Introduction • Biodeterioration, bioalternation, biocorrosion, biodecay • Biodeteriogen • Building stone • Cyanobacterial and algal growths • The habitat concept • Floristic studies • Taxonomy • Ecology and autecology • Assemblages • Biodeterioration of stone associated with cyanobacteria and algae • Remarks on control and removal of algae Biodeterioration • (Bio)alternation, (bio)corrosion, (bio)deterioration, (bio)decay • An exchange of material and energy • Two heterogenous open dynamic system: The solid substrate and its atmospheric (indoor and autdoor), aquatic or chthonic environment • The natural limit of turnover activities is given by the penetration depths and speeds of physical gradients, gases, solutions, and organisms (including their extracellular products) into mineral material. Biodeteriogen • We could not debar any organism from being an actual or potential biodeteriogen because of no special criteria for biodeteriogen. • Therefore the range of potential biodeteriogens is huge. • The deteriogenic effects are mainly a minor incidental part of their activities. • The biodeteriogenic organism usually inhabit secondarily an artificial man-made surface and participate on cycling of elements (Winkler, 1994). Building Stone • Stone is the oldest entity of Earth • Characterised by a wide range of mineral compositions, textures, and rock structures • Creates ecosystem which includes also environmental factors (light, nutrition, climate, pollution) • Significant deterioration of monuments has been begun since years 1870-1880, this phenomenon has not been possible to stop since 1950 Cyanobacterial and algal growths • Primarily concerned with the cyanobacteria and algae growing on the exposed surfaces of buildings constructed of man-made materials • Special consideration is given to those obscuring coloured stains on building stone • This account includes the tasks caused by them and the method used for their investigation, control and elimination The Habitat Concept • Most of the cyanobacteria and algae are ubiquitous • Cyanobacterial and algal taxa growing on man-made surfaces • Exactly, it is difficult to define the basis of habitat • Growing in non-aquatic environments have been termed terrestrial, aerial or subaerial forms • Man-made subaerial environment form stressing factors - temperature, dessication, high or low pH, toxicants (pollutants), high or very low irradiance Floristic Studies • More than 200 algal genera had been recorded world wide in floristic studies • The investigation of colonisations of stone by algae • Most species lists include a number of unidentified organisms • Difficult to make exact comparisons of the floristic data • Differences in the homogeneity of the surface sampled, the intensity of sampling • The experiences of collector • The method used for the identification of samples, inconsistencies in the taxonomy of some groups, and in the competence and special interests of the phycologist identifying the algae Taxonomy • subaerial cyanobacteria and algae are of a simple organization, exhibit a simple unicellular or filamentous habit • Cyanobacteria and green algae dominate on subaerial habitat • No systematic effort has been made to distinguish which genera are truly subaerial • Many taxonomic problems Ecology and Autecology • Main question are the adaptations to the subaerial environment • The capacity to tolerance desiccation is a feature of many cyanobacteria and algae of both aquatic and terrestrial origin • The most fundamental feature of all truly aubaerial algae is the ability to resist extremes of temperature, high light intensity and considerable water loss • Many opportunists in subaerial habitats, colonising and growing very fast over wetter or aquatic periods Assemblages • In subaerial habitats some cyanobacteria and algae were typically much more abundant than others • The assemblages were named by using the names of the most abundant forms: the blue-green assemblage, the Trentepohlia assemblage, Desmococcus assemblage, Prasiola assemblage, Stichococcus assemblage Biodeterioration of stone associated with cyanobacteria and algae • Excessive biofilm accumulation in porous substrata changes or reduces their properties or effectiveness • The pedogenic significance of cyanobacteria and algae is very important by biogeophysical weathering of stone surfaces • Mechanical damage caused by active penetration of euendoliths and chemical damage caused by substances and products of metabolism and of extracellular matrix • Aesthetic biodeterioration or soiling Remarks on control and removal of algae • Indirect methods • Reduction of humidity, firing, cover, isolation... • Direct methods • Eliminate crusts, patinas; UV-rays, X-rays, biocide chemicals... The Aims • Floristic screening and taxonomic study of microalgae causing the biological attack on building stone; • Which factor of urban environment (substrata, pollution, humidity, etc.) have an influence on the composition of the subaerial taxa; • To set a growth inhibition test on subaerial microalgae; • To test the microalgae causing actively biodeterioration which potentially damage the building stone. Individual studies forming the results • Gravestones in Cemetery • Tombstone in a historic cemetery • Subterranean Jewish cemetery • The walls of building in Bratislava (SK) • The walls of building in Murcia (ESP) • The walls of building in Brno (CZ) • Growth inhibition test on subaerial microalgae Materials and methods • Research area • The Bratislava City • The Murcia Region • The Brno Region • Sampling • Laboratory cultivation • Analysis for determination • Ultrastructural analysis (SEM) Research area • Bratislava - capital city is located in West Slovakia (48°10´N, 17°10´E) • Murcia - This region is located in South-east Spain (the Murcia City - 36°56´N, 1°07´W) • Brno - city in south Moravia eastern part of Czech Republic Sampling • Different substrata - horizontal distribution • One substrata - vertical distribution • North and south exposition on one building • Spatial distribution in a city Sampling Schema In Laboratory Stone Substrata Underground Historical Monuments Tombstone Laboratory Cultivation • Zehnder medium (Zehnder in Staub, 1961) • BG11 medium (Rippka et al., 1979) • BG11[0] (Rippka, 1988) • BBM (Smith & Bold, 1966) • either liquid or agarised at 20-22 °C • The unialgal strains were used for detail taxonomic study to obtain the data of whole life cycle Analysis for Determination • Type of stone substrata • 10 scores for each stone sample • Structure of algal assemblage • The phenotype features • Original drawings made by camera lucida from field and from cultured material • Olympus BH2 Photomicrography system and Nikon DXM1200 (F) colour camera by LUCIA imaging system (Y/C and composite) – image analysis software developed for image capture, archiving and analysis (© 2004 Laboratory Imaging s.r.o.) Ultrastructural Analysis (SEM) • Pieces of stone substrata were taken from outer walls of monuments and buildings in urban sites. • Samples were fixed, dehydrated, coated with gold as described previously (Garty and Delarea, 1987) • Examined by a JEOL JXA 840A scanning electron microscope (SEM) operating at 200 kV General Results and Discussion • Cca 115 taxa of microalgae • The competitive „first come, first serve“ relationships • Cyanobacteria 35% • Chlorophytes 42% • Chrysophytes 17% • Charophytes 8% • The north-facing or humid side of buildings Cyanobacteria Rhodophyta and Heterokontophyta Chlorophyta Chlorophyta Charophyta Life history SEM-micrographs Original drawings • Life cycle • Vegetative stages • Generative stages • Young habit • Adult habit • Old habit Coccal Cyanobacteria Trichal Cyanobacteria Heterokontophytes Green Algae Charophytes The New Growth Inhibition Test • To develop the techniques of new growth inhibition test • The biotest used on subaerial phototrophic microorganisms (autochtonous, isolated from deteriorated monuments) • A type of growth inhibitor was used white powder called in Spain “el estuco” Experiment Scheme Experiment Realisation The View of The Realisation Results • This material Calcium Ruthenium Oxide Hydrate totally inhibited and eradicated growths of algae • Active interests in developing miniaturized algal bioassays, algal biosensors and development of cultivation tests • Work on subaerial algae caused biodeterioration has highlighted their practical potencial Acknowledgement • Financial support for this study was provided by the Funding of Ministry of Education of Czech Republic; project No. MSM0021622416 • Prof. Dr. František Hindák • Prof. Dr. Marina Aboal Sanjurjo • Dr. Ľubomír Kováčik Thank you for your attention!