Diversity and bioactivity of fungi associated with the marine sea cucumber Holothuria poli: disclosing the strains potential for biomedical applications.

AIMS: Identification of the mycobiota associated to the marine echinoderm Holothuria poli and investigation of cytotoxic and pro-osteogenic potential of isolated strains. METHODS AND RESULTS: Fungal strains were isolated from the animal's body-wall, intestine and faeces. The species identification was based on DNA barcoding and morphophysiological observations. Forty-seven species were identified, all are Ascomycota and mainly belonging to Aspergillus and Penicillium genera. Sixteen strains were grown on three media for chemical extraction. Cytotoxic activity was tested on a hepatic cancer cell line (HepG2), the cells viability was evaluated after treatment using a resazurin based assay (AlamarBlue). Pro-osteogenic activity was tested on human Mesenchymal stem cell, differentiation was measured as the alkaline phosphatase production through reaction with p-nitrophenylphosphate or as the cells ability to mineralize calcium using a colorimetric kit (StanBio). Cytotoxic activity was recorded for four fungal species while five of 48 extracts highlighted bioactivity towards human mesenchymal stem cells. CONCLUSIONS: The presence of relevant animal-associated mycobiota was observed in H. poli and selected strains showed cytotoxic potential and pro-osteogenic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Our work represents the first report of a Mediterranean Sea cucumber mycobiota and highlights the isolates potential to synthetize compounds of pharmaceutical interest for regenerative medicine.

First Isolation of Pseudogymnoascus destructans, the Fungal Causative Agent of White-Nose Disease, in Bats from Italy.

White-nose disease, caused by the dermatophyte Pseudogymnoascus destructans, is a devastating pathology that has caused a massive decline in the US bat populations. In Europe, this fungus and the related infection in bats have been recorded in several countries and for many bat species, although no mass mortality has been detected. This study reports for the first time the presence of P. destructans in Italy. The fungus was isolated in the Rio Martino cave, a site located in the Western Alps and included in the Natura 2000 network. Twenty bats, belonging to five different species, were analysed. The fungus was retrieved on eight individuals of Myotis emarginatus. The allied keratolytic species P. pannorum was observed on two other individuals, also belonging to M. emarginatus. Strains were isolated in pure culture and characterized morphologically. Results were validated through molecular analyses. Future work should be dedicated to understand the distribution and the effects of the two Pseudogymnoascus species on Italian bats.

The culturable mycobiota associated with three Atlantic sponges, including two new species: Thelebolus balaustiformis and T. spongiae.

Covering 70 % of Earth, oceans are at the same time the most common and the environment least studied by microbiologists. Considering the large gaps in our knowledge on the presence of marine fungi in the oceans, the aim of this research was to isolate and identify the culturable fungal community within three species of sponges, namely Dysidea fragilis, Pachymatisma johnstonia and Sycon ciliatum, collected in the Atlantic Ocean and never studied for their associated mycobiota. Applying different isolation methods, incubation temperatures and media, and attempting to mimic the marine and sponge environments, were fundamental to increase the number of cultivable taxa. Fungi were identified using a polyphasic approach, by means of morpho-physiological, molecular and phylogenetic techniques. The sponges revealed an astonishing fungal diversity represented by 87 fungal taxa. Each sponge hosted a specific fungal community with more than half of the associated fungi being exclusive of each invertebrate. Several species isolated and identified in this work, already known in terrestrial environment, were first reported in marine ecosystems (21 species) and in association with sponges (49 species), including the two new species Thelebolus balaustiformis and Thelebolus spongiae, demonstrating that oceans are an untapped source of biodiversity.