Cave Soil Research Articles

Microbially Induced Calcite Precipitation Employing Environmental Isolates.

In this study, five microbes were employed to precipitate calcite in cohesionless soils. Four microbes were selected from calcite-precipitating microbes isolated from calcareous sand and limestone cave soils, with Sporosarcina pasteurii ATCC 11859 (standard strain) used as a control. Urease activities of the four microbes were higher than that of S. pasteurii. The microbes and urea–CaCl2 medium were injected at least four times into cohesionless soils of two different relative densities (60% and 80%), and the amount of calcite precipitation was measured. It was found that the relative density of cohesionless soils significantly affects the amount of calcite precipitation and that there is a weak correlation between urease activity and calcite precipitation.

Domibacillus antri sp. nov., isolated from the soil of a cave.

A Gram-reaction-positive, strictly aerobic, capsule-forming, motile and rod-shaped bacterium, designated strain XD80T, was isolated from the soil of a native cave in Lichuan, Hubei province, China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain XD80T was most closely related to Domibacillus iocasae CCTCC AB 2015183T (98.66 % sequence similarity), followed by Domibacillus robiginosus DSM 25058T(97.83 %), Domibacillus tundrae KCTC 33549T (97.70 %), Domibacillus enclensis CCTCC AB 2011121T (97.21 %) and Domibacillus indicus DSM 28032T (96.96 %). Levels of DNA-DNA relatedness between strain XD80T and D. iocasae CCTCC AB 2015183T, D. robiginosusDSM 25058T, D. tundrae KCTC 33549T and D. enclensis CCTCC AB 2011121T were 37.4 %, 53.8 %, 53.6 % and 52.7 %, respectively. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, aminophospholipid and two unknown phospholipids. The predominant fatty acids (>5 %) were iso-C15 : 0 (37.3 %), anteiso-C15 : 0 (10.8 %), C16 : 0 (10.4 %), iso-C17 : 0 (10.3 %), C16 : 1ω11c (9.6 %) and anteiso-C17 : 0 (7.4 %). MK-6 (86.4 %) was the major respiratory quinone. The DNA G+C content was 46.4 mol%. The cell-wall peptidoglycan contained meso-diaminopimelic acid (type A1γ). Ribose and glucose were the major whole-cell sugars. In addition, strain XD80T showed differential physiological characteristics from most members of the genus Domibacillus, encompassing hydrolysis of starch, acid production from inositol and raffinose, and production of valine arylamidase. The results of this polyphasic study indicated that strain XD80T represents a novel species of the genus Domibacillus, for which the name Domibacillus antri sp. nov. is proposed. The type strain is XD80T (=CCTCC AB 2015053T=KCTC 33636T).

Proposal of Sphaerimonospora cavernae gen. nov., sp. nov. and transfer of Microbispora mesophila (Zhang et al., 1998) to Sphaerimonospora mesophila comb. nov. and Microbispora thailandensis (Duangmal et al., 2012) to Sphaerimonospora thailandensis comb. nov.

The actinomycete strain N74T, isolated from cave soil, was studied using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain N74T formed a stable, distinct lineage cluster together with Microbispora thailandensis NN276T (99.3% similarity) and Microbispora mesophila JCM 3151T (97.5%). Strain N74T was observed to produce single spherical spores on aerial mycelium as reported for M. mesophila JCM 3151T and M. thailandensis NN276T but different from other known species of the genus Microbispora, which are characterized by pairs of spores on aerial hyphae. Multilocus sequence analyses based on concatenated partial gyrB, rpoB, atpD, recA and 16S rRNA gene sequences showed a clear distinction of strain N74T, M. mesophila JCM 3151T and M. thailandensis NN276T from other members of the genus Microbispora, although the chemotaxonomic characteristics of strain N74T were similar to the genus Microbispora; the cell wall contained meso-diaminopimelic acid and the whole-cell hydrolysate contained madurose as the diagnostic sugar. The major menaquinone was MK-9(H4). The fatty acid profile contained iso-C16:0. On the basis of morphological, chemotaxonomic and phylogenetic evidence, strain N74T is assigned to a novel species of a new genus, for which the name Sphaerimonospora cavernae gen. nov., sp. nov. is proposed. The type strain of Sphaerimonospora cavernae is N74T (=BCC 77604T=NBRC 111481T). It is also proposed that M. mesophila and M. thailandensis be transferred to this genus as Sphaerimonospora mesophila comb. nov. (type strain JCM 3151T=NBRC 14179T=DSM 43048T) and Sphaerimonospora thailandensis comb. nov. (type strain NN276T=BCC 41490T=NBRC 107569T), respectively.

Theoretical Analyses and Predictions of Indoor Thermal Environment for Cave Dwelling in Northwest of China

The population living in loess plateau is about 108 million, but so far there are about 27.8% of them still living in cave dwelling, which is a represented traditional residence in northwest China. Whether traditional cave dwelling should be inherited or replaced is gradually received extensive attention at home and abroad which has different characteristics, for example, adjusting measures to local conditions, low-cost, warm in winter and cool in summer with the rapid development of China's urbanization. Just as other vernacular architectures in China, the traditional cave dwellings are renovated in recent years. To compare the indoor thermal environment performance of cave dwellings and other local typical inhabitancy style, the cave-dwelling, courtyard, new cave-dwelling, general residence was selected as the research object being carried out indoor thermal environment of survey and questionnaires during July 4 ∼14, 2014, which is four typical architectural forms in Qingyang of Gansu province in loess plateau zones. The calculated value of prediction evaluation index PMV is generally higher than heat Sensation Vote value TSV through measuring. It turned out that the cave dwelling have a better thermal environment, but this test only based on one point temperature. However, in order to know about temperature field distribution, cliff soil cave dwelling, strengthening soil cave dwelling and new cave dwelling is regarded as objects through establishing physical model and heat transfer model at the same time. This article being based on different building structures deduces indoor temperature distribution. The result show that the temperature exponential decay along the cave dwelling depth direction. In addition, the airflow distribution inside cave dwelling are analyzed in theory using the application of plume model under the effect of hot-pressing ventilation. The article also get cave dwelling air velocity distribution of different interface.

Bacillus cavernae sp. nov. isolated from cave soil

A Gram-stain-positive, spore-forming, motile, strictly aerobic, rod-shaped bacterium, designated strain L5T, was isolated from soil of Tenglong cave, China. 16S rRNA gene sequence analysis showed that strain L5T was related most closely to Bacillus asahii MA001T (96.5 %) (the highest 16S rRNA gene sequence similarity), Bacillus kribbensis BT080T (96.4 %) and Bacillus deserti ZLD-8T (96.2 %). The DNA G+C content of strain L5T was 45.6 mol%. The major menaquinone was MK-7. The major fatty acids were iso-C14 : 0, anteiso-C15 : 0 and iso-C16 : 0, and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The diagnostic diamino acid in the cell-wall peptidoglycan was meso-diaminopimelic acid. In addition, strain L5T had different characteristics compared with the other Bacillus strains such as pink colony colour, low growth temperature and low nutrient requirement. The results indicate that strain L5T represents a novel species of the genus Bacillus, for which the name Bacillus cavernae sp. nov. is proposed. The type strain is L5T ( = KCTC 33637T = CCTCC AB 2015055T).

Paenibacillus cavernae sp. nov., isolated from soil of a natural cave.

A Gram-reaction-variable, strictly aerobic, motile, rod-shaped bacterium, designated strain C4-5T, was isolated from soil of a natural cave. Cells were oxidase- and catalase-positive and formed endospores in sporangia. The 16S rRNA gene sequence comparison showed that the organism formed a distinct clade within the genus Paenibacillus and was most closely related to Paenibacillus contaminans CKOBP-6T ( = KCTC 13623T) (96.85 % 16S rRNA gene sequence similarity) followed by Paenibacillus doosanensis CAU 1005T ( = KCTC 33036T) (94.82 %). The following chemotaxonomic features of strain C4-5T are typical for the genus Paenibacillus: meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, MK-7 as the predominant menaquinone, the major fatty acids of anteiso-C15 : 0, iso-C16 : 0 and C16 : 0 and the DNA G+C content of 54.8 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and four unidentified aminophospholipids. The phenotypic and phylogenetic data presented support that strain C4-5T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus cavernae sp. nov. is proposed. The type strain is C4-5T ( = KCTC 33652T = DSM 100100T).

Hymenobacter mucosus sp. nov., isolated from a karst cave soil sample.

A novel Gram-stain-negative, non-motile, rod-shaped and watermelon-red-pigmented aerobic bacterial strain, designated YIM 77969T, was isolated from a soil sample of Jiuxiang cave, a tourism cave located in Yiliang county, Yunnan province, south-west China. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 77969T belongs to the genus Hymenobacter, and was closely related to Hymenobacter tibetensis XTM003T (96.58 %), Hymenobacter gelipurpurascens Txg1T (96.02 %) and Hymenobacter xinjiangensis X2-1gT (95.80 %). Growth of strain YIM 77969T occurred at 5-35 °C, at pH 5.0-9.0 and in the presence of 0-1 % (w/v) NaCl. The predominant menaquinone was MK-7. The major fatty acids were iso-C15 : 0, C16 : 1ω5c and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The polar lipid profiles consisted of the major compound phosphatidylethanolamine, two unknown aminolipids, three unknown aminophospholipids, one glycolipid and one unknown polar lipid. Pigment analysis showed that the pigment belonged to the plectaniaxanthin series of carotenoid pigments. The genomic DNA G+C content was 55.2 mol%. On the basis of phylogenetic, phenotypic and chemotaxonomic characteristics, strain YIM 77969T is considered to represent a novel species of the genus Hymenobacter, for which the name Hymenobacter mucosus sp. nov. is proposed. The type strain is YIM 77969T ( = KCTC 32567T = DSM 28041T).

Changes in the storage and sink of carbon dioxide in subsurface atmospheres controlled by climate-driven processes: the case of the Ojo Guareña karst system

A comprehensive environmental monitoring program was conducted in the Ojo Guarena cave system (Spain), one of the longest cave systems in Europe, to assess the magnitude of the spatiotemporal changes in carbon dioxide gas (CO2) in the cave–soil–atmosphere profile. The key climate-driven processes involved in gas exchange, primarily gas diffusion and cave ventilation due to advective forces, were characterized. The spatial distributions of both processes were described through measurements of CO2 and its carbon isotopic signal (δ13C[CO2]) from exterior, soil and cave air samples analyzed by cavity ring-down spectroscopy (CRDS). The trigger mechanisms of air advection (temperature or air density differences or barometric imbalances) were controlled by continuous logging systems. Radon monitoring was also used to characterize the changing airflow that results in a predictable seasonal or daily pattern of CO2 concentrations and its carbon isotopic signal. Large daily oscillations of CO2 levels, ranging from 680 to 1900 ppm day−1 on average, were registered during the daily oscillations of the exterior air temperature around the cave air temperature. These daily variations in CO2 concentration were unobservable once the outside air temperature was continuously below the cave temperature and a prevailing advective-renewal of cave air was established, such that the daily-averaged concentrations of CO2 reached minimum values close to atmospheric background. The daily pulses of CO2 and other tracer gases such as radon (222Rn) were smoothed in the inner cave locations, where fluctuation of both gases was primarily correlated with medium-term changes in air pressure. A pooled analysis of these data provided evidence that atmospheric air that is inhaled into dynamically ventilated caves can then return to the lower troposphere as CO2-rich cave air.

Draft Genome Sequence of Jiangella alkaliphila KCTC 19222T, Isolated from Cave Soil in Jeju, Republic of Korea.

We report the draft genome sequence of Jiangella alkaliphila KCTC 19222T, isolated from cave soil in Jeju, Republic of Korea. This genome sequence, together with the previously sequenced J. gansuensis strain DSM 44835T, identified from a desert environmental source, will give us a better understanding of the school of “evolutionary taxonomy.”

Xiakemycin A, a novel pyranonaphthoquinone antibiotic, produced by the Streptomyces sp. CC8-201 from the soil of a karst cave.

Xiakemycin A, a novel pyranonaphthoquinone antibiotic, produced by the Streptomyces sp. CC8-201 from the soil of a karst cave

Examination of several Oklahoma bat hibernacula cave soils forPseudogymnoascus destructans,the causative agent of White-Nose Syndrome

White-nose syndrome is an often lethal fungal infection of bats that is caused by the fungus Pseudogymnoascus destructans, formerly Geomyces destructans. The fungal spores can persist for extended periods of time in the soil and on surfaces in caves where it might be found even after the bats depart. In 2010, a single bat, Myotis velifer, from a western Oklahoma gypsum cave, was initially diagnosed by the U.S. Geological Survey National Wildlife Health Center, as “suspect white-nose syndrome.” Based on this, we decided to examine soil samples from various bat caves across Oklahoma for the presence of P. destructans. We used Real-time Polymerase Chain Reaction to analyze 83 soil samples from 17 caves in Oklahoma. None of the soil samples were found to contain genetic material from P. destructans. We postulate that P. destructans has not yet reached Oklahoma because of the negative Real-time Polymerase Chain Reaction results, because a National Wildlife Health Center reexamination of the original bat suggested the bat was negative for P. destructans, and subsequent analyses of bat nose and wing swabs, bat tissues, and physical examination of hibernating bats have all been negative for white-nose syndrome.

Oscheius onirici sp. n. (Nematoda: Rhabditidae): a new entomopathogenic nematode from an Italian cave.

Oscheius onirici sp. n. (Nematoda: Rhabditidae) was isolated from a karst cave soil of Central Italy. Molecular and morphological analyses were performed. Total DNA was extracted from individual nematodes and the mitochondrial COI, the ITS containing region, the D2-D3 expansion domains of the 28S rRNA gene and the 18S rRNA gene were amplified and sequenced. BLAST search at NCBI by using all molecular markers revealed that this taxon is similar to Oscheius species. Phylogenetic trees of ITS, 28S and 18S rDNA revealed that O. onirici sp. n. belongs to Dolichura-group. Oscheius onirici sp. n. is characterized by small body size and stoma rhabditoid type. Female reproductive system is amphidelphic. Males are rare with peloderan bursa, spicules slender and small, nine pairs of papillae of different lengths, arranged in a 1+1+1/3+3 pattern. Entomopathogenicity bioassay revealed that this nematode is capable of infecting larvae of Galleria mellonella and Tenebrio molitor.

Bio-economics of melanin biosynthesis using electromagnetic field resistant Streptomyces sp.-EF1 isolated from cave soil

Moving charges due to electric current create electromagnetic waves, which generate electromagnetic fields (EMFs). The low frequency EMFs are unlikely to cause adverse effects on living organisms; however, health-related incidents have prompted researchers to investigate the biochemical characteristics in microorganisms. We hypothesized that a diverse variety of microorganisms, i.e., bacteria, will respond to EMF and bio-stimulate microbial metabolic products of commercial significance. An EMF generator that produces a 10-mT-strength magnetic field was built in-house to investigate the magnetic response of microbial communities from cave soil samples. 16S rRNA sequencing was employed to identify three EMF-responsive isolates: Streptomyces sp., Arthrobacter sp., and Lysinibacillus sphaericus, designated as EF1, EF2, and EF3, respectively. Streptomyces sp.-EF1 showed a dark brown pigment identified as melanin on nutrient agar plates supplemented with 0.4 % L-tyrosine and L-DOPA. We also investigated the bio-economics of melanin biosynthesis, which revealed that waste shredded paper ($33.29 /g melanin) followed by starch ($256.03 /g melanin) were among suitable substrates for melanin biosynthesis under non-EMF condition. The most time-efficient melanin formation was achieved by starch under submerged fermentation (3.52 h/μg/mL) conditions. Shredded, laser-printer paper was noticed to be a cost-efficient process ($33.29 /g melanin) for melanin biosynthesis.

Natural causes of changes in marine environment: Mercury speciation and distribution in anchialine caves

Mercury speciation and distribution were assessed, for the first time, from the water, sediment, rock, soil and air of anchialine caves. We evaluated the origin and distribution of four mercury species (total (THg), reactive (RHg), dissolved gaseous mercury (DGM) and monomethylmercury (MeHg)) in water from Bjejajka cave (Bjejajka) and Lenga Pit (Lenga) in the Croatian Adriatic Sea from 2006 to 2011.Concentrations of all mercury species were elevated at both sites compared to adjacent seawater, which had concentration range of 0.5–2.4 ng L−1 for THg, 0.06–0.2 ng L−1 for RHg, 0.02–0.07 ng L−1 for DGM, and 0.01–0.02 ng L−1 for MeHg. In Bjejajka, the maximum THg concentration (350 ng L−1) in the middle layer (3–7 m) of the 12 m deep stratified water column was up to 3 orders of magnitude greater than in adjacent seawater. During and after the rainy sampling periods in January 2009 and 2010, mixing of Bjejajka water column resulted in elevated concentrations, up to 3700 ng L−1 of THg (4 orders of magnitude higher compared to values in nearby seawater) in the bottom water layer. MeHg concentrations in the Bjejajka water column were also considerably elevated (0.7–6.3 ng L−1) compared to the surrounding seawater (0.01–0.02 ng L−1). The vertical distribution of MeHg concentrations followed that of THg, however the ratio of MeHg/THg above the Bjejajka halocline was drastically higher (up to 57%) compared to MeHg proportion (1–2 %) below the halocline, which was similar to that of surface seawater. In sediment of Bjejajka, THg concentrations were up to 3.3 mg kg−1, considerably above concentrations in unpolluted Adriatic marine sediment (0.1–0.3 mg kg−1). The highest THg amounts found in soil and air were inside and in close proximity to Bjejajka, while THg in rock (≤0.01 mg kg−1) were below reported values for unaltered carbonates.Laboratory experiments indicate that bat guano was the major source of elevated mercury concentrations in the water column and sediment of Bjejajka. Concentrations of THg in bat guano were up to 0.45 mg kg−1, above the range of THg concentrations found in unpolluted marine sediment. Seventy-two days after the addition of bat guano in solution, a 100-fold higher Hg concentration was observed, from 2.5 up to 252 ng L−1. In Lenga bat guano was not spotted and THg water and sediment concentrations were drastically lower, mostly below 7 ng L−1 and 0.8 mg kg−1, respectively.

Long-Term Persistence of Pseudogymnoascus destructans, the Causative Agent of White-Nose Syndrome, in the Absence of Bats.

Wildlife diseases have been implicated in the declines and extinctions of several species. The ability of a pathogen to persist outside its host, existing as an "environmental reservoir", can exacerbate the impact of a disease and increase the likelihood of host extinction. Pseudogymnoascus destructans, the fungal pathogen that causes white-nose syndrome in bats, has been found in cave soil during the summer when hibernating bats had likely been absent for several months. However, whether the pathogen can persist over multiple years in the absence of bats is unknown, and long-term persistence of the pathogen can influence whether hibernacula where bats have been locally extirpated due to disease can be subsequently recolonized. Here, we show that P. destructans is capable of long-term persistence in the laboratory in the absence of bats. We cultured P. destructans from dried agar plates that had been kept at 5°C and low humidity conditions (30-40% RH) for more than 5years. This suggests that P. destructans can persist in the absence of bats for long periods which may prevent the recolonization of hibernation, sites where bat populations were extirpated. This increases the extinction risk of bats affected by this disease.

Assessment of Site Conditions for Karst Areas with Complex Lithology: A Case Study of a Site for a Candidate Power Plant

Construction of a project at a proposed site requires understanding of the geological and hydrogeological conditions for the purpose of engineering design and environmental impact assessment. However, investigation and assessment of the site conditions may be challenging for limestone areas due to the irregular occurrence and development of karst. This paper presents a demonstration of the site investigation and assessment for the karst hydrogeological conditions through a case study of a site with complex lithology. A joint on-site surveying and mapping, drilling, geophysical exploration, supplemented by in-situ and laboratory tests, was implemented in the assessment. Results indicate the site area presents a multiplicity in lithology with steep attitude stratum. Northern part of the site distributed shales suffering different degree of weathering, while the rest areas were developed with carbonate rocks. Fracture zones with north-west and north-east trend were developed at the site, which are prone to form strong runoff zone of the groundwater. No shallow buried river, hall-or corridor-type karst caves were detected at the site, neither of large-scale well-connectivity karst channels. However, many small karst and soil caves were developed at the site, which suggest a post treatment of engineering measures for the purpose of engineering and environmental safety. Based on the distribution of karst ratio calculated from boreholes, the proposed site area can be divided into 3 zones of very-strongly developed zone with a karst ratio around 71.2%, strongly developed zone with a ratio around 26.3% and intermediate-slightly developed zone with a ratio of 15.0%. Characteristics of each sub-zones are analyzed regarding to the general layout and later engineering design of the power plant project. This study provides an example of a joint study for site condition assessment at karst areas with complex lithology. Such an assessment is crucial to the site selection, project layout, engineering design and to later environmental impact assessment.

Research on Detection Standard of Karst Roadbed Grouting Effect Based on the Geophysical Methods

To study the grouting detection standard of Karst roadbed based on resistivity method, transient Surface wave method and electromagnetic wave CT, the typical test areas in the Karst roadbed of NANNING-GUANGZHOU railway are selected, and the field test research of grouting detection standard of Karst roadbed was studied by the above three geophysical methods. The grouting qualified Karst roadbed was determined by multiple methods of borehole coring and field water pressure after grouting, and the test results of Karst roadbed grouting qualified was confirmed effective sample. The difference value of resistivity, surface wave velocity and electromagnetic wave absorption coefficient of limestone layer, clay, pebble soil and Karst cave was statistical analyzed based on the effective test data, and preliminarily obtained the detection standard of Karst roadbed grouting effect based on the above three geophysical methods.

Generalized cyclic p–y curve modeling for analysis of laterally loaded piles

Owing to their simplicity and reasonable accuracy, Beam on Nonlinear Winkler Foundation (BNWF) models are widely used for the analysis of laterally loaded piles. Their main drawback is idealizing the soil continuum with discrete uncoupled springs representing the soil reactions to pile movement. Static p–y curves, obtained from limited full-scaled field tests, are generally used as a backbone curve of the model. However, these empirically derived p–y curves could not incorporate the effects of various pile properties and soil continuity. The strain wedge method (SWM) has been improved to assess the nonlinear p–y curve response of laterally loaded piles based on a three-dimensional soil–pile interaction through a passive wedge developed in front of the pile. In this paper, the SWM based p–y curve is implemented as the backbone curves of developed BNWF model to study the nonlinear response of single pile under cyclic lateral loading. The developed nonlinear model is capable of accounting for various important soil–pile interaction response features such as soil and pile yielding, cyclic degradation of soil stiffness and strength under generalized loading, soil–pile gap formation with soil cave-in and recompression, and energy dissipation. Some experimental tests are studied to verify the BNWF model and examine the effect of each factor on the response of laterally loaded pile embedded in sand and clay. The experimental data and computed results agree well, confirming the model ability to predict the response of piles under one-way and two-way cyclic loading. The results show that the developed model can satisfactorily simulate the pile stiffness hardening due to soil cave in and sand densification as observed in the experiment. It is also concluded from the results that the gap formation and soil degradation have significant effects on the increase of lateral pile-head deflection and maximum bending moment of the pile in cohesive soils.

Novelties from a cave lace bug (Heteroptera, Tingidae): first nymphal description with a discussion on the morphological patterns of Vianaidinae immatures

A nymph of Thaumamannia Drake & Davis, 1960, a rare vianaidine genus known only from adults, is described by the first time. The specimens were collected in the soil and walls of caves located in the state of Pará, Brazil. This is the first record of Vianaidinae for Brazil. The T. vanderdrifti fifth instar is compared with the same stadium of Anommatocoris coleopteratus (Kormilev, 1955), the only other vianaidine nymph described so far, and patterns between them and within the family are also discussed.

The genus Hylebainosoma Verhoeff, 1899 (Diplopoda, Chordeumatida, Haaseidae): redescription of Hylebainosoma tatranum, description of a new troglobiont species and notes to the Hylebainosoma-Romanosoma species group.

A revision of the taxonomy, distribution and ecology of the millipedes of the genus Hylebainosoma Verhoeff, 1899 in the Carpathians is presented. Based on the study of numerous specimens, we consider the subspecies of the species Hylebainosoma tatranum Verhoeff, 1899 described in the past as overrated, reflecting simply intrapopulational variation attributable to the rather extended area of species distribution, different habitats of its occurrence (surface and cave habitats, soil, litter and stony debris) and wide altitudinal range from the forest zone in foothills to the alpine zone above timberline. Besides the redescription of males, the morphology of female vulvae is presented for the first time. Hylebainosoma tatranum is endemic to the extensive area of the Western and Eastern Carpathians, involving Slovakia, Hungary, the Czech Republic, Poland and highly probably Ukraine. A new troglobiont species from the karst area near the Tisovec Town (Muránska planina Mts., Central Slovakia), Hylebainosoma gulickai n. sp. is described, and is considered as stenoendemic for this small karstic region, with occurrence in few nearby caves. Hylebainosoma gulickai represents the first troglobiont chordeumatid millipede found in the northern territories of the Carpathians and the northernmost troglobiont in Central Europe in general. Morphological characteristics of both males and females are presented.Taxonomic relationships between the closely related genera Hylebainosoma and Romanosoma Ceuca, 1967 are discussed and replacement of the species Hylebainosoma cavernicola (Ceuca, 1967) n. comb., Hylebainosoma oltenica (Ceuca, 1967) n. comb. and Hylebainosoma odici (Ceuca, 1979) n. comb. into the genus Hylebainosoma is proposed. The fourth species described as Romanosoma (?) birtei Ceuca, 1967 we consider as invalid taxon. Romanosoma becomes a junior subjective synonym of the genus Hylebainosoma.