The environmental conditions of caves shape microbiota. Within caves’ microbial communities, actinomycetes are among the most abundant bacteria. Cave actinomycetes have gained increasing attention during the last decades due to novel bioactive compounds with antibacterial, antioxidant and anticancer activities. However, their potential role in soil environments is still unknown. This review summarises the literature dealing with actinomycetes from caves, underlining for the first time their potential roles in soil environments. We provide an overview of their diversity and biotechnological properties, underling their potential role in soil environments applications. The contribution of caves’ actinomycetes in soil fertility and bioremediation and crops biostimulation and biocontrol are discussed. The survey on the literature show that several actinomycetes genera are present in cave ecosystems, mainly Streptomyces, Micromonospora, and Nocardiopsis. Among caves’ actinomycetes, Streptomyces is the most studied genus due to its ubiquity, survival capabilities, and metabolic versatility. Despite actinomycetes’ outstanding capabilities and versatility, we still have inadequate information regarding cave actinomycetes distribution, population dynamics, biogeochemical processes, and metabolisms. Research on cave actinomycetes needs to be encouraged, especially concerning environmental soil applications to improve soil fertility and health and to antagonise phytopathogens.

The terrestrial subsurface offers privileged sites both to search for microbial life and to observe still mostly unknown characteristic lithologies. In particular, caves represent natural laboratories to investigate unique minerogenetic processes and biotic interactions, connected to these phenomena. Manganese mineralization in cave environments provides a window to understand the complex Mn cycle and the development of microbial communities in special conditions, such as low constant temperature, absence of light and, in particular, low-energy environments. In the current study, we isolated and characterized Mn-samples taken from the cave “Grotta Grande dei Cervi,” L’Aquila, Central Italy, and we used a multidisciplinary approach to characterize them, with the purpose of understanding the biogeochemical processes in extreme environments. A chemical characterization of the samples was done by EDS; further investigations are underway with other multidisciplinary methodologies to understand whether the Mn laminae are related to biological processes. SEM investigations revealed microbial imprints, showing cell-like structures and suggesting that the cell-like shapes occur within internal laminae. A culture-independent approach was used to assess the possibility that biotic factors may be involved in the production of these mineralizations and to investigate the nature of the microbial community in these materials. A molecular approach was the first step to investigate the role of microorganisms in forming manganese oxides associated with water bearing rocks. DNA from the black deposits was extracted and sequence analyses of specimens were performed. Our data support the hypothesis that microorganisms may contribute to the mineralizations of manganese in this environment, providing new encouraging insight into the role of microorganisms in the Mn cycle and the processes of energy acquisition in unfavorable conditions, with relevant implications for astrobiology.

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Caves are dark subsurface environments with relatively constant temperatures that allow studying bio-mineralization processes and paleoenvironmental or climate changes in optimal conditions. In the extreme and oligotrophic cave environment, manganese patinas having stromatolite-like features are uncommon. Here we provide the first detailed mineralogical, geochemical, and microbiological investigation of fine-grained and poorly crystalline MnFe stromatolite-like wall patinas formed in a deep-cave environment in Italy. These mineralizations, about 3 mm thick, consist of an alternation of Mn-layers and Fe-lenses. We show that the microbial communities' composition is dominated by Mn-oxidizing bacteria, such as Bacillus, Flavobacterium, and Pseudomonas. Our multidisciplinary investigation, integrating data from different analytical techniques (i.e., optical microscopy, SEM-EDS, μXRF, XRPD, FT-IR, Raman spectroscopy, and DNA sequencing), revealed peculiar chemical, mineralogical, and biological features: 1) A cyclical oscillation of Mn and Fe along the growth of the patinas. We propose that this oscillation represents the shift between oxic and suboxic conditions related to different phases occurring during paleo-flood events; 2) A typical spatial distribution of mineralogy and oxidation state of Mn, bacterial imprints, detrital content, and stromatolite-like morphologies along the Mn-layers. We propose that this distribution is controlled by the local hydraulic regime of the paleo-floods, which, in turn, is directly related to the morphology of the wall surface. Under less turbulent conditions, the combination of clay mineral catalysis and biological oxidation produced vernadite, a poor-crystalline phyllomanganate with a low average oxidation state of Mn, and branched columnar stromatolite-like morphologies. On the other hand, under more turbulent conditions, the sedimentation of clay minerals and microbial communities' development are both inhibited. In this local environment, a lower oxidation rate of Mn2+ favored the formation of todorokite and/or ranciéite, two compounds with a high average oxidation state of Mn, and flat-laminated or columnar stromatolite-like morphologies.

This study aimed to estimate the green formation lampenflora of “Stiffe” caves in order to evaluate their suitability as an isolation source of cyanobacteria useful for the production of polyhydroxyalkanoates (PHAs). The cave system was chosen as the sampling site due to its touristic use and the presence of high-impact illuminations. The biofilms and the mats of the illuminated walls were sampled. Samples were investigated by 16S rRNA gene analysis and culturable cyanobacteria isolation. The isolated strains were then screened for the production of PHAs under typical culturing and nutritional starvation. Cultures were checked for PHA accumulation, poly-β-hydroxybutyrate (PHB) presence (infrared spectroscopy), and pigment production. The 16S rRNA gene metabarcoding. Highlighted a considerable extent of the pressure exerted by anthropogenic activities. However, the isolation yielded eleven cyanobacteria isolates with good PHA (mainly PHB)-producing abilities and interesting pigment production rates (chlorophyll a and carotenoids). Under normal conditions (BG110), the accumulation abilities ranged from 266 to 1,152 ng mg dry biomass–1. The optimization of bioprocesses through nutritional starvation resulted in a 2.5-fold increase. Fourier transform infrared (FTIR) studies established the occurrence of PHB within PHAs extracted by cyanobacteria isolates. The comparison of results with standard strains underlined good production rates. For C2 and C8 strains, PHA accumulation rates under starvation were higher than Azospirillum brasilense and similar to Synechocystis cf. salina 192. This study broadened the knowledge of the microbial communities of mats and biofilms on the lightened walls of the caves. These findings suggested that these structures, which are common in tourist caves, could be used to isolate valuable strains before remediation measures are adopted.

The analysis of copepod behaviour gained an increasing impetus over the past decade thanks to the advent of computer-assisted video analysis tools. Since the automated tracking consists in detecting the animal’s position frame by frame and improving signals corrupted by strong background noise, a crucial role is played by the length of the video recording. The aim of this study is to: (i) assess whether the recording time influences the analysis of a suite of movement descriptive parameters; (ii) understand if the recording time influences the outcome of the statistical analyses when hypotheses on the effect of toxicants/chemicals on the freshwater invertebrate behaviour are tested. We investigated trajectory parameters commonly used in behavioural studies—swimming speed, percentage of activity and trajectory convex hull—derived from the trajectories described by the inbenthic–interstitial freshwater copepod Bryocamptus pygmaeus exposed to a sub-lethal concentration of diclofenac. The analyses presented in this work indicate that the recording time did not influence the outcome of the results for the swimming speed and the percentage of activity. For the trajectory convex hull area, our results showed that a recording session lasting at least 3 min provided robust results. However, further investigations are needed to disentangle the role of concurrent factors, such as the behavioural analysis of multiple individuals simultaneously, whether they are of the same or opposite sex and the implications on sexual behaviour, competition for resources and predation.

This study revealed how Bacteria and Archaea communities and their metabolic functions differed between two groups of black deposits identified in gorge and cave environments. Scanning electron microscopy coupled with energy dispersive spectroscopy was used to analyse the presence of microbial biosignatures and the elemental composition of samples. Metabarcoding of the V3–V4 regions of 16S rRNA was used to investigate Bacteria and Archaea communities. Based on 16S rRNA sequencing results, PICRUSt software was used to predict metagenome functions. Micrographs showed that samples presented microbial biosignatures and microanalyses highlighted Mn concretions and layers on Al-Si surfaces. The 16S rRNA metabarcoding alpha-diversity metrics showed similar Simpson's and Shannon indices and different values of the Chao-1 index. The amplicon sequence variants (ASVs) analysis at the different taxonomic levels showed a diverse genera composition. However, the communities of all samples shared the presence of uncultured ASVs belonging to the Gemmatales family (Phylogenesis: Gemmataceae; Planctomycetes; Planctomycetota; Bacteria). The predicted metagenome functions analysis revealed diverse metabolic profiles of the Cave and Gorge groups. Genes coding for essential Mn metabolism were present in all samples. Overall, the findings on structure, microbiota, and predicted metagenome functions showed a similar microbial contribution to epigean and hypogean black deposits Mn metabolism.

Abundance–Biomass Comparison (ABC) models, first described for marine benthic macrofauna, have been applied successfully to other marine and terrestrial/freshwater fauna but never to ecotonal communities. In particular, to our knowledge, ABC models have not been applied to hyporheic communities. This study represents the first application of ABC models to hyporheic assemblages. We aimed at testing the effectiveness of ABC models in describing the perturbation of hyporheic communities subjected to an existing/known disturbance. To this end, we applied the models to the hyporheic community of an Apennine creek, where the hyporheic waters of the upstream stretch were uncontaminated, whereas those of the downstream stretch were contaminated by ammonium. We also tested separated models for the summer and winter periods to account for potential variability due to season. ABC models provided a satisfactory description of the hyporheic community changes due to ammonium by showing the abundance dominance curve overlying that of the biomass in the downstream stretch contrarily to what was observed in the upstream stretch. However, ABC models did not highlight any significant seasonal effects. Our results showed that ABC models have the potential to be used as assessment tools for ecological quality of hyporheic zones in temperate regions.