The biology of cave‐dwelling species is a fascinating area, yet it is understudied due to logistic constraints, especially in regard to aquatic organisms. The olm (Proteus anguinus) is the largest ...known troglobiont vertebrate, showing extreme life‐history characteristics such as a possible lifespan of over 100 years and a reproductive cycle of over 12 years. However, most studies carried out on the species to date are based on laboratory studies, resulting in a severe lack of ecological data from natural populations studied in their original habitat. We applied a capture–mark–recapture approach on an Eastern Herzegovinian population for eight years to reveal its spatial strategy and general movement patterns. We found that P. anguinus is sedentary, can often be found within a few square metres over several years, and their moving distance is uncorrelated with the time elapsed between recaptures. Previous laboratory studies constrained to confined space and artificial environments have already suggested site fidelity and our results from a natural population confirm this. The low reproductive activity of the species together with the reported extreme site fidelity makes this top predator of aquatic cave communities highly vulnerable and a sensitive bio‐indicator of habitat‐changing human activities.
This is the first capture–mark–recapture study on a natural population of the obligate aquatic cave‐dwelling olm (Proteus anguinus), known for its extreme life‐history characteristics (lifespan over 100 years, reproductive cycle over 10 years). We observed extreme site fidelity; individuals moved typically less than 15 m over years.
Vitellogenin (Vtg) is a precursor protein of egg yolk proteins in oviparous and ovoviviparous vertebrates. Except in a case of exposure to estrogenic endocrine disruptors, Vtg is a female-specific ...protein and could be used as a molecular marker for sex identification. This would be especially useful in the case of the endangered European cave salamander Proteus anguinus in which sexes are indistinguishable according to external morphology, which hinders the establishment of a successful captive breeding program. Here we describe the identification, partial characterization, and purification of Vtg from P. anguinus. Vtg was identified in the plasma of a vitellogenic proteus female with visible oocytes. The identification of this protein was accomplished by mass spectrometry analysis. Two-dimensional gel electrophoresis revealed proteus Vtg as a mix of 190 kDa isoforms with isoelectric points in the pH range 5.3–6.0. Vtg was purified from proteus blood by gel filtration followed by anion-exchange chromatography. Using specific staining of SDS-PAGE gels, the Vtg was found to be phosphorylated and lipidated. Unlike the case in some other aquatic vertebrates, in P. anguinus, Vtg was not present in detectable amounts in cutaneous mucus. Degradation of oocytes in the captive vitellogenic female was accompanied by simultaneous decrease of Vtg concentration. Over a period of 10 months, the concentration of Vtg dropped from maximal to sub-detectable. Our results show that Vtg is a promising molecular marker for sex identification and ovary maturation in P. anguinus, which could contribute to the development of a viable program for captive reproduction of this unique species.
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•In Proteus anguinus, sex detemination is indistinguishable by outer morphology, sex -specific loci or sex chromosomes.•Vitellogenin, a female-specific protein, was identified in the plasma of Proteus anguinus.•Vitellogenin is a 190 kDa phospholipoprotein with an isoelectric point of 5.3–6.0.•In a captive female, the concentration of vitellogenin dropped from maximum to sub-detectable over a period of 10 months.
Emerging infections add to existing threats to the survival of amphibians worldwide. The olm (Proteus anguinus) is a vulnerable, troglobiont urodele species with a small European range and restricted ...to underground karstic systems. Population declines to emerging threats like the chytrid fungus Batrachochytrium salamandrivorans, are likely to go unnoticed due to inaccessibility of the species' habitat. We here studied the interaction between olms and B. salamandrivorans. Experimental inoculation of olms resulted in low-level, asymptomatic but persistent infections, with limbs as predilection sites. The lack of exponential fungal growth in the olms' epidermis correlated with limited fungal proliferation and dampened virulence gene expression after exposure to olm skin compounds. The olm is one of few western Palearctic urodeles that is tolerant to B. salamandrivorans infection and may act as a subterranean disease reservoir, yet costs of subclinical infection may compromise olm fitness on the long term.
Theories of extreme lifespan evolution in vertebrates commonly implicate large size and predator-free environments together with physiological characteristics like low metabolism and high protection ...against oxidative damages. Here, we show that the ‘human fish’ (olm, Proteus anguinus), a small cave salamander (weighing 15–20 g), has evolved an extreme life-history strategy with a predicted maximum lifespan of over 100 years, an adult average lifespan of 68.5 years, an age at sexual maturity of 15.6 years and lays, on average, 35 eggs every 12.5 years. Surprisingly, neither its basal metabolism nor antioxidant activities explain why this animal sits as an outlier in the amphibian size/longevity relationship. This species thus raises questions regarding ageing processes and constitutes a promising model for discovering mechanisms preventing senescence in vertebrates.
The European cave salamander Proteus anguinus is a charismatic amphibian endemic to the concealed and inaccessible subterranean waters of the Dinaric Karst. Despite its exceptional conservation ...importance not much is known about its ecology and interactions with the groundwater microbiome. The cutaneous microbiota of amphibians is an important driver of metabolic capabilities and immunity, and thus a key factor in their wellbeing and survival. We used high-throughput 16S rRNA gene sequencing based on seven variable regions to examine the bacteriome of the skin of five distinct evolutionary lineages of P. anguinus and in their groundwater environment. The skin bacteriomes turned out to be strongly filtered subsamples of the environmental microbial community. The resident microbiota of the analyzed individuals was dominated by five bacterial taxa. Despite an indicated functional redundancy, the cutaneous bacteriome of P. anguinus presumably provides protection against invading microbes by occupying the niche, and thus could serve as an indicator of health status. Besides conservation implications for P. anguinus, our results provide a baseline for future studies on other endangered neotenic salamanders.
In situ ecological studies on obligate cave-dwelling aquatic animals are scarce at best. This is particularly true for capture-mark-recapture (CMR) studies that form the basis of understanding ...population structure and dynamics. Here, we report on the in situ underwater application of the Visible Implant Elastomer (VIE) tagging system on the olm, Proteus anguinus, an obligate cave-dwelling aquatic amphibian. We tagged seven adult individuals and monitored the population during 31 dive transects during four years. We found that VIE tagging is applicable underwater. Based on our recaptures, the tags were recognisable after four years and the recaptured three individuals exhibited extreme site-fidelity. Our results indicate that CMR studies are feasible in underwater cave ecosystems without even temporarily removing individuals. In situ underwater tagging also holds great potential for studies of other aquatic ecosystems, where removing animals from water, their habitat or territory is problematic for ethical, logistic or scientific reasons.
Acanthocephalus balkanicus Batchvarov et Combes, 1974 was incompletely described from the northern crested newt, Triturus cristatus (Laurenti) (Amphibia: Salamandridae), a possible synonym of the ...Balkan crested newt, Triturus ivanbureschi Arntzen et Wielstra, from a pond in village of Pesnopoy, southern Bulgaria. We provide a full description of adult males and females of the same taxon from the olm, Proteus anguinus Laurenti (Amphibia: Proteidae), the only exclusively aquatic cave-dwelling vertebrate in Europe, captured in Postojna-Planina Cave System in Slovenia. Cystacanths were also collected from the cave ecomorph of Asellus aquaticus (Linnaeus) (Crustacea: Asellidae) in the same location. Molecular analysis of specimens from Slovenia revealed that they are genetically almost identical to those of Acanthocephalus anguillae (Müller, 1780), a common parasite of European freshwater fishes. We propose to recognise the morphological and host differences by describing A. balkanicus as a new subspecies of A. anguillae. Acanthocephalus anguillae balkanicus is rather small and cylindrical with cylindrical proboscis having 10 rows of 6 hooks with simple roots each, long neck, large balloon-shaped lemnisci, small spherical anterior testis, and 6 club-shaped cement glands in 3 pairs. SEM images reveal more morphological details and the X-ray scans of gallium cut hooks shows considerably higher levels of phosphorus and calcium in adult hooks than in cystacanth hooks, especially in basal areas. Sulfur levels were higher in the arch and basal area of cystacanth hooks than adult hooks. Considering that both definitive and intermediate hosts of the Slovenian population of this acanthocephalan are bound to cave life, it is possible that its entire life cycle is uniquely completed underground.
Abstract
Background
Lightless caves can harbour a wide range of living organisms. Cave animals have evolved a set of morphological, physiological, and behavioural adaptations known as ...troglomorphisms, enabling their survival in the perpetual darkness, narrow temperature and humidity ranges, and nutrient scarcity of the subterranean environment. In this study, we focused on adaptations of skull shape and sensory systems in the blind cave salamander, Proteus anguinus, also known as olm or simply proteus—the largest cave tetrapod and the only European amphibian living exclusively in subterranean environments. This extraordinary amphibian compensates for the loss of sight by enhanced non-visual sensory systems including mechanoreceptors, electroreceptors, and chemoreceptors. We compared developmental stages of P. anguinus with Ambystoma mexicanum, also known as axolotl, to make an exemplary comparison between cave- and surface-dwelling paedomorphic salamanders.
Findings
We used contrast-enhanced X-ray computed microtomography for the 3D segmentation of the soft tissues in the head of P. anguinus and A. mexicanum. Sensory organs were visualized to elucidate how the animal is adapted to living in complete darkness. X-ray microCT datasets were provided along with 3D models for larval, juvenile, and adult specimens, showing the cartilage of the chondrocranium and the position, shape, and size of the brain, eyes, and olfactory epithelium.
Conclusions
P. anguinus still keeps some of its secrets. Our high-resolution X-ray microCT scans together with 3D models of the anatomical structures in the head may help to elucidate the nature and origin of the mechanisms behind its adaptations to the subterranean environment, which led to a series of troglomorphisms.
The European cave salamander, Proteus anguinus, or proteus, is the largest obligate cave animal in the world. It is an endangered and charismatic species of high conservation importance for ...subterranean waters. Conservation genetic studies are hampered by the extreme size and repetitiveness of its nuclear genome.
The aim of the study was to develop and characterize the first microsatellite markers for proteus, and test their informativeness at the level of individuals, populations and between populations in the Postojna and Planina caves in Slovenia.
Twenty‐three novel polymorphic microsatellite markers were amplified in 201 individuals from both caves using three multiplex reactions. The number of alleles per locus varied from three to nine. The loci are largely unlinked and conform to Hardy–Weinberg genotype frequencies. Genetic equilibrium and an FST value of 0.0024 suggest a nearly panmictic population in both caves separated by some 10 km of subterranean river course, while Bayesian clustering detected weak genetic structure.
The microsatellites described fill the gap of urgently needed nuclear markers in Proteus that can be applied in genetic mark–recapture studies, population monitoring and identification of management units to assist conservation efforts.
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