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A Collection of Living Strains at ACUF (Naples, Italy) Ecological Questions 29 (2018) 3: 63–74 http://dx.doi.org/10.12775/EQ.2018.023 Microorganisms from harsh and extreme environments: a collection of living strains at ACUF (Naples, Italy) Luigi D’Elia1,2, Angelo Del Mondo1*, Mariano Santoro1,3, Antonino De Natale1, Gabriele Pinto1, Antonino Pollio1 1Department of Biology, University of Naples “Federico II”, Via Cinthia 26, 80126 Naples, Italy e-mail: [email protected] 2Department of Chemistry, University of Naples “Federico II”, Via Cinthia 26, 80126 Naples, Italy 3InBioS-Centre for Protein Engineering, University of Liège, Sart Tilman, 4000 Liège, Belgium Received: 28 April 2018 / Accepted: 22 July 2018 Abstract: The Algal Collection at the University Federico II (ACUF) is a bioresource center where over 800 live microalgal strains are maintained, mainly belonging to Cyanobacteria, Chlorophyta, Rhodophyta, and Bacillariophyceae. The extremophilic algae main- tained at ACUF include thermo-acidophilic and acidotolerant strains, mostly belonging to the Cyanidiophyceae isolated from Eu- ropean and extra-European sites, and also terrestrial isolates from bare rocks and monuments. The main target of the ACUF Center is the study and preservation of the diversity of extremophylic microalgae. This collection is used as a resource for studies about biochemical and evolutionary strategies as well as mechanisms involved in cell functioning under harsh environmental conditions. These organisms can be also useful sources for the production of chemical compounds or other biological products with potential biotechnological applications. Keywords: Culture collection, extreme environments, microalgae, biodiversity, biotechnology The ACUF collection article is to present the most significant features of ACUF collection, examples of applied research carried out by the Extreme environments are typically characterized by harsh ACUF staff, strains of interest and future directions in the conditions determined by spatial gradients of chemical and preservation and study of biodiversity. Table 1 provides physical factors, consisting of strong variations in tempera- a complete list of the strains maintained in the ACUF col- tures, humidity, salinity, and pH. They are usually repre- lection classified according to Division, Class, and Order. sented by hot and cold deserts, hot springs, salt lakes, vol- canic and thermal areas, sulfide mines near deep-sea vents as well as terrestrial environments exposed to desiccation History and aims and sharp variations of temperature, as bare rocks, but also building facades and monuments. Extremophilic microor- The culture collection of algae at the Federico II University ganisms are exposed to hostile conditions and are catego- of Naples was started in 1973 by professor Roberto Tad- rized on the basis of their ability to thrive in a specific dei, and was initially planned as a collection of Cyanidium type of niche (Rampelotto, 2013). During the past 45 years caldarium Geitler (sensu lato) strains from different acid- ACUF collection based at the Federico II University of ic-thermal sites of Italy (De Luca et al., 1973) and other Naples (Italy) has supported the study of photoautotrophic countries (De Luca et al., 1977). Further investigations on microorganisms dwelling those habitats. The aim of this acidic and thermal sites of Italy (Fig. 1a and b) led to the 64 Luigi D’Elia, Angelo Del Mondo, Mariano Santoro, Antonino De Natale, Gabriele Pinto, Antonino Pollio Table 1. Taxonomical distribution of the strains presently maintained in the ACUF Collection isolation of acido-resistant species belonging to Chloro- In these explorations, strains from Yellowstone Nation- phyta, Bacillariophyta and other divisions (Pinto and Tad- al Park, Playon de Auachapan (El Salvador), Los Azufres dei, 1977). In the late 1970’s the C. caldarium collection and Cerro Prieto (Mexico) were collected (Gambardella et was expanded to similar sites of different countries, with al., 1980). Following the American explorations, an expe- a focus on Central and North America. dition to Mount Lawu in Java (Indonesia) allowed to col- Microorganisms from harsh and extreme environments: a collection of living strains at ACUF (Naples, Italy) 65 lect Cyanidiophyceae in the type locality of C. caldarium a phenomenon described as desiccation tolerance (Holzinger (De Luca et al., 1981). Afterwards, in 2006 and at a later DQG.DUVWHQ 0LFURELDOELRILOPGHYHORSPHQWFDQEH stage in 2011, Pinto and Ciniglia increased considerably observed on virtually all kinds of stone monuments such the number of Cyanidiophycean strains in the ACUF col- as castles, caves, churches/cathedrals, fountains, temples, lection with the explorations of thermal and acidic sites in tombs/catacombs, etc. (Fig. 1d, e and f), and can be associ- Iceland (Fig. 1c) (Ciniglia et al., 2014) and Turkey. ated with problems of conservation. Several types of auto- trophic and heterotrophic microorganisms such as bacteria, Microalgae from thermoacidic habitats fungi, algae and lichens, are usually observed on stone mon- uments. The rock substratum provides harsh environmental During the years, ecophysiological and ultrastructural stud- conditions. Temperature may vary by several tens of degrees ies were conducted on chlorophycean microalgae isolated during a day, and can be accompanied by rapid desiccation from low pH environments, as Chlamydomonas pitschman- (or freezing); all that implies a limited availability of water nii Ettl (Pollio et al., 2005), Stichococcus bacillaris Nägeli (e.g., Walker and Pace, 2007). The ACUF team character- (Pollio et al., 1997), Pseudococcomyxa simplex (Mainx) ized and studied biofilm communities on stone monuments Fott (Albertano et al., 1990) and Auxenochlorella proto- in the archaeological sites of Campania (Italy), namely thecoides .UJHU .DOLQDHW3XQFRFKiURYi $OEHUWDQRDQG Pompeii, Herculanuem, Oplontis, Cumae and Nola among Taddei, 1984) together with the newly described species others. Microorganisms retrieved on monuments were ana- Viridiella fridericiana Albertano, Pollio, Taddei (Albertano lysed in their native biofilm structure by CLS-microscopy, et al., 1991). Also the chrysophyceaen Ochromonas vulca- isolated, characterized, and subsequently used as models to nia Gromov (Albertano et al., 1994) and the bacillariophy- perform in vitro experiments for understanding the patterns cean Pinnularia obscura .UDVVNH &LQLJOLD HW DO of microbial colonization of stone materials (Marasco et al., thriving in acidic ponds of Southern Italy were collected 2016; Del Mondo et al., 2017). and studied. A major focus was dedicated to Cyanidiophyceae. Biotechnological applications and toxicity tests Unicellular terrestrial Rhodophyta have been considered as a single species for a long time, and described for the Microalgae can be employed in a wide number of bio- first time as Coccochloris orsiniana by Meneghini (1839). chemical and biotechnological applications. In recent Almost a century later, this species had been placed in years, wastewater treatment, biodiesel production, biopoly- a number of already described genera, falling either to the mers and nutraceutical science have become hot topics. Cyanophyta or Chlorophyta, until Geitler assigned to this In response to the uprising energy crisis, climate change species the generally accepted binomial Cyanidium cal- and depletion of natural sources, advantages in the use of darium. The taxon was officially recognized as part of microalgae for biotechnological applications are represent- the Rhodophyta by Hirose (1958), who demonstrated the ed by their capability to grow on non-arable lands nearly presence of several characteristic rhodophycean features. all year long, thus non competing with conventional agri- Later, De Luca et al. (1978) described the new species culture. As a matter of fact, they only require freshwater, Cyanidioschyzon merolae De Luca, Taddei, Varano, fea- non-organic nutrients, and atmospheric CO2. Moreover, tured by its characteristic size and shape, whereas Merola their cultivation may be coupled to industrial processing, et al. (1981) differentiated Galdieria sulphuraria (Galdieri) i.e. reducing overall carbon dioxide emission or for waste- Merola from C. caldarium based on its ability to grow water treatment. Given this premise, collections of extre- in the dark. Currently, macroevolutionary studies integrate mophilic microorganisms allow the selection of strains that aspects of biogeography and geography in a phylogenetic produce high value compounds with specific features as context to answer questions related to the world diffusion thermo-resistance or cryo-resistance; this may also prevent of Cyanidium, Galdieria, and Cyanidioschyzon. Molecular environmental microbial contaminants that would not sur- phylogenetic studies suggest that the Cyanidiales represent vive the selected growth conditions (Ruiz et al., 2016). one of the most ancient groups of algae, having diverged Biofuels production by microalgae appeared promising about 1.3 billion years ago at the base of the Rhodophyta since the end of the last century: bio-oil extracted from (Müller et al., 2001; Yoon et al., 2002b). microalgae may be adopted as crude fuels or may be trans- esterified to biodiesel. In a study conducted by Olivieri Algal biofilms and cultural heritage et al. (2010) the strain ACUF 158 S. bacillaris has been selected as a promising candidate for biofuel production; Although most green algae typically thrive in aquatic en- in fact, this strain is characterized by
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