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.

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.

Growth of the Black Soldier Fly Hermetia illucens (Diptera: Stratiomyidae) on Organic-Waste Residues and Its Application as Supplementary Diet for Nile Tilapia Oreochromis niloticus (Perciformes: Cichlidae).

The black soldier fly, Hermetia illucens (BSF, Diptera: Stratiomyidae) is an insect with high protein value and a potential feed agent for animals aimed for human consumption. The growth parameters of BSF larvae reared on four substrates-restaurant-waste, fruit-waste, fish-waste, and commercial tilapia food-for 41 days before processing for inclusion into Oreochromis niloticus (Perciformes: Cichlidae, Nile tilapia) commercial fry diets at 30% (70:30) were determined. On fly larvae, the food substrate based on restaurant waste yielded the greatest larval weight and length. BSF larvae fed a fish-waste diet showed the shortest developmental time. The fruit-waste diet induced the lowest weight and length in the fly larvae/pre-pupae (immature stage). The pre-pupal protein values were similar to commercial food. On fry-fish, the diets with pre-pupae grown on fish waste showed the greatest yields regarding weight (biomass), length, and nutritional content. These results suggest the BSF has the potential to be used in fish feed and provides an alternative for commercial cultivation.

Microorganisms as a Potential Source of Molecules to Control Trypanosomatid Diseases.

Trypanosomatids are the causative agents of leishmaniasis and trypanosomiasis, which affect about 20 million people in the world's poorest countries, leading to 95,000 deaths per year. They are often associated with malnutrition, weak immune systems, low quality housing, and population migration. They are generally recognized as neglected tropical diseases. New drugs against these parasitic protozoa are urgently needed to counteract drug resistance, toxicity, and the high cost of commercially available drugs. Microbial bioprospecting for new molecules may play a crucial role in developing a new generation of antiparasitic drugs. This article reviews the current state of the available literature on chemically defined metabolites of microbial origin that have demonstrated antitrypanosomatid activity. In this review, bacterial and fungal metabolites are presented; they originate from a range of microorganisms, including cyanobacteria, heterotrophic bacteria, and filamentous fungi. We hope to provide a useful overview for future research to identify hits that may become the lead compounds needed to accelerate the discovery of new drugs against trypanosomatids.

Bactericidal Activity of Chrysomya rufifacies and Cochliomyia macellaria (Diptera: Calliphoridae) Larval Excretions-Secretions Against Staphylococcus aureus (Bacillales: Staphylococcaceae).

The inhibitory effect of Chrysomya rufifacies (Macquart) and Cochliomyia macellaria (Fabricius) larval excretions-secretions (ES) on Staphylococcus aureus was determined using a portable colorimetric method without the need for any dedicated spectral instrument. Polystyrene 96 well microplates were used and 100 µl of the bacterial inoculum (5 × 105 CFU/ml) plus 100 µl of the dipteran exosecretions at different concentrations were added to each well. Subsequently, 50 µl of a 1% solution of the triphenyl tetrazolium chloride stain was added to each well to determine the bacterial viability. The color development in each well was measured with the ImageJ software S. aureus was exposed to different concentrations of the ES of both species individually. At a concentration of 800 ppm ES of C. rufifacies or Co. macellaria, bacterial growth was inhibited 97.45 ± 1.70% and 82.21 ± 1.88%, respectively. As expected, exposure to a lower concentration (i.e., 50 ppm) was less inhibitory (C. rufifacies ES, 77.65 ± 4.25% and Co. macellaria ES, 43.54 ± 4.63%). This study demonstrates for the first time the bactericidal activity of C. rufifacies and Co. macellaria ES against S. aureus. This finding is promising as it could result in the identification and synthesis of proteins capable of suppressing pathogen development in wounds. Additionally, the proposed method can simplify the use of expensive laboratory instruments for antimicrobial activity determination.

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.

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.

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.

Synthesis, in vitro and in vivo giardicidal activity of nitrothiazole-NSAID chimeras displaying broad antiprotozoal spectrum.

We designed and synthesized five new 5-nitrothiazole-NSAID chimeras as analogues of nitazoxanide, using a DCC-activated amidation. Compounds 1-5 were tested in vitro against a panel of five protozoa: 2 amitochondriates (Giardia intestinalis, Trichomonas vaginalis) and 3 kinetoplastids (Leishmania mexicana, Leishmania amazonensis and Trypanosoma cruzi). All chimeras showed broad spectrum and potent antiprotozoal activities, with IC50 values ranging from the low micromolar to nanomolar order. Compounds 1-5 were even more active than metronidazole and nitazoxanide, two marketed first-line drugs against giardiasis. In particular, compound 4 (an indomethacin hybrid) was one of the most potent of the series, inhibiting G. intestinalis growth in vitro with an IC50 of 0.145µM. Compound 4 was 38-times more potent than metronidazole and 8-times more active than nitazoxanide. The in vivo giardicidal effect of 4 was evaluated in a CD-1 mouse model obtaining a median effective dose of 1.709µg/kg (3.53nmol/kg), a 321-fold and 1015-fold increase in effectiveness after intragastric administration over metronidazole and nitazoxanide, respectively. Compounds 1 and 3 (hybrids of ibuprofen and clofibric acid), showed potent giardicidal activities in the in vitro as well as in the in vivo assays after oral administration. Therefore, compounds 1-5 constitute promising drug candidates for further testing in experimental chemotherapy against giardiasis, trichomoniasis, leishmaniasis and even trypanosomiasis infections.

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.

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.

Succession of fungi colonizing porous and compact limestone exposed to subtropical environments.

Little is known about the dynamics of succession of fungi on limestone exposed in subtropical environments. In this study, the colonization of experimental blocks of compact and porous limestone by a fungal community derived from natural biofilms occurring on Structure X from the archaeological site of Becán (México), was studied using a cultivation-dependent approach after short-term (9 m) exposure in order to provide a preliminary insight of the colonization process under seminatural conditions. Microbial growth seen as the change of colour of stone surfaces to black/dark green was more abundant on the porous limestone. There was a fairly clear difference in microbial colonization between the onset of the experiment and the 6th month for both limestone types, but no significant increase in the colonization of coupons occurred between months 6 and 9. This could be related to the low rainfall expected for this period, corresponding to the dry season. A total of 977 isolates were obtained. From these, 138 sterile fungi were unidentified, 380 could only be assigned to the order Sphaeropsidales; the remaining isolates (459) were grouped into 27 genera and 99 different species. Nearly all detected fungal species belonged to the Ascomycota (90 %). Rare taxa (species represented by one to three isolates) included the recently described genus Elasticomyces, several species of genera Hyalodendron, Monodyctis, Papulospora, Curvularia, and Septoria. Other taxa were Minimedusa and Gliomastix luzulae, which have not been previously described for stone environments. Abundant fungi included several species of the common genera Cladosporium, Alternaria, and Taeniolella typical for a range of habitats. Succession of populations was observed for certain taxa, this shift in the composition of fungal communities was more evident in porous limestone. After 6 m of exposure, species of the genera Scolecobasidium, Hyalodendron, and Taeniolella were predominant, while after 9 m, the predominant species belonged to the genera Curvularia and Alternaria, particularly on porous stone. These results suggest that Curvularia and Alternaria replaced other fungi, due to a higher tolerance towards low levels of available water during the dry season. Higher levels of water within the porous stone, keep longer periods of microbial activity, minimizing the impact of desiccation. This study contributes to understand the diversity of fungal communities in stone surfaces in subtropical settings and the dynamics of colonization on limestone.

Culturable fungi associated with urban stone surfaces in Mexico City

Background: Urban surface stones in Mexico City are exposed to a temperate climate and a range of atmospheric conditions ranging from mildly impacted to heavily polluted areas. In this study, we focused on the characterization of the cultivable fungal component of selected biological patinas in the surrounding area of Chapultepec castle, a historic monument in Mexico City. Thirty four representative fungal isolates selected based on distinctive differential macroscopic characteristics out of a total of 300 fungi, were characterized using morphological and molecular approaches. Results: This identification strategy based on the combination of phenotypic- and molecular-based methodologies allowed us to discriminate the fungal community in some cases down to the species level. Conclusions: The characterization of this mycoflora revealed the presence of a complex fungal community mainly represented by filamentous fungi belonging to the genera Fusarium, Trichoderma, Aspergillus, Cladosporium, Alternaria, Mucor, Penicillium, Pestalotiopsis, and the dimorphic fungus Aureobasidium, along with the yeast Rhodotorula. A specific distribution of fungi could be observed based on the type of biological patina analyzed.

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.

Antifouling activity of sessile bacilli derived from marine surfaces.

Marine biofilms are a virtually untapped source of bioactive molecules that may find application as novel antifoulants in the marine paint industry. This study aimed at determining the potential of marine biofilm bacteria to produce novel biomolecules with potential application as natural antifoulants. Nine representative strains were isolated from a range of surfaces and were grown in YEB medium and harvested during the late exponential growth phase. Bacterial biomass and spent culture medium were extracted with ethanol and ethyl acetate, respectively. Extracts were assayed for their antifouling activity using two tests: (1) antimicrobial well diffusion test against a common fouling bacterium, Halomonas marina, and (2) anti-crustacean activity test using Artemia salina. Our results showed that none of the ethanolic extracts (bacterial biomass) were active in either test. In contrast, most of the organic extracts had antimicrobial activity (88%) and were toxic towards A. salina (67%). Sequencing of full 16 S ribosomal DNA analysis showed that the isolates were related to Bacillus mojavensis and Bacillus firmus. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) profiling of ethyl acetate extracts of culture supernatants showed that these species produce the bioactive lipopeptides surfactin A, mycosubtilin and bacillomycin D.

Biofilm colonization dynamics and its influence on the corrosion resistance of austenitic UNS S31603 stainless steel exposed to Gulf of Mexico seawater.

Viable bacterial counts, chemical markers, phospholipid fatty acid analysis (PLFA), and Fourier-transformed infrared spectroscopy (FTIR), together with electrochemical methods, were used to study biofilm dynamics and its impact on the corrosion resistance of UNS S31603 stainless steels exposed to the Gulf of Mexico seawater. Biofilms progressively accumulated, peaking on day 20, but finally detached. The extracellular polysaccharide (EPS)/cellular biomass ratio remained low most of the time, but reached its highest level (4.2+/-1.9) also on day 20. Viable bacterial cells reached their highest abundance earlier (approximately 800 CFU/cm2), on day 15. Biofilms were seen covering the stainless steel surfaces heterogeneously and were composed mainly of gram-negative rods, presumably EPS-producing bacteria. Despite the different levels of biofilm biomass and attachment state, field-exposed steel coupons ennobled significantly and showed more active pitting potentials (approximately +500 mVSCE) than on the abiotic control (+650 mVSCE), where no significant ennoblement occurred. These results suggest that the heterogeneous distribution of biofilms, as opposed to the quantity of surface-associated biomass, promotes formation of differential aeration cells, and that this in turn contributes to the ennoblement of these steels.

Cyanobacterial diversity and ecology on historic monuments in Latin America.

Cyanobacterial biofilms are complex communities of microorganisms that cause damaging activity on historic monuments. A combined molecular approach shows that cyanobacteria belonging to the order Pleurocapsales are the main colonizers at the Mayan site of Uxmal, Mexico, confirming previous microscopic and culture-based reports. An important, previously unrecognized non-cyanobacterial community comprising Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes has also been found in Uxmal. Cyanobacterial communities in Palenque were composed of over 10 species, mainly coccoid forms. A novel PCR method designed to directly amplify DNA from uncultured cyanobacterial cells on historic buildings in Brazil indicated that the identified cyanobacteria sequenced corresponded to their appropriate morphological groups (as defined by both the bacterial and botanical codes). However, their homologies with deposited sequences were, in general, low. Terrestrial cyanobacteria from stone surfaces in Brazil, again mainly coccoid, formed a distinct population that differed from the better-studied aquatic members. Overall, results here show demonstrate that coccoid cyanobacteria are the main colonizers on Latin American monuments under tropical and subtropical conditions and the assessment of their potential deteriogenic activity requires the further development of rapid molecular techniques. Polyphasic studies are essential to increase our knowledge of the diversity of terrestrial biofilms and of global microbial diversity.