Benjamín Otto Ortega-Morales

Analysis of Salt-Containing Biofilms on Limestone Buildings of the Mayan Culture at Edzna, Mexico

Biofilms on the salted ceiling of a limestone tunnel at the Mayan site of Edzna, Mexico, were characterised using SEM, EDS, chemical analysis, light microscopy, culture and pigment analysis. Major superficial biomass was pigmented, scytonemin-rich Subsection II cyanobacteria. Main endolithic phototrophs were Subsection I and II cyanobacteria and the alga Nanochlorum. Bacteria and actinomycetes of the Geodermatophilus, nocardioform, and streptomycete groups were present at all levels. Salt crystals, mainly sulfates, were found throughout the stone. Microbial ion transport, metabolite production, chelation, and water retention caused degradation and salt deposits. Exfoliation is the major erosion process.

Antifungal Coatings Based on Ca(OH)2 Mixed with ZnO/TiO2 Nanomaterials for Protection of Limestone Monuments

The presence and deteriorating action of microbial biofilms on historic stone buildings have received considerable attention in the past few years. Among microorganisms, fungi are one of the most damaging groups. In the present work, antimicrobial surfaces were prepared using suspensions of Ca(OH)2 particles, mixed with ZnO or TiO2 nanoparticles. The antimicrobial surfaces were evaluated for their antifungal activity both in the dark and under simulated natural photoperiod cycles, using Penicillium oxalicum and Aspergillus niger as model organisms, and two limestone lithotypes commonly used in construction and as materials for the restoration of historic buildings. Both Ca(OH)2-ZnO and Ca(OH)2-TiO2 materials displayed antifungal activity: ZnO-based systems had the best antifungal properties, being effective both in the dark and under illumination. In contrast, TiO2-based coatings showed antifungal activity only under photoperiod conditions. Controls with coatings consisting of only Ca(OH)2 were readily colonized by both fungi. The antifungal activity was monitored by direct observation with microscope, X-ray diffraction (XRD), and scanning electron microscopy (SEM), and was found to be different for the two lithotypes, suggesting that the mineral grain distribution and porosity played a role in the activity. XRD was used to investigate the formation of biominerals as indicator of the fungal attack of the limestone materials, while SEM illustrated the influence of porosity of both the limestone material and the coatings on the fungal penetration into the limestone. The coated nanosystems based on Ca(OH)2-50%ZnO and pure zincite nanoparticulate films have promising performance on low porosity limestone, showing good antifungal properties against P. oxalicum and A. niger under simulated photoperiod conditions.

Valuable processes and products from marine intertidal microbial communities

Microbial communities are ubiquitous in marine intertidal environments. These communities, which grow preferentially as biofilms on natural and artificial surfaces, carry out key processes contributing to the functioning of coastal environments and providing valuable services to human society, including carbon cycling, primary productivity, trophic linkage, and transfer and removal of pollutants. In addition, their surface-associated life style greatly influences the integrity and performance of marine infrastructure and archaeological heritage materials. The fluctuating conditions of the intertidal zone make it an extreme environment to which intertidal biofilm organisms must adapt at varying levels. This requirement has probably favored the development and spread of specific microorganisms with particular physiological and metabolic processes. These organisms may have potential biotechnological utility, in that they may provide novel secondary metabolites, biopolymers, lipids, and enzymes and even processes for the production of energy in a sustainable manner.

2-acylamino-5-nitro-1,3-thiazoles: preparation and in vitro bioevaluation against four neglected protozoan parasites.

The 2-acylamino-5-nitro-1,3-thiazole derivatives (1-14) were prepared using a one step reaction. All compounds were tested in vitro against four neglected protozoan parasites (Giardia intestinalis, Trichomonas vaginalis, Leishmania amazonensis and Trypanosoma cruzi). Acetamide (9), valeroylamide (10), benzamide (12), methylcarbamate (13) and ethyloxamate (14) derivatives were the most active compounds against G. intestinalis and T. vaginalis, showing nanomolar inhibition. Compound 13 (IC50=10nM), was 536-times more active than metronidazole, and 121-fold more effective than nitazoxanide against G. intestinalis. Compound 14 was 29-times more active than metronidazole and 6.5-fold more potent than nitazoxanide against T. vaginalis. Ureic derivatives 2, 3 and 5 showed moderate activity against L. amazonensis. None of them were active against T. cruzi. Ligand efficiency indexes analysis revealed higher intrinsic quality of the most active 2-acylamino derivatives than nitazoxanide and metronidazole. In silico toxicity profile was also computed for the most active compounds. A very low in vitro mammalian cytotoxicity was obtained for 13 and 14, showing selectivity indexes (SI) of 246,300 and 141,500, respectively. Nitazoxanide showed an excellent leishmanicidal and trypanocidal effect, repurposing this drug as potential new antikinetoplastid parasite compound.

[27] Extracellular polymers of microbial communities colonizing ancient limestone monuments

Publisher Summary This chapter discusses the extracellular polymers of microbial communities colonizing ancient limestone monuments. Biofilms are layered microbial communities growing on inert and livingsurfaces in a variety of terrestrial and aquatic environments, including submerged artificial substrata, invertebrate teguments, sediments, soils, and rock surfaces. The exopolymers, mainly composed of polysaccharides, are an important component of such biofilm communities and their functional roles in terms of attachment, nutrient absorption, and protection against desiccation and antimicrobial agents. Given the anionic substituents that these compounds may have, they can interact actively with dissolved ions in aqueous environments and with framework elements in mineral matrices. The implications of such interactions among exopolysaccharides (EPS) and minerals may also include cation mobilization in soils, solubilization of toxic metals, and the dissolution of minerals. On other hand, studies carried out in Mayan archaeological sites (Yucatan, Mexico) have shown that thick microbial biofilms dominated by cyanobacterial populations contributed to the biodegradation of these buildings by supporting growth of organic acid-producing microorganisms and through active boring. These microbial exopolymers may contribute directly to the deterioration processes by interacting through their polysaccharide fraction with metal cations, which could lead to the complexation of soluble calcium, sequestering it from the limestone matrix. This chapter describes procedures to extract and characterize the polysaccharidic fraction of naturally occurring microbial exopolymers associated with epilithic biofilms.

Emulsifying Activity and Stability of a Non-Toxic Bioemulsifier Synthesized by Microbacterium sp. MC3B-10

A previously reported bacterial bioemulsifier, here termed microbactan, was further analyzed to characterize its lipid component, molecular weight, ionic character and toxicity, along with its bioemulsifying potential for hydrophobic substrates at a range of temperatures, salinities and pH values. Analyses showed that microbactan is a high molecular weight (700 kDa), non-ionic molecule. Gas chromatography of the lipid fraction revealed the presence of palmitic, stearic, and oleic acids; thus microbactan may be considered a glycolipoprotein. Microbactan emulsified aromatic hydrocarbons and oils to various extents; the highest emulsification index was recorded against motor oil (96%). The stability of the microbactan-motor oil emulsion model reached its highest level (94%) at 50 °C, pH 10 and 3.5% NaCl content. It was not toxic to Artemia salina nauplii. Microbactan is, therefore, a non-toxic and non-ionic bioemulsifier of high molecular weight with affinity for a range of oily substrates. Comparative phylogenetic assessment of the 16S rDNA gene of Microbacterium sp. MC3B-10 with genes derived from other marine Microbacterium species suggested that this genus is well represented in coastal zones. The chemical nature and stability of the bioemulsifier suggest its potential application in bioremediation of marine environments and in cosmetics.

Bioweathering Potential of Cultivable Fungi Associated with Semi-Arid Surface Microhabitats of Mayan Buildings

Soil and rock surfaces support microbial communities involved in mineral weathering processes. Using selective isolation, fungi were obtained from limestone surfaces of Mayan monuments in the semi-arid climate at Yucatan, Mexico. A total of 101 isolates representing 53 different taxa were studied. Common fungi such as Fusarium, Pestalotiopsis, Trichoderma, and Penicillium were associated with surfaces and were, probably derived from airborne spores. In contrast, unusual fungi such as Rosellinia, Annulohypoxylon, and Xylaria were predominantly identified from mycelium particles of biofilm biomass. Simulating oligotrophic conditions, agar amended with CaCO3 was inoculated with fungi to test for carbonate activity. A substantial proportion of fungi, in particular those isolated from mycelium (59%), were capable of solubilizing calcium by means of organic acid release, notably oxalic acid as evidenced by ion chromatography. Contrary to our hypothesis, nutrient level was not a variable influencing the CaCO3 solubilization ability among isolates. Particularly active fungi (Annulohypoxylon stygium, Penicillium oxalicum, and Rosellinia sp.) were selected as models for bioweathering experiments with limestone-containing mesocosms to identify if other mineral phases, in addition to oxalates, were linked to bioweathering processes. Fungal biofilms were seen heavily covering the stone surface, while a biomineralized front was also observed at the stone-biofilm interface, where network of hyphae and mycogenic crystals was observed. X-ray diffraction analysis (XRD) identified calcite as the main phase, along with whewellite and wedellite. In addition, lower levels of citrate were detected by Attenuated Total Reflectance-Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Overall, our results suggest that a diverse fungal community is associated with limestone surfaces insemi-arid climates. A subset of this community is geochemically active, excreting organic acids under quasi-oligotrophic conditions, suggesting that the high metabolic cost of exuding organic acids beneficial under nutrient limitation. Oxalic acid release may deteriorate or stabilize limestone surfaces, depending on microclimatic dynamics.

Biomass and taxonomic richness of epilithic cyanobacteria in a tropical intertidal rocky habitat

Abstract The epilithic biofilms colonizing the intertidal and splash zones of a rocky shore in the southern Gulf of Mexico were studied by phase contrast and scanning electron microscopy (SEM), along with phytopigment analyses. These biofilm communities comprised a mixed assemblage of phototrophs, dominated by cyanobacteria, with occasional diatoms and other eukaryotic microalgae. Eight out of nine cyanobacterial genera identified were filamentous. A Stigonema species, a cyanobacterium that rarely occurs in marine habitats, was found in low intertidal samples. Microscopy analysis (phase contrast and SEM) showed that more diverse cyanobacterial populations (morphotypes) occurred on intertidal rocks than in splash zone biofilms, probably owing to the desiccation and thermal stress to which the latter are exposed. However, the phycobiliprotein content, as indicative of cyanobacterial biomass, was similar at both tidal levels. The numerical dominance of Pleurocapsa species and its wide distribution among intertidal and splash zone rocks may be related to its ability to withstand environmental stress by producing moisture-retaining external capsules.

Interactions between abundant fungal species influence the fungal community assemblage on limestone

The assembly of fungal communities on stone materials is mainly influenced by the differential bioreceptivity of such materials and environmental conditions. However, little is known about the role of fungal interactions in the colonization and establishment of fungal species. We analyzed the effects of intra- and interspecific interactions between 11 species of fungi in oligotrophic and copiotrophic media and on limestone coupons. In a previous study, these species were the most frequently isolated in the epilithic biofilms of limestone walls exposed to a subtropical climate. In the culture media, we found a greater frequency of intra- and interspecific inhibitory effects in the oligotrophic medium than in the copiotrophic medium. On the limestone coupons, all fungi were able to establish; however, the colonization success rate varied significantly. Cladosporium cladosporioides had a less extensive colonization in isolation (control) than in dual interactions (coexistence) with other species. Phoma eupyrena exhibited the highest colonization success rate and competitive dominance among all tested species. X-ray diffraction (XRD) and scanning electron microscope (SEM) analyses revealed that Pestalotiopsis maculans and Paraconiothyrium sp. produced calcium oxalate crystals during their growth on coupon surfaces, both in isolation and in dual interactions. Our results demonstrate that interactions between abundant fungal species influence the fungal colonization on substrates, the biomineralization and the fungal community assemblage growing in limestone biofilms.

Antimicrobial Properties of Nanomaterials Used to Control Microbial Colonization of Stone Substrata

Nanoparticle-based materials are applied in the conservation of cultural heritage for their consolidating and self-cleaning abilities. Recently, nanoparticles (NPs) have been found to possess inherent antimicrobial activity, which has stimulated their application in the control of microbial colonization of stone and other mineral materials. A literature survey shows diverse testing procedures and limited research on the antimicrobial effectiveness of nanomaterials under real conditions. Most research reports laboratory-scale studies, employing either mono- or dual species (two organisms) assays over short-term incubation of days or weeks. Antimicrobial effectiveness is often assessed using microbiological, microscopy-based methods and surface colorimetry. There is a potential adverse ecotoxicological impact of NPs after release from treated surfaces. This chapter covers the antimicrobial properties of NPs and their limitations and advantages for application on built cultural heritage.