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ISSN (Online) 2603-3925
Published by Pensoft Homepage  [58 journals]
  • Factors affecting soil invertebrate biodiversity in agroecosystems of the
           Po Plain area (Italy)

    • Abstract: ARPHA Conference s 5: e95808
      DOI : 10.3897/aca.5.e95808
      Authors : Matteo Montagna, Matteo Brunetti, Alberto Spada, Alex Cussigh, Sumer Alali, Paola Cremonesi, Flavia Pizzi, Giulia Magoga, Pietro Marino Gallina : Soil is a fundamental component of the biosphere, whose properties and quality are affected by human activities, such as agriculture. Soil health is fundamental for different ecosystem services and soil biota has a crucial role in maintaining it. Elucidating how different crops and agricultural practices affect soil invertebrates communities is of relevance. In the present study, a DNA metabarcoding approach was adopted to evaluate the effects of different biotic and abiotic factors, including agricultural practices, on the composition and diversity of soil invertebrate communities of different agro-ecosystems (Po Plain-Italy). At this aim, the DNA markers and the more effective primers in retrieving soil metazoan communities were established. Bulk soil samples from different agro-ecosystems (i.e., cornfield, alfalfa, paddy fields, vineyard, stable meadow, woodland) were collected and, processed for obtaining 18S rRNA and coi sequences (raw reads analyzed using QIIME2 and R). Soil physical and chemical parameters were measured for each soil sample (e.g., pH, carbon-nitrogen ratio, texture, porosity) and metadata on farms management were retrieved. The most efficient primer pairs in recovering soil metazoans were M620F/M1260R for 18S rRNA, and mlCOIintF/jgHCO2198R for coi gene. Soil communities resulted dominated by Nematoda, Arthropoda, Annelida, Rotifera and Tardigrada. The most diverse invertebrate communities have been found in the soil of stable meadows and woodlands, while cornfields showed the lowest level of diversity. The diversity of soil invertebrate communities (Hill numbers) was positively correlated with the level of porosity and carbon-nitrogen ratio, while it was negatively correlated with the phosphate abundance. This pattern probably reflects the negative effect of excessive fertilization with phosphates on soil fauna, while the abundance of organic matter and microhabitats were found to enhance the presence of more complex communities. Other soil properties were correlated only with specific taxa (e.g., pH was negatively correlated with the diversity of Annelida and Rotifera). HTML XML PDF
      PubDate: Thu, 29 Sep 2022 12:15:00 +030
  • The response of aquatic fauna to variable environmental conditions in
           Ghețarul de la Vârtop cave (Apuseni Natural Park, Romania)

    • Abstract: ARPHA Conference s 5: e90725
      DOI : 10.3897/aca.5.e90725
      Authors : Orest Sambor, Karina Battes, Ana Camacho, Mirela Cîmpean, Constantin Marin, Aurel Perșoiu, Bianca Șarcani, Alin Tudorache, Sanda Iepure : Caves with permanent and temporary ice formations exists at mid-to-high northern latitudes, at elevations between 0 and>3000 meters above sea level. In such caves, rare and endemic species or glacial relicts strictly tied to cold microclimates habitat conditions have evolved and the negative temperatures have contributed significantly in shaping the structural pattern of both terrestrial and aquatic communities. Aquatic dwellers inhabiting ice caves are likely to show resistance and have special physiological adaptation to cope with constantly low air and water temperatures. Ghețarul de la Vârtop cave is a short (340 m) cave located in the Apuseni Natural Park (northwest Romania) that hosts temporary ice formations near its entrance. In 2021, we have initiated a study aiming to understand how low temperatures in the cave are shaping the structure of underground fauna, along a temperature gradient through the cave. The sampling design imply monthly monitoring of air temperature (hourly measurements using data loggers), water physical and chemical characteristics and the structure of aquatic fauna communities. In this paper we present preliminary data on the environmental conditions and aquatic invertebrate communities present in percolation water and associated gours from the cave. Aquatic fauna is represented by nematodes, oligochaetes and several crustacean species the majority stygobites, of which at least three are potentially new to science. The surprising presence of Acanthocyclops reductus, considered a Tertiary relict living in warm water raises discussions on its tolerance to highly variable temperatures but also on the colonisation history of the caves on a regional scale. Investigations of cave aquatic fauna in caves with permanent and temporary ice offer hints to understand the ecology of the fauna, and also to further assess the mechanisms involved in adaptations of species to cope with constantly low-water temperatures. HTML XML PDF
      PubDate: Mon, 1 Aug 2022 10:00:00 +0300
  • Evolutionary insights and constraints from the nervous systems and
           behavior of cavefish

    • Abstract: ARPHA Conference s 5: e90267
      DOI : 10.3897/aca.5.e90267
      Authors : Daphne Soares : Caves and other subterranean habitats represent one of the most challenging environments on the planet. Bony fishes are one of the few vertebrate groups that have successfully colonized and are completely restricted to those habitats. Despite being known to science for over 150 years, only recently have cavefishes become model systems for evolutionary studies pertaining the nervous system. Several cavefishes, such as the Mexican characid Astyanax mexicanus, have provided valuable insights into how fishes have evolved to cope with life in perpetual darkness. Here, I summarize the current understanding of nonvisual sensory modalities and divergent social behavior in the Mexican Astyanax mexicanus, the Brazilian Eigenmannia vicentespelea and the Ecuadorian Astroblepus pholeter. Only future comparative studies nested within well-resolved phylogenies will clarify the sensory adaptation of fishes to subterranean habitats. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • The cave salamander Proteus anguinus in captivity in Moulis, France for
           the scientific research.

    • Abstract: ARPHA Conference s 5: e89484
      DOI : 10.3897/aca.5.e89484
      Authors : Olivier Guillaume : Proteus anguinus fascinated the most famous savants like Lamarck, Cuvier and Darwin. However, the knowledge about this species was improved little because very few specimens were available, even less alive. To face this situation, The French National Center for Scientific Research built in 1948 the cave-laboratory at Moulis and started to breed in P. anguinus. The first hatchings date from 1959 (Vandel and Bouillon 1959, Juberthie et al. 1991). Since then, more than 160 egg-layings producing more than 4,600 eggs has occurred. The availability of a certain number of specimens and the continuity of this breeding for more than 60 years have made possible to better understand the biology of this species, for instance, the development (sexual maturity reached at 15 years old, lifespan estimated from 68 to 100 years old, see Voituron et al. 2010). The aim of this presentation is to make a review of the latest and prospective research conducted with partners on captive P. anguinus in Moulis. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • Overcoming shortfalls and impediments in subterranean biology: a challenge
           for the future

    • Abstract: ARPHA Conference s 5: e90209
      DOI : 10.3897/aca.5.e90209
      Authors : Fabio Stoch, Jean-François Flot : Since the discovery and description of the first specialized cave species, subterranean fauna stimulated the scientific research of several generations of speleobiologists especially after the publication of Racovitza's classification of cave species, which is still used today, amended for non-karstic areas and groundwaters. More than 28,000 obligate subterranean species are known worldwide; however, these figures are likely to be underestimated since species richness is highly correlated with research effort (Stoch and Galassi 2010). Subterranean ecosystems are very rich in strict endemic species and taxonomic efforts are still quite low (i.e., the so-called "taxonomic impediment"), while several environments are very difficult to be explored (i.e., the "Racovitzan impediment: Ficetola et al. 2018).Furthermore, several paradigms were debated for years. Albeit constrasting hypotheses were conceived to explain the colonization of subsurface habitats, their importance is still debated (i.e., climate relicts vs. adaptive shift in colonization and speciation, dispersal vs. vicariance in shaping distributional patterns, and selective vs. neutral hypotheses in explaining regressive evolution). Moreover, the paradigm of a "truncated functional diversity" of subterranean ecosystems (Gibert and Deharveng 2002) lasted for years but was recently challenged by the discovery of chemoautotrophic ecosystems in hypogenic and anchialine caves, and the recognition that caves are not isolated environments, but they are highly interconnected with surface ecosystems. The increased importance of their conservation (like in the case of GDEs, Groundwater Dependent Ecosystems) was recently recognized, together with their provision of important ecosystem services (Boulton 2020).A deeper knowledge is required to assess biodiversity hotspots as well as to plan efficient monitoring surveys (Mammola et al. 2020). In the last decades, a growing amount of molecular data has been obtained for subterranean species, allowing some of the classical debates on colonization, evolution, and dispersal to be revisited (Bauzà-Ribot et al. 2012); moreover, novel promising techniques like metabarcoding and environmental DNA were applied in field surveys and monitoring efforts.Unfortunately, after more than one century of research in subterranean biology, large gaps remain in our knowledge of phylogeny, richness, and distribution of subterranean fauna (formalized in the so-called Darwinian, Linnean and Wallacean shortfalls), preventing the definition of large-scale sound management and protection plans. It is proposed that data from recent biomolecular techniques coupled with remotely sensed data may enhance biodiversity mapping and conservation and are promising approaches to fill our knowledge gaps. Perhaps this is the greatest challenge that tomorrow's subterranean biologists will face. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • A Feulgen glimpse into genome evolution during range expansion: a case
           study of the subterranean amphipod Niphargus schellenbergi

    • Abstract: ARPHA Conference s 5: e90203
      DOI : 10.3897/aca.5.e90203
      Authors : Mohammed M.Tawfeeq, Dieter Weber, Jean-François Flot : In 1923, Robert Feulgen designed a quantitative approach to measure the relative DNA content of a species (its so-called “C-value”, which is defined as half of the DNA content of a somatic cell and is expressed in pg). The Feulgen approach has several advantages: it can be used on ethanol-preserved samples, it is relatively inexpensive, and it does not require sophisticated equipment (Hardie et al. 2002, Jeffery and Gregory 2014, Kasten 2003). Gathering DNA content information is a key preliminary step for whole-genome sequencing of non-model species and also yield important information for ecological and evolutionary studies as well as for conservation (Jeffery et al. 2013), since genome size has been shown to be strongly correlated to extinction risk in animals and plants (Vinogradov 2004, Vinogradov 2003).Among invertebrates, Niphargidae is among the largest families of freshwater subterranean amphipods in the world, and more than 450 niphargid species have been described (Horton et al. 2021). Thus, it is considered an ideal model for evolutionary and ecological studies. Using a new, improved Feulgen protocol, we generated novel genome size estimates for 26 samples of Niphargus schellenbergi collected from central and western European niphargids. The obtained C-values ranged from 2.75 to 5.25 pg. Such a two-fold intraspecific variation is unusual but not unheard of: it occurs also in the monogonont rotifer Brachionus asplanchnoidis (Stelzer et al. 2021). Further research will be required to find out the mechanisms responsible for this variation, which may result from copy-number variation, polyploidy and/or various amounts of transposable elements. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • Occurrence of Li in groundwaters and plants from Dobrogea karst area,

    • Abstract: ARPHA Conference s 5: e90172
      DOI : 10.3897/aca.5.e90172
      Authors : Anamaria Iulia Torok, Ana Moldovan, Erika Levei, Oana Cadar, Claudiu Tănăselia, Oana Teodora Moldovan : A positive association of Li consumption with the potentially protective and beneficial for the human health was reported (Barjasteh-Askari et al. 2020). Drinking water, grains, or vegetables can be a significant Li source for humans. Microdoses of Li intake may have antisuicidal, mood-stabilizing, antidepressive, and antimanic effects (Knudsen et al. 2017; Ng et al. 2019). The assessment of naturally occurring Li concentrations in water and food sources in different regions may present a high interest in the wellbeing of locals. In this study, a versatile quantitative ICP-MS method for Li quantitative determination in water and plant samples was optimized, and the relationship between Li, macroelements (Na, Mg, Al, K, Ca, Fe, Mn), and microelements (Cr, Co, Ni, Cu, Zn, Pb, Sr, Ba, V, As, Sr, Cd, Pb) concentration was assessed. Contents of Li, and micro- and macroelements were measured in groundwater (Praporgescu-GWR27, Closca-GWR28, Sipote-GWR29, and Tufani-GWR30) and plant samples (ryegrass—Lolium sp., nettles—Urtica sp., and mint—Mentha sp.) collected from Dobrogea karst area, Romania. The results indicated an acceptable precision in all studied matrixes and a reproducibility between 2.46 and 4.22% of the developed method. In the case of water, the highest Li concentration was measured in GWR27 followed by GWR28 (12.2 and 5.6 µg/L), while in the case of the plant’s samples, Lolium sp. collected from GWR28 and GWR27 (11.1 and 8.8 mg/kg DW) had the highest Li concentration. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • Stressed out underground': Illuminating the genomic response to heat
           stress in surface and subterranean predaceous diving beetles

    • Abstract: ARPHA Conference s 5: e90170
      DOI : 10.3897/aca.5.e90170
      Authors : Perry Beasley-Hall, Terry Bertozzi, Tessa Bradford, Charles Foster, Karl Jones, William Humphreys, Andrew Austin, Steven Cooper : Subterranean realms possess high environmental stability with respect to light levels, temperature, and humidity. The transition to a subterranean lifestyle can therefore cause massive shifts in a species’ biology. How does the colonisation of these habitats affect the thermal tolerance of an organism' Past studies demonstrate species in extremely stable environments might lose the ability to mount a heat shock response, which involves the expression of heat shock proteins to remediate misfolded or denaturing proteins as a result of heat stress. Such organisms might therefore be at particular risk of decline in the face of climate change. However, similarly robust data are limited for subterranean organisms. To help address this knowledge gap, here we compared the heat-shock response of a surface-dwelling and subterranean species of predaceous diving beetle (Dytiscidae, Hydroporinae), Paroster nigroadumbratus and P. macrosturtensis, the latter of which is restricted to a single groundwater aquifer in the Yilgarn region of Western Australia. Though P. macrosturtensis is able to survive warmer conditions than it encounters in nature based on past survival experiments, it is nonetheless less robust in the face of temperature extremes compared to surface-dwelling relatives; the genomic basis of this difference is unknown. By sequencing transcriptomes of experimentally heat-shocked individuals we demonstrate both species can indeed mount a heat shock response at high temperatures (35ºC). However, the genes involved in these responses differ and a far greater number are differentially expressed in the surface species, including those involved in remediating oxidative stress, which might explain its more robust response to heat stress. In contrast, the subterranean species significantly upregulated a heat shock protein gene under conditions it encounters in nature, suggesting it is far more sensitive to ambient stressors. These findings have conservation implications for P. macrosturtensis and contribute to a growing narrative concerning weakened thermal tolerances in obligate subterranean organisms at the molecular level. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • Is the gut microbiome involved in adaptation of beetles to caves'

    • Abstract: ARPHA Conference s 5: e90169
      DOI : 10.3897/aca.5.e90169
      Authors : Oana Teodora Moldovan, Paul Adrian Bulzu, Erika Levei, Ruxandra Nastase-Bucur, Cristian Sitar, Catalina Haidau : Cave beetles are endemic to one or few caves and live in a stable subterranean microclimate and permanent darkness. However, not much is known about the food sources of cave beetles and the share of autotrophic sources in their diet. Seven cave beetles were sampled seasonally from five Romanian caves together with a sampling of sediments and climatic measurements. Gut was extracted from the sampled specimens, and Illumina's 16S amplicon-based metagenomics sequencing protocol was applied to identify the gut microbiome. About 50% of the most abundant bacteria genera in the studied sediments are unknown taxa. The core elements of the sediments' microbiome were wb1-P19, Pseudomonas, and Lysobacter, all producers and decomposers of organic matter. Only Acinetobacter was shared for sediments and all cave beetles. The core bacteria in the gut of Leiodidae beetles of all five species were Vagococcus, Dysgonomonas, Candidatus Soleaferrea, and two unknown phylotypes. The cave Carabidae gut was dominated by Carnobacterium in one species and Enhydrobacter in the other, showing different food regimes within the Duvalius genus. Vagococcus was present in the gut microbiome of Leptodirini and Duvalius, a genus which might be involved in adaptation to life in caves, where food is scarce and autochthonous productivity is low. Higher body weight in cave beetles is another helpful strategy to cope with an infrequent food supply, as demonstrated by the presence of obesity-related gut microbiota representatives, Lactococcus and Serratia. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • The ground beetles (Coleoptera, Carabidae) from the René Jeannel
           collection of the Babeș-Bolyai University Zoological Museum, Romania

    • Abstract: ARPHA Conference s 5: e90094
      DOI : 10.3897/aca.5.e90094
      Authors : Cristian Sitar, Geanina Iacob, Marius Kenesz, Oana Teodora Moldovan : 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. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • The genus Protopholeuon (Coleoptera, Leptodirini): distribution,
           morphological, ultratructural and genetic details

    • Abstract: ARPHA Conference s 5: e90130
      DOI : 10.3897/aca.5.e90130
      Authors : Cristian Sitar, Ruxandra Bucur, Marius Kenesz, Oana Teodora Moldovan : 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. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • Biology Students Association as the base for establishing future experts
           and cave fauna exploration

    • Abstract: ARPHA Conference s 5: e90136
      DOI : 10.3897/aca.5.e90136
      Authors : Jana Bedek, Mia Šepčević, Dora Kermek, Lea Okićki, Nikolina Kuharić, Tvrtko Dražina : Biology Students Association – BIUS (BIUS) from Zagreb, Croatia is a non-government and non-profit association founded in 1999 by biology students from the Faculty of Science, University of Zagreb. Aiming to develop future experts in different fields of biology, BIUS is composed of groups that specialize in various subject areas. Besides holding diverse science lectures, workshops, and field trips, the main BIUS' activities are annual student research camps with the participation of up to 130 members.An integral part of BIUS from the very beginning is the Biospeleology group (Group). During these first 23 years of the Group's existence, many members discovered their interest in cave biology through field research, cabinet work, and educational workshops. The Group organized two student research camps and participated in 16 others. In addition, the Group organized seven independent projects. Throughout all of these projects, cave fauna was collected from over 150 caves and pits at different localities of Croatian karst. The Group's long-term existence is facilitated by the guidance of mentors from the Croatian Biospeleological Society. Altogether, the Group provides numerous skills and valuable opportunities to students and is important for developing future scientific experts in cave biology in Croatia.So far, 12 Group members have written their master thesis and four have their doctoral thesis in the field of cave biology. Former members currently work at universities, research institutes, government, and civil sector, or as entrepreneurs on topics closely related to cave biology and karst research. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • IUCN red-listing of subterranean invertebrates: problems, gaps and the

    • Abstract: ARPHA Conference s 5: e90134
      DOI : 10.3897/aca.5.e90134
      Authors : Louis Deharveng, Ana Komericki, Sonia Khela : Red-listing is a widely used approach to globally evaluate the threats that affect a species, according to a set of standardized criteria. The IUCN Red List of Threatened Species is the world's most comprehensive inventory of the global conservation status of species. It is an international reference in this field, and a powerful tool for conservation of endangered species, sites and habitats.A number of subterranean organisms of various taxonomical groups have been assessed from a few decades, and are currently being assessed, revealing serious difficulties and uncertainties linked to the special characteristics of subterranean ecosystems and species. The most critical problems, that are linked to distribution, dispersion, life cycle and sensitivity to disturbance, and some ways to overcome them, are discussed in this paper.Partly linked to this, redlisting of subterranean species has so far achieved moderate results with regard to the diversity and vulnerability of subterranean species - with the well-known exception of bats. Data drawn from IUCN online database of redlsted species reveals that subterranean invertebrates are hugely under-covered at geographical and taxonomic levels. Moreover, among red-listed species that are connected to subterranean habitats, most are stygophiles and troglophiles, i.e. not obligate cave dwellers. In contrast, very few of the species strictly linked to subterranean life (stygobionts and troglobionts) have been red-listed so far, in spite of the exceptional proneness to endemicity of subterranean invertebrate groups.On an other hand, assessments are extremely uneven across countries and across taxa. Many regions with significant cave fauna do not have any assessed subterranean species. The richest countries in subterranean diversity are not those which have the highest number of assessed species. The zoological invertebrate groups that dominate cave fauna in diversity (beetles, springtails, microcrustacea...) have contrasted proportion of redlisted species: higher for snails, very low for microcrustaceans, beetles and springtails, with rare local exceptions.Assessment difficulties underlined above, low coverage of cave fauna in available assessments, limited progress in number of red-listed species in the face of the high number of concerned species, and the limited human resources to do the job led us to favour a more pragmatic approach to subterranean invertebrates red-listing for the coming years. Really threatened subterranean species are actually not numerous, because subterranean habitats are protected from most critical disturbance that devastate so many surface habitats. The aim will be therefore to focus on the most threatened species and sites in the world, with a particular attention on mining, water pollution and large scale surface habitats destruction. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:10:00 +030
  • Speleomics: omics insights into subterranean ecosystems

    • Abstract: ARPHA Conference s 5: e90095
      DOI : 10.3897/aca.5.e90095
      Authors : Jean-François Flot : The vocable 'omics' encompasses genomics, transcriptomics, proteomics and metabolomics. All these subfields deal with what used to be called "big data" that describe complex interactions. What can be the use of such highly sophisticated approaches when dealing with caves and other subterranean environments' Are subterranean ecosystems omics-worth'Throughout this presentation, I will discuss recent, ongoing and planned studies when I used, use or will be using omics approach to shed light on the biology and biodiversity of cave ecosystems, as well as on the often invisible interspecific interactions they help reveal. Far from replacing traditional taxonomy and naturalist approaches to cave studies, I propose to blend them with omics to gain a more holistic understanding of subterranean ecosystem. I dub this brave new field 'speleomics'. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Identification of sulfur–oxidizing Thiothrix bacteria on
           microcrustaceans from the sulfidic groundwaters of Mangalia (southeastern

    • Abstract: ARPHA Conference s 5: e90001
      DOI : 10.3897/aca.5.e90001
      Authors : Filip Boancă, Claire Chauveau, Jean-François Flot, Maria Pop, Sanda Iepure : Movile Cave, located in southeastern Romania close to the Black Sea, is one of the most remarkable diversity hotspots worldwid with 52 species of invertebrates of which 37 are endemic (Brad et al. 2021). Due to the high concentration of hydrogen sulfide in its water, the primary production of organic matter in Movile Cave relies almost entirely on the chemoautotrophic activity of microorganisms, notably sulfur-oxidizing gammaproteobacteria belonging to the genus Thiothrix. In the presence of oxygen, these filamentous bacteria can oxidize hydrogen sulfide and reduce it into various sulfidic compounds, generating energy in the process. In sulfidic ecosystems, Thiothrix bacteria are frequently found free-living but also as epibionts or ectosymbionts growing on other organisms, such as amphipods (Flot et al. 2014). However, it is unclear whether Thiothrix bacteria also grow on microcrustaceans such as copepods or ostracods, of which several species are known from the sulfidic mesothermal aquifer of Mangalia, where Movile Cave is located. To find it out, we combined DNA sequencing using the reference bacterial 16S ribosomal RNA gene with morphological observations (including fluorescence microscopy). Our results reveal that Thiothrix bacteria are indeed present on microcrustaceans from Movile Cave and surrounding wells in the Mangalia region, highlighting the versatility of Thiothrix-crustacean associations in sulfidic ecosystems. This is the first report of an association between Thiothrix and groundwater microcrustaceans, and the second report of an association between Thiothrix and a nonmarine ostracod (Khalzov et al. 2021). HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The microbiome of phosphate-rich deposits in Muierilor Cave, South-Western

    • Abstract: ARPHA Conference s 5: e89863
      DOI : 10.3897/aca.5.e89863
      Authors : Catalina Haidau, Ruxandra Nastase-Bucur, Paul Bulzu, Ionut Mirea, Luchiana Faur, Silviu Constantin, Oana Moldovan : Muierilor Cave is one of the most important caves in Romania from paleontological, biological, and archaeological points of view. A newly discovered chamber, with unique yellow calcite crystals, fine-grained crusts, and black sediments, is connected to the upper levels that contains fossil bones and a large pile of guano. Samples were taken from this chamber and another passage to investigate the diversity of microorganisms related to the substrates and identify potential pathogenic taxa for humans and animals. Chemical, mineralogical, and whole community 16S rRNA gene-based metabarcoding analyses were undertaken, and the base of the guano deposit was radiocarbon dated. Metabarcoding of the analyzed samples found that ~16% of the identified bacteria are potentially pathogenic to humans. Moreover, more than 87% of the identified genera were not previously reported in caves. We identified bacteria involved in the phosphate cycle that can only originate from the organic deposits inside the cave, such as the bats' guano in the touristic, upper level, or the fossil bones. Our study indicate also the guano deposit as the possible source of potentially pathogenic microorganisms. The results on the microbiome of different deposit types unravel the diversity of microorganisms and indicate the potentially pathogenic taxa for humans and animals. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Insights into the Eucyclops graeteri species complex (Copepoda,
           Cyclopidae) — the case of sulphidic groundwaters of Mangalia (Southern
           Dobrogea, Romania)

    • Abstract: ARPHA Conference s 5: e89798
      DOI : 10.3897/aca.5.e89798
      Authors : Maria Pop, Andrei Ștefan, Filip Boancă, Fabio Stoch, Sanda Iepure : Members of the Eucyclops graeteri species complex are widely distributed in European groundwaters and share some morphocharacters such as:a long aesthetasc on the ninth segment of female antennula,short or very reduced spinule row ("serra") on furcal rami,lateral hairiness of 5th thoracic somite strongly reduced, andfew or no hairs on the distal margin of the intercoxal sclerite of 4th legs pair.All these characters are subject to strong selective pressure in the underground environment; therefore, the study of molecular markers is needed to better define the taxonomy and phylogeny of this species group. The Southern Dobrogea region in Romania conceals dark, isolated, and sulfide-rich aquifers, accessible through artificial wells, springs, and the Movile Cave. The chemoautotrophically-based Movile Cave hosts a member of the E. graeteri complex, described several years ago as a subspecies (E. graeteri scythicus Plesa, 1989). The morphological and molecular analyses performed on specimens of this species complex collected from Southern Dobrogea revealed new records for E. graeteri scythicus both in sulfidic and non-sulfidic artificial wells dug near the cave as well as a species putatively new to science collected in three sulfidic wells from Mangalia. Mitochondrial markers allocated the Movile Cave population in a distinct clade within the genus Eucyclops, Eucyclops serrulatus s.l. being the closest taxon. The putative new species belongs to a different clade within the subfamily Eucyclopinae, raising doubts on the monophyly of this species complex. Furthermore, our investigations revealed the occurrence of ectosymbiont sulfur-oxidizing Thiothrix bacteria associated with E. graeteri scythicus suggesting that this putative symbiosis could allow this taxon to better cope with sulphidic-rich environments. These unique sulfidic groundwater environments therefore provide an interesting model system in which to study the taxonomy of copepods and their adaptation to very harsh environmental conditions. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Molecular phylogeny of Alpine groundwater Niphargidae (Crustacea,
           Amphipoda) reveals the effects of Quaternary climate fluctuations

    • Abstract: ARPHA Conference s 5: e89760
      DOI : 10.3897/aca.5.e89760
      Authors : Alice Salussolia, Jean-François Flot, Fabio Stoch : The groundwater amphipod genus Niphargus comprises over 425 described species mainly distributed in the Western Palearctic. Representatives of the genus are quite rare in areas formerly occupied by Quaternary glaciers. Although a couple of species were previously discovered in high-elevation caves and karstic springs, it is only recently that the genus was found to be widely distributed along the Alpine chain (Fišer et al. 2018; Stoch et al. 2020). Using a molecular phylogeny approach, we tested the hypothesis of multiple origins of Alpine Niphargus species followed by dispersal versus allopatric speciation in glacial refugia. We sampled all type localities of species present in the Alps and collected at several new sites, both in formerly glaciated areas and in areas very close to the last Quaternary glacial border. All sequences of Alpine species stored in GenBank were downloaded and included in the analysis as well. Our results show that several phyletic lineages independently colonized the Alpine chain at different periods. We found examples of preglacial colonization, with species surviving as glacial relicts with a very limited range expansion, as well as of recent post-glacial dispersal allowing colonization of very large areas on the northern slope of the Alps. In the Southern Limestone Alps as well as in the Western Alps, most species have a distribution restricted to small karstic massifs south of the area occupied by glaciers during the Last Glacial Maximum (Stoch et al. 2022), except for a single putative relict. Three clades show a relict distribution with highly isolated species found also in caves more than 2,000 m above sea level. The reconstructed time-calibrated phylogeny strongly supports the hypothesis that the evolutionary history of most clades is pre-Pleistocenic on the southern slope of the Alps, while on the northern slope both phylogenetic relicts and recent dispersers co-exist. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • A new species of Pseudomoraria from an alpine spring of Picos de Europa,
           North of Spain

    • Abstract: ARPHA Conference s 5: e89757
      DOI : 10.3897/aca.5.e89757
      Authors : Paulo Corgosinho, Sanda Iepure, Aleksandr Novikov : Pseudomoraria triglavensis was described by Brancelj (1994) from specimens collected in a high-alpine reservoir at the Triglav National Park (NW Slovenia) at an altitude of 1690 m a.s.l. During an expedition in the Picos de Europa National Park (N Spain) in 2018, a new species of Pseudomoraria was collected in an epikarst spring (Fuente Escondido), below the ice cave of Altaiz at an altitude of 2112 m a.s.l. The new species can be easily distinguished from Pseudomoraria triglavensis by the presence of an inner seta on the edopodite 1 of female and male pereopod 1; absence of the outer spine on the exopodite 2 of the third and fourth female pereopods; four, instead of five setae on the second endopodite of the fourth female pereopod; and the female furca lacks the ventral distal patch of spinules characteristic of P. triglavensis. In the male, the new species differs from its congeners by the position and shape of the apophysis of the second edopodite of the pereopod 3, which is positioned in the outer, instead of the inner margin, and is bent around the distal inner spine; the second exopodite of the pereopod 4 has spines/setae, lacking the innermost seta which is present in P. triglavensis. The female armature of the pereopod 5 is highly variable, with 4-6 setae/spines on the exopod and 4-6 setae on the baso-endopod. According to Brancelj (1994), P. triglavensis could not be included in any genus known at the time and concluded, based on the armature of the pereopod 5 of both sexes, that it would be most closely related to Moraria. We disagree with this author and propose a close relationship of this genus with the genus Hypocamptus Chappuis, 1929 sharing similar male pereopod segmentation and armature. In addition, the species belonging to both genera are a characteristic for the fauna of the alpine water bodies. It is not yet clear the phylogenetic relationship between the two genera, but evidences suggest that Pseudomoraria may be a junior synonym of Hypocamptus. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Study of the Invertebrate diversity in Prometheus Show Cave (Georgia,

    • Abstract: ARPHA Conference s 5: e89721
      DOI : 10.3897/aca.5.e89721
      Authors : Eter Maghradze, Shalva Barjadze, Arnaud Faille, Zezva Asanidze : Prometheus Cave is one of the largest caves in Georgia among the local six show caves. Before opening the cave as a tourist attraction, no research was conducted on the cave to study the invertebrate community living there, despite the cave's status as a natural monument. Before our study, only 22 species of invertebrates were known from Prometheus Cave, while none of the invertebrate species have been reported from adjacent non-touristic Datvi and Melouri caves.Cave invertebrate fauna was monitored monthly from 2018 to 2021 in the Prometheus cave and adjacent non-touristic Datvi and Melouri caves. The sampling was conducted on at least ten sites per cave each month. The abundance and incidence of the invertebrates were recorded on each site during field surveys. We ordinated the invertebrate diversity of the studied caves using Principal Component Analysis (PCA). After intensive investigations for three years, the number of species in Prometheus Cave increased from 22 to 47. Besides, ten species in Datvi and 11 species in Melouri caves were found for the first time. Our results suggest the existence of a significant difference in the diversity of the cave invertebrate fauna between the touristic part of Prometheus show Cave and its non-touristic part - Alpinist's hall, Datvi, and Melouri caves. The primary factor differentiating Prometheus show Cave from the others is the lower abundance of the species. Hurlbert's PIE and rarefaction analysis of the diversity explains that the diversity is also most unevenly distributed in this cave. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Decipher groundwater food web interactions by means of stable isotope and
           gut microbiome analyses

    • Abstract: ARPHA Conference s 5: e89712
      DOI : 10.3897/aca.5.e89712
      Authors : Christian Griebler, Clemens Karwautz, Moritz Grabner, Ayac Jimenez Salvador, Felix Puff, Rajiv-Raphael Kumar, Mona Lauritz, Lina Vinkovic, Julian Kiralyhidi, Gertraud Steniczka, Andreas Fuchs : The traditional perception of groundwater food webs is that of short and poorly structured food chains extending to only few trophic levels. This is mainly attributed to the lack of light and the shortfall of substantial primary production as well as the shortage of dissolved organic carbon and nutrients. It is frequently argued that microbes form the basis of groundwater food webs. In reality, the obvious lack of comprehensive biofilms may point to detritus as the main common food source and an opportunistic and omnivorous feeding strategy with most groups and species. Also, the relative stability of groundwater environments may have led to low trait variability. However, recent research also contrasts the paradigm of poorly structured food chains against emerging convoluted mechanisms sustaining a great range of biotic diversity and functional complexity. In summary, understanding of the ecological and functional linkages among stygofaunal groups, and their interaction with the microbial community, is still in its infancy. Stygofauna sampled from three spots of a shallow alluvial aquifer in the vicinity of the Danube River in the city of Vienna, Austria, has been analyzed by means of bulk stable carbon and nitrogen isotope and gut microbiome analysis to disentangle food web interactions. Our preliminary data revealed the following pattern:plant material, terrestrial insects, and detritus show a narrow range in carbon (-32 to -25δ13C) and a wide range in nitrogen (-1.5 to 6.5 δ15N) isotope signatures. In terms of nitrogen isotope values, the next tropic level is represented by isopods and gastropods, followed by ostracods, oligochaetes and amphipods. The broader range of stygofauna in carbon isotope signatures indicate an additional food source; most probably microbes;gut microbiome analyses indicate differences with oligochaetes, gastropods, ostracodes, isopods and amphipodes, however, with a few individuals from the later three groups sharing a very similar gut microbiome;within the amphipods, there is obviously a difference in gut microbiome composition with species and sampling sites;carbon and nitrogen isotope signatures underline amphipod-species differences;microbiome diversity in all cases was less divers in the animal guts when compared to environmental water, sediment and detritus samples.Finally, we first collected evidence for anaerobic processes taking place in animal guts living in an oxic groundwater environment. In light of our data, putative groundwater food web interactions in the shallow alluvial aquifer are discussed. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Phylogenetic diversity of water scorpions (Nepa spp., Insecta, Hemiptera)

    • Abstract: ARPHA Conference s 5: e89707
      DOI : 10.3897/aca.5.e89707
      Authors : Andrei Ștefan, Jean-François Flot, Elena Iorgu, Luis Popa, Steven Keffer, Fabio Stoch, Șerban Sârbu : Water scorpions belonging to the genus Nepa are predatory freshwater aquatic insects of the order Hemiptera. There are currently five accepted species, with a Holarctic distribution: N. cinerea (Eurasia and Northern Africa), N. sardiniensis (Sardinia and Corsica), N. anophthalma (endemic to Movile Cave, Romania), N. apiculata (North America) and N. hoffmanni (Eastern Asia). Mitochondrial and nuclear genetic markers indicate a correlation between genetic diversity and geographic distribution. Analyses also reveal a cryptic diversity in the Western Mediterranean basin, with specimens of the cinerea clade being assigned to the sardiniensis clade. The cave-adapted N. anophthalma is genetically closest to N. cinerea, suggesting surface populations of N. cinerea as possible ancestors. N. cinerea samples from three cave systems in Italy indicate various degrees of gene flow between surface and subterranean populations and an instance of a possible incipient speciation event. Despite the overlap in distribution range in mainland Eastern Asia between N. cinerea and N. hoffmanni, there is a high genetic distance between the two species. A similar value of genetic distance is found between N. cinerea and N. apiculata, but also between N. hoffmanni and N. apiculata, suggesting the assignment of N. hoffmanni and N. apiculata to different genera. The single species currently considered as stygobiotic, N. anophthalma, deserves more research to understand the timing of colonization of Movile groundwater by its surface ancestors. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Quantitative microbial risk assessment as a tool for groundwater
           monitoring. A case study in the rural communities of Romania

    • Abstract: ARPHA Conference s 5: e89699
      DOI : 10.3897/aca.5.e89699
      Authors : Zamfira Stupar, Erika Levei, Emilia Neag, Oana Teodora Moldovan : Karst springs are an essential source of drinking water in rural communities in North-Western Romania. Due to the fractured dolomite aquifers, dolines, or blind valleys, these springs are vulnerable to contamination by human and zoonotic pathogens. In this study, we assessed the human health risk associated with the contamination of six springs’ water with gastrointestinal pathogens. The data on the occurrence of E. coli and estimated Campylobacter and Rotavirus for a year of monitoring from twenty-four samples were inputted into a Quantitative Microbial Risk Assessment (QMRA) model (Machdar et al. 2013). The QMRA was conducted following steps: hazard identification, dose-response assessment, exposure assessment, and risk characterization (Haas C N et al. 2014; World Health Organization 2016). In order to predict the total cases of illnesses for the spring water consumers, the probability of infection and illness was calculated for adults and children. Results have shown that the estimated probability of infection for enteropathogenic E. coli, Campylobacter, and Rotavirus ranged between 1.1 × 10-3 and 1.0 × 100, values much higher than the acceptable limit of 10-4 infections/person/year established by WHO (2016). The estimated probability of illness due to the pathogenic E. coli and Campylobacter infection was high, with values between 8.8 × 10-2 and 3.5 × 10-1 for five out of six groundwater sources. The estimated probability of illness due to the Rotavirus infection ranged between 4.0 × 10-4 and 6.4 × 10-2 for both adults and children. The finding suggests that consuming contaminated spring water could lead to severe acute, chronic, or sometimes fatal health consequences for the locals. This study provides valuable QMRA information on the contaminated karst groundwater, with important implications for human health and groundwater resources management. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Groundwater contamination and human health risk assessment in the main
           karst areas of Romania

    • Abstract: ARPHA Conference s 5: e89698
      DOI : 10.3897/aca.5.e89698
      Authors : Ana Moldovan, Anamaria Torok, Eniko Kovacs, Ionut-Cornel Mirea, Erika Levei, Oana Teodora Moldovan : Long-term monitoring reveals the temporal evolution of groundwater chemistry and the potential human health risk posed by the use of contaminated waters (Giri and Singh 2015). Groundwater aquifers are an important source of drinking water in Romania. This study was conducted to appraise the groundwater chemistry and the potential non-carcinogenic risks for human health associated with the groundwater consumption through oral and dermal pathways. In order to achieve this aim, 193 samples were collected from 29 groundwater sources in the main karst areas of Romania during 2019-2021. A total of 15 parameters were analyzed (F-, Cl-, SO42-, NO3, Ca, Na, K, Mg, Fe, Mn, Cu, Zn, Ni, Cr, and As) and compared with the World Health Organization standards for drinking water (WHO 2017). The heavy metals pollution index showed that no groundwater samples had detectable levels of metal contamination, while the heavy metal evaluation index revealed that 4 out of 29 groundwater sources were classified as contaminated: 3 sources with a medium level of pollution and 1 source with a high level of pollution, indicating a potential risk for human health. The human health risk for oral exposure indicated a potential non-carcinogenic risk only in the karst area from South Dobrogea. The non-carcinogenic risk posed by nitrates is higher than that posed by metals in the aquifers from South Dobrogea. Therefore, control and remedial actions should be taken to ensure appropriate water quality for the locals using the contaminated water sources. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Different components of interconnected karst environment select for highly
           distinct microbial communities

    • Abstract: ARPHA Conference s 5: e89679
      DOI : 10.3897/aca.5.e89679
      Authors : Anusha Singh, Tanja Shabarova, Paul Bulzu : Karst environments, formed by dissolution of rocks, present a unique opportunity to study groundwater and surface water as a single dynamic system. We aimed to explore the hydrological connections and identify shared microbiome between different components of this complex landscape (terrestrial, surface-subsurface freshwater networks and two different lake strata) on example of karst landscape at north shore of Lake Thun (CH).52 samples from soil and 87 from the freshwater network were collected during low hydrological conditions. Additionally, 43 samples from 13 different lakes across Europe were included in the analysis. A 16S rRNA gene amplicon analysis revealed highly homogenous lake communities dominated by typical lake microbes represented by members of Alphaproteobacteria, Gammaproteobacteria and Actinobacteria. On the contrary, communities in soils and surface and subsurface water were observed to be highly heterogenous within the karst landscapes. Soil samples built two robust groups that corresponded to alkaline and acidic pH values associated with two types of bedrocks. Alkaline soils had higher abundance of Gammaproteobacteria and Alphaproteobacteria, while acidic soils were dominated by Acidobacteria. Alkaline cluster was better connected with aquatic environments and shared genotypes from the families Nitrosomonadaceae and Nitrospiraceae with cave sediments and lake hypolimnia, that implied ammonia and nitrite oxidation as important chemotrophic processes in light depleted environments. Rhodoferax, Limnohabitans and Sediminibacterium were shared within subsurface network and lakes. Single Rhodoferax genotype was detected in all aquatic environments. Additionally, we observed that lineages from the families Pirellulaceae and Gemmataceae (phylum Planctomycetota) were alternatively distributed in both soil clusters and lakes. A similar pattern was observed in subsurface aquatic components and deeper lake strata within family Methylophilaceae, where Methylotenera was extensively replaced by Methylopumilus. This indicates that despite high hydrological connectedness between different karst landscape components, deterministic selective factors seem to prevail in the community assemblies. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Heat below the City – Is temperature a key driver in urban
           groundwater ecosystems'

    • Abstract: ARPHA Conference s 5: e89677
      DOI : 10.3897/aca.5.e89677
      Authors : Constanze Englisch, Eva Kaminsky, Cornelia Steiner, Christine Stumpp, Gregor Götzl, Christian Griebler : Living in an urban groundwater ecosystem comes with many challenges for its inhabitants. In Central Europe, groundwater fauna has adapted to a dark, cold, and typically energy-poor habitat for thousands of years, making these highly specialised animals susceptible to short-term (years to decades) changes in environmental conditions. In urban areas, many anthropogenic pressures like surface sealing, subsurface infrastructures, organic pollution, or accumulation of toxicants impact the groundwater ecosystem. On top, the urban subsurface is heating up with an enormous speed, characterized by the formation of subsurface urban heat islands. With urban groundwater temperatures continuously increasing way beyond natural background values, a loss of biodiversity and ecosystem functioning with ultimately a deterioration of water quality is predicted. In the current research project ‘Heat below the City’, we target the urban groundwater ecosystems of the city of Vienna, Austria. 150 groundwater wells distributed all over the city have been sampled twice, in autumn 2021 and spring 2022. Groundwater samples are analysed for a large set of physical-chemical, microbiological and faunistic variables, aiming to identify main drivers of groundwater ecosystem biodiversity and functioning. The focus lies on temperature induced cascading effects that may lead from ecologically intact oxic habitats to anoxic zones lacking fauna with deteriorated water quality. First findings show mean groundwater temperatures in Vienna to be at 14°C, about 3°C above the natural background, with anthropogenic heat sources having a main impact on the distribution and degree of warming. The absence of dissolved oxygen (DO) and NO3- as well as the presence of Fe2+, S2- and CH4 hint at zones of reduced groundwater below the city. First data on physical-chemical conditions, microbiological and faunal communities will be introduced. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • How much we know about Brazilian subterranean aquatic fauna

    • Abstract: ARPHA Conference s 5: e89658
      DOI : 10.3897/aca.5.e89658
      Authors : Tamires Zepon, Jonas Gallão, Maria Elina Bichuette : The first study concerning Brazilian subterranean fauna was the description of the phreatobitic fish Phreatobius cistenarum Goeldi, 1905, followed by the stygobitic fish Pimelodella kronei (Miranda Ribeiro, 1907). After that, biospeleological studies mainly focused on terrestrial taxa description, and in 1972 the first stygobitic invertebrate, Aegla cavernicola Türkay, 1972 (Decapoda, Aeglidae), was described. Here, we provided an overview of the Brazilian subterranean aquatic fauna knowledge. To date, ca. 270 troglobitic species are described, of which 64 are aquatic (22 fishes; 42 invertebrates) and 13 have amphibian habits (all isopods). Considering aquatic species, 15 species were described between 1905 and 1999, 13 in the 2000s, 27 in the 2010s, and 9 in the 2020s. Furthermore, new stygobitic species were confirmed by experts but not published due to the deficit of trained specialists. Brazil has the second higher world diversity in stygobitic fishes, and data about habitat, biology, and ecology are available. Most of them are generalist carnivorous occurring in alluvial sediments, shallow base-level streams, flooded caves, lakes in the water table, upper vadose tributaries, or epikarst aquifers. Usually, they have small populations, but some species have large population sizes for the subterranean patterns. Some species have regression of the agonistic, cryptobiotic, and photophobic behaviors and regression of the circadian rhythms compared to their epigean relatives. Moreover, some species have morphological and behavioral intraspecific differences. Around 50 fish species have stygophilic populations, and some studies compared biological and morphological aspects of epigean and hypogean populations. There are available data about the habitat and natural history of the described stygobitic invertebrates, of which most are amphipods and planarians. Few phylogenetic relicts and epikarstic species were described. Usually, stygobites have restricted distribution in a micro-basin or are endemic within a unique cave and have small populations. Morphological and populational studies are available for few taxa (e.g., decapods and gastropods), as well as behavior studies. Community studies discuss species richness and distribution related to abiotic variables, lithology, and responses to disturbances; only a study is about epikarstic communities. For stygophilic species, there are mainly studies on distribution patterns and new records. Concerning amphibian species, five species were described in the 2010s and eight in the 2020s, with notes on their natural history and habitat. The main threats to the subterranean aquatic fauna are pollution by pesticides, decreasing aquifer levels due to deforestation and water exploitation for irrigation, headwaters of rivers outside protected areas, suppression of habitats by mining and hydroelectrical activities, allied to the current dismantling and attacks to the Brazilian environmental policies. To date, 46 stygobites (16 fishes and 30 invertebrates) and five troglobitic amphibians are on the Brazilian Red List. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • On the conservation of subterranean ecosystems

    • Abstract: ARPHA Conference s 5: e89655
      DOI : 10.3897/aca.5.e89655
      Authors : Stefano Mammola : Subterranean ecosystems harbor a broad diversity of specialized organisms that are of interest from both a conservation and evolutionary perspective. These species are often short-range endemics, and some represent ancient faunas that disappeared from surface habitats. Thus, they account for a unique fraction of the global taxonomic, phylogenetic, and functional diversity that is currently imperiled by human activities, including the destruction of subterranean habitats, pollution, and climate change. Furthermore, subterranean ecosystems and landscapes deliver critical nature contributions to people, most notably the provisioning of potable water. To emphasize the importance and urgency of protecting subterranean biodiversity, the year 2021 was elected the International Year of Caves and Karst (later extended to 2022). Yet, the subterranean biome is still systematically overlooked in global biodiversity targets and climate change agendas. For example, only 6.9% of known subterranean ecosystems overlap with protected areas globally. Importantly, most of these subterranean ecosystems are protected simply because they occur within protected areas established for surface species or habitats, and therefore not designed to account for their vertical dimension and efficiently protect subterranean biota. Given this scenario, the time is ripe to provide a quantitative assessment of solution-based approaches to safeguard subterranean biota and associated habitats. During this lecture, I will discuss the available evidence for the effectiveness of conservation interventions in subterranean ecosystems and future directions concerning subterranean conservation. I will emphasize the importance of making conservation efforts in subterranean systems more practical, cost-effective, and long-lasting. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Preliminary data on the bacterial diversity of Dobšinská and
           Demänovská ice caves (Slovakia)

    • Abstract: ARPHA Conference s 5: e89620
      DOI : 10.3897/aca.5.e89620
      Authors : Cătălina Haidău, Alena Nováková, Mark Cunningham, Alexander Allenby, Deepak Kumaresan, Alexandra Hillebrand-Voiculescu : Our work brings preliminary data on the diversity of microorganisms in Dobšinská and Demänovská ice caves (Slovakia). The caves are located at about 80 km from each other. Demänovská Ice Cave is part of the Demänovské caves system, located in the Demänovská Valley in the Low Tatras National Park. Dobšinská Ice Cave (969m asl), is part of the Stratená Cave System, located on the south-western edge of the Slovak Paradise National Park in the Spiš-Gemer karst. Both caves have been opened for the public during the last decades of the 19th century and are presently intensively visited by tourists [in 2014 alone, Demänovská ice cave was visited by 70,769 tourists (Nudziková 2018)].There are few reports on the biocenosis of these two caves, the available data referring mainly to the invertebrate fauna (Papáč et al. 2019). In what concerns the microorganisms, up to our investigation, in focus have been only the fungi. Thus, in Dobšinská Ice Cave was reported the presence of species such as Botrytis cinerea and Aspergillus fumigatus isolated from bat guano (Nováková 2006) and the abundance of fungi outside and inside the cave was compared while from Demänovská Ice Cave, based on ITS sequences, Ogórek et al. (2018) reports on the phenotypic and genotypic diversity of airborne fungal spores.Our work complements these two studies adding data on the bacterial diversity from different parts of the two caves and depending on the substrate types (ice blocks and/or sediments).Using Sanger sequencing of the 16S amplicons from DNA isolated from pure bacterial cultures obtained from ice and/or sediments of Demänovská and Dobšinská Ice Caves, we identified 7 and respectively 12 species of bacteria. Surprisingly, only two species where common to both caves. In both Demänovská Ice Cave and Dobšinská Ice Cave the bacterial communities were dominated by Acinetobacter and Bacillus. Beside the identification of the main bacterial groups present in the underground habitats, Illumina sequencing enabled us to build genus heatmaps for the caves, allowing for comparision between various segments of each cave in relation with the specific climate features. When compared, the different types of samples (water, sediment, ice) showed different abundance of bacteria, ice diversity being greater than sediment/water diversity in both Dobšinska and Demänovská ice caves. The chemical composition of the samples (ice and water) is also discussed.This work is the first to bring insights on the bacterial diversity of Demänovská and Dobšinská Ice Caves and we appreciate what it opens new directions of research not only on the evolution of the microbial communities in relation to the climate changes occurred in past times, but also on how intense visiting affects the structures of the microorganisms communities. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Jugatala cribelliger (Berlese, 1904) a new oribatid mite species for the
           Romanian fauna

    • Abstract: ARPHA Conference s 5: e89438
      DOI : 10.3897/aca.5.e89438
      Authors : Ioana Nae, Augustin Nae, Mark Maraun : Jugatala cribelliger (Berlese, 1904) = Mycobates (Calyptozetes) cribelliger Subias, 2004 was found for the first time in the Romanian fauna. The species was identified using Electronic Microscope images. The species has an unknown habitat (Fischer and Schatz 2013) and is distributed in Northern Italy—Prov. Bolzano, Trento; Austria, Switzerland—Grisons; Iberian Peninsula. The specimen was collected form MSS (Mesovoid Shallow Substratum) from Piatra Craiului Mountains, Central Carpathians, Romania, in drillings used for sampling the alpine scree from Marele Grohotis, at an altitude of 1579 m. The combination of following characters of adults is considered as diagnostic features for Jugatala: rostrum rounded; lamella narrow to wide, with cuspides and translamella; lamellar cuspides without lateral and median dens or with minute lateral dens; bothridium cup shaped; sensillus with capitate or clavate head, rounded distally; tutorium narrow, with cusp pointed distally; notogaster with pteromorphs curved ventrally, line of desclerotization absent; lenticulus present or absent; 10 or 11 pairs of notogastral setae, dp present or absent; four to seven pairs of notogastral porose areas; six pairs of genital setae; all legs heterotridactylous; tibia I with dorsodistal apophysis bearing solenidion φ2; seta l” of tibiae and genua I–IV thick, heavily barbed (Bayartogtokh and Schatz 2008).With a distribution in Central and south-western Europe, Jugatala cribelliger (Berlese, 1904) is first recorded in the Carpathians, from Piatra Craiului Mountains in MSS, a subterranean environment. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Groundwater ecosystems in changing times

    • Abstract: ARPHA Conference s 5: e89199
      DOI : 10.3897/aca.5.e89199
      Authors : Susanne Schmidt, Miroslava Svátková, Vít Kodeš, Tanja Shabarova : Waterbodies worldwide undergo changes and this influences the ecosystems with the resident communities. Groundwater is no exception. However, few studies have so far focused on how the combined effects from catchment management and a changing climate impact the communities living in the groundwater. In 2019-2021 in southwestern Czech Republic, we sampled fauna and microorganisms in 37 wells that had been monitored by CHMI for up to 40 years and that varied in the trends in chemical and physical properties. The wells tapped the shallow quaternary and deeper aquifers of seven major hydrogeological zones. As expected, more quaternary than deeper wells showed - stronger - trends in physical and chemical properties of the groundwater. The chemical property changing significantly in the highest proportion of wells, i.e. 100% of the wells sampled for fauna, was silica. The trend of silica was increasing in all cases. Faunal numbers were lowest in the wells with the highest silica values. This may be an indirect effect of the interaction between changing agricultural practices and climate change, and may have implications for ecosystem functions, such as the use of groundwater for drinking water production. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Can the Karaman-Chappuis method significantly enhance understanding of
           stygofauna ecology'

    • Abstract: ARPHA Conference s 5: e89165
      DOI : 10.3897/aca.5.e89165
      Authors : Juliana Pille Arnold, Syngeon Rodman, Michael Curran, Dean Main : Comprehensive surveys on subterranean fauna are an emerging need for assessing species diversity patterns and for developing biodiversity conservation strategies considering anthropogenic changes to the environment (e.g., mineral and water extraction, climate change). Subterranean habitats comprise mainly terrestrial and aquatic aspects of caves and fissures, as well as interstitial waters. Sampling stygofauna in Northern Western Australia is different to other parts of the world due to the predominance of meso- and micro-caverns, and thus methods are largely constrained to net hauling or pumping of either drill holes or bores installed for mineral or groundwater exploration. Water is also encountered at shallower depths beneath water courses (the interstitial or hyporheic zone) and although it is known to share fauna with subterranean systems, is largely excluded in stygofauna studies. Interstitial fauna can be sampled with the well-known Karaman-Chappuis (KC) method, which involves digging a hole in saturated alluvial deposits, letting interstitial water accumulate, and use of a scoop net to collect invertebrates. However, the application of the KC as a standard complementary method for stygofauna surveys has to date been largely unexplored. In collaboration with Rio Tinto, we included the KC method alongside traditional subterranean fauna sampling at five study areas in the Hamersley Range of Pilbara region, Western Australia. We aimed to determine its utility to better understand the ecology of subterranean fauna recorded, and therefore improve conservation outcomes. Here, we summarise the faunal groups collected using the KC method (43 samples) from surveys conducted between 2016 and 2019. We found that the stygofaunal groups more frequently collected by the KC method were Ostracoda (30.4%), Cyclopoida (28.0%), Haplotaxida (12.5%) and Amphipoda (10.7%), but other groups – Harpacticoida, Syncarida, Aphanoneura, Isopoda, Enchytraeida and Trombidiformes – were also collected in lower frequency (total of 4.9% of the records). Notably, the KC method also collected troglofaunal groups as by catch. Pauropoda (28.1%), Symphyla (21.9%), Hemiptera (12.5%), Diplura (12.5%), Zygentoma (9.4%) and Chilopoda (6.3%) were the groups most frequently collected using the KC method, but Isopoda, Palpigradi and Polyxenida were also recorded (total of 9.3% of the records). Additionally, for one project where low level taxonomic data was available (based on morphological and molecular identification), we also assessed differences in species composition of stygofauna sampled with the KC method (10 samples) compared to the traditional haul net sampling method (139 samples). We found significant differences in stygofauna species composition between the KC and haul net methods, with 13 species (out of 78 species) collected exclusively with the KC method. However, there was also a considerable overlap of species collected by the two methods (11 species). Despite the current limited data, our findings demonstrate that the KC method could be complementary to traditional stygofauna sampling methods. It also allows sampling of potentially connected groundwater habitats to occur beyond the areas drilled for minerals or water. While this method usually targets stygofauna, the collection of troglofauna as by catch is significant and provides a potential way to evaluate the ecology of troglobitic taxa. Further study is needed to assess the KC method as a valuable addition to sampling methods aiming to produce comprehensive inventories of stygofauna that integrate species found in interstitial subterranean environments. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Maintaining laboratory cultures of cave-dwelling and surface-dwelling
           isopod crustaceans

    • Abstract: ARPHA Conference s 5: e87807
      DOI : 10.3897/aca.5.e87807
      Authors : Nikolina Kuharić, Marko Lukić, Jana Bedek, Lada Jovović, Tin Rožman, Daniel Fong, Helena Bilandžija : Many aspects of cave animal biology cannot be understood through field observations alone. Access to some subterranean habitats is often technically challenging, rendering long-term monitoring of cave-dwelling species or investigating some aspects of their biology difficult. Also, contemporary research questions and methods require sampling individuals at certain life stages, and most cave adapted species have lengthy life spans or are not accessible in their natural habitats year-round. Therefore, we have established an animal facility in our laboratory where we keep cave-dwelling and related surface-dwelling crustacean species under controlled conditions. This allows us to study physiological and behavioral adaptations and the role of phenotypic plasticity in the evolution of these traits in cave dwellers. We are culturing freshwater, marine, and terrestrial isopods from three different families (Asellidae, Sphaeromatidae, and Trichoniscidae), among others. To date, we have established breeding colonies of several different cave and surface species pairs of aquatic asellids, and our most successful colonies have lived for two years and three generations. We have also established colonies of cave (freshwater) and surface (marine) sphaeromatids with varying degrees of success. Among terrestrial trichoniscids, colonies of cave species have persisted for more than two years, but without successful reproduction. Our repeated attempts to maintain surface species of Trichoniscus have been unsuccessful. Here we present our experiences to provide guidance for other researchers making similar attempts. We discuss general care practices such as decontamination of equipment and tools, quarantine, good and bad practices for small aquarium and terrarium design, housing, water treatment and conditioning, various food sources, and breeding approaches. We also review some of the problems we have encountered with our colonies and describe our attempts to solve them with varying degrees of success. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Culturing cave mollusks in the laboratory: strategies and troubleshooting

    • Abstract: ARPHA Conference s 5: e87764
      DOI : 10.3897/aca.5.e87764
      Authors : Marko Lukić, Magdalena Grgić, Tin Rožman, Nikolina Kuharić, Lada Jovović, Robert Weck, Helena Bilandžija : Objective obstacles such as high water levels that prevent access to caves at certain times of the year or the need to use complex tools and skills such as cave diving make biological research in caves extremely complex and occasionally dangerous. Moreover, to study physiological and behavioral adaptations of cave animals, they must be kept under controlled laboratory conditions. For this reason, we have established an animal facility in our laboratory. The available literature on invertebrate setup, operation, and care was sparse and unsuitable for the species we were studying. Fortunately, many cave biologists provided us with advice and support during the initial phase. Here we present our experiences to provide some guidelines for other researchers undertaking similar efforts. Among other animal groups, we are culturing several morphotypes of the cave snail Physella sp. and two cave and one surface species pair of bivalves: Congeria spp. and Dreissena polymorpha. We discuss good and bad practices in mollusk care - housing, water treatment, different types of food, and small aquarium design. Special attention is given to problems we have encountered with our colonies and changes we have made to address these problems, both successful and unsuccessful. We also address the general protocols required for keeping multiple species in a single facility, including procedures for decontamination of equipment and tools, and quarantine. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Spatial genetic structure of Dinaric cave-dwelling spiders

    • Abstract: ARPHA Conference s 5: e87717
      DOI : 10.3897/aca.5.e87717
      Authors : Martina Pavlek, Jérémy Gauthier, Miquel Arnedo, Vanina Tonzo, Julia Bilat, Nadir Alvarez : Caves put strong constraints on organisms living there, which results in acquisition of variety of troglomorphic traits, most obvious ones being depigmentation and degradation of visual system (Christiansen 2012). Cave habitats are considered fragmented, and the animals living there face limited connectivity between potential habitats (Barr 1967, Bregović and Zagmajster 2016, Trontelj 2018). Additionally, once adapted to underground conditions, cave-dwellers are thought to be unable to use the surface for dispersal, thus demonstrating population dynamics similar to those of the species found in islands (Barr and Holsinger 1985). We compared the population genetic structure of several cave spider species with different biology and ecology from different caves in the Dinarides. Namely, depigmented and anophthalmic Dysderidae species which were never collected outside caves, and several species from the genus Troglohyphantes, which exhibit different levels of troglomorphisms and dependency on cave habitat (some, like T. excavatus and T. kordunlikanus, can be occasionally found in surface habitats). Additionally, Dysderidae are wandering active hunters, while all Troglohyphantes species spin webs. In our study, we applied a hybridization-capture approach, i.e., HyRAD (Suchan et al. 2016) to extract and capture DNA from historical samples. We compared the population genetic structures among the species, and studied isolation by distance, and identified contrasted genetic structures related to the biology and ecology of each species. We identified a deeper population structuring and more pronounced patterns of isolation by distance in the highly troglomorphic species compared to the less troglomorphic ones. Species that can occasionally be found in surface habitats showed less structured populations compatible with higher dispersal ability. Additionally, we detected several common breaks in gene flow for most studied species, some of which can be explained by geographic and geologic features. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Arthropod communities and drivers of their species diversity and
           composition in caves

    • Abstract: ARPHA Conference s 5: e87555
      DOI : 10.3897/aca.5.e87555
      Authors : Raluca Ioana Băncilă, Augustin Nae, Andrei Giurginca, Ioana Nae, Eugen Nitzu, Popa Ionut, Stefan Cӑtӑlin Baba, Rodica Plăiaşu : Understanding the drivers of diversity patterns in ecological communities remains a major challenge in ecology. Moreover, few studies have considered invertebrate or subterranean communities and little is known about which factors are responsible for structuring arthropod communities in caves. In this study we:assessed the abundance, richness, and composition of arthropod communities from eight caves located in Banat and Mehedinti region, Romania; anddetermined whether altitude, cave physical characteristics (temperature, relative humidity, light intensity) and features (cave extension, hight, width, heterogeneity, substrate (wall versus floor)) influence cave arthropod communities along the horizontal development of the cave (i.e. from the cave entrance, through the intermediate zone, to the aphotic zone).Overall, 89 species were found, of which three were obligate subterranean dwellers and 17 were endemic species. Species abundance distributions showed evident variation in species numbers among cave zones and between the wall and floor substrate. Species abundance was influenced by cave extension, relative humidity and light intensity. Species richness was related to cave extension while relative humidity and cave heterogeneity affected the species diversity. The results also indicated differences in species abundance, richness and diversity among cave zones and between substrate types. The arthropod community structure and composition was determined by altitude, relative humidity, temperature and substrate. The floor and wall substrate had different species composition while the species composition of the three cave zones largely overlapped. We conclude that several biotic and abiotic factors explain the diversity and composition of cave arthropods communities and that our results may be relevant to other types of island-like habitats. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The use of baited stygofauna traps as a complimentary sampling method for
           sampling groundwater bores in the Pilbara, Western Australia

    • Abstract: ARPHA Conference s 5: e87515
      DOI : 10.3897/aca.5.e87515
      Authors : Syngeon Rodman, Juliana Pille Arnold, Michael Curran, Dean Main : Stygofauna sampling predominantly comprises active sampling methods, such as net hauling or hand netting, which although highly effective at collecting specimens, can cause significant specimen damage and is unlikely to collect vagrant species. Studies assessing the efficacy of using stygofauna traps (both baited and unbaited) have been conducted with varying levels of success. Thus, the development of a robust and effective stygofauna trap design may be advantageous. Between 2021 and 2022, in conjunction with Rio Tinto, Biologic began developing and trialling a stygofauna trap designed to sample groundwater bores or drill holes intersecting groundwater, with the aim of evaluating the following hypotheses:The stygofauna trap design and method used would successfully collect stygofauna in groundwater bores and drill holes intersecting groundwater.Some taxa collected in stygofauna traps may differ from those collected in net haul samples, due to differences in ecological habit of fauna (crawling taxa colonising traps) or vagrant stygofauna (fauna travelling between suitable habitats).Specimens collected in traps will be in better condition than those collected in net haul samples. This has positive implications for taxonomic and molecular identification, as well as the collection of voucher specimens.Based on a fish funnel or box trap, the stygofauna traps consist of a 90mm cylindrical PVC frame, with a 150µm mesh inner lining. Both ends of the trap are capped with a removable funnel, allowing fauna to enter. In September 2021, a preliminary trial was conducted on sites intersecting the Robe and Bungaroo aquifers, in the coastal Pilbara region of Western Australia. These shallow aquifers are characterised by a highly abundant and rich stygofauna assemblage, typically dominated by crustaceans. Stygofauna traps (one trap per site) were deployed in seven groundwater bores or drill holes intersecting groundwater. The traps, baited with algae pellets, were set at the bottom of the bore, and collected between 12 and 24 hours of deployment. For comparison, net haul sampling was conducted at each site immediately prior to trap deployment. Results from the initial trial were promising, prompting a two season (wet and dry) study. This study commenced in March 2022, where traps were deployed in 11 sites across both aquifers. Two baited traps per site were deployed, one within 1m of the groundwater level and one at the end of the hole, for 72 hours, to determine if a longer deployment and an additional trap would yield better results. The final phase of sampling is planned for August 2022.Results from both the preliminary trial and first (wet) season of the 2022 study were very promising, with stygobitic taxa collected in four (of the seven) and nine (of eleven) trap sites respectively. The traps recorded a rich assemblage of stygofauna comprising Amphipoda, Copepoda, Gastropoda, Isopoda, Ostracoda and Thermosbaenacea. Compared to the respective net haul samples, the stygofauna traps yielded lower abundances and taxa richness. However, specimens collected in traps were in excellent morphological condition. At the conclusion of this study, taxonomic composition and abundance of stygofauna from the traps and net hauls will be analysed to provide further contextualisation of results. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Preliminary study on differences characterizes the populations of olm
           (Proteus anguinus) in cave and those found in epigean environment

    • Abstract: ARPHA Conference s 5: e87442
      DOI : 10.3897/aca.5.e87442
      Authors : Benedetta Barzaghi, Francesco Gentile Ficetola, Edgardo Mauri, Marco Restaino, Bianca Lombardi, Raoul Manenti : The olm (Proteus anguinus) is considered the most representative example of all the stygobiont fauna. Nevertheless, it is also reported in some spring habitats of Friuli Venezia Giulia (North-eastern Italy). Springs are intriguing habitats that border two strongly distinct environments: surface and underground. They may provide distinct environmental pressures to stygobiont animals promoting new adaptations.The aim of this study is to verify if there are differences in the structural morphology of the head and in the presence of melanin in the skin between the olms that are found in caves compared to the individuals found in the external environment.During 2021, 28 olms, 18 from underground habitats and 10 from external sites, were photographed in standard conditions with a metric reference. For each individual, only the cephalic region has been considered. The pictures were processed using TPSdig software. For each picture we placed 2 landmark homologues, matching the emergence of the gills branches, and 48 semi-landmark to define the contour of the head. With regard to melanin 2 mm of skin were removed from the tail of the olm and analyzed through a spectrophotometer. The results indicate a significant effect of the environment on the shape of the olm’s head, in fact the shape of the head appears more elongated in the front and narrower in the parietal region in the olms coming from the hypogean environment. In addition, the olms that use the epigean environment show higher melanin levels in their skin.These findings provide a first evidence of differences occurring between olm populations/sub-populations living in caves and those exploiting surface habitats. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Assessment of biological diversity of underground anthropic habitats in
           cultural landscapes

    • Abstract: ARPHA Conference s 5: e87429
      DOI : 10.3897/aca.5.e87429
      Authors : Rodica Plăiaşu, Raluca Băncilă, Andrei Giurginca, Augustin Nae, Eugen Nitzu [ Nițu ], Ionut Popa, Stefan Baba, Valerică Toma, Alexandru Petculescu : Subterranean habitats of anthropic origin (e.g. tunnels, mines, rupestrian settlements) have similar characteristics of natural voids, such as caves. These anthropic habitats have been shown to provide habitats for wildlife and harbor high species richness. In particular, rupestrian underground shelters have both natural and cultural importance. However, their assessment assumes the cultural value as significant whilst the natural importance is often ignored. Despite the contribution of subterranean anthropic habitats to underground biodiversity, few studies investigated invertebrate fauna of these habitats. We studied invertebrate communities structure and composition and the environmental factors that affect them using transects, following light intensity in the underground shelters, and in the surrounding surface habitats, in the rupestrian settlement Alunis-Bozioru, from Buzau County, Romania. The diversity of the rupestrian settlement was studied both at community-level and species-level (for Collembola, Diplopoda, Araneae, Opiliones and Coleoptera). The results showed an overlap in the taxa composition of invertebrate assemblages between the underground shelters and the surrounding surface habitats. The most abundant species in the underground shelters and surrounding surface habitats were among:Araneae Achaearanea tepidariorum, Tegenaria sp., Lepthyphantes leprosus, and Pardosa sp.;Diplopoda Cylindroiulus boleti, andOpiliones Lacinius dentiger.The underground shelters harbored a diverse invertebrate community dominated by taxa associated with dark habitats. The main factors affecting taxa composition were humidity and vegetation cover in the surface habitats. Overall, our results indicated that rupestrian underground shelters could provide substitute habitats and might play an important role in conservation of subterranean biodiversity in the context of increasing human pressures on subterranean ecosystems. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The role of phenotypic plasticity in the evolution of cave-dwelling

    • Abstract: ARPHA Conference s 5: e87225
      DOI : 10.3897/aca.5.e87225
      Authors : Helena Bilandžija : The surface ancestors of subterranean species are often hypothesized to possess pre-adaptations that enable them to successfully initiate colonization of caves. Nocturnal habits and the use of microhabitats such as the underside of rocks or leaf litter are two simple examples. However, there are many exceptions, and a mechanistic explanation for the evolution of a whole range of troglomorphic adaptations has yet to be found. To shed light on the phenotypic and molecular changes that accompany the early stages of cave colonization, we experimentally exposed the most closely related surface species or population of several cave-dwelling species including fish and crustaceans, from the earliest embryonic stage possible, to constant darkness, the single environmental condition characteristic of all subterranean habitats. Our results show that darkness induced a surprisingly large number of phenotypic changes, many of which resembled troglomorphic adaptations found in their respective cave-dwelling relatives (for example changes in fat content). However, some changes were contrary to what is considered adaptive in darkness, such as increased body pigmentation and enhancement in visual system, opposite to the two iconic features of troglomorphy. Overall, we conclude that phenotypic plasticity is an important mechanism for the rapid initiation of changes that may be favored or targeted by natural selection during critical initial steps in the evolution of cave-dwelling species. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Activity of stygobionts in spring habitats: behavioral, ecological and
           evolutionary insights

    • Abstract: ARPHA Conference s 5: e87217
      DOI : 10.3897/aca.5.e87217
      Authors : Veronica Zampieri, Filippomaria Cassarino, Giulia Pacinotti, Benedetta Barzaghi, Mattia Falaschi, Edgardo Mauri, Marco Restaino, Valentina Balestra, Danilo Borgatti, Matteo Galbiati, Stefano Lapadula, Valeria Messina, Magdalena Gajdošová, Roberta Pennati, Francesco Gentile Ficetola, Raoul Manenti : Cave-dwelling animals might be regarded as dead-end points when it comes to their evolution and habitat exploitation. However, in the past different observations of the olms’ typical troglomorphic populations have been reported for springs of Venetia Giulia, along with a community rich in stygobionts (Bressi et al. 1999; Stoch 2017). The aim of this work is to point out the non-random active use of surface habitats by this community, providing a comparison with the occurrence observed in caves and performing an assessment of factors favouring ecotone habitats exploitation. Since 2020 we started multiple day and night surveys of olms in both 71 springs and 11 caves. Each spring and cave habitat monitored has been characterised by respect to abiotic and biotic features, including planktonic and benthic prey availability. Additionally, cave and spring populations of Troglocaris sp. were tested to determine whether they showed any behavioral difference regarding their response to light stimuli and predatory cues, as potential adaptations to the different conditions found in the two habitats. We used 34 individuals sampled from three springs and 24 from two caves of the Italian karst.We detected the olm at least once in 12 springs, with a maximum of 9 individuals occurring together. Detection probability in springs and caves was similar. Spring habitats provided higher density of potential prey available. Olms seem to prefer springs without predator fish and temporary hydroperiod. We recorded in one spring a larva of 3.5 cm which could be the smallest ever recorded in the field. Preliminary results of the behavioral experiments show no susceptibility to light neither for spring nor cave populations of Troglocaris sp., as for chemical cues we expect a difference in reactivity depending on the habitat of origin of the shrimp, consistently with the top predator present.We suggest that epigean habitats and borders with surface may have an underestimated importance for animals adapted to subterranean environments (Manenti and Piazza 2021), including the olm. Further studies focusing on the ipogean community which actively exploits spring habitats are underway, with the aim of determining the diel occupation of these complex ecotones. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The morphology of the colonizers of the springs from the down-under

    • Abstract: ARPHA Conference s 5: e87202
      DOI : 10.3897/aca.5.e87202
      Authors : Anna Biró, Gergely Balázs, Žiga Fišer, Gábor Herczeg, Cene Fišer : Transition from surface to subterranean environment generally results in easily detectable morphological changes, generally referred as troglomophism. Although the cave colonisation history of species can differ considerably, in certain cases there is a possibility to directly examine the cave-induced morphological and life history changes, as some of the troglobiont and stygobiont species still have closely related surface species. A less common and therefore less known phenomenon is the invasion in the opposite direction, when cave-adapted species establish stable populations (and later, new species) in surface or surface-connected (ecotonal) habitats. A great model taxon to study if morphological changes occur in such cases is the Niphargus genus, which primarily comprises subterranean species, although some species successfully colonised surface or ecotonal habitats.To get insight of morphological changes and whether these changes were affected by sex, we measured 15 functional morphological traits on 488 individuals from both sex of eight Niphargus species, out of which four inhabit ecotonal (spring) habitats, while the other four are exclusively subterranean. We analysed our data trait-by-trait with linear mixed models. We found that colonising the “new” habitat did not demonstrably affect morphology. In contrast habitat dependent sexual dimorphism was found in case of the 2nd gnathopods’ coxa. Besides the aforementioned habitat dependent sexual dimorphism, we found sexual dimorphism in case of 11 out of the 15 traits. Based on our result we can assume that morphological changes might happen in the course of the colonisation of the novel habitat, but they are not as unambiguous as when surface species colonise caves. It is also possible that although we consider springs as ecotone or even surface habitats, they might be feasible for subterranean species without striking morphological changes as they do not meet selection forces strikingly different from those present in caves. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Phylogenomic insights into the evolution of subterranean Coleoptera

    • Abstract: ARPHA Conference s 5: e87194
      DOI : 10.3897/aca.5.e87194
      Authors : Pau Balart-Garcia, Slavko Polak, Perry Beasley-Hall, Tessa Bradford, Ignacio Ribera, Steven Cooper, Rosa Fernández : Subterranean specialization is often accompanied by dramatic phenotypic changes epitomized by regressive evolution (e.g. loss or reduction of eyes and pigmentation). Nevertheless, the genetic underpinnings underlying these changes have been largely unexplored. The beetle tribes Leptodirini (Leiodidae), Hydroporini and Bidessini (Dytiscidae) represent ideal systems for exploring the genomic basis of adaptation to life in subterranean habitats, as both are represented by epigean and hypogean species including numerous lineages that independently colonized terrestrial and freshwater underground habitats respectively (Cooper et al. 2002, Ribera et al. 2010). We investigated gene repertoire evolutionary dynamics in both surface-dwelling and subterranean Coleoptera in lineages that underwent six independent underground colonization events. We generated highly complete transcriptomes for eight aquatic and fourteen terrestrial beetles including epigean and hypogean species and explored the evolution of their gene repertoire through a phylogenomic approach. Our results indicated that gene loss was a major force facilitating adaptation to an underground lifestyle. In contrast, we also observed that gene gain and duplication were also remarkable drivers for subterranean adaptation. Gene families experiencing contractions were involved in carbohydrate metabolism, response to starvation, wing disk development, rhodopsin and ommochrome biosynthetic processes and response to hypoxia, among many other examples. Conversely, gene families significantly expanded in subterranean lineages including those related to the regulation of apoptosis, alcohol metabolism, cell redox homeostasis, chitin-based cuticle development, larval instar development, oogenesis, and negative regulation of TORC1 signaling. Moreover we found that some gene families had experienced a more complex evolutionary dynamic encompassed by both expansion and contraction events, such as those involved in regulation of transcription, nervous system development, lipid metabolism, eye development, DNA repair and chemosensation, indicating that these gene families underwent an in-depth reshaping throughout the evolutionary time. At a lineage-specific level, we did not observe many differences between the gene repertoire of the hypogean and epigean Leptodirini species in terms of gene gain and loss. Nonetheless, Hydroporini and Bidessini stygobitic species showed more disparity in their gene repertoire compared to their surface-dwelling relatives. Our results thus indicate that genomic exaptation may have facilitated underground colonization in Leptodirini prior to the diversification of the tribe, while in Hydroporini and Bidessini recent gene family turnover (dominated by gene loss) may have been guiding the surface-to-groundwater transition. These findings shed light towards understanding how the genomic toolkit has been reshaped in hypogean fauna from a macroevolutionary perspective. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Groundwater invertebrates and droughts: resistance in stygobiont isopods
           and planarians

    • Abstract: ARPHA Conference s 5: e87190
      DOI : 10.3897/aca.5.e87190
      Authors : Stefano Lapadula, Benedetta Barzaghi, Roberto Della Toffola, Raoul Manenti : In surface freshwater habitats water level can be strongly variable and benthic invertebrates are usually adapted to cope with hydrological variability. Groundwater habitats are usually more stable, even if in sites at the interface with the vadose zone, in epikarst streams and dripping pools hydroperiod may vary. Adaptations to survive droughts are thus likely to have been developed also by groundwater-dwelling animals. However very few studies have been performed to assess stygobiont resistance to dryness. They involve mainly the amphipods of the genus Niphargus and Stygobromus; one case is reported for a triclad of the genus Atrioplanaria.Here we describe cases of resistance to drying in stygobiont planarians of the genus Dendrocoelum and the in the isopod Monolistra pavani.Since 2016 we performed multiple surveys in 53 caves in Italy and Switzerland, sampling different stable and unstable groundwater environments. Stygobionts were searched visually and by distressing the substrate. When we detected stygobionts in dry sectors of a cave, we observed their features and behaviours, we pictured them, and we tried to rinse them. We rinsed M. pavani individuals in small plastic tanks and recorded the time of reactivation.During the surveys we recorded a Dendrocoelum planarian encapsulated in a thick mucus layer on the substrate of a dry subterranean stream. When placed in water the planarian started gliding slowly and reached a length of 18 mm. During the drought of January-April 2022, we detected individuals of M. pavani in dry areas of two different caves. 72% of the individuals were able to reactivate. Time to reactivate ranged between 0 s to 30 s. We recorded living M. pavani individuals that were able to reactivate themselves even after 39 days of drought.Our observations provide new insights into the natural history of groundwater-dwelling invertebrate taxa which global climatic changes increase the risk of drought occurrence in subterranean environments. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Comparative analysis of lampenflora in two show caves in Serbia

    • Abstract: ARPHA Conference s 5: e87188
      DOI : 10.3897/aca.5.e87188
      Authors : Slađana Popović, Nataša Nikolić, Marija Pećić, Ana Anđelković, Gordana Subakov Simić : Nowadays, caves are increasingly being transformed into tourist attractions by constructing paths, electricity, water infrastructure, and the introduction of light, thereby becoming affected by various disturbances that alter their inherent equilibrium. Conservation of these sensitive environments should be a priority from the moment of their opening. All parameters, including the potential changes around artificial lights, such as lampenflora, should be monitored. Lampenflora in show caves in Serbia has been done for several years in the Lazar and Resava caves, of which the Lazar cave was monitored for over 6 years. This work aimed to compare lampenflora of these two caves, considering samples taken in the year 2021 at the beginning and the end of the official tourist season. Artificial light created favorable conditions for the proliferation of phototrophs, and lampenflora developed in both caves. Considering the community type, epiliths and endoliths were present in both localities. The difference is that endoliths in the Lazar Cave covered more significant areas of substratum, constantly spreading, thus posing a more significant threat to cave substratum and structures. One of the problematic genera was Chlorella, which can switch from an autotrophic to a mixotrophic, and finally to a heterotrophic lifestyle, thus making lampenflora dangerous and hard to deal with. Lampenflora in the Lazar Cave was developed over a large area of the cave substratum, while in Resava Cave, it was usually localized near artificial lights. When caves were compared based on phototrophic microorganism composition, using multivariate analysis, all morphological groups of Cyanobacteria (coccoid, simple trichal, and heterocystous) were more related to the Resava cave and Chlorophyta and Bacillariophyta to the Lazar Cave (based on diversity and number of sampling sites on which they are found). In the Lazar and Resava cave, Cyanobacteria dominated at the beginning of the tourist season and Chlorophyta at the end. Mosses were present, too, with the difference that the moss protonema was more characteristic of Lazar Cave in many places. In contrast, developed mosses were frequently found in Resava Cave near reflectors. In addition to the analysis of phototrophic microorganisms, ecological parameters, primary production, and biofilm parameters were measured, showing seasonal or sampling site variations. We hope that monitoring will be performed more regularly in the future and that it will include a more significant number of show caves for their conservation. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The roles of landscape of fear and light in allowing the exploitation of
           spring habitats by subterranean amphipods: an experimental and field

    • Abstract: ARPHA Conference s 5: e87144
      DOI : 10.3897/aca.5.e87144
      Authors : Matteo Galbiati, Raoul Manenti, Martina Forlani, Benedetta Barzaghi, Andrea Melotto, Francesco Gentile Ficetola, Stefano Lapadula : Border habitats such as interfaces and ecotones promise research targets from an evolutionary and zoological point of view. Springs are typical ecotones that border two strongly distinct environments: surface and underground. They are exploited by both subterranean and surface species for which they may provide specific environmental pressures promoting phenotypic plasticity and local adaptations.The aim of this study is to understand how the landscape of fear (LOF) and physical constraints, like light occurrence, affect springs' exploitation by both a subterranean (Niphargus thuringius) and a surface crustacean amphipod species (Echinogammarus stammeri).From March to May 2021, we surveyed 15 springs, divided into 25 plots according to their distance to the border, and both day and night, we recorded amphipods activity and LOF levels for them. In a subterranean laboratory, we also reared 80 N. thuringius and 80 E. stammeri in safe and risky conditions with constant darkness and diel light variation assessing their activity and survival for 30 days. Risky conditions were represented by meso-predators (four fire salamander larvae) alone or with a top-predator (a dragonfly larva of the species Cordulegaster boltonii).While in field conditions, the activity of N. thuringius seemed negatively affected by the number of active predators, in laboratory experiments, the main role was played by the light treatment; activity was significantly higher in constant darkness conditions.E. stammeri activity in the field was higher in surface plots, while in laboratory conditions was affected by LOF. Predation risk negatively affected the survival of both amphipods.Our findings reveal that while light conditions seem to shape activity patterns of stygobionts strongly, predators have a lower effect on activity, even though predators have negative effects on survival. Moreover, physical constraints, such as light exposure, can affect antipredator responses of subterranean organisms, thus representing selective pressures for the exploitation of surface environments. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Large sulfur oxidizing bacteria of the Thiovulaceae (Campylobacterota)
           thriving in the sulfidic groundwater of Movile Cave, in Romania

    • Abstract: ARPHA Conference s 5: e87133
      DOI : 10.3897/aca.5.e87133
      Authors : Traian Brad, Mina Bizic, Danny Ionescu, Lucian Barbu-Tudoran, Joost Aerts, Radu Popa, Luca Zoccarato, Jessica Ody, Jean-François Flot, Scott Tighe, Daniel Vellone, Serban Sarbu : Life in Movile Cave (Romania) relies entirely on carbon fixation by bacteria oxidizing sulfide, methane and ammonia, using oxygen, nitrate, sulfate, and ferric iron as electron acceptors. There, our attention was drawn by a white veil-like structure at the water surface. Microscopic analysis of the veil revealed a dense population of microorganisms of various shapes and sizes, some of which being much larger than the other spirilla, rods and cocci that could be observed. We studied these larger microorganisms with optical and electron microscopy techniques, sequenced their genome and analyzed the main physiological abilities. Their shape is spherical to ovoid, 12-16 µm in diameter, and their cytoplasm is rich in intracellular sulfur globules. They are present in densities of up to 5.5×103 cells/ml and they are very motile. These cells were identified as Thiovulum sp. (Campylobacterota), forming a separated cluster from marine Thiovulum sp., consisting mostly of cave bacteria. The Thiovulum microhabitat is located at the water surface in the lower, partially submerged, level of Movile Cave. Here, H2S is brought in by diffusion and convection, and some water flow is only present deep below the water surface. Frequent attachment is observed between cells, which is consistent with other reports of Thiovulum sp. being clustered in dense, veil like, aggregates attached to polysaccharide matrices. Nevertheless, the Movile Cave strain occurs in the water and is not attached to any rocky surface. The Movile Cave Thiovulum genome is small at 1.72 Mbp, contains 1804 coding sequences, 3 rRNA operons, and has a GC content of 28%. The genome suggested that the Movile Cave Thiovulum strain can switch between aerobic and anaerobic sulfide oxidation using O2 and NO3- as electron acceptors, respectively. In the latter case, NO3- is likely reduced by Thiovulum to NH3 via dissimilatory nitrate reduction, thereby contributing to the complete nitrogen cycle in this environment. Additionally, coupling the genomic analysis, with new electron microscopy images, we suggest that in absence of motor-like structures along the membrane, the short peritrichous filamentous structures, typical to Thiovulum, are pili, likely of type IV, for which genes were found in all 6 available Thiovulum genomes. These pili may play a role in veil formation, connecting adjacent cells and support the exceptionally fast swimming behavior of these bacteria. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The fascinating biology of stinky caves

    • Abstract: ARPHA Conference s 5: e87132
      DOI : 10.3897/aca.5.e87132
      Authors : Serban Sarbu : The discovery of Movile Cave in 1986, similarly to the discovery of the deep-sea vents 9 years earlier, showed that, when redox interfaces are present in subsurface ecosystems, diverse and abundant biological communities can thrive based primarily on autochthonous food produced in situ by chemosynthesis. Additional caves, partially flooded by sulfidic water have been discovered, i.e. the Frasassi caves in Italy, Ein-Nur Cave and Ayyalon Cave in Israel, Melissotrypa Cave in Greece, Tashan Cave in Iran, caves in the Sharo-Argun Valley in the Caucasus Mountains, etc. All of these are inhabited by numerous endemic cave-adapted organisms present in dense populations sustained by a rich food base generated within the subterranean environment by microorganisms that use atmospheric oxygen to oxidize the reduced chemical compounds present in the water (H2S, NH4+, CH4). Stable isotope data has shown that the biological communities found in sulfidic caves are not dependent on carbon fixation in green plants that use light energy in the process of photosynthesis. Special adaptations have been identified in some of these inhabitants that allow them to cope with the adverse environmental conditions such as toxicity of H2S, hypoxia, and extreme pH values. Symbiotic associations between crustaceans, protozoa, and sulfide oxidizing bacteria were discovered in several sulfidic caves. New species of microorganisms have also been described from these unusual subsurface environments.Unusual microbiomes have recently been discovered in dry caves that contain chemoclines between heavy volcanic gasses dominated by CO2, but also containing H2S and CH4, and atmospheric air that floats on top of the volcanic gas emissions. Unique sulfur mineralization, microbial biofilms containing new species of bacteria that thrive at extremely pH values, symbiotic relationships between bacteria and fungi have been discovered in Sulfur Cave in Romania, Bosoletto Cave in Italy, and Sousaki caves in Greece. The extreme life conditions encountered in these caves are of special interest to astrobiologists using these caves as a model for better understanding conditions that life may be present in on other planets and celestial bodies in our Solar System and beyond. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Convergent behaviours in subterranean species

    • Abstract: ARPHA Conference s 5: e87105
      DOI : 10.3897/aca.5.e87105
      Authors : Enrico Lunghi, Stefano Mammola, Alejandro Martinez, Thomas Hesselberg : The specialised subterranean fauna is often described as an iconic example of convergent evolution driven by environmental constraints, representing therefore an ideal model system for eco-evolutionary studies. During the colonization of subterranean environments, behavioural plasticity likely plays a fundamental role, as the quick behavioural response of individuals to the new environment is a key process enabling their long-term establishment. However, scientific research on the behavioural adaptations of subterranean organisms has lagged behind and is mostly biased towards a few model species. By reviewing the available literature, we aim to assess whether a convergent evolution of behavioural traits among subterranean species exists. We considered four different types of behaviour that are commonly studied in subterranean species: the explorative behaviour (i.e., how much individuals move), the diet (i.e., the variability of consumed prey), the social behaviour (i.e., type of intraspecific interactions) and the anti-predator response (i.e., if individuals adopt specific behaviours to reduce predation risk). We analysed>130 papers (both scientific and grey literature) published in the period 1909–2021, in which these four specific behaviours were described. We attributed species to one of the three main ecological classifications for subterranean species [(stygo-)trogloxene, (stygo-)troglophile and (stygo-)troglobite] according to the information reported in the literature. We collected data on the behaviour of more than 135 species belonging to> 75 different taxonomic families, including both vertebrates and invertebrates. From our preliminary analyses, we observed a lower movement in trogloxenes. We detected a significant increase in the trophic spectrum in troglophile species, while trogloxenes showed the narrowest trophic niche. We observed a higher occurrence of anti-predatory behaviours in trogloxenes, as well as an increase in intraspecific antagonistic interactions. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Average telomere length in cave vs surface Astyanax mexicanus

    • Abstract: ARPHA Conference s 5: e87044
      DOI : 10.3897/aca.5.e87044
      Authors : Enrico Lunghi, Helena Bilandžija : Telomeres are specialized and highly repetitive noncoding DNA structures at the end of linear chromosomes that are essential for maintaining genomic integrity. Each time a cell divides, telomeres are not fully replicated and the resulting cells have shorter telomeres than the progenitor cells. This incomplete replication of telomeres (i.e., shortening) is considered one of the major mechanisms of aging. Furthermore, telomeres are not only shortened by cell divisions, but multiple environmental stressors can also reduce their length (known as somatic redundancy). Studies of telomere length that include a comparison between subterranean and surface species can make an important contribution to understanding the role of these DNA structures in aging and in the ability of individuals to cope with environmental stressors. We conducted a preliminary assessment of the potential divergence of average telomere length in the Mexican tetra Astyanax mexicanus. This fish has surface and subterranean populations (i.e., ecomorphs), each characterized by specific adaptations to its environment. The study of telomere length in conspecific ecomorphs can provide valuable information on the effects of telomeres on the lifespan and longevity of individuals, as well as the role of various environmental stressors on telomere lengths. By adopting a single-species model, it is possible to reduce the potential variability due to the highly divergent evolutionary history or genetics of different species. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Mechanisms underlying albinism as a consequence of ommochrome deficiency
           in cave isopods

    • Abstract: ARPHA Conference s 5: e87043
      DOI : 10.3897/aca.5.e87043
      Authors : Lada Jovović, Florent Figon, Helena Bilandžija : Albinism, or loss of pigment, is a typical adaptation to living in caves and occurs in numerous cave dwellers. To understand the evolution of this trait, one must identify the type of biological pigment present in the phylogenetically closest surface relatives. The absence of pigmentation has been studied primarily in terms of melanin deficiency, while the exact mechanism of ommochrome loss in any cave dwellers is largely unknown. Ommochromes are tryptophan derivatives and generally belong to a less studied class of biological pigments previously described in protostomes such as isopod crustaceans. Using a simple method for extraction and detection of ommochromes in conjunction with liquid chromatography coupled with mass spectrometry (UPLC-MS), we characterized the nature of pigments in surface isopods. We detected, for the first time and to the best of our knowledge, ommochromes in three groups of surface isopods: the families Sphaeromatidae (Flabellifera) and Trichoniscidae (Oniscidea) and the genus Proasellus (Asellota). To determine which step of the ommochrome production pathway is disrupted in cave isopods, we quantified the precursors of ommochrome synthesis in closely related surface and cave species. Our results suggest that the disruption of ommochrome synthesis is preferentially located at the beginning of this anabolic cascade, as we observed an accumulation of the precursor tryptophan in albino species. To address this problem at the molecular level, we are trying to determine the exact genes and mutations that may be involved in the loss of ommochromes. We performed RNA-seq on two representatives of surface and cave species of the genus Proasellus and analyzed the data for expression changes in candidate genes. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • A glimpse into the biosynthetic potential and resistome of microbial
           communities inhabiting sulfidic, chemoautotrophic Movile Cave ecosystem

    • Abstract: ARPHA Conference s 5: e86928
      DOI : 10.3897/aca.5.e86928
      Authors : Iulia Chiciudean, Diana Bogdan, Oana Moldovan, Horia Banciu : Background: Microbial secondary metabolites (SM), especially those produced by soil microorganisms, have been a valuable source of antibiotics, antitumor agents, pigments, growth-promoting substances, etc., with tremendous market potential. These molecules are encoded by biosynthetic gene clusters (BGCs) within the bacterial genome. Their synthesis confers survival advantages by facilitating chemical defense, interspecies communication, and adaptation to well-defined ecological niches.Caves, particularly Movile Cave (Romania) - a sulfidic autotrophic-based ecosystem - can be considered extreme environments suited to investigate and discover novel bioactive microbial molecules. Here, low nutrient availability can lead to resource competition and, consequently, antimicrobial production to deter nearby microbial competitors.Aim: Our study focused on highlighting the biosynthetic gene clusters (BGCs) and biosynthetic potential of the sediment-associated microbiome in Movile Cave.Methods: Over 100 high-quality metagenome-assembled genomes (MAGs) were retrieved by whole-community shotgun-sequencing of 7 sediment samples collected in different Movile Cave’s galleries (Chiciudean et al. 2022). Detected MAGs were then analyzed for the presence of BGCs by antiSMASH (v. 6.1.1) (Blin et al. 2021) whereas antibiotic resistance genes were predicted by ResFinder (v. 4.1) (Florensa et al. 2022). The statistical analysis of BGCs data was performed by Past (v. 4.03).Results: We detected 637 BGCs across 106 high-quality MAGs that were affiliated to 22 phyla. The diversity of predicted BGCs varies across sediment samples with no apparent correlation to the number of analyzed MAGs per sample. The MAGs recovered from the sulfidic water-sediment interface (sample code PMV4) had the lowest alpha BGCs diversity among all sampled locations and it was clearly distinct in BGCs composition and abundance (β-diversity) from dry gallery samples (PMV7 and PMV8). The most abundant BGCs predicted in Movile Cave metagenomic dataset encode for terpenes, non-ribosomal peptides (NRPs) and ribosomally synthesised and post-translationally modified peptides (RIPPs). Acidobacteriota and Chloroflexota- affiliated MAGs were most enriched in BCG containing 20 and 23 BGCs per MAG, respectively, in contrast with the candidate phylum Ca. Patescibacteria-related MAGs that showed no SM biosynthetic capabilities. Two antimicrobial resistance (AR) genes (ole(C), oqxB) encoding resistance to antibiotics (i.e. oleandomycin, chloramphenicol, ciprofloxacin, trimethoprim) and disinfectants were identified in MAGs affiliated with the class Actinomycetia and Gammaproteobacteria.Based on the analyzed data, the biosynthetic potential of Movile Cave is significant compared with other microbiomes (Donia et al. 2014) and has a pronounced degree of novelty whereas the resistome (that is the genetic potential for antibiotic resistance) is reduced. Considering the uncommon futures of Movile Cave environment, future in-depth analysis of the identified BGCs might lead to the discovery of novel bioactive compounds. Additionally, the seclusion of this environment may provide an exciting opportunity for surveying the occurrence and environmental drivers of natural AR traits. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The laboratory of subterranean biology “Enrico Pezzoli”: a new
           underground facility for zoological research.

    • Abstract: ARPHA Conference s 5: e86916
      DOI : 10.3897/aca.5.e86916
      Authors : Raoul Manenti, Andrea Melotto, Benedetta Barzaghi, Mauro Villa : Since the beginning of subterranean biological research, the development of underground laboratories has provided scientists with protected places to perform long-term studies and to collaborate with others across subjects. However, aided by advances in technology for research centres and universities and by concerns for the conservation of natural subterranean environments, mots of past subterranean laboratories went under the radar of the institutions that owned them.Here we describe a new underground facility that was built in Italy by the Regional Park of “Monte Barro” (Lecco, Lombardy) and dedicated to the memory of Enrico Pezzoli who was strongly active in the study of spring and cave biodiversity.The laboratory has been established by repurposing an ancient draining gallery of the Park; it has been conceived to study local fauna, especially that of local aquifers. Removable equipment has been set stretching along the gallery, from the beginning to deep sections, creating distinct sectors with different conditions allowing to rear, study and compare both troglophile and troglobiont species. In every sector of the laboratory independent blocks with different controlled conditions of light and biotic features can be easily placed and used to rear individuals of both invertebrates and amphibians. Experimental activity does not prevent the exploitation of the gallery by local fauna, with spiders, crustaceans and amphibians usually occurring.The first experiences performed in the new resource suggest that subterranean laboratories, if appropriately conceived, may represent sustainable facilities thanks to their low energetic requirements and their strong potential in integrating research and education. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Caves as evolutionary dead end' The journey of the stygobiont isopod
           Monolistra pavani toward sunlight

    • Abstract: ARPHA Conference s 5: e86915
      DOI : 10.3897/aca.5.e86915
      Authors : Raoul Manenti, Giorgio Scarì, Benedetta Barzaghi, Silvia Mercurio, Stephanie Sherpa, Andrea Melotto, Roberta Pennati, Gentile Ficetola : Monolistra are stygobiont isopods spread in numerous karst areas from the south-eastern parts of Dinaric karst to south-eastern Switzerland. They inhabit different typologies of groundwater habitats from lentic to lotic ones where they can be particularly important in terms of biomass. M. pavani is a steno-endemic species that was reported only for a single cave system in Lombardy (Italy).From October 2018 to March 2022, we performed multiple surveys in the subterranean system and in its surrounding springs recording abundance and distribution of the species. In December 2018 we discovered three spring populations/subpopulations of M. pavani in which pigmented individuals also occurred. We performed UV and infrared spectroscopy to evidence potential differences in the signals related to C-C bonds of the aromatic ring of the melanin molecules between cave and spring individuals. In 2020, we also compared circadian rhythm and responses to light stimuli of cave and spring individuals.Occurrence of M. pavani in springs was regular and associated to specific spring microhabitats; individuals were active both during day and during night. Nor spring and cave individuals avoid light stimuli. Circadian rhythm analyses revealed that individuals from springs are more active than those from cave, showing higher activity during day than during night. The 20% of spring individuals are pigmented, and eumelanin occurs in dark individuals.Our results suggest the possible ongoing adaptation to surface habitats at the border with groundwater in a species considered as a strict stygobiont. Molecular analyses on the population relationships are currently ongoing. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • The pleiotropic effects of melanin loss in cave snails Physella sp.

    • Abstract: ARPHA Conference s 5: e86885
      DOI : 10.3897/aca.5.e86885
      Authors : Magdalena Grgić, Robert Weck, Gaj Keresteš, Helena Bilandžija : A traditional explanation for the loss of pigmentation in cave dwellers is the absence of negative selection acting on surface species to remove all albinos from the population. Recently, however, evidence has been accumulating that albinism in the cavefish Astyanax mexicanus has several beneficial pleiotropic effects. Albino cavefish have higher catecholamine levels due to a loss-of-function mutation in the oca2 gene, higher resistance to anesthesia, and sleep less than their pigmented, surface-dwelling conspecifics. All of these traits are beneficial to cavefish, given the scarcity of food and the availability of mates in caves. To understand whether pleiotropy might be a general mechanism for the loss of melanin pigmentation in cave animals, we use Physella sp., a freshwater snail from SW Illinois, USA. Within one cave Physella sp. occurs in several different morphs, ranging from completely albino to fully pigmented individuals. Using melanogenic substrate assay, we demonstrated that the first step of melanin synthesis is blocked in Physella sp. To investigate the pleiotropic effects of this blockade, we tested anesthesia resistance (AR). Similar to A. mexicanus, albino morphs of Physella sp. exhibited higher AR levels compared to pigmented morphs, with AR being controlled by the noradrenergic system in both morphs. AR and the noradrenergic system are linked to melanin synthesis via a common precursor L-tyrosine, which can be redirected between melanin and catecholamine pathways. Using RNA sequencing of albino and pigmented morphs of Physella sp. we aim to gain further insight into other pleiotropic effects of melanin loss. Our results suggest that pleiotropy may be involved in the evolution of albinism in both cave vertebrates and invertebrates. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Proasellus thrive in the dark – plasticity enabling cave

    • Abstract: ARPHA Conference s 5: e86909
      DOI : 10.3897/aca.5.e86909
      Authors : Jana Bedek, Helena Bilandžija : Darkness is the hallmark of all subterranean habitats and, to some degree, of many surface habitats. Many surface animals that live in semi-dark habitats, such as freshwater benthos, are photophobic. Yet, few of them can colonize caves and form subterranean populations. We hypothesize that phenotypic plasticity induced by darkness enables surface crustaceans to colonize caves, as has been previously shown for the cavefish Astyanax mexicanus.Our target organisms were surface species that form distinct subterranean populations with different adaptations to caves and have closely related cave species. Proasellus proved to be a good model system, and we selected the surface species Proasellus coxalis and its cave relative P. anophtalmus to learn what adaptations evolve in subterranean forms. We exposed a randomly selected group of both species to complete darkness (DD) where we could establish breeding colonies. Control breeding colonies are exposed to a 12:12 h light-dark photoperiod (LD). Other conditions are kept as constant and equal as possible, including conditioned Alnus glutinosa leaves used for shelter and food.We examined the molecular phenotypes of both Proasellus species, animals originally collected from the wild and preserved after four months under the experimental conditions DD and LD. The number of observed differentially expressed genes (DEGs) was significantly higher between cohorts DD and LD of P. coxalis compared to P. anophtalmus. The direction of regulation of DEGs also showed the opposite effect, with the cohort of P. coxalis DD showing predominant upregulation and the cohort of P. anophtalmus DD showing predominant downregulation relative to their cohorts LD. A greater number of DEGs in both species were associated with various metabolic processes such as lipid regulation. In contrast, DEGs related to growth, cell division, rhythmic processes, sensory perception of taste, and sexual reproduction were observed only in P. coxalis.We examined the changes at the phenotypic level in the first generation born in the experiment. Surprisingly, P. coxalis showed better fitness under DD conditions (growth rate, fecundity, survival). Other phenotypes were altered in both adaptive and non-adaptive directions i. e. towards vs. opposite of cave adaptations. We conclude that darkness is not a limiting factor in cave colonization and suggest that it may even be a key factor in the evolution of troglomorphy via phenotypic plasticity. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Invertebrate fauna in municipal wells in Kraków, Poland

    • Abstract: ARPHA Conference s 5: e86699
      DOI : 10.3897/aca.5.e86699
      Authors : Elzbieta Dumnicka, Joanna Galas : This study of fauna in municipal wells continues our previous investigations performed in rural areas (Dumnicka et al. 2017). The invertebrate fauna of city wells was rarely studied: by Vejdovský (1882) and Řehačkova (1953) in Prague, Jaworowski (1893) in Kraków and recently by Koch et al. (2020) in Karlsruhe.In 2019-2020 studies of fauna composition and water chemistry were done twice (spring-summer and autumn period) in 91 wells situated in various parts of the Kraków city. This work evaluated fauna distribution and diversity with patchy bedrock geological structure and local water pollution.Water temperature and conductivity were measured in situ using a portable Elmetron CX-742 pH meter. In contrast, chemical analyses were done in the laboratory with DIONEX ICS 2000 unit and a DIONEX ICS 5000 unit, equipped with a DIONEX AS18 anion column and a DIONEX CS16 cation column. For fauna studies, 100 l of water was filtered by a plankton net (50 μ m mesh size), and invertebrates were sorted in vivo under a stereoscopic microscope with 10x magnification. The material was determined to family level, except for Rotifera.Our results showed that water chemistry in Kraków wells varied strongly spatially and between sample collection times – this was also stated by various authors and summarized by Chowaniec et al. (2007). The depth of the wells varied from 5.0 to 100 m. Water temperature was relatively high (average value 12.95 ±1.4 o C) and pH almost neutral (7. 13 ±0.34). Water conductivity values were very high (1324.3±572 µS) due to high concentrations of calcium (170.3±66 mg L-1), sulfates (151.8±81.4 mg L-1), and less frequently chlorides (155.5±143.7 mg L-1). In many wells, ammonium ion was present (0.354±0.764 mg L-1), whereas, in other samples, nitrate concentrations varied strongly from 0.1 to 130.5 mg L-1 (average value 21.92±27 mg L-1).The occurrence of invertebrates (at least from one taxonomic group) was stated in 74 wells. Copepoda (family Cyclopidae) were found most frequently (in 40 wells) and in highest numbers (it means that in some wells several dozens of individuals were caught). Nematods and annelids were stated in 19 and 17 wells respectively, but in small number. The remaining groups, such as turbellarians (Catenulida and Rhabdocoela), rotifers and ostracods were present in a few wells only, while amphipods (Niphargus tatrensis morphospecies) exclusively in two wells. Moreover, dipterans larvae (from families: Anisopodidae, Chironomidae, Culicidae, Empididae, Limoniidae, Psychodidae and Rhagionidae) were found in 13 wells, whereas Collembola, considered to be terrestrial fauna, were found in 49 wells. In the majority of cases all springtails and fly larvae were alive. The presence of Diptera and Collembola inside the wells might result from leak of pump mechanisms therefore these invertebrates could just survive for some time inside pumps.The occurrence of invertebrates (at least from one taxonomic group) was started in 74 wells. Copepoda (family Cyclopidae) was found most frequently (in 40 wells) and in the highest numbers (it means that in some wells, several dozens of individuals were samples). Nematodes and annelids were stated in 19 and 17 wells, respectively, but in small numbers. The remaining groups, such as turbellarians (Catenulida and Rhabdocoela), rotifers, and ostracods, were present in a few wells, while amphipods (Niphargus tatrensis morphospecies) exclusively in two wells. Moreover, dipterans larvae (from families: Anisopodidae, Chironomidae, Culicidae, Empididae, Limoniidae, Psychodidae, and Rhagionidae) were found in 13 wells, whereas Collembola, considered to be terrestrial fauna, were found in 49 wells. In the majority of cases, all springtails and fly larvae were alive. The presence of Diptera and Collembola inside the wells might result from the leak of pump mechanisms; therefore, these invertebrates could just survive for some time inside pumps.Statistical analysis (Student t-test) revealed that the presence of fauna was not dependent on the depth of the wells (p ˂ 0.12). The effect of water pollution (expressed as increased concentrations of ammonium and chloride ions) on the presence of fauna was also not statistically significant, except for sulfide ions (p ˂ 0.058).In wells situated close to small running waters located on karstic bedrock or alluvial sediments, the presence of stygobionts (Niphargus tatrensis morphospecies, Trichodrilus cernosvitovi and Typhlocypris cf. eremita) was stated, whereas in wells situated along Vistula River no aquatic fauna was found probably due to the impermeable clay layer, which isolated wells water from water table of the river.The only study of fauna in dug municipal wells in Kraków made by Jaworowski (1893) showed higher water pollution than was found in this study: the crustaceans were found rarely, whereas ciliates were abundant and diversified.Collected material demonstrated the occurrence of relatively rich invertebrate fauna in subterranean city waters, but the effect of various parameters on its distribution has not been firmly established. 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      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Environmental Hypoxia is a Driver of Sonic Hedgehog-dependent
           Troglomorphic Evolution in Astyanax Cavefish

    • Abstract: ARPHA Conference s 5: e86667
      DOI : 10.3897/aca.5.e86667
      Authors : William Jeffery : Astyanax mexicanus cavefish (CF) have evolved numerous troglomorphic traits since their divergence from surface fish (SF) ancestors about 20,000 years ago. A large number of these traits are under the control of Sonic Hedgehog (Shh), a signaling morphogen that is overexpressed along the midline in CF relative to SF embryos. The Shh-dependent traits include eye degeneration and increases in the olfactory lobes, hypothalamus, jaws, and oral taste buds. Eye degeneration and tastebud enhancement are antagonistic tradeoffs governed by the pleiotropic shh gene. CF have also evolved multiple changes in circulatory system development, highlighted by larger hematopoietic domains, more erythrocytes, and reversals in normal heart asymmetry, which are also controlled by Shh signaling. Although Shh signaling is an important factor in CF troglomorphism, the shh gene is not associated with any of the many QTL identified by genetic analysis, suggesting that upstream processes, rather than mutation of the shh locus itself, may be important in Shh regulation. Due to the absence of primary productivity, cave pools harboring Astyanax CF are deficient in dissolved oxygen. Accordingly, the Hypoxia Inducing Factor (HIF) transcription factors and their downstream targets are constitutively active during CF development. To determine if oxygen deficiency is an upstream regulator of Shh signaling, we subjected early SF embryos to 8 or 16 hours of hypoxia in the laboratory and assayed the effects on shh expression, erythrocyte formation, heart asymmetry, and eye development. The results indicated that hypoxia can cause shh overexpression, increase erythropoiesis, reverse heart asymmetry, and induce eye degeneration in SF embryos. We conclude that environmental hypoxia may be responsible for the evolution of troglomorphic traits in Astyanax CF through hyperactivation of the Shh signaling pathway. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Metabolic scaling and thermal acclimation of the cave asellid Proasellus

    • Abstract: ARPHA Conference s 5: e85959
      DOI : 10.3897/aca.5.e85959
      Authors : Tiziana Di Lorenzo, Ana Sofia P.S. Reboleira : Metabolism is the set of life-sustaining chemical reactions in organisms. It scales allometrically to the body mass in ecto- and endotherm organisms, implying that larger individuals are more economical in terms of energy requirements. Within the limited range of “biologically relevant” temperatures (from 0° to 40°C), the allometric scaling with a factor between 0.66 and 0.75 has been consistently observed in unicellular microbes and plentiful multicellular plants and animals (Gillooly et al. 2001). The allometric scaling factor does not vary when organisms acclimate to changing temperatures where thermal acclimation represents a phenotypic response with essential implications for coping with global climate change. Cave fauna, which live in thermally-stable habitats, are expected to conform to allometric scaling though showing a substantial inability to acclimate to increasing temperatures rapidly. We measured the individual oxygen consumption (as a proxy of metabolic response to temperature variation) of Proasellus lusitanicus, a cave asellid (Crustacea Isopoda: Asellidae) endemic of the Estremenho karst massif in central Portugal (Reboleira et al. 2011). We measured the thermal acclimation ability of this species in a thermal ramp-up experiment over a range of temperatures (from 17 ° to 22.5 C), also assessing the scaling of oxygen consumption with body mass. We found out that P. lusitanicus shows low thermal plasticity, likely inadequate to protect this species from the temperature increase expected in the next 90 years. Furthermore, we observed that the metabolism of P. lusitanicus does not scale allometrically with the body mass. Rather, the metabolism of this species does not vary with body mass, similarly to two other subterranean species (Di Lorenzo et al. 2014), and contrarily to other epigean animals. The lack of metabolic scaling is likely to be a novel trait in subterranean ecosystems where the temporal unavailability of food and oxygen can be relevant and protracted. Our study sheds light in understanding regarding the phenotypical adaptations of cave animals to subterranean environments and also highlighted that narrowly-distributed cave species may be at extinction risk because of temperature increase to climate change. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • History and significance of tropical cave biology

    • Abstract: ARPHA Conference s 5: e85878
      DOI : 10.3897/aca.5.e85878
      Authors : Francis Howarth : “Nothing could possibly live there!” They said. Indeed, until recently, few specialized cave-adapted animals were known from volcanic, tropical, or oceanic island caves, and plausible theories had been put forward to explain their absence. But assume nothing in science! One must illuminate, explore, and survey habitats before declaring them barren. Our understanding of cave biology changed dramatically about 50 years ago following the serendipitous discovery of cave-adapted insects and other terrestrial arthropods in lava caves on the young oceanic islands of the Galapagos and Hawai‘i. The discovery and subsequent studies on the evolutionary ecology of cave animals have revealed a remarkable hidden fauna and created new sub-disciplines within biospeleology. Biological surveys of caves in other regions have confirmed the results developed in Hawai‘i. We now predict that, rather than being relicts trapped in caves by changing climate, animals actively colonized caves and adapted to exploit food resources wherever there were suitable subterranean voids. The physical environment in caves can be determined with great precision because the habitat is buffered by rock. Furthermore, the bizarre adaptations displayed by obligate cave animals are similar across many taxonomic groups. These two characteristics make caves nearly ideal natural laboratories for studying evolution and ecology. However, to the untrained researcher, caves can appear hostile and dangerous, and in fact, fieldwork in caves requires a unique marriage of athletic ability and science. In other words, it is exciting! Join me on a virtual tour of exploration, discovery, and research in caves during the past half-century. HTML XML PDF
      PubDate: Thu, 14 Jul 2022 10:00:00 +030
  • Genetic diversity as a paramount factor determining wheat adaptation to
           drought stress

    • Abstract: ARPHA Conference s 5: e89829
      DOI : 10.3897/aca.5.e89829
      Authors : Mariyana Georgieva, Anna Dimitrova, Valya Vassileva : Wheat (Triticum aestivum L.) is a globally important crop for food and nutritional security that requires sufficient water supply for optimal production. All stages of wheat growth and development are adversely affected by reduced water supply, which limit productivity across large areas of Central and Southern Europe. Since drought episodes are expected to occur more frequently and with a greater intensity, the research efforts are devoted to identification of key traits associated with drought resistance that could be used as direct and indirect selection criteria for wheat drought tolerance. We evaluated and quantified several key genetic and molecular features of wheat genotypes with contrasting drought tolerance under severe dehydration triggered by the treatment of wheat with 250 mM sorbitol in hydroponic systems. Plant status was evaluated non-destructively by measuring leaf chlorophyll index, nitrogen content, and leaf relative water content. The drought-tolerant genotype displayed higher chlorophyll and nitrogen content, when grown under drought and benign conditions. Since root architecture appears to be a key trait for breeding against dehydration, we analysed the members of the DEEPER ROOTING (DRO) gene family that allow the root to penetrate deeper into soil, which could increase the yield even upon water shortage. Expression patterns of the DRO members showed genotype-specific changes under drought stress, which correlated well with plant drought tolerance. Our experiments suggest that genes affecting root system architecture could contribute to development of wheat cultivars with higher drought adaptability. HTML XML PDF
      PubDate: Tue, 5 Jul 2022 17:26:00 +0300
  • Genetic diversity in high-mountain Thymus species in Bulgaria revealed by
           ISSR genetic markers

    • Abstract: ARPHA Conference s 5: e89720
      DOI : 10.3897/aca.5.e89720
      Authors : Petar Zhelev, Georgi Bonchev, Ina Aneva : High mountain populations of the plant species possess particular interest from both evolutionary and conservation points of view. The mode of distribution, limited gene flow and severe environmental conditions act as evolutionary forces shaping the level and distribution of genetic diversity within and among populations. The paper reports results of a study on four populations of Thymus praecox aggr., including the taxa T. vandasii and T. jankae. Two populations are located in Rila Mts (Belmeken dam and Yastrebets), one – in Pirin (Vihren hut) and one – in the Rhodopes (Perelik hut).  Eleven inter simple sequence repeat (ISSR) markers were applied to document the genetic diversity within and among populations. The level and distribution of the diversity correspond to the values reported in other studies on the species of the genus Thymus and other species with similar life-history characteristics. The populations from Rila and Pirin were genetically closer to each other, while the population from the Rhodopes was the most differentiated. The results are discussed in the light of the conservation and sustainable use of the species resources.   HTML XML PDF
      PubDate: Tue, 5 Jul 2022 17:26:00 +0300
  • Phylogenetic identification of Balkan endemic Stachys species and genomic
           stability during ex vitro conservation 

    • Abstract: ARPHA Conference s 5: e89656
      DOI : 10.3897/aca.5.e89656
      Authors : Desislava Mantovska, Georgi Bonchev, Miroslava Zhiponova, Zhenya Yordanova : The genus Stachys is one of the largest in the Lamiaceae family. Representatives of the genus are among the most ancient medicinal plants used in the ethnomedicine. The Balkan endemic species S. thracica, S. bulgarica and S. scardica are included in The Red Data Book of Bulgaria and due to their endangered status are scarcely studied. The aim of the present work was to examine the genetic status of these three endemic Stachys species during the process of their ex situ conservation. To gain information about their taxonomic position in the genus Stachys, we applied the DNA barcoding approach. Nuclear (ITS) and plastid (rbcL, matK and trnH‑psbA) DNA barcodes were generated and aligned with accessions available in the data base. In the constructed phylogenetic trees S. thracica was placed in a cluster together with S. alpina, S. germanica and S. cretica, while S. bulgarica and S. scardica were clustered with S. officinalis. The ex situ conservation was achieved by the initiation of in vitro shoot cultures and their subsequent adaptation in ex vitro conditions. To check the genomic stability of the plants during the acclimatisation from in vitro conditions to ex vitro, analysis by sequence-related amplified polymorphism (SRAP) markers was performed. No difference was detected between the SRAP profiles of in vitro cultivated and ex vitro adapted S. thracica and S. scardica plants. In S. bulgarica, only 0.4% fragment difference was detected. The obtained results indicated that the three Stachys species preserved their genetic stability during the process of in vitro multiplication, which is a prerequisite for conserved bioactive capacity. HTML XML PDF
      PubDate: Tue, 5 Jul 2022 17:26:00 +0300
  • First data on DNA barcoding of representatives of the genus Cеntaurea
           s.l. (Asteraceae) from Bulgaria

    • Abstract: ARPHA Conference s 5: e89469
      DOI : 10.3897/aca.5.e89469
      Authors : Svetlana Bancheva, Vladimir Vladimirov, Irina Boycheva, Georgi Bonchev : The genus Centaurea s.l. is one of the richest and most taxonomically complex of the Asteraceae family. It includes between 400 and 700 species, of which more than 70 are found in Bulgaria – a territory considered as one of the secondary centers of speciation of the genus. There is a large number of endemics (Bulgarian and Balkan) with endemism reaching up to 50% in some groups, such as Cyanus gr. Due to the ongoing active speciation in the Balkans, the boundaries between closely related taxa cannot be easily established based entirely on morphological features. DNA barcoding is cost-efficient and reliable approach for identifying and retrieving previously known species with the potential to accelerate the discovery of new plant species. Despite the huge potential of the method, no any Bulgarian population from this genus has been barcoded so far. The present study presents the first DNA barcoding data of 11 species of the genus Centaurea s.l. from 12 populations based on four regions – ITS, matK, rbcL and trnH-psbA. The first three DNA barcodes are promising whereas the trnH-psbA has somewhat lower resolution. The preliminary results suggest that the ‘Cyanus’ group is well separated from the ‘Centaurea s.str.’ group which corresponds well to their treatment as different subgenera or genera. Within ‘Cyanus’, grouping of taxa corresponds well to the morphology of the species. Within ‘Centaurea s.str.’, although a relatively low number of species has been included, grouping of taxa in most cases is congruent with the morphological characters. However, there are some incongruent tree topologies which should be investigated further both by repeating the extraction and sequencing of the samples, and by addition of new, presumably closely related species in the study. HTML XML PDF
      PubDate: Tue, 5 Jul 2022 17:26:00 +0300
  • Molecular identification and phylogenetic relationships of Colletotrichum
           isolates pathogenic on cultivated plants in Bulgaria

    • Abstract: ARPHA Conference s 5: e89434
      DOI : 10.3897/aca.5.e89434
      Authors : Vasilissa Manova, Zornitsa Stoyanova, Irina Boycheva, Rossitza Rodeva, Georgi Bonchev : Fungi of the genus Colletotrichum are causal agents of plant diseases with constantly growing economic importance. Accurate pathogen identification is a significant prerequisite for effective disease control. The aim of the present investigation was to clarify the species affiliation of Colletotrichum isolates obtained from different hosts in Bulgaria and to determine the phylogenetic relationships between them by applying DNA barcoding. Thirty-five fungal isolates obtained from five botanical families (Solanaceae, Rosaceae, Musaceae, Cucurbitaceae and Caryophyllaceae) were morphologically characterized and subjected to molecular analysis based on four fungal barcode markers – the primary ITS barcode and the secondary marker regions ACT, EF-1a and TUB2 (Fig. 1). Three of the barcodes (ITS, ACT and TUB2) showed complete success rate of PCR amplification and sequencing and proved efficient for reliable identification at species level. BLAST analyses identified eleven Colletotrichum species assigned to five different complexes – C. coccodes, C. acutatum, C. gloeosporioides, C. dematium and C. spaethianum. The resolution power of ITS region was  not sufficient to discriminate interspecies variations within C. coccodes, C. dematium and C. spaethianum complexes confirming the requirement for secondary barcodes in order to resolve the genetic variability of the Colletotrichum isolates. DNA barcoding analyses revealed that the highest species variation was observed among the isolates from pepper (Capsicum annuum). Interestingly, an isolate from the same host identified as C. truncatum on the basis of morphological characters appeared to be C. circinans when applying DNA barcode markers. According to our knowledge, this species has not been reported as a causal agent of pepper anthracnose. Data obtained in this study improve our understanding of the genetic diversity within the Colletotrichum population pathogenic on cultivated plants in Bulgaria. HTML XML PDF
      PubDate: Tue, 5 Jul 2022 17:26:00 +0300
  • Identification of fungal taxa with pathogenic potential in soil samples
           from Perunika Glacier’s newly formed forefields - Livingston Island,

    • Abstract: ARPHA Conference s 5: e87573
      DOI : 10.3897/aca.5.e87573
      Authors : Yordan Hodzhev, Borislava Tsafarova, Vladimir Tolchkov, Pavel Stoev, Rostislav Bekchiev, Mario Langourov, Niya Toshkova, Stanimira Deleva, Nedko Nedyalkov, Ognyan Mikov, Gancho Slavov, Rumyana Nenova, Docho Dochev, Miroslav Tsviatkov, Nikolay Simov, Stefan Panaiotov : Antarctica peninsula periphery islands undergo one of the most dramatic ecological changes due to ongoing global warming. The front fields of the Antarctic glaciers are extreme environments and pioneering sites for ecological succession. Rising temperatures lead to deglaciation in the Antarctic habitats, and the new terrain is subjected to the process of soil formation and microbial colonization. In the present study, we investigated the formation of pathogenic fungal soil microbiomes as an effect of forefield deglaciation. Soil samples were taken from two different forefields, one formed several years ago and the other freshly uncovered. Both habitats were in the vicinity of the Perunika Glacier situated in the northeastern direction of Hurd Peninsula, Livingston Island, the second largest island from the South Shetland Archipelago, about 100 km north of the Antarctic Peninsula. Total DNA was extracted and targeted ITS amplicon sequencing was applied. The ITS marker sequences were then taxonomically identified. The abundance of the fungal taxa was calculated. Alpha and Beta diversity analyses to obtain fungal richness in samples were performed. Our results showed that soil habitat formation, initiated by deglaciation, was such that:In the newly deglaciated forefield, there was almost no fungal DNA, which prevented further analysis; At older drier fields metagenome content was much higher; Further analysis showed that the most abundant genera were Pseudogymnoascus, Simplicillium, Hanseniaspora, Mycothermus, and Malassezia.It is known that Pseudogymnoascus and Malassezia species have pathogenic keratinolytic activity. In conclusion, the phylum Ascomycota, which dominated the core microbiome showed much higher ecological diversity and abundance, i.e. potential for colonization of the glacier forefields. In contrast, the phylum Basidiomycota appeared to be less fit for these conditions. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • Detection and monitoring of invertebrate non-indigenous species through
           DNA metabarcoding in a recreational marina of the Northwest of Portugal

    • Abstract: ARPHA Conference s 5: e87557
      DOI : 10.3897/aca.5.e87557
      Authors : Fábio Amaral, Ana Sofia Lavrador, Sofia Duarte, Filipe Costa, Pedro Vieira : In marine and coastal ecosystems, the proliferation of non-indigenous species (NIS) is among the top causes of biodiversity loss. Thus, it is essential to prevent the entrance and proliferation of NIS by detecting their presence as early as possible at the most susceptible locations, such as ports and marinas. Molecular techniques, like DNA metabarcoding, may be faster and more effective in the early detection of NIS. Its high sensitivity allows the detection of species at any stage of their life cycle and when present at very low densities. However, the detection power can be affected by the methodologies employed through the DNA metabarcoding analytical chain. This study aimed to assess the seasonal effects, sampling method (hard and artificial substrates, zooplankton and eDNA) and genetic marker (COI and 18S) in the recovery of NIS through DNA metabarcoding. Sampling was conducted in 3 seasons (spring, autumn, and winter of 2020/2021) in the “Porto Atlântico” marina, which is located near a major commercial port of the North of Portugal (Leixões). Overall, 626 species were identified through DNA metabarcoding, with the dominance of Crustacea, Ascidiacea and Mollusca. The sampling method and season significantly affected the marine invertebrate species recorded, for both molecular markers, with a high proportion of species detected exclusively using one method (7 to 20%). Overall, 31 NIS were detected, 7 of which are considered invasive and 6 are presumed new records in mainland Portugal. Zooplankton sampling and winter were, respectively, the method and the season where the highest number of NIS was detected, but the maximum number was only attained when pooling the results from all sampling methods and seasons. Our study highlights that different sampling methods and genetic markers, over different seasons, are needed to detect NIS in marine habitats, since the number of species shared by these factors is low. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • Testing COI primers for ichthyoplankton metabarcoding and their capability
           to assess local mesozooplankton communities

    • Abstract: ARPHA Conference s 5: e87495
      DOI : 10.3897/aca.5.e87495
      Authors : André O. Ferreira, Cristina Barroso, Joana Cruz, Sofia Duarte, Conceição Egas, Pedro Gomes, Cláudio Oliveira, Pedro E. Vieira, A. Miguel Piecho-Santos, Filipe O. Costa : DNA metabarcoding is particularly helpful for monitoring taxonomically complex communities and hard to identify morphologically, such as several zoo and ichthyoplankton, which contain eggs and larval stages of unknown species. However, the efficiency of metabarcoding in diversity recovery is dependent on the targeted genetic markers and primers employed. In this work, we compared the performance of three different primer pairs from cytochrome oxidase subunit I (COI) genetic marker in species detection from marine mesozooplankton samples and its potential to be implemented in biomonitoring programs. We employed the mlCOIintF/LoboR1 primer combination targeting marine metazoans, and two newly designed fish-specific primer cocktails for targeting the ichthyoplankton. Mesozooplankton samples were collected at 4 locations on the Portuguese coast – 1 in the northwest (Viana do Castelo) and 3 in the south (coastal lagoons of Ria de Alvor and Ria Formosa, and in the river Guadiana estuary). Bulk community DNA was extracted using a non-destructive protocol and amplicon libraries produced for the 3 primers combinations. After quality-filtering bioinformatic steps, we obtained 3.04 x 105 usable sequences, of which 76.26% were clustered into OTUs (operational taxonomic units) and 46.30% were identified at species level - corresponding to 103 taxa from 8 different metazoan Phyla. The most diverse classes were Malacostraca, Actinopterygii, and Copepoda. As expected, the generic primer pair for marine metazoa (mlCOIintF/LoboR1) retrieved a higher number of species (94) compared with the fish-specific primer cocktails (30). Nevertheless, 9 % of the total species were identified exclusively by the cocktails, of which 42% were fish. These results confirmed the potential of metabarcoding as a tool for profiling zooplankton communities and to assess ichthyoplankton diversity. Multiple primers pairs increased species detection from different taxonomic groups, being the protocol optimization for fish-specific primer cocktails, the next step for its implementation in fish stock assessments.  HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • Assessing the seasonal dynamics of zooplankton in a recreational marina of
           the northwest of Portugal through multi-marker DNA metabarcoding

    • Abstract: ARPHA Conference s 5: e87492
      DOI : 10.3897/aca.5.e87492
      Authors : Jorge Moutinho, Ana S. Lavrador, Pedro E. Vieira, Filipe O. Costa, Sofia Duarte : The monitoring of larvae in plankton samples in recreational marinas, ports, or the close vicinities, may provide key information about non-indigenous species (NIS) introduction status or detect their presence at an earlier stage. DNA metabarcoding is a powerful method to assess biodiversity but its efficiency is dependent on the methodologies employed along the analytical chain, namely the targeted genetic markers. Therefore, we aim to analyze the ability of two genetic markers - the mitochondrial cytochrome C oxidase subunit I gene (COI) and the variable region V4 of the nuclear small subunit ribosomal gene (18S) - to assess the seasonal dynamics of zooplankton communities, including NIS, through DNA metabarcoding, and further to compare it with a compiled list of zooplanktonic species identified via morphology that occur in the Lima Estuary. To this end, we sampled zooplankton communities, spanning three consecutive seasons (spring, autumn, and winter 2020/2021) in three sampling points in the recreational marina of Viana do Castelo, located in the Lima Estuary, North of Portugal. Globally, metabarcoding recovered 157 species belonging to 19 phyla, with a dominance of Annelida, Crustacea, and Mollusca that represent 64.3% of all species. Even though our sampling area poorly represents the Lima Estuary, this approach allowed the detection of 6 NIS, which were not yet reported by traditional methods of identification. The composition of the zooplankton communities recovered was greatly affected by the markers employed, and to a lesser extent by the season. Overall 18S detected more species and fewer species were recovered in winter. NIS were predominantly detected during spring and autumn, but only 5.1% of all species were detected in all seasons. This study shows the DNA metabarcoding efficiency in assessing the dynamics of zooplankton communities and highlights the interest in using a multi-marker and seasonal approach to species detection, particularly to target NIS. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • Full chloroplast sequencing using genome skimming for novel plant DNA
           barcode discovery in Amaranths

    • Abstract: ARPHA Conference s 5: e87349
      DOI : 10.3897/aca.5.e87349
      Authors : Leonardo Galindo Gonzalez, Marie-Claude Gagnon, Sarah Kyte, Andréanne Charron, Adam Colville : DNA barcoding has been established as an efficient, sensitive and reliable methodology for plant identification. However, in spite of efforts to find a universal DNA plant barcode, some taxa are not sufficiently resolved by typical plant barcoding genes like matK or rbcL. We have used a technique known as genome skimming, which relies on the empiric low coverage sequencing of a full plant genome, resulting in high coverage of the high copy genome fractions such as chloroplast and rDNA. Phylogenetic studies show that these regions are a reservoir of variability which could be further exploited for DNA barcode discovery. We sequenced eight amaranth species using Illumina Next Generation Sequencing technology to test the feasibility of this technique. Amaranths were chosen due to their increasing impact as invasive species bearing multiple herbicide resistance mechanisms. Our results showed that complete chloroplast genomes could be assembled for all of the eight species tested. We obtained an average of 47 million reads for each one of the amaranth nuclear genomes, which range in size between 400-700Mb approximately. These reads provide an average theoretical coverage of 10-15X for each nuclear genome, but resulted in an average chloroplast genome coverage in the range of 500-8000X due to multiple chloroplast genome copies per cell. Alignment of the eight chloroplast genomes shows variability in the single copy regions (Fig. 1), especially on intergenic sections (Fig. 2). Additional preliminary analyses also show variation among different populations of the same species, demonstrating the importance of studying both inter and intraspecific diversity to design reliable and accurate DNA barcodes that can be used in species identification. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • Catmint (Nepeta nuda L.) phylogenetics defined by nuclear and chloroplast
           DNA barcodes 

    • Abstract: ARPHA Conference s 5: e87343
      DOI : 10.3897/aca.5.e87343
      Authors : Georgi Bonchev, Miroslava Zhiponova, Anita Tosheva, Valya Vassileva : Nepeta nuda L. (Lamiaceae) is a medicinal plant with a wide distribution in Europa and Asia. In Bulgaria, N. nuda is also known as “naked (or hairless) catmint”, which likely refers to the naked or sparse short hairy stem and leaves. This study aims to generate DNA barcodes for precise genetic discrimination and determination of phylogenetic position of the plant among other Nepeta species. To achieve this goal, we applied the DNA barcoding technique based on conserved nuclear (internal transcribed spacer/ITS) and chloroplast (rbcL, matK, trnH) DNA regions. The generated DNA barcode sequences were submitted to the Barcode of Life Data System (BOLD) database (https://www.boldsystems.org; accession number BUL002-22). The obtained N. nuda DNA barcodes and corresponding sequences of other Nepeta species available in the BOLD and NCBI database were utilized for taxonomic classification. ITS-derived sequences for Nepeta species were the most enriched in the database, and DNA fragments matching 47 Nepeta species were selected for construction of phylogenetic tree. The results show that N. nuda is clustered in a subclade together with N. sheilae, N. deflersiana, N. isaurica, N. congesta, N. heliotropifolia, N. schiraziana and N. cataria. The information in BOLD was also retrieved for rbcL and matK, and corresponded to 15 and 10 different Nepeta species, respectively. For trnH, only NCBI sequences corresponding to 6 different Nepeta species were found. Тhe three chloroplast markers highlighted the close relation of N. nuda to N. italica, N. tuberosa, N. cataria, N. grandiflora and N. hemsleyana. In conclusion, we suggest a DNA barcode system for genetic discrimination of N. nuda, which could assist its accurate taxonomic characterization. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • First results on the genetic variation of the Bulgarian populations of
           Morimus asper funereus (Mulsant, 1862) (Coleoptera, Cerambycidae)

    • Abstract: ARPHA Conference s 5: e87224
      DOI : 10.3897/aca.5.e87224
      Authors : Rumyana Kostova, Aneliуa Bobeva, Rostislav Bekchiev : There is an ambiguity about the taxonomic status of Morimus asper funereus (Mulsant, 1862), a saproxylic longhorn beetle, widespread in Bulgaria and other Balkan countries. It is a protected species under the Habitats Directive (Annex II) and under the Bulgarian Biodiversity Act. A recent molecular study, based on the cytochrome C oxidase subunit I (COI) and the second internal transcribed spacer (ITS2) gene se­quences, found that all European and Turkish populations of Morimus should be referred to M. asper (Sulzer, 1776) (Solano et al. 2013). The territory of Bulgaria and some other Balkan countries remain as an unstudied gap in that research. Here we present preliminary results on the genetic divergence and diversity of Morimus populations on the Balkans based on COI gene sequences using material from Bulgaria and Albania and the haplotypes’ sequences of Solano et al. (2013) obtained from GenBank. The material collected from Bulgaria аnd two localities in Albania was identified based on morphology characteristics as M. asper funereus (44 samples), M. verecundus bulgaricus Danilevski et all., 2016 (1 sample from the type locality), Morimus orientalis Reitter, 1894 (1 sample from Strandzha Mts.) and Lamia textor (Linnaeus, 1758) (1 sample) used as an outgroup. All obtained sequences were analyzed and haplotype diversity was estimated. The results show that Bulgarian populations of Morimus demonstrate relatively high haplotype diversity in correspondence to the diversity of the other European populations. Also, a phylogeographical hypothesis of the linkage between Bulgarian and other Balkan populations was developed. Four main lineages of divergence were identified. In addition, our results support the assumption that M. verucundus is a tentative subspecies of the morphologically and genetically variable M. asper. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • Let’s talk about the (lady)birds and the bees: how insects can
           whisper a multitude of stories

    • Abstract: ARPHA Conference s 5: e85529
      DOI : 10.3897/aca.5.e85529
      Authors : Physilia Chua, Petra Korlevic, Lyndall Pereira-da-Conceicoa, Cameron Ferguson, Leia Zhao, Mara Lawniczak : If you have watched A Bug’s Life, you would have seen that insects come in an assortment of colours, shapes, and sizes. They are the perfect organism that could be used to describe the myriad diversity of all life on earth. These six-legged creatures are one of the most diverse groups of species, accounting for more than 80% of all documented living animals Ødegaard 2008. This huge diversity also makes it extremely time- and labour-intensive to carry out large-scale monitoring of insects. High-throughput sequencing technologies and the use of DNA barcodes for species identification have paved the way for the rapid biomonitoring of insects Hebert et al. 2003. However, millions of insect species are not well-represented in DNA reference databases, making species-level identification challenging for molecular research. Projects such as the Darwin Tree of Life (DToL) aim to cover this gap and generate DNA barcodes for all eukaryotic species found in the UK Blaxter et al. 2022. At the Wellcome Sanger Institute, the BIOSCAN UK for Flying Insects project has two main aims;Documenting the diversity of UK flying insectsDiscovering insect-cobiont interactionsTo meet these aims, we will be documenting the diversity of one million malaise-caught insects from 100 sites across the UK in the next five years. We will be using a non-destructive DNA extraction technique to preserve insect specimen integrity for museum collections or educational purposes Korlević et al. 2021. COI barcoding will be carried out using ONT and/or PacBio long-read technology to identify each insect specimen. To tease apart insect conbiont interactions, we will carry out mini barcoding using primers targeting microbes, parasites, vertebrates, invertebrates, and plants Fig. 1. Together, this molecular dataset consisting of one million specimens collected over space and time in the next five years will allow us to discover how insects interact with the ecosystem and advance insect biomonitoring research in the UK. HTML XML PDF
      PubDate: Fri, 17 Jun 2022 11:00:00 +030
  • First data about cave fauna from the Albanian Alps

    • Abstract: ARPHA Conference s 5: e85375
      DOI : 10.3897/aca.5.e85375
      Authors : Joanna Kocot-Zalewska, Jakub Bienias, Robert Rozwałka, Paweł Sienkiewicz, Andrzej Woźnica : In September of 2021, a Polish expedition took place in the Valbona Valley in the Albanian Alps (Kocot-Zalewska et al. 2021) During the expedition, a project study of invertebrate cave fauna had begun. The main purpose of this presentation is to show the preliminary results of a study on the invertebrate fauna in the Albanian Alps. Within 14 days, arthropods were collected in subterranean habitats between 1500 and 2300 m a.s.l. The study was led on the left and right sides of the Valley. On the left side of the Valley, we explored bauxite mines and Spella Huxhise Cave; on the right side, the Ice Cave, Spella de Akullt, Spella e Valbones, and Spella Sportive caves were explored. Mainly, the direct searching method was used. However, pitfall traps with baits were used in one cave and one tunnel. Altogether, from five caves and a complex of four tunnels, 102 arthropods belonging to insects (beetles, flies, and crickets), millipedes, spiders, harvestmen, and springtails, have been collected. Flies and beetles are the most numerous groups. The majority of the collected specimens have been determined to the species level. Among these, many are new to Albanian fauna. Particular attention should be paid to the species highly related to the subterranean environment and those found in ice caves. This project will continue in the year 2022. We plan to explore the next caves in the studied area and re-examine the previously visited sites. HTML XML PDF
      PubDate: Tue, 26 Apr 2022 09:00:00 +030
  • The effect of climate on age specific survival and senescence in a Hazel
           Dormouse (Muscardinus avellanarius) population in Lithuania, across

    • Abstract: ARPHA Conference s 5: e85540
      DOI : 10.3897/aca.5.e85540
      Authors : Thomas Berg, Fernando Colchero, Owen Jones, Lene Sanderhoff, Rimvydas Juškaitis : It is well known that survival and the dynamics of wild populations are affected by environmental factors. Recent research has found that, among some species of mammal, differences in environmental conditions between populations of the same species translate into changes in infant and juvenile mortality, but not in the rate of senescence (‘rate of aging’). This has been confirmed among primates and some species of carnivores, but has not been tested on other taxonomic groups, such as rodents. Here, we analyse age-specific survival and mortality on the most extensive capture-mark-recapture data set on Hazel Dormouse available from Lithuania. We used Bayesian survival trajectory analysis (BaSTA) and tested different models of age-specific mortality. Since the Hazel Dormouse population is in decline across its northern distribution, potentially in response to climate change, we divided the data into three periods to assess changes in survival over time. Regional climate data were obtained from the NOAA data service to test the effect of climatic factors on survival during winter and summer respectively. Our results show that, during all three periods, male life expectancies were longer than those of females. We found that the overall level of mortality was high for all three periods, with lowest mortality during the period 1999  - 2004. We found large differences in juvenile mortality and age-independent mortality between the three periods, but not in adult mortality or in the rate of senescence. This is consistent with previous findings on other mammals, supporting the invariant rate of aging hypothesis. HTML XML PDF
      PubDate: Tue, 26 Apr 2022 09:00:00 +030
  • Detecting Hazel Dormice in the UK: what are we missing'

    • Abstract: ARPHA Conference s 5: e84871
      DOI : 10.3897/aca.5.e84871
      Authors : Simone Bullion, Alison Looser : Multiple techniques can be used to detect the presence of Hazel Dormice and as a result, there is reasonable confidence that lack of detection means that dormice are probably absent. In the UK, ancient woodlands are frequently considered as the reservoir of Hazel Dormouse populations and are a usual starting point for surveys. However, we present evidence that this may be one of the most difficult habitats in which to detect the species. This is largely due to many of these woodlands being unmanaged, with a poorly developed shrub layer due to heavy shading. This reduces the probability of finding natural field signs such as nests and chewed hazel nuts. Similarly, because detection devices (nest boxes, nest tubes and footprint tunnels) are typically positioned no more than two metres from the ground for ease of study, there is an assumption that Hazel Dormice will leave the dense canopy to use them. We predict that there is a higher risk of false negatives in this habitat type and that more research is needed to provide better certainty of detection when surveying in high canopy woodland. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Ecology and biology of the Roach’s Mouse-tailed Dormouse (Myomimus
           roachi, Bate 1937)

    • Abstract: ARPHA Conference s 5: e85302
      DOI : 10.3897/aca.5.e85302
      Authors : Nedko Nedyalkov, Ivailo Raykov, Lisa Hesse, Anna Staneva : The Roach’s Mouse-tailed Dormouse (Myomimus roachi, Bate 1937) is endemic to the Western Palearctic (occurring in SE Bulgaria, Turkish and Greek Thrace, and western Anatolia) and is one of the less known mammals within the region. There is no information about its biology and ecology in the wild. In the course of the past 3 years (2019-2021), we conducted intensive research on a small population situated in southeast Bulgaria (Sakar mountain). The population was regularly monitored during the active time of the species. To study its ecology and biology we used a combination of nest boxes, live traps and camera traps. During this period, we collected 259 records of sex, age, and weight. Those bigger than 10 g. were microchipped. The active period for the Roach’s Mouse-tailed Dormouse lasts between April and September. Males emerged first from hibernation at the end of April (24th of April - the earliest registration). During this short active period, the dormouse breeds only once, the litter size being between 5-9 young. The first young appeared at the end of June. We observed a summer dormancy (estivation); the adult males became less active and went in the estivation first. The last active dormice were observed in the first ten days of September.Here we also present data about the population numbers and dynamics, age, and sex structure of Roach’s Mouse-tailed Dormouse in the study area. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Status and distribution of dormice (Mammalia, Gliridae) in Bulgaria

    • Abstract: ARPHA Conference s 5: e85301
      DOI : 10.3897/aca.5.e85301
      Authors : Nedko Nedyalkov, Ivaylo Raykov, Yordan Koshev : Here we summarize all available data about dormice (Gliridae) in Bulgaria, published one and our own observations, collected in the last 20 years. Four species are found in Bulgaria - the Forest Dormouse (Dryomys nitedula), the Edible Dormouse (Glis glis), the Hazel Dormouse (Muscardinus avellanarius) and the Roach’s Mouse-tailed Dormouse (Myomimus roachi). Altogether we collected 1200 records from 283 locations. A database has been created in which every single record is kept. The most common and widespread species are the Edible (G. glis) and Forest Dormouse (D. nitedula), found in 212 and 139 locations respectively. They inhabit mainly the deciduous and mixed forest in the country. The Hazel Dormouse (M. avellanarius) is found in 77 locations, the bulk of them are in the mountains, but there are some locations along the Danube and southeast Bulgaria. It prefers woods with a well-developed understorey. The Roach’s mouse-tailed dormouse (M. roachi) is the rarest one - found in 26 locations in southeast Bulgaria. In contrast to the other dormice, it avoids woods and lives in semi-open habitats with shrubs, tree lines, and islands in grasslands and agricultural fields. There are several anecdotal reports of the Garden Dormouse (Eliomys quercinus) but no hard proof exists for the presence of this species. We discussed the patterns of distribution, conservation status, and coexistence of Bulgarian dormice. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • How to deal with Hazel Dormice affected by large infrastructure projects

    • Abstract: ARPHA Conference s 5: e85238
      DOI : 10.3897/aca.5.e85238
      Authors : Lisa Höcker, Hanna Voll, Oliver Wild, Markus Dietz : Hazel Dormice (Muscardinus avellanarius) are protected by national implementation of European law. Protection comprises of the prohibition of physical harm to individuals and the destruction of breeding or nesting sites. Due to Germany’s policy goal of changing from a fossil fuel dominated energy supply to more sustainable energy sources, large infrastructural projects became crucial to, for example, redistribute energy from Northern and Eastern Germany to Bavaria. Within the context of the inevitable building projects, the protection goals for species like the Hazel Dormouse need to be maintained.The layout of such a project with a linear design stretching across hundreds of kilometers and intersecting different landscapes and habitats poses new challenges. Aspects of the construction phase of underground cables need to be considered in planning as well as the design of the protective strip above the finished cable. In order to plan mitigation and compensation measures, species presence and absence data need to be collected effectively.A total of 223 sample sites along a 207 km long corridor were equipped with 5,499 nest-tubes and monitored regularly. For habitat potential, biotope types were assessed and mapped within the corridor. All types of potential Hazel Dormouse habitat were selected considering regional differences. Based on previous research, we established guidelines for the transferability of presence and absence data.We present the procedure for data collection spanning a vast area using nest-tubes, proposed guidelines for the transferability of results, as well as mitigation and compensation measures necessary to ensure protection of the Hazel Dormouse. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Protecting local populations of Hazel Dormice (Muscardinus avellanarius)
           by translocation: a long-term case study from North Rhine-Westphalia

    • Abstract: ARPHA Conference s 5: e85236
      DOI : 10.3897/aca.5.e85236
      Authors : Lisa Höcker, Katja Weiß, Markus Dietz : As part of the mitigation measures adopted in the context of preparatory clearcutting for surface mining, Hazel Dormice have been translocated since 2011. The project is situated in the geographical region of Lower Rhine Basin (North Rhine-Westphalia, Germany). To date, Hazel Dormice have been translocated from an old-growth deciduous forest to four other sites. Three release sites are re-cultivation areas which were planted with trees and shrubs roughly 10 to 30 years ago. The other release site is an old-growth deciduous forest with a well-developed understory. Nest boxes were placed in the source forest and checked between April and October. Any dormice that were found were individually marked and released, inside their nest boxes, at the new sites. Two or three additional nest boxes were placed within the surroundings of each translocated box.Between 2011 and 2018 we translocated 1,840 individuals. We verify the success of translocation by monitoring the release sites in June and September of the first, second, fifth and eighth year after translocation. Additionally, a sample of 200 nest boxes were checked each month between April and October for five years. Finally, we examined groups of nest boxes shortly after translocation to gather information on the translocated animals immediately after release. Individual records reveals that translocation can be a successful conservation measure to secure local populations if their former habitat is damaged. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Settling in: space utilization behaviour of translocated Hazel Dormice
           (Muscardinus avellanarius)

    • Abstract: ARPHA Conference s 5: e85235
      DOI : 10.3897/aca.5.e85235
      Authors : Lisa Höcker, Markus Dietz, Joanna Fietz : The Hazel Dormouse (Muscardinus avellanarius) is protected under the EU’s Habitat Directive and its national implementation, whereby physical harm to individuals as well as the destruction of breeding and nesting sites is prohibited. Within the context of inevitable environmental impacts these protection goals are not always maintained. Therefore, mitigation and compensation measures to secure a population’s viability are required.For sedentary and less mobile species like the Hazel Dormouse, mitigation often includes the translocation of individuals to a different habitat patch. In 2017 and 2018 a total of 108 individuals were translocated from an old growth oak-hornbeam forest to an area that was re-forested 40 years ago. Prior to translocation, individuals were equipped with a radio transmitter and they were followed for three consecutive nights and one control night, during which bearings were taken every five minutes using the triangulation technique. A control group of established Hazel Dormice was followed in the same manner. Nests and resting sites were searched for during the daytime.Twelve male Hazel Dormice were radio-collared and between 234 and 427 bearings were taken per individual. Resting sites of translocated dormice were mainly located within the forest canopy (33.3%). Furthermore, individuals were often found active during the daytime. Mean activity range size (MCP) and resource search areas (LoCoH 95 %) were larger in translocated dormice, whereas the mean core habitat size (LoCoH 50 %)  was larger in established dormice. The activity range during all of the monitored nights overlapped significantly more in established dormice, demonstrating exploratory behaviour by translocated individuals. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Forests for the Forest Dormouse: Building conservation guidelines with and
           for land managers

    • Abstract: ARPHA Conference s 5: e85194
      DOI : 10.3897/aca.5.e85194
      Authors : Birgit Rotter, Robin Sandfort, Christine and Stefan Resch, Claudia Kubista : The Forest Dormouse (Dryomys nitedula) is native to a variety of Austrian forest landscapes, although recordings of the species are very rare. In a 3-year nationwide search we are trying to collect data on abundance, habitat preferences, and response to forest management practices. Nest box occupancy is recorded at 20 study sites. In an extensive citizen science initiative, we are also looking at data from footprint tunnels, camera traps and casualties from cat predation. Additionally, audio detectors (AudioMoths) are tested against other methods as a new monitoring approach.First year results indicate a surprising accumulation of Forest Dormice near or in anthropogenically shaped areas, such as settlements and pastures. Distribution of the species in Austria seems to be much more concentrated on some alpine areas than previously thought.The study results, together with data on land management, will form the basis for management guidelines which will be developed together with foresters and county administrations. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Bridging environment, physiology and life history: stress hormones in the
           Edible Dormouse (Glis glis)

    • Abstract: ARPHA Conference s 5: e84811
      DOI : 10.3897/aca.5.e84811
      Authors : Nadine Havenstein, Franz Langer, Joanna Fietz : The performance of wild animals is impacted by diverse challenges imposed by the environment and distinct life history stages such as breeding and hibernation. Glucocorticoids (GCs) are hormonal mediators that reflect the response to these challenges, exerting their far-reaching effects on numerous processes such as energy allocation, immunity and behavior. Whereas short term elevations of GC levels are crucial for survival, by activation of the emergency life history stage, chronically increased GCs are capable of impairing various body functions and ultimately hamper survival and reproduction. The aim of this study was to disentangle the effects of reproductive activity, prolonged food limitation and hibernation on stress hormone levels in the Edible Dormouse (Glis glis) and to link them to formerly observed survival rates. We therefore measured urinary cortisol levels in wild Edible Dormice in South-western Germany during their active season (2012-2014). Results of our study revealed that reproductive activity was associated with high cortisol levels. During the mating season, particular individuals with a low body mass had high stress hormone levels. Elevated levels of cortisol were also measured during pre-hibernation fattening and were increased in females that had formerly invested in reproduction. Thus, reproduction represents a demanding, potentially stressful, event for both sexes and is linked to distinctly lowered survival rates occurring during years of high reproductive activity. Prolonged food limitation occurring during years of mast failure, did not affect stress levels and were not associated with increased mortality, demonstrating the ability of dormice to predict and cope with food restriction. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • The Benefit of Healthy Hedgerows

    • Abstract: ARPHA Conference s 5: e84975
      DOI : 10.3897/aca.5.e84975
      Authors : Ian White, Megan Gimber : There is an interest in hedgerows because they are home to both plants and animals and provide habitat connection corridors to enable wildlife can move around the landscape. They also deliver a number of additional benefits to farmland, beyond that of a field boundary. These can include:• shelter for livestock• regulation of water supply for crops• improvements in animal health• reduction in soil erosion• reduction in the need for pesticide use• sustaining pollinator communities which support productive farming• providing a sustainable source of wood fuel• reducing the risk of bovine tuberculosis in cattle herds• reducing the rate of climate change through carbon storage• reducing pollution  HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Hedgerow Management Cycle

    • Abstract: ARPHA Conference s 5: e84976
      DOI : 10.3897/aca.5.e84976
      Authors : Ian White, Megan Gimber : Hedgerows are as dynamic as the plants that make them, which means it would be fighting a losing battle if we tried to keep them the same size and shape forever. Instead, work out where the hedge is on the management cycle and work according to that. The ultimate goal in hedge management it to create a thick, dense hedgerow. These are the hedges that are most beneficial to landowners as well as for nature.When we cut hedges at the same point year after year, it will produce fewer flowers and fruits for wildlife, it will lose its lower branches and the hedge will become tall and at risk of invasion, opening up gaps and eventually even failure. Trimming to a slightly higher and wider point each year will help prevent this, and the hedge can be re-shaped when needed.However the hedge is managed each year, at some time the lower parts of the hedge vegetation will become thin and the hedge will need more dramatic action like laying or even coppicing. This only needs to be done every 40+ years to keep the hedge healthy and valuable for our wildlife. This is shown in more detail on the hedgerow management cycle plan. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Diel activity patterns of Garden Dormice (Eliomys quercinus) assessed by
           camera trap data

    • Abstract: ARPHA Conference s 5: e85026
      DOI : 10.3897/aca.5.e85026
      Authors : Hendrik Queckenstedt, Sven Büchner, Hermann Ansorge, Johannes Lang : Garden Dormouse populations have been decreasing in range and declining in number throughout Europe. Understanding, and therefore preventing, this decline is urgently needed. To devise effective conservation strategies, it is essential to gain knowledge on activity patterns of the target species. Diel activity patterns of Garden Dormice (Eliomys quercinus) were investigated at 25 different sites in Germany. Through an intensive camera-trap survey from March to October 2020 a total of 192,136 pictures were recorded in over 3,682 camera-trap days. The association between activity and environmental parameters, such as habitat (urban vs. natural), the lunar phase and sunrise and sunset, was investigated. Garden Dormice were found to be predominantly nocturnal, but were occasionally active during the day in the summer months. The peak of activity at all sites was shortly before midnight. Garden Dormice in this study did not arrange their activity according to the lunar phase. Daytime activities could be related to the presence of young or the food supply. Weather conditions and cloud cover could explain the small influence of the lunar phase on the animals’ activity. This study revealed the effectiveness of camera trapping to gather information on activity of an elusive small rodent with minimum disturbance to the animals. The data was useful for analyzing and understanding diel activities in different habitats. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Genetic assessment of the Forest Dormouse Dryomys nitedula (Pallas, 1773)
           in Latvia

    • Abstract: ARPHA Conference s 5: e84997
      DOI : 10.3897/aca.5.e84997
      Authors : Dainis Ruņģis, Inese Gavarāne, Linda Bankovska, Valdis Pilāts, Zanda Segliņa, Digna Pilāte : The Latvian population of the Forest Dormouse is small and isolated, situated far from the main distribution range of this species. The population is located in the south-eastern region of Latvia, close to the Belarusian border. Sixty-three nest-boxes were located throughout this distribution area and captured forest dormouse individuals were sampled for genetic analysis. Animal age, sex, and coordinates from place of capture were recorded. DNA was extracted from 120 individuals using the QIAamp DNA Mini Kit (Qiagen, Germany). RAPD-PCR polymorphisms at 65 loci were used to examine the genetic diversity and differentiation of the Forest Dormouse population in Latvia. 89% of loci were polymorphic, average expected heterozygosity over all loci was 0.321, average polymorphism information content (PIC) was 0.246. The program ‘Structure’ was used to perform Bayesian assignment of individuals to a predefined number of clusters. The deltaK method identified that the most likely number of clusters within the Latvian Forest Dormouse population was three. Assignment of individuals to a particular cluster mostly corresponded with the location where the individual was captured. Analysis of molecular variance (AMOVA) indicated that there was significant genetic differentiation between the three clusters with pairwise Phipt (a modification of Fst) values ranging from 0.118 to 0.180. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Structural re-design of the Animex Wildlife Bridge for the Hazel Dormouse
           (Muscardinus avellanarius): Lessons Learnt from Two Connectivity
           Mitigation Case Studies in England

    • Abstract: ARPHA Conference s 5: e84977
      DOI : 10.3897/aca.5.e84977
      Authors : Ian White, Darrelle Moffat, Steve Béga : The Hazel Dormouse is an arboreal mammal present in Europe and Asia Minor. The population is declining in the United Kingdom (UK), partially due to habitat fragmentation caused by the development of roads and other linear infrastructure. In 2016, we designed and tested an arboreal bridge in Britain that proved to be effective for Hazel Dormice. Subsequently, the bridge materials were upgraded to meet the technical standards of UK road agencies, so that it could be approved and implemented as mitigation on projects throughout the country. In the UK, each bridge must be technically certified by the relevant road authority and, as the bridge is a unique structure, this can pose challenges on projects. Two installation designs were created: the ‘standalone’ bridge and the ‘retrofit’ bridge. Several bridges have now been installed above roads in the UK. We discuss the development and implementation process from two case studies: a 40-metre retrofit bridge to an underpass at St Athan in Wales and two 76+ metre bridges retrofitted to an overbridge and underpass on the M1 motorway at Gayhurst, Buckinghamshire. This poster discusses what we have learned from these projects, how they have influenced the future designs of the Animex wildlife bridge, and the production of best practice guidance to provide a summary of the entire process of implementing a bridge project. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Developing an environmental DNA protocol for Hazel Dormouse surveys

    • Abstract: ARPHA Conference s 5: e84962
      DOI : 10.3897/aca.5.e84962
      Authors : Emma Cartledge, Paula Stockley : Recording Hazel Dormouse (Muscardinus avellanarius) presence allows conservation and mitigation efforts to be effectively directed towards populated sites. As it is a protected species, licences are required for survey methods which disturb the animals or their habitat. Therefore, reliable low disturbance survey techniques are desirable.In this study, we tested the potential of detecting environmental DNA (eDNA) from soil as a survey method for dormice. Firstly, we designed species-specific primers and the associated PCR protocol using tissue samples from Hazel Dormice. To ensure specificity, we also checked the protocol against tissue samples from non-target species. Next, we collected soil samples from occupied sites in England to test the methodology with environmental samples. This included testing site-level heterogeneity of dormouse DNA distribution and localised level DNA distribution surrounding occupied nest boxes. Finally, we tested the soil for DNA presence changes over time, after removal from the site.Using eDNA for dormouse surveys is promising. Our results indicate that a usable survey methodology can be developed, providing a relatively low cost and time-efficient tool when compared to existing dormouse survey methods. Future work should aim to further understand the changes of dormouse eDNA through time. Through identifying populated sites, eDNA could support mitigation projects and help to direct dormouse conservation efforts. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • 30 years of Dormouse Monitoring

    • Abstract: ARPHA Conference s 5: e84932
      DOI : 10.3897/aca.5.e84932
      Authors : Ian White, Nida Al-Fulaij, Laura Bower : The People's Trust for Endangered Species (PTES) owns a private woodland on the Isle of Wight (IoW) that is managed predominantly for woodland bats, Red Squirrels and Hazel Dormice. Dormice are considered to be widespread across the island in suitable habitat. The IoW is different from the mainland as it has no wild deer species. PTES has been monitoring dormice in its woodland since 1992, when nest boxes were first put up by Paul Bright. However, in spite of appropriate woodland management for dormice at the reserve, dormouse numbers there appear to be declining. This raised the question: “Is the apparent decline in dormice recorded in nest boxes, real or perceived'” If the decline was real, it may be necessary to reconsider management advice that we give for dormice. If the decline was perceived, then it may be necessary to reconsider advice that we give for monitoring dormice. The first challenge was to identify what the woodland may have looked like 30 years ago and identify why high numbers of dormice were recorded. We could then apply the known woodland management that was done in the intervening years, to determine why dormouse nest box occupancy changed by varying amounts in different parts of the wood. We were able to check some of our ideas using data from footprint tunnels and this work is ongoing in 2022. This talk will discuss woodland state, woodland management and dormouse next box occupancy in a dormouse hotspot in southern England. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • The Great British Hedgerow Survey

    • Abstract: ARPHA Conference s 5: e84931
      DOI : 10.3897/aca.5.e84931
      Authors : Ian White, Megan Gimber : In 2013, when the dormouse reintroduction protocol was revised, a national hedge survey methodology was available. This survey enabled individual hedge quality to be assessed and from that, it was possible to quantify the quality of landscape links between reintroduction sites. There was an opportunity to build on this survey so that it could be used more widely and have more useful outcomes. The People's Trust for Endangered Species (PTES) acquired the existing data from the national (DEFRA) survey and started to develop a survey methodology that would be compatible with it, be simple to use, provide a report on the existing state of the hedgerow and give management advice on how to maintain the hedgerow in good condition. As part of the survey development, posters were designed. One of these asks: ‘What have hedgerows ever done for us’, to demonstrate the benefit of hedgerows in the landscape. The other shows ‘The hedgerow management cycle’ to highlight the dynamic nature of hedgerow condition. There are two different hedgerow surveys available, each with a slightly different focus and outcomes. Healthy Hedgerows is a rapid hedgerow survey designed for landowners that want to create a hedge management plan. The Great British Hedgerow Survey generates more detailed feedback on hedgerow health and offers management advice. It is particularly suited to volunteer groups. This talk will discuss the development of the surveys and their part in hedgerow protection in the UK in the future. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • The Dormouse Reintroduction Program in the UK

    • Abstract: ARPHA Conference s 5: e84930
      DOI : 10.3897/aca.5.e84930
      Authors : Ian White : Hazel Dormice have been lost from 17 English Counties over the past 150 years and the population has declined by 51% since the year 2000. The dormouse reintroduction program has been running since 1993 with the aim of restoring dormice to their native range in the UK. In the early years, a single dormouse population was released in a suitable woodland with the hope that it would occupy all available habitats in the release site and then start to disperse into the wider countryside. While this approach did have some successes, it also had some failures. The program was reviewed in 2014 and some amendments were proposed to the original protocol. A key amendment was to identify at least two woodlands at a site, in relatively close proximity, each suitable for a dormouse release and to undertake at least two reintroductions in an area. Further to the dormouse reintroductions, a landscape project would be established to improve connectivity between the woodlands. The longer-term aim is now to facilitate the creation of a dormouse metapopulation in an area rather than just hoping it would happen. This talk discusses the advantages and difficulties of this new approach and its successes and failures. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Number and length of whiskers in dormice

    • Abstract: ARPHA Conference s 5: e84929
      DOI : 10.3897/aca.5.e84929
      Authors : Eva Marie Famira-Parcsetich, Holger Meinig, Sven Büchner, Johannes Lang : Whiskers (vibrissae) are slender, curved and tapered hairs present in most mammals, which transmit vibrotactile information. In contrast to pelage hair, whiskers are longer and thicker and they possess highly innervated follicles. So called ‘whisking’, a symmetrical, cyclic movement of whiskers can be observed in some tactile specialists. It is used to scan the environment especially during locomotion and foraging. Animals showing the ability of whisking are mainly small, nocturnal and arboreal species like dormice. However, nearly everything we know about whiskers and whisking derives from just a handful of species, including laboratory rats and mice. Until now, information on whiskers and whisking in dormice has been scarce. We describe for the first time the number, length and arrangement of whiskers in the Garden Dormouse, the Edible Dormouse and the Hazel Dormouse.We counted and measured macro- and micro-vibrissae of five cadavers of the three species and calculated median values for each species. Anatomical studies on whiskers provide data for comparative studies between different species. Moreover, knowledge about whiskers and whisker-use can guide us in designing enriched enclosures for captive mammals. This is especially useful for small, nocturnal species that use whiskers for orientation; including a range of textures and climbing frames in their enclosures is important. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Looking up to the sky: using high resolution remote sensing to
           characterise hibernaculum locations of the Hazel Dormouse

    • Abstract: ARPHA Conference s 5: e84812
      DOI : 10.3897/aca.5.e84812
      Authors : Leonardo Gubert, Fiona Mathews, Jon Bennie, Robbie McDonald, Robert Wilson, Ruud Foppen, Pim Lemmers, Maurice La Haye : The Hazel Dormouse is predominantly an arboreal species that moves down to the ground to hibernate in the autumn in temperate parts of its distributional ranges at locations not yet well understood. In this study, we tested whether environmental characteristics surrounding Hazel Dormouse hibernacula can be identified using high-resolution remote sensing and data collected in situ. We modelled remotely sensed variables, including canopy height and cover, topographic slope, sky view, solar radiation and cold air drainage around 83 dormouse hibernacula in England (n=62) and the Netherlands (n=21), and identified environmental characteristics that may be favoured by pre-hibernating dormice. We also collected and analysed data on leaf litter depth, temperature, canopy cover and distance to the nearest tree collected in situ at hibernaculum locations in England. We found that remotely sensed data were effective in identifying attributes surrounding the locations of dormouse hibernacula and, when compared to in situ information, provided more conclusive results. Our study suggests that remotely sensed topographic slope, canopy height and sky view have an influence on animals choosing suitable locations to hibernate; whilst in situ data suggested that average daily mean temperature at the hibernaculum may also have an effect. Remote sensing proved capable of identifying localised environmental characteristics in the wider landscape that may be important for hibernating dormice. We also propose that this method can provide a novel progression from habitat modelling to conservation management for the Hazel Dormouse, as well as other species using habitats where topography and vegetation structure influence fine-resolution favourability. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Comparing methods to detect Garden Dormice (Eliomys quercinus) in
           mountainous forest habitats

    • Abstract: ARPHA Conference s 5: e84916
      DOI : 10.3897/aca.5.e84916
      Authors : Sarah Beer, Klaus Hackländer, Johannes Lang : Garden Dormouse populations show a severe and ongoing decline all over Europe. The drivers for this process are still unknown, as well as the exact distribution of the remaining populations. An evaluation of the occupied habitats could give information on important habitat parameters. Therefore, it is essential to improve detection methods and to evaluate the efficiency of available techniques. In this study, three different methods were tested on ten transects in the Fichtel Mountains, Germany. On each transect we installed two camera traps (CT), one autonomous sound recording unit (AudioMoth, AM) and 25 footprint tunnels (FT), which were checked weekly from the beginning of June until the end of October. AMs did not record a single call, FT detected Garden Dormice in seven and CT detected them in all ten transects. FT worked best during July, when they provided evidence of Garden Dormouse on six transects. CT worked best in August and September, confirming Garden Dormice in all ten transects. In every month, CT outperformed FT in the number of positive findings, and in the time interval until initial detection. CT worked most efficiently in this type of habitat, whereas FT produced false absence data. As Garden Dormice inhabit a variety of different habitats, this study should be repeated to see if the results are transferable. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • A 24-year investigation of the reproductive ecology of the Edible Dormouse
           (Glis glis)

    • Abstract: ARPHA Conference s 5: e84743
      DOI : 10.3897/aca.5.e84743
      Authors : Sarah Brooks, Roger Trout, Davina Hill : The Edible Dormouse is a non-native rodent introduced to the Chilterns, UK, in 1902. It shows extreme annual variation in breeding; either having one litter per year or, in some years, failing to breed at all. Despite this slow reproductive rate compared to other rodents, its population is increasing, and has spread beyond the anticipated landscape barriers. Communal nesting is sometimes observed. This study aims to characterise the reproduction parameters over time, including from communal breeding.Based on a population of>11,000 PIT-tagged individuals, 600 litters and 4000 nestlings at our Chilterns study site over 24 years, we show the best fit of the number of new-borns is an increasing exponential. Monthly weight of nestlings also increased over the 24 years, while annual reproduction rate, litter size and nestling growth rates remained unchanged. Litter size decreased from 6.6 new-borns in August to 4.3 in October, indicating an average death rate of 37% in the first two months of life. Preliminary analysis suggests a small decrease in death rate over the years. Average growth rate of nestlings was c.100% of birth weight per week in their first month, but then slowed. We also examined evidence of kinship between nest sharers and investigated whether this, or communal breeding in general, promotes offspring survival. Our results will provide a deeper insight into the factors that drive the reproductive success of this invasive species and enable investigation of the effect of climatic variables on the number, survival and growth rate of nestlings. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Size variation in the Edible Dormouse and its environmental correlates

    • Abstract: ARPHA Conference s 5: e82818
      DOI : 10.3897/aca.5.e82818
      Authors : Boris Kryštufek, Franc Janžekovič : Intraspecific variation in size along spatial and environmental gradients has been documented in many studies, and different hypotheses have been proposed to explain these patterns of geographic variation. We explored size variation in 4 island and 35 mainland samples of the Edible Dormouse (Glis glis Linnaeus, 1758) from the western Balkans in Slovenia, Croatia, Bosnia and Herzegovina, Montenegro, and Serbia. Samples comprised 4 to 207 (mean ± SD=19.0±32.9; total n=740) adult skulls (age>1 year) of both sexes. To remove phylogenetic effects, all samples were from a widespread European lineage. Condylobasal length of skull was used as a proxy for overall size. Dormice were the smallest along the southern margin of the Pannonia Plain, the largest were from beech forests of the Dinaric Alps, and intermediate in hilly parts of Serbia and on the Adriatic islands. The largest skulls (from Mt. Zelengora, Bosnia and Herzegovina) were on average 25% longer than the smallest, from the population occurring near Belgrade in northern Serbia. Size correlated with longitude (R2=0.262; p=0.0009) and the 1st Climatic Principal Component (R2=0.687; p
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Places to be: the Garden Dormouse (Eliomys quercinus) in south-western

    • Abstract: ARPHA Conference s 5: e84892
      DOI : 10.3897/aca.5.e84892
      Authors : Harald Brünner, Joanna Fietz, Franz Langer, Rieke Vorderbrügge, Ulrich Weinhold : In Germany the Garden Dormouse (Eliomys quercinus) is assumed to have one of its main distributional areas in the south-western federal state of Baden-Württemberg. However, recent and detailed information from this area was lacking. In order to describe the actual distribution of the Garden Dormouse, and to characterize suitable habitats in this region, a monitoring study was conducted within the national biodiversity project “In Search of the Garden Dormouse” in 2021. Footprint tunnels and wildlife cameras were used to systematically investigate the occurrence of the species on 136 plots distributed over Baden-Württemberg. The data set was combined with Garden Dormouse records from a citizen science online reporting tool, registered between 2019 and 2021. The Garden Dormouse was found most frequently in the mountainous, conifer-rich Black Forest and in the settlements of the Mannheim-Heidelberg region. Another type of habitat was the dry deciduous forests bordering the river Rhine in the Southern Upper Rhine Valley. Only two isolated occurrences were detected in the eastern part of Baden-Württemberg. The results of this study show, that the distribution of the Garden Dormouse in South-West Germany is more limited than previously assumed. However, the diversity of the detected habitats is considerable, ranging from coniferous forests in higher altitudes to urban areas and almost sub-mediterranean deciduous forests. Further studies will focus on the ecology and biology of this species in the latter. For Germany this is an exceptional, but also threatened, habitat which may serve as a stepping stone to recolonize suitable forest habitats in the Rhine valley. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • Dormouse acoustics: The use of artificial intelligence to detect and
           identify European dormouse species

    • Abstract: ARPHA Conference s 5: e84742
      DOI : 10.3897/aca.5.e84742
      Authors : Jennifer MacIsaac : Passive acoustic monitoring (PAM) is becoming increasingly popular for wildlife surveying due to the benefits it offers in terms of non-invasiveness, reduced field time and spatial scalabilty. Acoustic surveying protocols are well established for bat species, but the use of PAM for non-volant small mammals has been largely unexplored. Some dormouse species are highly vocal and are therefore potentially good candidates for acoustic surveying.A landscape-scale acoustic survey is currently being carried out in Polesia, a vast lowland region in Eastern Europe. Three dormouse species have been recorded in this region: Dryomys nitedula, Glis glis and Muscardinus avellanrius, with a fourth species Eliomys quercinus considered rare or locally extinct. Acoustic surveys at this scale generate may hours of recordings, however data analysis can be automated using species classifiers. Classifiers have achieved varying levels of accuracy, but recent advances in artificial intelligence, specifically deep learning has led to the development of acoustic classifiers with high levels of accuracy.In this study, a convolutional neural network classifier was developed to identify D. nitedula, M. avellanarius and nine additional small mammal species. Overall, the classifier achieved 0.944 training accuracy and 0.943 test accuracy. During testing, the classifier achieved D. nitedula and M. avellanarius identification accuracies of 100% and 98% respectively. These preliminary results suggest that PAM could provide an effective, non-invasive method to survey dormice in large geographic areas. Further work will include expansion of the classifier to include G. glis and E. quercinus when sufficient acoustic data are obtained. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
  • The gastrointestinal tract histology of Eliomys quercinus and Glis

    • Abstract: ARPHA Conference s 5: e84817
      DOI : 10.3897/aca.5.e84817
      Authors : Mia Jakopović, Romana Gračan, Duje Lisičić : The Garden Dormouse (Eliomys quercinus) and the Edible Dormouse (Glis glis) differ in their omnivorous diet, with the first being more carnivorous and readily consuming small vertebrates and even poisonous invertebrates, while the latter consumes more plant-based foods. These differences could lead to histological differences in their gastrointestinal tract, which we wanted to test. Six animals of each species were caught in nature and fed the same diet for two weeks to diminish the impact of the short-term external factors. Tissue samples were taken of middle oesophagus, glandular stomach, middle small intestines and large intestines. They were stained with H&E for measuring the thickness of layers in oesophagus gastric mucosa and epithelial cell height in the intestines. Alcian blue - PAS stain was used for goblet cell count and type identification in the intestines, differentiating blue (acidic), lilac (neutral) and purple (mixed) cells. Results showed Eliomys quercinus having greater variance in oesophageal layer thicknesses, significantly thicker muscularis mucosae and thinner tunica muscularis in the oesophagus, and thicker gastric mucosa compared to Glis glis. Eliomys quercinus also had a significantly higher total goblet cell count in both intestines and a higher count of each type in the large intestine, although Glis glis had far more mixed and neutral goblet cells in the small intestine. Both showed intriguingly low numbers of neutral goblet cells in the small intestine. These results indicate interesting differences in gastrointestinal histology and goblet cell distribution in comparison with their distinct diets. HTML XML PDF
      PubDate: Fri, 15 Apr 2022 10:45:00 +030
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