF), and X-ray Photoelectron Spectroscopy (XPS). The results show that using solid films of blood yields composition, which can be reproducibly measured by IBA, XPS and XRF to varying degrees. XPS’s depth of analysis, limited to ∼5 nm, probes a small fraction of the HTSF, but provides insights into the range of thickness for homogeneous compositions in HTSFs. Statistical and error analysis help establish whether measurements taken in sets of three typically used in lab fall below the medically accepted error threshold (<10%) for each technique and element detected. Measurements are repeated and taken at various locations and on different HTSFs to establish reproducibility. XRF is of particular interest, because it is fast, accurate, portable and can be conducted in air, making it ideal for areas with limited resources.

We propose a numerical method for the simulation of a quasi-linear parabolic biofilm model that exhibits three non-linear diffusion effects: (i) a power law degeneracy, (ii) a super diffusion singularity and (iii) non-linear cross-diffusion. The method is based on a spatial Finite Volume discretisation in which cross-diffusion terms are formally treated as convection terms. Time-integration of the resulting semi-discretised system is carried out using an error-controlled, time-adaptive, embedded Rosenbrock–Wanner method. We compare several variants of the method and two variants of the model to investigate how details such as the choice cross-diffusion coefficients, and specific variants of the time integrator affect simulation time.

We derive a macroscopic model for biofilm formation in a porous medium reactor to investigate the role of suspended bacteria on reactor performance. The starting point is the mesoscopic one-dimensional Wanner–Gujer biofilm model. The following processes are included: hydrodynamics and transport of substrate in the reactor, biofilm and suspended bacteria growth in the pore space, attachment of suspended cells to the biofilm, and detachment of biofilm cells. The mesoscopic equations are up-scaled from the biofilm scale to the reactor scale, yielding a stiff system of balance laws, which we study numerically. We find that suspended bacteria and attachment can have a significant effect on biofilm reactor performance.

Actinobacillus pleuropneumoniae is a Gram-negative bacterium that belongs to the family Pasteurellaceae. It is the causative agent of porcine pleuropneumonia, a highly contagious respiratory disease that is responsible for major economic losses in the global pork industry. The disease may present itself as a chronic or an acute infection characterized by severe pathology, including hemorrhage, fibrinous and necrotic lung lesions, and, in the worst cases, rapid death. A. pleuropneumoniae is transmitted via aerosol route, direct contact with infected pigs, and by the farm environment. Many virulence factors associated with this bacterium are well characterized. However, much less is known about the role of biofilm, a sessile mode of growth that may have a critical impact on A. pleuropneumoniae pathogenicity. Here we review the current knowledge on A. pleuropneumoniae biofilm, factors associated with biofilm formation and dispersion, and the impact of biofilm on the pathogenesis A. pleuropneumoniae. We also provide an overview of current vaccination strategies against A. pleuropneumoniae and consider the possible role of biofilms vaccines for controlling the disease.

In this paper, we describe the development of an International Space Station experiment, BioRock. The purpose of this experiment is to investigate biofilm formation and microbe–mineral interactions in space. The latter research has application in areas as diverse as regolith amelioration and extraterrestrial mining. We describe the design of a prototype biomining reactor for use in space experimentation and investigations on in situ Resource Use and we describe the results of pre-flight tests.

Cellular elasticity is frequently measured to investigate the biomechanical effects of drug treatment, diseases, and aging. In light of the cellular viscosity property exhibited by filament actin networks, this study investigates the viscoelasticity alterations of the human hepatocellular carcinoma (SMMC-7721) cell subjected to fullerenol treatment by means of creep tests realized by atomic force microscopy indentation. An SMMC-7721 cell was first modeled as a sphere and then as a flattened layer with finite thickness. Both Sneddon’s solutions and the Dimitriadis model have been modified to adapt to the viscoelastic situation, which are used to fit the same indentation depth–time curves obtained by creep tests. We find that the SMMC-7721 cell’s creep behavior is well described by the two modified models and the divergence of parameters determined by the two models is justified. By fullerenol treatment, the SMMC-7721 cell exhibits a significant decrease of elastic modulus and viscosity, which is presumably due to the disruption of actin filaments. This work represents a new attempt to understand the alternation of the viscoelastic properties of cancerous cells under the treatment of fullerenol, which has the significance of comprehensively elucidating the biomechanical effects of anticancer agents (such as fullerenol) on cancer cells.

This Research Communication describes the study of in vitro biofilm formation of mastitis causing pathogens. Biofilms are communities of bacteria that are attached to a surface and to each other and are embedded in a self-produced matrix of extracellular polymeric substances. Biofilm formation is an important virulence factor that may result in recurrent or persistent udder infections and treatment failure through increased resistance to antibiotics and protection against host defences. In the present study 252 bacterial isolates from milk samples from bovine udder quarters with intramammary infections were examined with Congo Red agar (CRA) method and tube method (TM) for their ability to form biofilms. Both tests revealed a high number of biofilm-positive strains. Literature reports that the cure rates for Staphylococcus aureus infected udders are lower (27%) in comparison to cure rates of Streptococcus uberis (64–81%) or coagulase-negative staphylococci (CNS) mastitis (80–90%). The findings of the present study suggest that biofilm formation is not the main factor for the differences in cure rates of the various bacteria genera, because all tested pathogen groups showed a similarly high proportion of biofilm formation. Further research is needed to detect microbial biofilms on bovine udder epithelia.

The biofilm-associated protein (Bap) of Staphylococcus aureus is a high molecular weight cell-wall-anchored protein involved in biofilm formation, first described in bovine mastitis strains from Spain. So far, studies regarding Bap were mainly based on the Spanish strain V329 and its mutants, but no information on the genetic variability of bap-positive Staph. aureus strains is yet available in the literature. The present study investigated the molecular characteristics of 8 bap-positive Staph. aureus strains from subclinical bovine mastitis, isolated in 5 herds; somatic cell counts (SCC) of milk samples were also registered. Strains were characterised using MLST, SPA typing and microarray and the results were compared with V329. All isolates from this study and V329 were assigned to ST126, t605, but some molecular differences were observed. Only herd A and B strains harboured the genes for β-lactams resistance; the leukocidin D/E gene, a type I site-specific deoxyribonuclease subunit, 3rd locus gene and serin-protease A and B were carried by all strains, but not by V329, while serin-protease E was absent in V329 and in another isolate. Four isolates and V329 harboured the fibronectin-binding protein B gene. SCC showed the highest value in the milk sample affected by the only strain carrying all the virulence factors considered. Potential large variability of virulence was evidenced among V329 and all bap-positive Staph. aureus strains considered: the carriage of fnb could enhance the accumulation of biofilm, but the lack of lukD/E and splA, B or E might decrease the invasiveness of strain.