Permanent artificial lighting systems in tourist underground environments promote the proliferation of photoautotrophic biofilms, commonly referred to as lampenflora, on damp rock and sediment surfaces. These green-colored biofilms play a key role in the alteration of native community biodiversity and the irreversible deterioration of colonized substrates. Comprehensive chemical or physical treatments to sustainably remove and control lampenflora are still lacking. This study employs an integrated approach to explore the biodiversity, eco-physiology and molecular composition of lampenflora from the Pertosa-Auletta Cave, in Italy. Reflectance analysis showed that photoautotrophic biofilms are able to absorb the totality of the visible spectrum, reflecting only the near-infrared light. This phenomenon results from the production of secondary pigments and the adaptability of these organisms to different metabolic regimes. The biofilm structure mainly comprises filamentous organisms intertwined with the underlying mineral layer, which promote structural alterations of the rock layer due to the biochemical attack of both prokaryotes (mostly represented by Brasilonema angustatum) and eukaryotes (Ephemerum spinulosum and Pseudostichococcus monallantoides), composing the community. Regardless of the corrosion processes, secondary CaCO3 minerals are also found in the biological matrix, which are probably biologically mediated. These findings provide valuable information for the sustainable control of lampenflora.

The dispersal of Homo sapiens across Eurasia during MIS 3 in the Late Pleistocene is marked by technological shifts and other behavioral changes, known in the archaeological record under the term of Initial Upper Paleolithic (IUP). Bacho Kiro Cave in north Bulgaria, re-excavated by us from 2015 to 2021, is one of the reference sites for this phenomenon. The newly excavated lithic assemblages dated by radiocarbon between 45,040 and 43,280 cal BP and attributed to Homo sapiens encompass more than two thousand lithic artifacts. The lithics, primarily from Layer N1-I, exist amid diverse fauna remains, human fossils, pierced animal teeth pendants, and sediment with high organic content. This article focuses on the technological aspects of the IUP lithics, covering raw material origin and use-life, blank production, on-site knapping activities, re-flaking of lithic implements, and the state of retouched lithic components. We apply petrography for the identification of silicites and other used stones. We employ chaîne opératoire and reduction sequence approaches to profile the lithics techno-typologically and explore the lithic economy, particularly blade production methods, knapping techniques, and artifact curation. Raw material analysis reveals Lower Cretaceous flints from Ludogorie and Upper Cretaceous flints from the Danube region, up to 190 km and 130 km, respectively, from Bacho Kiro Cave, indicating long-distance mobility and finished products transport. Imported lithic implements, were a result of unidirectional and bidirectional non-Levallois laminar technology, likely of volumetric concept. Systematic on-anvil techniques (bipolar knapping) and tool segmentation indicate re-flaking and reshaping of lithic implements, reflecting on-site curation and multifaceted lithic economy. A limited comparison with other IUP sites reveals certain shared features and also regional variations. Bacho Kiro Cave significantly contributes to understanding the technological and behavioral evolution of early Homo sapiens in western Eurasia.

Dimba Cave is a large array of natural galleries in limestone mountains of the Democratic Republic of the Congo that contains highly valued pre-historic archaeological artifacts. The cave attracts a high number of tourists every year and is used by local populations as a water supply source. The main objective of the research undertaken in Dimba Cave consisted of assessing the quality of water and sediments from Dimba Cave ponds through evaluating contamination by heavy metals (15 elements analyzed, including As, Cd, Pb, and Hg) and by microbial populations (including Escherichia coli and total coliforms) in order to estimate the ecotoxicological risk to humans and to non-human biota. All water samples collected in the cave ponds showed very high metal concentrations exceeding the internationally recommended limits for drinking water, particularly for Cr, Mn, As, Pb, and Hg. Most sediment samples from cave ponds also displayed high heavy metal concentrations. The calculated pollution parameters, such as the enrichment factor (EF), and ecological risk parameters, such as the ecological risk index (Eri), indicated that the sediment may be toxic to aquatic biota. Furthermore, the microbiological analysis of pond waters indicated a widespread contamination with bacteria such as Escherichia coli, Enterococcus spp., total coliforms, and Pseudomonas spp., probably from anthropogenic and/or animal sources. Therefore, the consumption of Dimba Cave water as a drinking water represents a threat to public health. Urgent management measures should be enforced to protect public health and the cave ecosystem.

Cyanobacteria are adaptable and dominant organisms that exist in many harsh and extreme environments due to their great ecological tolerance. They produce various secondary metabolites, including cyanotoxins. While cyanobacteria are well studied in surface waters and some aerial habitats, numerous other habitats and niches remain underexplored. We collected 61 samples of: (i) biofilms from springs, (ii) aerial microbial mats from buildings and subaerial mats from caves, and (iii) water from borehole wells, caves, alkaline, saline, sulphidic, thermal, and iron springs, rivers, seas, and melted cave ice from five countries (Croatia, Georgia, Italy, Serbia, and Slovenia). We used (q)PCR to detect cyanobacteria (phycocyanin intergenic spacer-PC-IGS and cyanobacteria-specific 16S rRNA gene) and cyanotoxin genes (microcystins-mcyE, saxitoxins-sxtA, cylindrospermopsins-cyrJ), as well as amplicon sequencing and morphological observations for taxonomic identification. Cyanobacteria were detected in samples from caves, a saline spring, and an alkaline spring. While mcyE or sxtA genes were not observed in any sample, cyrJ results showed the presence of a potential cylindrospermopsin producer in a biofilm from a sulphidic spring in Slovenia. This study contributes to our understanding of cyanobacteria occurrence in diverse habitats, including rare and extreme ones, and provides relevant methodological considerations for future research in such environments.

In cave environments, stable conditions devoid of light-dark cycles and temperature fluctuations sustain circadian clock mechanisms across various species. However, species adapted to these conditions may exhibit disruption of circadian rhythm in locomotor activity. This study examines potential rhythm loss due to convergent evolution in five semi-aquatic troglobitic isopod species (Crustacea: Styloniscidae), focusing on its impact on locomotor activity. The hypothesis posits that these species display aperiodic locomotor activity patterns. Isopods were subjected to three treatments: constant red light (DD), constant light (LL), and light-dark cycles (LD 12:12), totaling 1656 h. Circadian rhythm analysis employed the Sokolove and Bushell periodogram chi-square test, Hurst coefficient calculation, intermediate stability (IS), and activity differences for each species. Predominantly, all species exhibited an infradian rhythm under DD and LL. There was synchronization of the locomotor rhythm in LD, likely as a result of masking. Three species displayed diurnal activity, while two exhibited nocturnal activity. The Hurst coefficient indicated rhythmic persistence, with LD showing higher variability. LD conditions demonstrated higher IS values, suggesting synchronized rhythms across species. Significant individual variations were observed within species across the three conditions. Contrary to the hypothesis, all species exhibited synchronization under light-dark conditions. Analyzing circadian activity provides insights into organism adaptation to non-cyclical environments, emphasizing the importance of exploring underlying mechanisms.

Fossil identification practices have a profound effect on our interpretation of the past because these identifications form the basis for downstream analyses. Therefore, well-supported fossil identifications are necessary for examining the impact of past environmental changes on populations and communities. Here we apply an apomorphic identification framework in a case study identifying fossil lizard remains from Hall\'s Cave, a late Quaternary fossil site located in Central Texas, USA. We present images and descriptions of a broad comparative sample of North American lizard cranial elements and compile new and previously reported apomorphic characters for identifying fossil lizards. Our fossil identifications from Hall\'s Cave resulted in a minimum of 11 lizard taxa, including five lizard taxa previously unknown from the site. Most of the identified fossil lizard taxa inhabit the area around Hall\'s Cave today, but we reinforce the presence of an extirpated species complex of horned lizard. A main goal of this work is to establish a procedure for making well-supported fossil lizard identifications across North America. The data from this study will assist researchers endeavoring to identify fossil lizards, increasing the potential for novel discoveries related to North American lizards and facilitating more holistic views of ancient faunal assemblages.

The mountain region of central Veracruz, Mexico hosts a large system of karst and volcanic caves that are unexplored. In particular, the vertebrates that inhabit these subterranean ecosystems are unknown. This study evaluated the diversity of mammals, birds, reptiles, amphibians, and fish in three environments (euphotic, disphotic, and aphotic) of 16 caves of different geological origin (12 karst caves and 4 volcanic caves) distributed along an altitudinal gradient (300-2400 m a.s.l.). We found a richness of 242 vertebrate species (184 birds, 30 mammals, 15 reptiles, 12 amphibians, and 1 fish) and an abundance of a total of 11,323 individuals (4,969 mammals, 6,483 birds, 36 reptiles, 27 amphibians, and 5 fish). The richness of all vertebrate classes was higher in karst than in volcanic caves. Vertebrate diversity was also higher at mid-altitudes between 600-899 m a.s.l. Diversity varied between environments, where bird and reptile richness was higher in the euphotic environment, while mammal and amphibian diversity was higher in the aphotic environment. The similarity in the composition of vertebrate species does not depend on the distance between karstic and volcanic caves. Volcanic and karst caves shared on average up to 70% and 55% of vertebrate species, which indicates that only 30% and 45% of species, respectively, is different in each cave type. Given the vulnerability and fragility of these subterranean ecosystems, as well as the important diversity that they contain, we recommend including the caves of the central region of Veracruz in the conservation agenda of local governments and communities. Community-based conservation can help ensure the presence of vertebrate species in the caves of this region.

It has been just over 10 years since the first fossils attributed to Homo naledi were recovered from the Rising Star Cave system in South Africa\'s Cradle of Humankind. The hominin fossil evidence for H. naledi displays a distinctive combination of primitive and derived morphology, yet for a time-averaged fossil sample it is remarkable for its relatively low level of variation. Thus-unusually for palaeoanthropology-there has been little pushback against the decision to recognize a single novel taxon for all of the material recovered from the Rising Star Cave system. However, almost everything else claimed about H. naledi-its age, burial context and behaviour-has been controversial. Here we examine the strength of the evidence for these claims.

Intense environmental pressures can yield both regressive and constructive traits through complex evolutionary mechanisms. Although regression is well-studied, the biological bases of constructive features are less well understood. Cave-dwelling Astyanax fish harbor prolific extraoral taste buds on their heads, which are absent in conspecific surface-dwellers. Here, we present novel ontogenetic data demonstrating extraoral taste buds appear gradually and late in life history. This appearance is similar but non-identical in different cavefish populations, where patterning has evolved to permit taste bud re-specification across the endoderm-ectoderm germ layer boundary. Quantitative genetic analyses revealed that spatially distinct taste buds on the head are primarily mediated by two different cave-dominant loci. While the precise function of this late expansion on to the head is unknown, the appearance of extraoral taste buds coincides with a dietary shift from live-foods to bat guano, suggesting an adaptive mechanism to detect nutrition in food-starved caves. This work provides fundamental insight to a constructive evolutionary feature, arising late in life history, promising a new window into unresolved features of vertebrate sensory organ development.

Anchialine systems are coastal groundwater habitats around the world which host a unique community of cave adapted species (stygobionts). Such communities are expected to be separated by haloclines into either fresh or saline groundwater communities, hence climate changes (e.g., eustatic sea level shifts) and anthropic driven changes (e.g., salinization) may have a great impact on these stygobiont communities. Here we used cave-restricted species of Typhlatya from the Yucatan Peninsula as models to identify physiological capacities that enable the different species to thrive in marine groundwater (T. dzilamensis) or fresh groundwater (T. mitchelli and T. pearsei), and test if their distribution is limited by their salinity tolerance capacity. We used behavior, metabolic rates, indicators of the antioxidant system and cellular damage, and lactate content to evaluate the response of individuals to acute changes in salinity, as a recreation of crossing a halocline in the anchialine systems of the Yucatan Peninsula. Our results show that despite being sister species, some are restricted to the freshwater portion of the groundwater, while others appear to be euryhaline.