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This paper reviews the ground beetles from René Jeannel collection which is preserved in Zoological Museum of Babeș-Bolyai University. The illustrious coleopterologist, René Jeannel was a French entomologist, who worked between 1920-1930 as Deputy Director of the Institute of Speleology founded by Emil Racovitza, the first institute of its kind in the world. He was also a professor of general biology at the Faculty of Sciences of the University of Cluj Napoca. As well, in that period he published numerous articles on the systematics, phylogeny, biogeography and evolution of cave and endogenous beetles. In 1931, R. Jeannel left Romania to start working at the Muséum National d'histoire naturelle, whose director he was between 1945 -1951. During the 10 years of activity in Romania, he created and organized an important collection of ground beetles (Coleoptera, Carabidae). The total number of prepared and labeled specimens is 1672. The material was collected between 1921-1927, mostly in Transylvania, Romania. The collection includes 182 species from 56 genera and contains rare or endemic species. Among the endemic species we list: Pterostichus bielzii (Fuss, 1858), endemic to the Apuseni mountains, is present in the collection from 13 different locations; Nebria (Nebria) transsylvanica Germar, 1823, an endemic species in the Romanian Carpathians and in the southern part of the Carpathians in Ukraine; Platynus (Batenus) banaticus (I.Frivaldszky von Frivald, 1865) present in the collection from 3 locations. For the genus Carabus, representative species are Carabus (Megodontus) planicollis Küster, 1846, endemic species in Romania, and the Natura 2000 species Carabus variolosus Fabricius, 1787 and Carabus (Procerus) gigas (Creutzer, 1799). Among the Trechinae the most representative are the genus Duvalius, with four species: Duvalius cognatus subsp. longicollis Jeannel, 1928, D. laevigatus (Bokor, 1913), D. budae subsp. dioszeghyi Mallasz, 1928 and D. hegeduesii subsp. jonescoi (Jeannel, 1919), and the genus Trechus with 7 species, of which 3 are endemic in Romania: T. bannaticus Dejean, 1831, T. biharicus Meixner, 1912, and T. marginalis Schaum, 1862. The collection is a very valuable one, both from a historical and scientific point of view, because it offers an overview of the diversity and distribution of carabids in the karst areas of Romania, at the beginning of the 20th century.

Protopholeuon is a monospecific genus of Leptodirini, endemic to Romania, represented by Protopholeuon hungaricum, described incompletely by Csiki in 1904. In 1923, René Jeannel published a more comprehensive morphological description of the species, along with the first drawing of male genitalia. The type locality of the species is Lucia Cave, in the Metaliferi Mountains (Apuseni Mountains). This was the only locality where the species was found and collected. In the last decade, representatives of the genus Protopholeon were found in other caves of the Metaliferi Mountains. These caves are: Grohot, Izbucul Topliței, Ponor, Rusești, Hodobana, Cizmei, Urșilor Bulzești. The aerial distance between Lucia Cave and the newly explored caves is about 20 km. The patchy geology and hydrology of the area can act as natural barriers that shape the species' geographic distribution and enhance the speciation process. We provide morphometric information for Protopholeuon specimens obtained from 80 individuals. We also present the pictures of Protopholeuon with morphological and ultrastructural details (SEM microscopy) for head, mandibles, antennae, elytra surface and aedeagus. All this completes the morphological description made in 1923 by Jeannel. We present the first data on the molecular analysis of a mitochondrial COI fragment on Protopholeuon all the mentioned caves. For every cave, we analyzed 2-3 specimens. Our working hypothesis was that considering the distribution of caves, P. hungaricum from Lucia Cave represents a distinct phylogenetic unit from the other sampled caves of the Metaliferi Mountains. As an outgroup for rooting the phylogenetic tree, we used sequences from Pholeuon, another genus of endemic Romanian Leptodirini. The molecular analysis revealed a unique clade for all samples from Metaliferi Mountains. Moreover, a single clade is built by all Metaliferi haplotypes, except Lucia Cave, with a high bootstrap value (85%). The genetic distance between ingroup and outgroup samples is 7-8%, while between the Lucia haplotype and the other haplotypes from Metaliferi Mountain the range is 3-4%, which agrees with other results from the group of Romanian Leptodirini. The distances between Metaliferi Mountain haplotypes (except Lucia Cave) are 0.1-2.8%, grouping in two distinct clades, with high statistical values. Still, the taxonomic status of samples from the Metaliferi Mountains should be further investigated by corroborating morphologic and genetic data.

In karst areas, anthropogenic contaminants reach the subsurface with detrimental effects on the groundwater ecosystem and downstream springs, which often serve as drinking water sources for the local human communities. We analyzed the water chemistry and microbial community composition in upstream and downstream locations of five hydrokarst systems (HKS) during four seasons. Conductivity and nitrates were higher in the downstream springs than in the pre-karst waters, whereas the concentration of organic matter, considered here as a pollution indicator, was lower. The microbial community composition varied largely between upstream and downstream locations, with multiple species of potentially pathogenic bacteria decreasing in the HKS. Bacteria indicative of pollution decreased as well when passing through the HKS, but potential biodegraders increased. This suggests that the HKS can filter out part of the polluting organic matter and, with it, part of the associated microorganisms. Nevertheless, the water quality, including the presence of pathogens in downstream springs, must be further monitored to control whether the water is appropriate for consumption. In parallel, the human populations located upstream must be advised of the risks resulting from their daily activities, improper stocking of their various wastes and dumping of their refuse in surface streams.

Human activities and natural factors determine the hydrogeochemical characteristics of karst groundwaters and their use as drinking water. This study assesses the hydrogeochemical characteristics of 14 karst water sources in the Apuseni Mountains (NW Romania) and their potential use as drinking water sources. As shown by the Durov and by the Piper diagrams, the chemical composition of the waters is typical of karst waters as it is dominated by HCO3− and Ca2+, having a circumneutral to alkaline pH and total dissolved solids ranging between 131 and 1092 mg L−1. The relation between the major ions revealed that dissolution is the main process contributing to the water chemistry. Limestone and dolostone are the main Ca and Mg sources, while halite is the main Na and Cl source. The Gibbs diagram confirmed the rock dominance of the water chemistry. The groundwater quality index (GWQI) showed that the waters are of excellent quality, except for two waters that displayed medium and good quality status. The quality of the studied karst waters is influenced by the geological characteristics, mainly by the water–rock interaction and, to a more limited extent, by anthropogenic activities. The investigated karst waters could be exploited as drinking water resources in the study area. The results of the present study highlight the importance of karst waters in the context of good-quality water shortage but also the vulnerability of this resource to anthropogenic influences.

(1) Background: Show caves are unique natural attractions and touristic traffic can trigger their degradation within a very short time. There are no universal solutions to counter the effects of the touristic impact upon cave environment and both protection protocols and management plans have to be established on a case-by-case basis; (2) Methods: The study includes four show caves from the Romanian Carpathians, where monitoring of the number of visitors, paralleled by the monitoring of the main physico-chemical parameters of the air and water (CO2, temperature, humidity, drip rate, conductivity, and pH) was implemented; (3) Results and Conclusions: The results of the study has: (i) established a monitoring protocol to be applied to each of the four show caves; (ii) established a set of basic principles to be enforced by the management of show caves; (iii) issued a set of preventive measures and instructions to be followed by the personnel and stakeholders of the caves.

In rural areas without centralized water supply systems, inhabitants often use groundwater of unknown quality as drinking water, without understanding the possible negative consequences on their health. Karstic spring waters from Dobrogea region in Romania were assessed for their potential to be used as drinking water source, according to their quality and seasonal variation. The physico-chemical parameters of waters were compared with the guideline values for drinking water established by the World Health Organization and the Directive 98/83/EC. The nitrate and Cr concentrations exceeded the guideline value in the springs from Southern Dobrogea, but met the quality criteria in those from Northern Dobrogea, thus, to be used as drinking water, the karstic springs located in Southern Dobrogea require treatment for nitrates removal. Heavy metals pollution indices showed low to medium cumulative heavy metal pollution in all springs, while the human health risk assessment by oral exposure indicated possible noncarcinogenic risks of nitrates, both for adults and children in springs from South Dobrogea. A rigorous monitoring of the water quality before human consumption is recommended for all four studied water sources.

Caves often hold valuable palaeoclimate archives including speleothems, fossil remains, and clastic sediments that complement each other. This paper presents a multi-archive interdisciplinary study of an extensive deposit of fossil mammals from the scientific reserve in the Muierilor Cave, Southern Carpathians, Romania. We present two new palaeontological excavations that indicate a high abundance and diversity of MIS 3–2 fossil mammals (carnivores, omnivores and herbivores) synchronous with the early modern humans known from this cave. Using geochronological and sedimentological methods, we present a general reconstruction of the cave evolution between ~120 kyr B.P. and the Holocene. The study is based on a combination of geochronological tools including OSL dating of sediments, U/Th dating of speleothems, and radiocarbon dating of fossil remains, with a total of 54 ages. Based on U/Th dating of speleothems from stratigraphically-relevant positions, we show that the MIS 3 assemblage of fossil mammals were massively reworked and deposited during the post-LGM deglaciation, slightly earlier than previously known for the Southern Carpathians. On the other hand, several young radiocarbon ages of cave bear samples suggest that the Southern Carpathians might have been functioning as a glacial refuge for this species as late as ~22 kyr B.P.

Introduction: Karstic springs are used worldwide by rural communities as sources of fresh water for humans and livestock. In Romania, one-third of the population has no direct access to a public water supply. The present study is part of a country-wide project to develop simple, quick and cheap methods for seasonal environmental and microbiological monitoring of karstic springs used as drinking water by rural populations. Critical steps for monitoring workflow consist of evaluating water quality and selecting suitable membrane filters to efficiently capture environmental DNA for further microbial diversity estimation using 16S rRNA gene-based metabarcoding. Methods: Several commercial membrane filters of different compositions and pore sizes were tested on the water sampled from three karstic springs in Romania, followed by water chemistry and whole community 16S rRNA gene-based metabarcoding analysis. Results: We found that different types of applied membrane filters provide varying recovery in diversity and abundance of both overall and pathogenic bacteria. Conclusions: The result of the experiment with different filters shows that mixed cellulose ester, cellulose acetate, and nitrate membranes of 0.20 and 0.22 &micro, m are the best for amplicon-based metabarcoding monitoring of karst springs.

The increasing human impact in Romanian caves raises the urgency of publishing a correct database of the strictly-adapted cave fauna. Previous attempts at indexing cave fauna and classifying caves by using their fauna opened many questions regarding the use of an incomplete list of cave species and mixed lists of troglobionts/stygobionts with troglophiles/stygophiles for ranking caves with priority for protection. It has also become obvious that there is a need to publish a list of Romanian cave species that are under threat. Cave species in Romania (and elsewhere) are endemic on small ranges, are unique and must be considered as important units for conservation. A cave must be equally protected if it has one or more rare and strictly endemic cave species. Although not exhaustive, we here provide the first checklist of Romanian troglobionts/stygobionts developed in the framework of the DARKFOOD and GROUNDWATERISK projects, coordinated by the “Emil Racovita” Institute of Speleology, Cluj-Napoca, Romania. The GIS application was used to complement the checklist of cave species with data on caves and surface environments above the caves. Until complete data on species diversity and population sizes are made available for each cave, measures of conservation can be implemented, based on the presence/absence of cave species, while classifications of caves for protection, based on the number of species, must be avoided. We also propose a list of Romanian caves with fauna that are under threat and a tentative Red List of Romanian troglobiont/stygobionts. This is the first database with identified troglobiont and stygobiont species of Romania, with a critical analysis of their distribution inside the country. A list of caves that need protection for their rare and unique species and a tentative Red List of Romanian cave fauna are also added. A total of 173 species were identified, of which 77 troglobionts and 96 stygobionts are currently registered in 366 caves. The database is divided into two parts, one part with a list of troglobionts, their revised systematic position, cave name, cave code and geographic region; and the second part with the same information on stygobionts. The database represents the contribution of many active researchers, who are the authors of this paper and of review publications of many other authors of the "Emil Racoviță" Institute of Speleology.

Air microbial pollution in touristic areas poses a risk for both the integrity of an ecosystem and human health. Microbiological monitoring together with environmental parameters monitoring allows for the assessment of the impacts and formulation of sound management decisions to protect humans and ecosystems. Four show caves from the Carpathian Mountains were selected for our study. The caves were sampled monthly to obtain an overview of the changes that occur over a yearly cycle. For the microbial monitoring, we used RIDA®COUNT test plates, while the environmental parameters were monitored with a variety of devices. Second and third generations of microbes extracted from the plates were grown on specific media for the purpose of 16S rRNA and 18S rRNA extraction and taxa identification. The bacterial communities identified in the air samples in the four investigated show caves were dominated by Staphylococcus, while regarding the fungi communities, Penicillium was more likely to occur in the touristic part of the caves and Cladosporium in the non-visited passages. Together with data on number of visitors, number of bats and radon levels we were able understand the impact of tourists on the cave environment and to generate microbiological risk maps for human health. This type of comprehensive study can be used not only to protect the integrity of a touristic area from the impacts caused by the introduction of allochthonous organic matter, but also for the protection of the tourists and guides from potential pathogenic taxa.

Spatial distribution analyses of fossil bones are often used in cave taphonomy for i) assessing directly the size of a given fossil population and ii) indirectly, for palaeo-environmental reconstructions. Urşilor Cave from northwestern Romania hosts one of the richest MIS 3 cave bear bone assemblages of Europe, and as the nature of the bone deposition was not settled yet, the study of the size of cave bear population and the distribution of the skeletal element was needed. More than 11,500 cave bear skeletal elements (NISPtotal = 11, 511), derived from 105 individuals (MNIcanines = 105) were extracted from the palaeontological excavation (ca. 9 m2) from the lower level of the cave and documented (photographed, mapped, and topographically measured). Almost 58% of all analyzed bones and teeth are concentrated in three quadrants of the excavation (C1, D1 and E2); the bones’ density decreases in the D4 and A1 quadrants. Based on the obtained results on the spatial distribution of the cave bear bones, we assume that the studied bone assemblage should be now regarded as a consequence of a cave trap (deposition in situ) while the hypothesis of fluvial transport (previously assumed) becomes questionable.

The aim of this study was to monitor factors affecting the microorganisms of water bodies in caves and apply the findings to improve their management. Four show-caves in Romania were sampled monthly to obtain an overview of the changes occurring in the fungal and bacterial communities of cave waters in relation to surface and underground impacts. Test plates were used for microbiological monitoring, alongside the recording of environmental parameters - physicochemical, drip rates, and number of both bats and tourists. A higher number of microbes were obtained during springtime and the summer tourist peak-season. None of the identified strains were cave-specific, and the presence of human-associated taxa was observed from all of the sites. A lack of significant correlation between the abundances of the microorganisms and the physicochemical parameters of the water emphasise the reliability of microorganisms as significant proxies for impacts on underground environments and their use in management decisions.

Carpathian Mountains were one of the main refuge areas during the climate changes of the Pleistocene and the Holocene in Europe and one of the richest regions in the world in subterranean (caves and associated habitats) endemic species. Nevertheless, the Carpathian Mountains subterranean fauna importance is underestimated especially due to dispersed information on its diversity and the scarcity of molecular studies in the area. Here, we present a first general view of the cave fauna hotspot represented by the Romanian Carpathians and the geological and historical processes that shaped the patterns of subterranean distribution and diversity at regional scale. The Carpathians are an amalgam of various geological units with complex paleogeographical evolution that is reflected in completely different species assemblages dominated by unit specific fauna groups. Phylogeography of Coleoptera and environmental parameters are adding to the general view at regional scale and offer additional explanation for this exceptional subterranean diversification in a non-Mediterranean region. We also use the example of the Carpathians cave fauna as proxy for past environmental changes in the area. Troglobionts are endemic on small areas and by studying their present distributions and phylogeny, past processes of landscape evolution on the surface can be better understood.

The Zăton Lake is a closed depressionlocated in the Ponoarele karst area (south-western Romania) formed along a tectonic-erosional window as a NE-SW corridor (250–450 m a.s.l.). In this area Mesozoic sedimentary rocks outcrop, along with the metamorphic basement of the Danubian Domain of the Southern Carpathians. The entire river basin is drained underground through the caves of Zăton (105 m in lenght) and Bulba (5 km long, developed on three levels). Flooding of the lake is temporary, during periods with high precipitation or the spring snow melting. A total of 34 samples were taken from one sedimentary section of the Zăton Lake and the diversity and abundance of fossil mites was assessed at different depths of the sediments. OSL (optical stimulated luminescence) datings and fossil mites’ determination were correlated with sediments geochemistry and rock magnetic properties and record the changes in temperature and vegetation in the area from present day to more than 2000 years ago.

The Ursilor Cave (NW Romania) is a famous cave bear paleontological site hosting an important Late Pleistocene faunal assemblage and subject to systematic excavation works. To better understand the origin of fossil assemblages, the sedimentary history of the cave must be reconstructed. We conducted a series of investigations on various cave deposits which included sedimentology and grain-size analyses, U-series dating of speleothems, OSL dating of sediments, and AMS radiocarbon dating of fossil remains. The results allowed for the identification of several major chronological controls for the evolution of the cave during the last 300,000 years. Five evolutionary stages or key-moments were dated, and a tentative speleogenetic scenario is presented. The combined numerical dates and sedimentological study show that the evolution of the cave was more complex than previously thought. In particular, the fossil accumulation was related to a succession of rapid flooding events at ∼47–40 ka. Alternating depositional and erosional phases have occurred since at least 210 ka complicating the sediment structures. The results suggest that the upper and lower levels of the cave may have been functioned occasionally as hydrologically separated karst systems and that the animal populations from what is now a single cave system may not necessarily be synchronous. This case study shows that reconstructing sedimentary history of a given cave is crucial for the correct understanding of its thanatocenosis.