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A special issue of Applied Sciences (ISSN 2076-3417). This special issue belongs to the section "Applied Biosciences and Bioengineering".

Deadline for manuscript submissions: closed (15 October 2020) | Viewed by 49419

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Dr. Filomena De Leo

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Guest Editor

Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, 98122 Messina, Italy
Interests: biodeterioration of cultural heritage; applied microbiology; black fungi; molecular microbiology
Special Issues, Collections and Topics in MDPI journals

Dr. Valme Jurado

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Guest Editor

Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, 41012 Sevilla, Spain
Interests: cultural heritage; environmental microbiology; molecular microbiology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

We are inviting manuscript submission for a Special Issue on “Microbial Communities in Cultural Heritage and Their Control”.

Cultural heritage plays a key role in understanding the history of humankind; therefore, adopting appropriate strategies for its conservation is essential. Microorganisms in particular environmental conditions cause serious problems in the conservation of cultural heritage, making the adoption of prevention and conservation strategies critical. Currently, most studies on microbial communities involved in the biodeterioration of cultural heritage rely on molecular identification and provide a list of species, in many cases without reference to their ecological characteristics or metabolic traits. For microbiologists involved in the protection of cultural heritage, this is particularly frustrating, because the identification of microorganisms is not sufficient to discern the origins and causes of a microbial attack, understand dispersion patterns, and prevent further undesired biodeterioration processes. Controlling microorganisms in cultural heritage is a major challenge for microbiologists. The search of appropriate methods to stop progressive microbial invasions without provoking secondary, and perhaps more dangerous, biological successions and biodeterioration is a necessity.

This Special Issue is aimed at publishing outstanding papers presenting cutting-edge advances in the field of cultural heritage and the control of its biodeterioration.

Dr. Filomena De Leo
Dr. Valme Jurado
Guest Editors

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Applied Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

Cultural heritage
Microbiology
Biodeterioration
Fungi
Actinomycetes
Biofilm
Microbial communities
Nanobiocides
Control of biodeterioration
Green chemistry
Conservation

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Related Special Issue

The Role of Fungi in Biodeterioration of Cultural Heritage: New Insights for Their Control in Applied Sciences (10 articles)

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Editorial

Jump to: Research, Review

4 pages, 186 KiB

Open AccessEditorial

Editorial for the Special Issue “Microbial Communities in Cultural Heritage and Their Control”

by Filomena De Leo and Valme Jurado

Appl. Sci. 2021, 11(23), 11411; https://doi.org/10.3390/app112311411 - 2 Dec 2021

Cited by 5 | Viewed by 1475

Abstract

This editorial focuses on the studies published within the present Special Issue presenting advances in the field of biodeterioration of cultural heritage caused by microbial communities with a particular focus on new methods for their elimination and control. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

Research

Jump to: Editorial, Review

14 pages, 7789 KiB

Open AccessArticle

Thymus vulgaris Essential Oil and Hydro-Alcoholic Solutions to Counteract Wooden Artwork Microbial Colonization

by Silvia Sparacello, Giuseppe Gallo, Teresa Faddetta, Bartolomeo Megna, Giovanna Nicotra, Beatrice Bruno, Belinda Giambra and Franco Palla

Appl. Sci. 2021, 11(18), 8704; https://doi.org/10.3390/app11188704 - 18 Sep 2021

Cited by 16 | Viewed by 2593

Abstract

Aromatic plants represent a source of natural products with medicinal properties, and are also utilized in the food and pharmaceutical industries. Recently, the need for eco-compatible and non-toxic products, safe for both the environment and human health, have been proposed for the sustainable conservation of historic–artistic artifacts. In this study, in order to counteract microbial colonization (Aspergillus sp., Streptomyces sp., Micrococcus sp.) on wooden artwork surfaces, Thymus vulgaris L. (Lamiaceae) essential oil (EO) and hydro-alcoholic (HA) solutions were applied in a polyphasic approach. The antimicrobial activities of EO and HA solutions were preliminarily assessed by agar disc diffusion (ADD) and well plate diffusion (WPD) in vitro methods, defining the specific concentration useful for bacterial and fungal genera, identified by optical microscopies, in vitro cultures (nutrient or Sabouraud agar), and DNA base molecular biology investigations. Specifically, the microbial patina was directly removed by a hydro-alcoholic solution (while evaluating the potential colorimetric change of the artwork’s surface) combined with exposure to EO volatile compounds, performed in a dedicated “clean chamber”. This study proposes, for the first time, the combined use of two plant extracts to counteract microbial development on wooden artworks, providing supplementary information on these products as bio-agents. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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18 pages, 3646 KiB

Open AccessArticle

High Dimension Granite Pavement Bio-Desalination Practical Implementation

by Pilar Bosch-Roig, Lourdes Pérez-Castro, Ángeles Fernández-Santiago and Ignacio Bosch

Appl. Sci. 2021, 11(14), 6458; https://doi.org/10.3390/app11146458 - 13 Jul 2021

Cited by 3 | Viewed by 2087

Abstract

Biocleaning technology is based on the use of safe environmental microorganisms for green cultural heritage (CH) restoration. Compared with traditional cleaning products, this biological technique is very specific, effective, and nontoxic. This innovative biotechnological application has been used for recovering diverse monuments and artworks. Most CH in situ surfaces that are treated with microorganisms are small areas; however, some important pathologies, such as salt contamination, can affect high dimension artistic surfaces. The purpose of this study is to analyze and overcome the problems and limitations of scaling up the bio-desalination protocol for in situ applications. Three water-based gel delivery systems and three heating systems were tested in situ and evaluated in terms of performance difficulty, efficacy, and costs. The tests were carried out on the salt contaminated granite pavement of Cristo Chapel of Sta Ma de Conxo in Santiago de Compostela (Spain). Ground agar 2% and a heating electric mat were selected as the best performing systems. The implemented protocol was applied for the bio-desalination of the 233 m² Chapel pavement. Conductivity, nitrate–nitrite measurements, biological monitoring, and digital image analysis were performed to determine the efficacy of the treatment. This research allowed for the development of an innovative and optimized in situ, high dimension bio-desalination application protocol transferable to other large scale, in situ biocleaning strategies. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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12 pages, 598 KiB

Open AccessArticle

An In Vitro Evaluation of the Biocidal Effect of Oregano and Cloves’ Volatile Compounds against Microorganisms Colonizing an Oil Painting—A Pioneer Study

by Lucrezia Gatti, Federica Troiano, Violetta Vacchini, Francesca Cappitelli and Annalisa Balloi

Appl. Sci. 2021, 11(1), 78; https://doi.org/10.3390/app11010078 - 24 Dec 2020

Cited by 28 | Viewed by 3835

Abstract

In this study, the biocidal activity of two plant derivatives (oregano and cloves’ essential oils—EOs) was evaluated, as a potential innovative and eco-friendly cleaning method for canvas paintings. The object of the study was the oil painting on canvas entitled “Studio di nudo” (Giovanni Maria Mossa, 1921), showing stains caused by microorganisms. The research focused on: (1) isolation and identification of microorganisms associated with discolorations on the obverse and reverse sides of the canvas; (2) evaluation of biocidal activity of selected EOs against fungal and bacterial collections. The phylogenetic identification was conducted with both cultivation and molecular methods. The canvas was mainly colonized by Penicillium, Aspergillus, and Cephaloteca fungal genera and by bacteria of the Bacillus genus. To evaluate the biocidal effect of the EOs’ volatile components only, an antibiogram assay (agar disc diffusion method) and a customized assay (named the contactless test) were conducted. Tested EOs showed antimicrobial activity on fungi and bacteria. However, compared to cloves, oregano EO exhibited a better inhibition activity both in contact and contactless tests. The work is pioneering for the use of EOs’ volatile compounds against oil painting biodeteriogens, and gives insights into possible extended, innovative and eco-friendly cleaning methods for painting control procedures. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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15 pages, 3119 KiB

Open AccessArticle

Varnishes with Biocidal Activity: A New Approach to Protecting Artworks

by Julio Romero-Noguera, Rafael Bailón-Moreno and Fernando Bolívar-Galiano

Appl. Sci. 2020, 10(20), 7319; https://doi.org/10.3390/app10207319 - 19 Oct 2020

Cited by 7 | Viewed by 2708

Abstract

Keeping agents that cause biodeterioration under control is a common task in restoring and conserving artworks. However, there are very few studies, specifically those concentrating on pictorial works, that provide solutions compatible with the complex mix of organic and inorganic materials to be found in such cultural assets. This study aims to use biocide products that are commonly used in this field (benzalkonium chloride, o-phenylphenol, and tributyltin naphthenate) and which are associated with natural varnishes with a long tradition in artistic practices, in order to protect paintings and polychrome sculptures from environmental fungi and bacteria without altering the original materials or the visual appearance of the artworks. The effectiveness or innocuousness of the treatments was tested chemically and visually via optical microscopy, gas chromatography–mass spectrometry (GC-MS), and a multidimensional scaling analysis (MDS); the treatments produced good results as regards the inhibition of microbial growth and scarce interaction with the artistic materials being studied. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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15 pages, 983 KiB

Open AccessArticle

Assessment of Bacterial Contamination of Air at the Museum of King John III’s Palace at Wilanow (Warsaw, Poland): Selection of an Optimal Growth Medium for Analyzing Airborne Bacteria Diversity

by Mikolaj Dziurzynski, Karol Ciuchcinski, Magdalena Dyda, Anna Szych, Paulina Drabik, Agnieszka Laudy and Lukasz Dziewit

Appl. Sci. 2020, 10(20), 7128; https://doi.org/10.3390/app10207128 - 13 Oct 2020

Cited by 13 | Viewed by 3167

Abstract

There is no standardized protocol for the assessment of microbial air contamination in museums and other cultural heritage sites. Therefore, most museums conduct such assessments based on their own guidelines or good practices. Usually, microbial air contamination is assessed using only classical microbiology methods with the application of a single growth medium. Therefore, this medium should be carefully selected to limit any selective cultivation bias. Metabarcoding, i.e., a next-generation sequencing (NGS)-based method, combined with classical microbiological culturing was used to assess the effectiveness of various media applications in microbiological screening at the Museum of King John III’s Palace at Wilanow (Warsaw, Poland). The obtained results indicated that when using a classical microbiology approach to assess the microbial air contamination at the museum, the selection of a proper growth medium was critical. It was shown that the use of rich media (commonly applied by museum conservators) introduced significant bias by severely underreporting putative human pathogens and the bacterial species involved in biodeterioration. Therefore, we recommend the use of other media, such as Frazier or Reasoner’s 2A (R2A) medium, as they could yield more diverse communities and recovered the highest number of genera containing human pathogens, which may be suitable for public health assessments. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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18 pages, 3458 KiB

Open AccessArticle

Frequent Microalgae in the Fountains of the Alhambra and Generalife: Identification and Creation of a Culture Collection

by Fernando Bolívar-Galiano, Clara Abad-Ruiz, Pedro Sánchez-Castillo, Maurizio Toscano and Julio Romero-Noguera

Appl. Sci. 2020, 10(18), 6603; https://doi.org/10.3390/app10186603 - 21 Sep 2020

Cited by 4 | Viewed by 4002

Abstract

Cyanobacteria, green algae and diatoms are significant factors in the biodeterioration of stone cultural heritage sites, and specifically fountain monuments, due to the constant presence of water. In this study, samples were taken from different fountains in the Alhambra and Generalife, which are among the Spanish monuments of greatest historical and artistic value and which together were declared a World Heritage Site by UNESCO in 1984. The aim was to identify which species of colonising microalgae are most frequent and to obtain monoalgal cultures from them. From a conservation point of view, it is interesting to identify which algae are growing in these fountains and how they behave in order to develop new methods to control their growth. The most abundant groups of algae in our samples were green algae and cyanobacteria. The most common genera in the former group were Bracteacoccus, Chlorosarcina, Chlorosarcinopsis, Apatococcus and Klebsormidium. As for cyanobacteria, the most abundant genera were Phormidium, Calothrix, Leptolyngbya, Chamaesiphon, Pleurocapsa and Chlorogloea. Using our collected samples, 10 genera of green algae and 13 genera of cyanobacteria were isolated, thereby constituting the base samples for the creation of a reference collection of living algae from the Alhambra and Generalife contexts, which can be used in subsequent studies to develop new types of treatment against biodeterioration. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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15 pages, 8295 KiB

Open AccessFeature PaperArticle

Biocidal Activity of Phyto-Derivative Products Used on Phototrophic Biofilms Growing on Stone Surfaces of the Domus Aurea in Rome (Italy)

by Lorenza Rugnini, Giada Migliore, Flavia Tasso, Neil Thomas William Ellwood, Anna Rosa Sprocati and Laura Bruno

Appl. Sci. 2020, 10(18), 6584; https://doi.org/10.3390/app10186584 - 21 Sep 2020

Cited by 17 | Viewed by 3143

Abstract

Hypogean or enclosed monuments are important cultural heritage sites that can suffer biodegradation. Many of the stone walls of the prestigious Domus Aurea are overwhelmed by dense biofilms and so need intervention. Room 93 was chosen as a study site with the aim to test the efficacy of phyto-derivatives as new biocides. Laboratory studies were performed comparing the effects of liquorice leaf extract (Glycyrrhiza glabra L.), lavender essential oil (Lavandula angustifolia Mill.) and a combination of both. In situ studies were also performed to test the effect of liquorice. The results were compared with those of the commonly used synthetic biocide benzalkonium chloride. The effects on the biofilms were assessed by microscopy along with chlorophyll fluorescence analysis. The phototrophs in the biofilms were identified morphologically, while the heterotrophs were identified with culture analysis and 16S gene sequencing. Results showed that the mixed solution liquorice/lavender was the most effective in inhibiting the photosynthetic activities of biofilms in the laboratory tests; while, in situ, the effect of liquorice was particularly encouraging as an efficient and low-invasive biocide. The results demonstrate a high potential biocidal efficacy of the phyto-derivatives, but also highlight the need to develop an efficient application regime. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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19 pages, 4213 KiB

Open AccessFeature PaperArticle

Biodeteriogens Characterization and Molecular Analyses of Diverse Funeral Accessories from XVII Century

by Zuzana Kisová, Matej Planý, Jelena Pavlović, Mária Bučková, Andrea Puškárová, Lucia Kraková, Magdaléna Kapustová, Domenico Pangallo and Katarína Šoltys

Appl. Sci. 2020, 10(16), 5451; https://doi.org/10.3390/app10165451 - 6 Aug 2020

Cited by 13 | Viewed by 3557

Abstract

A historical crypt offers us a particular view of the conditions of some buried materials (in this case textiles) and the various biogenic phenomena to which they were subjected over the centuries. In addition, significant knowledge can come by studying the DNA of buried objects which allows the recognition of materials, but also to reveal some practice of the funeral ceremony. In this study, the deteriorating microbial communities colonizing various funeral textile items were identified and characterized using microscopic observation, cultivation, polymerase chain reaction (PCR) and sequencing, hydrolytic tests; and culture-independent analysis (high-throughput sequencing, MinION platform). Different PCR assays and consequent sequencing of amplicons were employed to recognize the animal origin of bodice reinforcements and the type of plant used to embellish the young girl. The analysis of ancient DNA (aDNA from animal and plant) was also completed by the application of high-throughput sequencing through Illumina platform. The combination of all these techniques permitted the identification of a complex microbiota composed by dangerous degradative microorganisms able to hydrolyze various organic substrates such as fibroin, keratin, and cellulose. Bacteria responsible for metal corrosion and bio-mineralization, and entomopathogenic and phytopathogenic fungi. The analysis of aDNA identified the animal component used in bodice manufacturing, the plant utilized as ornament and probably the season of this fatal event. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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13 pages, 1652 KiB

Open AccessArticle

Fungal Community Analysis and Biodeterioration of Waterlogged Wooden Lacquerware from the Nanhai No. 1 Shipwreck

by Yin Jia, Liuyu Yin, Fengyu Zhang, Mei Wang, Mingliang Sun, Cuiting Hu, Zijun Liu, Yue Chen, Jie Liu and Jiao Pan

Appl. Sci. 2020, 10(11), 3797; https://doi.org/10.3390/app10113797 - 29 May 2020

Cited by 10 | Viewed by 3285

Abstract

To avoid the lacquerware of the Nanhai No. 1 shipwreck from being corroded by microorganisms and to improve the knowledge on microbial ecology of the wood lacquers, we conducted a series of tests on the two water samples storing the lacquerware and colonies on the surface of the lacquerware. The high-throughput sequencing detected dominant fungal communities. After that, the fungal strains were isolated and then identified by amplification of ITS- 18S rRNA. Then the activity of ligninolytic and cellulolytic enzymes was detected on potato dextrose agar (PDA) plates with 0.04% (v/v) guaiacol and carboxymethyl cellulose (CMC) agar plates. Finally, we tested the biocide susceptibility of these fungi. Penicillium chrysogenum (NK-NH3) and Fusarium solani (NK- NH1) were the dominant fungi in the sample collected in April 2016 and June 2017. What is more, both showed activity of ligninolytic and cellulolytic enzymes. Four biocidal products (Preventol^® D7, P91, BIT 20N, and Euxyl^® K100) inhibited the growth of the fungal species in vitro effectively. In further research, the microbial community and environmental parameters in the museum should be monitored to assess the changes in the community and to detect potential microbial outbreaks. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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13 pages, 2055 KiB

Open AccessFeature PaperArticle

Biological Control of Phototrophic Biofilms in a Show Cave: The Case of Nerja Cave

by Valme Jurado, Yolanda del Rosal, Jose Luis Gonzalez-Pimentel, Bernardo Hermosin and Cesareo Saiz-Jimenez

Appl. Sci. 2020, 10(10), 3448; https://doi.org/10.3390/app10103448 - 16 May 2020

Cited by 26 | Viewed by 3540

Abstract

Cyanobacteria and microalgae are usually found in speleothems, rocks and walls of show caves exposed to artificial lighting. These microorganisms develop as biofilms coating the mineral surfaces and producing aesthetic, physical and chemical deterioration. A wide number of physical, chemical and environmental-friendly methods have been used for controlling the biofilms with different results. Natural biological control has been suggested by some authors as a theoretical approach but without direct evidence or application. Here we report the finding of a natural biological control of phototrophic biofilms on the speleothems of Nerja Cave, Malaga, Spain. The formation of plaques or spots where the phototrophic microorganisms disappeared can be assumed on the basis of processes of predation of bacteria, amoebas and some other organisms on the phototrophic biofilms. This study aims at investigating the potentialities of the biological control of phototrophic biofilms in caves, but the originality of these data should be confirmed in future studies with a larger number of biofilm samples in different ecological scenarios. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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Review

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15 pages, 2005 KiB

Open AccessReview

A Review on Sampling Techniques and Analytical Methods for Microbiota of Cultural Properties and Historical Architecture

by Xinghua Ding, Wensheng Lan and Ji-Dong Gu

Appl. Sci. 2020, 10(22), 8099; https://doi.org/10.3390/app10228099 - 16 Nov 2020

Cited by 23 | Viewed by 5496

Abstract

World cultural heritage suffers from deterioration caused by both natural and anthropogenic processes, among which microorganisms are significantly involved. Among the key issues of this topic, sampling techniques and analytical methods for revealing the microbiome are fundamental to obtaining useful results for understanding the key players and processes involved, and also for effective protection and management of the cultural heritage for humanity. A non-invasive and non-destructive sampling method is required for sampling of cultural properties prior to further analysis of the microbiome. One example is illustrated in this article. For many years, culture-dependent methods had been used before the invention of polymerase-chain reaction (PCR) methods and, more recently, specifically high-throughput next generation sequencing (NGS). NGS reveals the whole microbial community composition and the active microorganisms from genomic DNA and RNA, respectively. The recovered environmental DNA and RNA from samples provide the information on microbial community and composition, and the active members and biochemical processes of the microbial attributes. It should be emphasized that the metabolically-active members of functional microflora in the biofilm or microbiome on cultural heritage must be determined and identified from the RNA-based analysis to gain a substantially important insight of the active biodeterioration processes and also the effectiveness of the conservation strategies. The importance of the culture-independent technique, based on NGS, is that it can be used in combination with the conventional culturing methods to guide the isolation and enrichment of new microorganisms to gain further biochemical insights to advance the role of the specific microbial groups for biodeterioration of cultural heritage. At the same time, effective restoration and maintenance strategies can be formulated for the protection of world cultural heritage. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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17 pages, 4305 KiB

Open AccessReview

Present and Future Perspectives for Biocides and Antifouling Products for Stone-Built Cultural Heritage: Ionic Liquids as a Challenging Alternative

by Sandra Lo Schiavo, Filomena De Leo and Clara Urzì

Appl. Sci. 2020, 10(18), 6568; https://doi.org/10.3390/app10186568 - 20 Sep 2020

Cited by 48 | Viewed by 4475

Abstract

This review offers an overview of the most recent research activities on counteracting the biodeterioration process of stone monuments, underlining all those aspects regarding eventual procedural drawbacks and compliance with sustainable criteria. For this purpose, the definition of “green conservation of cultural heritage” has been proposed. Its basics have been utilized in the text to highlight the issues arising from the most common conservative procedures as well as guidelines for the development of innovative technologies. The review then deals with the most innovative antimicrobial approaches, among which nano- and bio-technologies play a main role. Ionic liquids are a special class of salts, which can be prepared by applying Safe by Design concepts, to meet the Green Conservation criteria. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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25 pages, 2660 KiB

Open AccessReview

Biological Risk for Hypogea: Shared Data from Etruscan Tombs in Italy and Ancient Tombs of the Baekje Dynasty in Republic of Korea

by Giulia Caneva, Daniela Isola, Hyun Ju Lee and Yong Jae Chung

Appl. Sci. 2020, 10(17), 6104; https://doi.org/10.3390/app10176104 - 2 Sep 2020

Cited by 33 | Viewed by 4487

Abstract

Biological growth represents one of the main threats for the conservation of subterranean cultural heritage. Knowledge of the conditions which favour the various taxonomic groups is important in delineating their control methods. Combining our experience regarding hypogea in Italy and the Republic of Korea, we aim to perform a critical review and comparison of the Biodeterioration Patterns (BPs) found, the materials used, and the conservative treatments applied. For this purpose, we focused on Etruscan tombs (Italy, 7th to 3th century BC) and the ancient tombs of the Baekje Dynasty (Republic of Korea, 6th to 7th centuries AD), most of which have been designated UNESCO World Heritage Sites, collecting original and bibliographic data as well as official documents. Results highlight the rich biodiversity of the bacterial and fungal species. Phototrophs were observed only in niches with sufficient light and the development of roots was also detected. Changes in humidity and temperature, the nature of the soil, nutrient accumulation, and vegetation above the hypogea along with human activities explain the different BPs. The effects of biocide treatments are also discussed, such as the emergence of dangerous fungal species. The shared data also enhance the role of overlaying tumuli and vegetation as well as protective barriers to reduce biological risk. Full article

(This article belongs to the Special Issue Microbial Communities in Cultural Heritage and Their Control)

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