Influence of bacterial biopolymers on physical properties of experimental limestone blocks.

This article describes the consolidation effects of bacterial biopolymers synthesized by biofilm bacteria colonizing Mayan limestone buildings on the surface properties of limestone blocks, including disaggregation, hardness, and total color change at the laboratory level. The biopolymers evaluated, produced by bacterial isolates TM1B-488, TM1B-489, TM1B-349, and TM1B-464, influenced surface properties at different levels. 16S rRNA gene sequences analysis showed that isolate TM1B-349 was related with Psychrobacter sp. strain Marseille P-5312, TM1B-464 was related with Agrococcus terreus strain BT116, and isolates TM1B-488 and TM1B-489 were related with Xanthomonas citri pv. mangiferaeindicae strain XC01. Biopolymer A reduced the surface disaggregation of the material (26%) compared to the untreated control, as revealed by the peeling test, followed by biopolymer B (10%), while the remaining biopolymers had a negligible effect. The cactus biopolymer reduced disaggregation at higher levels (37%). On the other hand, there was a similar concomitant increase in surface hardness of limestone samples coated with biopolymer A (34%) and biopolymer B (32%), higher than biopolymers C (10%) and D (19%). Total color change for all treatments was below the threshold value of 5, indicating a non-significant color alteration. Partial chemical characterization of best-performing biopolymer (A) suggests its probable glycoprotein nature, whose constitutive acidic monosaccharides probably contributed to higher adherence to the limestone surfaces, contributing to surface stabilization, hardening the surface, and decreasing surface decohesion. These preliminary findings suggest its potential application in bioconsolidants, but further studies are required.

Epilithic and endolithic microorganisms and deterioration on stone church facades subject to urban pollution in a sub-tropical climate.

Weathering of two church facades in Rio de Janeiro was caused substantially by salts, mainly halite and gypsum, detected by SEM and chemical analyses, which cause physical stresses by deposition within the rock. Biofilm populations, determined by SEM and as operational taxonomic units (OTUs), degraded stone by penetration, solubilization and redeposition of minerals on their surfaces. Endolithic cyanobacteria were associated with gypsum deposits. Microbiomes were typical for high-stress environments, high salt, intense insolation, low water and low nutrients (eg halophilic Rubrobacter, Salinicola, Sterigmatomyces). The main colonizers on the church most affected by traffic (Nossa Senhora da Candelária - CA) were Actinobacteria; Gammaproteobacteria (chiefly Pseudomonas) were predominant on the site situated in a leafy square (São Francisco de Paula - SF). Major Gammaproteobacteria on CA were halophilic Halomonas and Rhodobacteriaceae. Fungal OTUs on both churches were principally dimorphic, yeast-like basidiomycetes. Many OTUs of thermophilic microorganisms (eg the Thermomicrobia class, Chloroflexi) were present. This is the first use of next generation sequencing (NGS) to study microbial biofilm interactions with metamorphic and granite buildings in an intensely urban, sub-tropical climate.

Microbiomes of Biofilms on Decorative Siliceous Stone: Drawbacks and Advantages of Next Generation Sequencing.

Next Generation Sequencing (NGS), using the Illumina® metabarcoding system, showed differences between biofilm communities on three degraded siliceous stone church façades in central Rio de Janeiro. Two church biofilms (on granite and augen gneiss) were dominated by Actinobacteria; the third (granite), surrounded by trees and further from intense vehicular traffic, by Gammaproteobacteria. Yeast-like forms of Basidiomycetes and Ascomycetes were major fungi on all facades, but 22.8% of Operational Taxonomic Units could not be assigned to any fungal taxon after DNA amplification with ITS primers and analysis with the UNITE database, indicating the need for more fungal NGS studies. The pipeline used in analysis of the V4 region of rRNA bacterial gene sequences influenced the taxa detected, with two major classes and many genera identified only by the pipeline using the Greengenes, and not the Silva, database. Principal Components Analysis separated façade biofilms into the appropriate three groups and indicated greater dissimilarity of the tree-surrounded church biofilm from the others, confirmed by Jaccard Similarity coefficients, suggesting that local environment influences community composition more than stone type. NGS allows rapid and detailed analysis of microbiomes, but results must be carefully assessed and must not be used as the sole indication of community composition.

Microbial colonization affects the efficiency of photovoltaic panels in a tropical environment.

Sub-aerial biofilm (SAB) development on solar panels was studied in São Paulo. After 6, 12 and 18 months' exposure, photovoltaic panels were covered by increasing proportions of organic matter (42%, 53% and 58%, respectively). Fungi were an important component of these biofilms; very few phototrophs were found. Major microorganisms detected were melanised meristematic ascomycetes and pigmented bacterial genera Arthrobacter and Tetracoccus. While diverse algae, cyanobacteria and bacteria were identified in biofilms at 6 and 12 months, diversity at a later stage was reduced to that typical for SAB: the only fungal group detected in 18 month biofilm was the meristematic Dothideomycetes and the only phototrophs Ulothrix and Chlorella. Photovoltaic modules showed significant power reductions after 6, 12 (both 7%) and 18 (11%) months. The lack of difference in power reduction between 6 and 12 months reflects the dual nature of soiling, which can result from the deposition of particulates as well as from SAB fouling. Although 12-month old SAB demonstrated an almost 10-fold increase in fungal colonization and a higher organic content, the larger non-microbial particles (above 10 µm), which were important for efficiency reduction of lightly-biofilmed panels, were removed by high rainfall just before the 12-month sampling.

Biodeterioration and Chemical Corrosion of Concrete in the Marine Environment: Too Complex for Prediction.

Concrete is the most utilized construction material worldwide. In the marine environment, it is subject to chemical degradation through reactions with chloride (the most important ion), and sulfate and magnesium ions in seawater, and to biodeterioration resulting from biological (initially microbiological) activities, principally acid production. These two types of corrosions are reviewed and the failure of attempts to predict the degree of deterioration resulting from each is noted. Chemical (abiotic) corrosion is greatest in the splash zone of coastal constructions, while phenomenological evidence suggests that biodeterioration is greatest in tidal zones. There have been no comparative experiments to determine the rates and types of microbial biofilm formation in these zones. Both chemical and microbiological concrete deteriorations are complex and have not been successfully modeled. The interaction between abiotic corrosion and biofilm formation is considered. EPS can maintain surface hydration, potentially reducing abiotic corrosion. The early marine biofilm contains relatively specific bacterial colonizers, including cyanobacteria and proteobacteria; these change over time, producing a generic concrete biofilm, but the adhesion of microorganisms to concrete in the oceans has been little investigated. The colonization of artificial reefs is briefly discussed. Concrete appears to be a relatively prescriptive substrate, with modifications necessary to increase colonization for the required goal of increasing biological diversity.

Endolithic phototrophs in built and natural stone.

Lichens, algae and cyanobacteria have been detected growing endolithically in natural rock and in stone buildings in various countries of Australasia, Europe and Latin America. Previously these organisms had mainly been described in natural carbonaceous rocks in aquatic environments, with some reports in siliceous rocks, principally from extremophilic regions. Using various culture and microscopy methods, we have detected endoliths in siliceous stone, both natural and cut, in humid temperate and subtropical climates. Such endolithic growth leads to degradation of the stone structure, not only by mechanical means, but also by metabolites liberated by the cells. Using in vitro culture, transmission, optical and fluorescence microscopy, and confocal laser scanning microscopy, both coccoid and filamentous cyanobacteria and algae, including Cyanidiales, have been identified growing endolithically in the facades of historic buildings built from limestone, sandstone, granite, basalt and soapstone, as well as in some natural rocks. Numerically, the most abundant are small, single-celled, colonial cyanobacteria. These small phototrophs are difficult to detect by standard microscope techniques and some of these species have not been previously reported within stone.

Resistance of cyanobacterial fouling on architectural paint films to cleaning by water jet.

Mortar panels painted with three different white acrylic coatings were exposed to the environment in urban (São Paulo) and rural (Pirassununga) sites in Brazil for 7 years. After this time, all panels were almost equally discoloured, and paint detachment was observed to only a small degree. The biofilms were composed mainly of cyanobacteria and filamentous fungi, principal genera being Gloeocapsa and Chroococcidiopsis of the cyanobacteria, and Cladosporium and Alternaria of the fungi. Two of the three paints in Pirassununga became covered by a pink film that contained red-encapsulated Gloeocapsa and clay particles. The third, an 800% elastomeric matt formulation, became discoloured with a grey, only slightly pink, film, although the same cyanobacteria were present. The levels of paint detachments from all films in both locations were low, with rating range of 0-1 of a maximum 5 (100% detachment). After high-pressure water jetting, paint detachments increased at both locations, up to 2 in Pirassununga and 3 in São Paulo. Discoloration decreased; L*A*B* analysis of surface discoloration showed that ΔE (alteration in colour from the original paint film) changed from 28-39 before cleaning to 13-16 afterwards. The pink coloration was not entirely removed from Pirassununga samples, suggesting that cyanobacterial cells are difficult to detach, and microscopic analysis of the biofilms confirmed that Gloeocapsa was still present as the principal contaminant on all surfaces, with Chroococcidiopsis being present as the second most common. Almost no fungi were detected after water jet application.

Microbial biofilms on the sandstone monuments of the Angkor Wat Complex, Cambodia.

Discoloring biofilms from Cambodian temples Angkor Wat, Preah Khan, and the Bayon and West Prasat in Angkor Thom contained a microbial community dominated by coccoid cyanobacteria. Molecular analysis identified Chroococcidiopsis as major colonizer, but low similarity values (<95%) suggested a similar genus or species not present in the databases. In only two of the six sites sampled were filamentous cyanobacteria, Microcoleus, Leptolyngbya, and Scytonema, found; the first two detected by sequencing of 16S rRNA gene library clones from samples of a moist green biofilm on internal walls in Preah Khan, where Lyngbya (possibly synonymous with Microcoleus) was seen by direct microscopy as major colonizer. Scytonema was detected also by microscopy on an internal wall in the Bayon. This suggests that filamentous cyanobacteria are more prevalent in internal (high moisture) areas. Heterotrophic bacteria were found in all samples. DNA sequencing of bands from DGGE gels identified Proteobacteria (Stenotrophomonas maltophilia and Methylobacterium radiotolerans) and Firmicutes (Bacillus sp., Bacillus niacini, Bacillus sporothermodurans, Lysinibacillus fusiformis, Paenibacillus sp., Paenibacillus panacisoli, and Paenibacillus zanthoxyli). Some of these bacteria produce organic acids, potentially degrading stone. Actinobacteria, mainly streptomycetes, were present in most samples; algae and fungi were rare. A dark-pigmented filamentous fungus was detected in internal and external Preah Khan samples, while the alga Trentepohlia was found only in samples taken from external, pink-stained stone at Preah Khan. Results show that these microbial biofilms are mature communities whose major constituents are resistant to dehydration and high levels of irradiation and can be involved in deterioration of sandstone. Such analyses are important prerequisites to the application of control strategies.

Biodeterioration of stone and metal - Fundamental microbial cycling processes with spatial and temporal scale differences.

Fundamental processes for the biodeterioration of stone and metal involve many of the same microbially mediated reactions - oxidation, reduction, acid dissolution and elemental cycling - resulting from the activities of many of the same groups of environmental microorganisms. Differences depend on the nature of the substratum - stone vs. metal - and the composition of the surroundings, whether terrestrial (stone) or aquatic (stone and metal). Reactions within surface-related biofilms dominate the biodeterioration of metals and contribute greatly to the biodeterioration of stone. In the latter, phototrophic organisms, and especially cyanobacteria, are important first participants, while metal biodeterioration is almost entirely associated with bacteria, archaea and fungi. Biofilms on metal surfaces can produce chemical and electrochemical responses. While electrochemical responses are absent in stone, extracellular electron transfer can be a biodeterioration mechanism in some iron-rich rocks. Microorganisms in biofilms can penetrate and create fissures or cracks in stone and metals. However, the most obvious differences in the reactions of built stone and metal structures are related to the definition of failure, length of time required for a defined failure of the substratum, the area over which the failure occurs and the consequences of failure. Time and space are, similarly, quite distinct for biological breakdown and mineral cycling of metal and stone, with stone/rock cycling potentially occurring over thousands of years and kilometers.

Biocide-containing varnish for the protection of sandstone: comparison of formulations and laboratory test methods.

Two formulations of acrylic varnish, with and without either of two dry film biocides--one a mixture of isothiazolinones and benzimidazole derivatives, and the other a carbamate--were tested in vitro for their activity against mixtures of filamentous fungi and cyanobacteria found on sandstone buildings. Growth on filter-paper squares coated with the varnishes was assessed semi-quantitatively by naked eye, quantitatively by image analysis and chemically by measurement of ergosterol and chlorophyll a. The lower solvent content (higher resin) varnish was more inhibitory to cyanobacteria than the higher varnish content, whilst the opposite was true for the fungal inoculum. The carbamate biocide was effective against cyanobacteria, unlike the isothiazolinone mixture, but the latter produced more inhibition of fungal growth. The three assay methods produced generally similar results, although visual observation was obviously the most imprecise. There was an anomaly in the ergosterol measurements, which was considered to be caused by the varying ergosterol content and unequal inhibition of the three fungal genera used in the inoculum. Fusarium sp. was shown to contain higher levels of this membrane component than Cladosporium sp. and Penicillium sp. For this reason, the most appropriate method overall, giving reliable quantitative results, was deemed to be the image analysis.

Effect of culture medium on biocalcification by Pseudomonas Putida, Lysinibacillus Sphaericus and Bacillus Subtilis.

The objective of this study is to investigate the efficiency of calcium carbonate bioprecipitation by Lysinibacillus sphaericus, Bacillus subtilis and Pseudomonas putida, obtained from the Coleção de Culturas do Instituto Nacional de Controle de Qualidade em Saúde (INCQS), as a first step in determining their potential to protect building materials against water uptake. Two culture media were studied: modified B4 containing calcium acetate and 295 with calcium chloride. Calcium consumption in the two media after incubation with and without the bacterial inoculum was determined by atomic absorption analysis. Modified B4 gave the best results and in this medium Pseudomonas putida INQCS 113 produced the highest calcium carbonate precipitation, followed by Lysinibacillus sphaericus INQCS 414; the lowest precipitation was produced by Bacillus subtilis INQCS 328. In this culture medium XRD analysis showed that Pseudomonas putida and Bacillus subtilis precipitated calcite and vaterite polymorphs while Lysinibacillus sphaericus produced only vaterite. The shape and size of the crystals were affected by culture medium, bacterial strain and culture conditions, static or shaken. In conclusion, of the three strains Pseudomonas putida INQCS 113 in modified B4 medium gave the best results precipitating 96% of the calcium, this strain thus has good potential for use on building materials.

Paint fragments as polluting microplastics: A brief review.

Paint particles are part of the increasingly important microplastics (MPs) pollution of our oceans. They contain polyurethanes, polyesters, polyacrylates, polystyrenes, alkyls and epoxies. In spite of their prevalence, paint fragments are often excluded from MP audits. This review, citing 127 references, discusses detection, characteristics, sources and ecological effects of paint fragments in our oceans, as well as the abundance of paint fragments in MP samples around the world and their colonization by marine microorganisms, which differs from that of non-paint MPs. Paint MPs arise from shipping and boating activities, road markings and external surfaces of buildings. Many paint fragments come from antifouling paints used on commercial vessels and leisure boats; these may be regarded as particular pollutants, not only containing but also leaching heavy metals and biocides. Some effects of antifouling paint particles on aquatic biota are caused by these toxins. Paint particles are an understudied portion of marine MP pollution.

Nanoplastics in aquatic systems - are they more hazardous than microplastics?

The fragmentation of plastic materials into nanoparticles of less than 1000 nm (secondary nanoplastics) and their possible accumulation in the environment is a recent matter of concern. There are still no suitable standard methods for determining the concentrations and chemical makeup of these particles in aquatic systems and the fate and effect of nanoplastics in the aquatic environment has been little explored, although there has been research using engineered nanoparticles as models. In this review, we give a summary of the (mainly laboratory-based) studies on the influences of nanoplastics. We aim to provide an updated overview of this emerging topic, reviewing the literature mainly from 2018 onwards and considering the effects of nanoplastics on ecosystems, their uptake and transport of polluting molecules, and the challenges that are faced by workers in this area. The review includes 119 references.

Plastic microfibre pollution: how important is clothes' laundering?

Plastic microfibre pollution produced by domestic and commercial laundering of synthetic textiles has recently been incriminated in the press and the scientific literature as the main source (up to 90%) of primary microplastics in the oceans. Polyethylene terephthalate (PET) is the most common microfibre encountered. This review aims to provide updated information on worldwide plastic microfibre pollution caused by textile laundering and some possibilities for its control. Release of microfibres during domestic washing and tumble drying, their fate in wastewater treatment plants (WWTPs) and the oceans, and their environmental effects on the aquatic biota are discussed, as well as potential control methods at the levels of textile modification and laundry procedures. Environmental effects on aquatic biota are important; as a result of their small size and length-to-diameter ratio, microfibers are more effectively incorporated by organisms than other plastic particle groups. Simulation laundering studies may be useful in the development of a Standard Test Method and modification of WWTPs may reduce microfibre release into aquatic systems. However, improvements will be necessary in textile design and appliance design, and recommendations should be made to consumers about reducing their personal impact on the environment through their laundering choices, which can include appliances, fabric care products and washing conditions. Official regulation, such as that introduced recently by the French government, may be necessary to reduce plastic microfibre release from clothes' laundering.

Comparing the concentrations of heavy metals on two bivalve species in Santos Bay, Brazil: Subsidies to understanding the assimilation dynamic of bivalve contaminants.

Contaminant substances consist of chemical elements that present the potential to adversely impact the ecology of the environment, thus representing a threat to local fauna and flora. In this context, heavy metals are critical agents that, depending on the nature and level, are potentially toxic to living organisms. In order to evaluate the bioaccumulation of heavy metals in the Santos estuary and to determine the potential influence of salinity gradient on the heavy metal bioconcentration, the present study measured the concentrations of As, Cd, Pb, Cu, Cr, Fe, Ni, and Zn in two bivalve species (Crassostrea rhizophorae and Perna Perna) sampled at different sites of Santos Bay, in the southeastern region of Brazil. Throughout the study, the "sentinel species" used were effective in bioaccumulating contaminants. In oysters, based on the Brazilian legislation, critical limits were exceeded for As, Zn, Cu, and Cr. In the case of mussels, on the other hand, only for As, Zn, and Cr, the critical limits were overcome. In the present study, obtained data suggested salinity as a determinant parameter in As incorporation processes of bivalve mollusks. PRACTITIONER POINTS: The present study presents important results for the development of environmental management policies in estuarine environments. The present study points out differences between different organisms as biomonitors, providing subsidies for the decision of an effective biomonitoring program. The present study discusses values of contaminants as a danger to public health in Santos Bay, which can be extrapolated to other similar environments around the world.

Occurrence of microplastics and heavy metals accumulation in native oysters Crassostrea Gasar in the Paranaguá estuarine system, Brazil.

The ubiquitous presence of contaminants in the marine environment is considered a global threat to marine organisms. Heavy metals and microplastics are two distinct classes of pollutants but there are interactions between these two stressors that are still poorly understood. We examined the potential relationship between heavy metals (Al, Cr, Mn, Fe, Ni, Cu, Zn, As, Cd, Ba, Hg, Pb) and microplastic particles in oysters sampled along the Paranaguá Estuarine System. The results suggested high levels of As and Zn in the bivalves, which are destined for human consumption. Microplastic particles were found in oysters from all sampled locations, demonstrating the spread of this pollutant in the marine environment and its ability to bioaccumulate in oysters. However, our data did not demonstrate a direct relationship between microplastics and heavy metals, suggesting that these particles are not the main route for heavy metal contamination of oysters in the Paranaguá Estuarine System.

Microbial deterioration of stone monuments--an updated overview.

Cultural heritage monuments may be discolored and degraded by growth and activity of living organisms. Microorganisms form biofilms on surfaces of stone, with resulting aesthetic and structural damage. The organisms involved are bacteria (including actinomycetes and cyanobacteria), fungi, archaea, algae, and lichens. Interactions between these organisms and stone can enhance or retard the overall rate of degradation. Microorganisms within the stone structure (endoliths) also cause damage. They grow in cracks and pores and may bore into rocks. True endoliths, present within the rock, have been detected in calcareous and some siliceous stone monuments and are predominantly bacterial. The taxonomic groups differ from those found epilithically at the same sites. The nature of the stone substrate and the environmental conditions influence the extent of biofilm colonization and the biodeterioration processes. A critical review of work on microbial biofilms on buildings of historic interest, including recent innovations resulting from molecular biology, is presented and microbial activities causing degradation are discussed.

Argemone mexicana (Papaverales: Papavaraceae) as an Alternative for Mosquito Control: First Report of Larvicidal Activity of Flower Extract.

Culex quinquefasciatus Say (Diptera: Culicidae), an arboviral and filarial vector, is one of the most widespread mosquitoes in the world. The indiscriminate use of synthetic chemical insecticides has led to the development of resistance in mosquito populations worldwide. The effect of continuous exposure to crude extracts of Argemone mexicana, the Mexican poppy, on the development and growth stages of second-instar larvae of the mosquito was studied, along with qualitative chemical analysis of the different plant parts. Inhibition, mortality, and larval and pupal duration phases were assessed. Second-instar mosquito larvae were exposed to crude ethanol extracts of flowers, stems, and seeds. Flower extract exhibited the strongest larvicidal activity with LC50 and LC90 values after 24 h of exposure of 18.61 and 39.86 ppm, respectively, and 9.47 and 21.76 ppm after 48 h. Extracts from stem and seeds were significantly less effective. The flower extract registered a Growth Inhibition Index of 0.01 at 25 ppm, with stems and seeds registering 0.05 and 0.08, respectively, at 100 ppm (control group 1.02). Qualitative chemical analysis by thin-layer chromatography showed characteristic spots indicating the presence of alkaloids and flavonoids and phytochemical screening showed the presence of alkaloids, anthraquinones, flavonoids, tannins, and terpenoids in the various crude extracts. This is the first report of the effectiveness of an ethanol flower extract of A. mexicana on Cx. quinquefasciatus; it can be considered a promising alternative control for this mosquito species.

Weathering of granitic gneiss: A geochemical and microbiological study in the polluted sub-tropical city of Rio de Janeiro.

Chemical and biological weathering were studied on two historic churches in the city of Rio de Janeiro, Brazil. The churches exhibited contour scaling, granular disintegration and black crust development. High levels of SO4 were found in façade stones of one church, São José, with significant levels of lead and copper. This suggests that vehicle emissions were important in stonework decay. Both gypsum and halite were detected, typical of buildings influenced by marine atmosphere and fuel-polluted environment. There was little bioweathering on this church, although the filamentous cyanobacterium Scytonema detected could be involved in black crust production. The other church (Nossa Senhora da Gloria) showed strong granular disintegration, with a green coloration on internal surfaces of some flakes removed for study. This church showed lower levels of geochemically important weathering compounds but was more colonized by fungi and cyanobacteria. The latter were shown by scanning electron microscopy to grow within the granitic gneiss stone and were probably involved in dissolution and redeposition of minerals. This church is rather far from the intense traffic of the centre of Rio de Janeiro and from Guanabara Bay, source of marine aerosols; it is located upon a green hill, where plant-associated fungi and cyanobacteria can readily gain access to the façade. The results of this study show the importance of local environment on the relative proportions of chemical and biological weathering of stone.

Biosorption of Cadmium by Non-Toxic Extracellular Polymeric Substances (EPS) Synthesized by Bacteria from Marine Intertidal Biofilms.

Cadmium is a major heavy metal found in polluted aquatic environments, mainly derived from industrial production processes. We evaluated the biosorption of solubilized Cd2+ using the extracellular polymeric substances (EPS) produced by Bacillus sp. MC3B-22 and Microbacterium sp. MC3B-10 (Microbactan); these bacteria were originally isolated from intertidal biofilms off the coast of Campeche, Mexico. EPS were incubated with different concentrations of cadmium in ultrapure water. Residual Cd2+ concentrations were determined by Inductive Coupled Plasma-Optic Emission Spectrometry and the maximum sorption capacity (Qmax) was calculated according to the Langmuir model. EPS were characterized by X-ray photoelectron spectroscopy (XPS) before and after sorption. The Qmax of Cd2+ was 97 mg g-1 for Microbactan and 141 mg g-1 for MC3B-22 EPS, these adsorption levels being significantly higher than previously reported for other microbial EPS. In addition, XPS analysis revealed changes in structure of EPS after biosorption and showed that amino functional groups contributed to the binding of Cd2+, unlike other studies that show the carbohydrate fraction is responsible for this activity. This work expands the current view of bacterial species capable of synthesizing EPS with biosorbent potential for cadmium and provides evidence that different chemical moieties, other than carbohydrates, participate in this process.