Seed dormancy, climate changes, desertification and soil use transformation threaten the Mediterranean endemic monospecific plant Petagnaea gussonei.

This study investigated the germination capacity (endogenous factor) of Petagnaea gussonei (Spreng.) Rauschert, an endemic monospecific plant considered as a relict species of the ancient Mediterranean Tertiary flora. This investigation focused also on the temporal trends of soil-use, climate and desertification (exogenous factors) across the natural range of P. gussonei. The final germination percentage showed low values between 14 and 32%, the latter obtained with GA3 and agar at 10 °C. The rising temperatures in the study area will further increase the dormancy of P. gussonei, whose germination capacity was lower and slower at temperatures higher than 10 °C. A further limiting factor of P. gussonei is its dormancy, which seems to be morpho-physiological. Regarding climate trends, in the period 1931-2020, the average temperature increased by 0.5 °C, from 15.4 to 15.9 °C, in line with the projected climate changes throughout the twenty-first century across the Mediterranean region. The average annual rainfall showed a relatively constant value of c. 900 mm, but extreme events grew considerably in the period 1991-2020. Similarly, the land affected by desertification expanded in an alarming way, by increasing from 21.2% in 2000 to 47.3% in 2020. Soil-use changes created also a complex impacting mosaic where c. 40% are agricultural areas. The effective conservation of P. gussonei should be multilateral by relying on germplasm banks, improving landscape connectivity and vegetation cover, and promoting climate policies.

A new species of terrestrial toad of the Rhinellafestae group (Anura, Bufonidae) from the highlands of the Central Cordillera of the Andes of Colombia.

The genus Rhinella (Bufonidae) comprises 92 species of Neotropical toads. In Colombia, Rhinella is represented by 22 recognized species, of which nine belong to the Rhinellafestae group. Over the past decade, there has been increasing evidence of cryptic diversity within this group, particularly in the context of Andean forms. Specimens of Rhinella collected in high Andean forests on both slopes of the Central Cordillera in Colombia belong to an undescribed species, Rhinellakumandaysp. nov. Genetic analyses using the mitochondrial 16S rRNA gene indicated that the individuals belong to the festae species group. However, they can be distinguished from other closely related species such as Rhinellaparaguas and Rhinellatenrec by a combination of morphological traits including the presence of tarsal fold, a moderate body size, and substantial genetic divergence in the 16S rRNA gene (> 5%). Through this integrative approach, the specimens from the Central Cordillera of Colombia are considered an evolutionary divergent lineage that is sister to R.paraguas, and described as a new species. Rhinellakumandaysp. nov. is restricted to the Central Cordillera of Colombia inhabiting both slopes in the departments of Caldas and Tolima, in an elevational range between 2420 and 3758 m. With the recognition of this new species, the genus Rhinella now comprises 93 species with 23 of them found in Colombia, and ten species endemic to the country.

Indigoferajintongpenensis (Fabaceae, Papilionoideae, Indigofereae), a new species from Yunnan, southwest China.

Indigoferajintongpenensis, a new species of the subfamily Papilionoideae of Fabaceae, is described and illustrated from Yunnan, southwest China. The new species is characterised by having a prostrate habit, flexible stems and branches, as well as spreading, sub-basifixed, asymmetrically 2-branched trichomes covering the entire plant, discoid calyx, and racemose inflorescences 6-8-flowered, short, 1-2 (-3.5) cm in length, apparently shorter than the leaf. A distribution map and comparison of morphological diagnostic characters with its morphologically similar species are provided. Additionally, a preliminary conservation assessment of I.jintongpenensis is proposed following IUCN criteria.

Contributions of a small collection of terrestrial microsnails (Pupilloidea, Hypselostomatidae) from Myanmar with description of three new species.

Land snails were collected for the project 'Conserving Myanmar's Karst Biodiversity' from the limestone karsts in Mon, Kayin, and Shan states and in the regions of Tanintharyi and Mandalay between 2015 and 2017, through cooperation with Fauna and Flora International (FFI) and the Forestry Department of Myanmar. Here, we report on a portion of the collection, and list 17 species from seven genera of the Hypselostomatidae microsnails. Three new species from two genera are described as Bensonellataiyaiorum Tongkerd & Panha, sp. nov., B.lophiodera Tongkerd & Panha, sp. nov., and Gyliotrachelaaunglini Tongkerd & Panha, sp. nov. All new species are known only from the type locality in Shan State (Bensonella) and Kayin State (Gyliotrachela). A new combination of Acinolaemusdayanum and three newly recorded species, namely A.cryptidentatus, B.anguloobtusa and G.hungerfordiana are discussed. The low morphological variability of the widely distributed G.hungerfordiana is discussed, and two species are proposed for formal synonymisation. Constituting the first records for Myanmar, five species of Bensonella and two species of Acinolaemus were collected.

Spatial heterogeneity of neo- and paleo-endemism for plants in Madagascar.

Madagascar is a biogeographically unique island with a remarkably high level of endemism. However, endemic taxa in Madagascar are massively threatened due to unprecedented pressures from anthropogenic habitat modification and climate change. A comprehensive phylogeny-based biodiversity evaluation of the island remains lacking. Here, we identify hotspots of taxonomic and phylogenetic plant diversity and neo- and paleo-endemism by generating a novel dated tree of life for the island. The tree is based on unprecedented sampling of 3,950 species (33% of the total known species) and 1,621 genera (93% of the total known genera and 69% of endemic genera) of Malagasy vascular plants. We find that island-endemic genera are concentrated in multiple lineages combining high taxonomic and phylogenetic diversity. Integrating phylogenetic and geographic distribution data, our results reveal that taxon richness and endemism are concentrated in the northern, eastern, and southeastern humid forests. Paleo-endemism centers are concentrated in humid eastern and central regions, whereas neo-endemism centers are concentrated in the dry and spiny forests in western and southern Madagascar. Our statistical analysis of endemic genera in each vegetation region supports a higher proportion of ancient endemic genera in the east but a higher proportion of recent endemic genera in the south and west. Overlaying centers of phylogenetic endemism with protected areas, we identify conservation gaps concentrated in western and southern Madagascar. These gaps should be incorporated into conservation strategies to aid the protection of multiple facets of biodiversity and their benefits to the Malagasy people.

Gentianamopanshanensis (Gentianaceae), a new species from Yunnan, southwest China.

Gentianamopanshanensis, a new species of the family Gentianaceae is here described and illustrated. This species is presently known only from the Mopanshan Mountain, Yunnan Province, southwest China. Phylogenetic analysis based on ITS sequence data has shown that this new species is a member of the series Fimbriatae of the section Chondrophyllae. Morphologically, it mostly resembles G.mairei and G.panthaica, but differs clearly from the latter two species in the shape and size of the leaves, and the characters of the corolla throat and plicae.

A new characid species with remarkable sexual dimorphism (Characiformes: Characidae: Stevardiinae) from the upper Guayabero River, Orinoco basin, Colombia.

A new species of characid with remarkable sexual characteristics is described from the upper Guayabero River drainage from the Orinoco basin in Colombia. The new species is included in the genus Monotocheirodon by sharing most of the previously proposed diagnostic features of this genus. It differs from all Stevardiinae by the combination, in adult males, of an enlarged urogenital papilla in contact with the first anal-fin unbranched ray and a highly modified anal fin with enlarged and distally elongated first and second branched anal-fin rays, forming a gonopodium-like structure. In addition, it differs from congeners by the presence of an adipose fin, an incomplete lateral line, an ascending process of the premaxilla dorsally oriented, and a long snout. The new species was discovered from a poorly sampled region in Colombia and is an unexpected new record given its disjunct geographic distribution from other species of the genus. Monotocheirodon species were previously known from piedmont drainages in Bolivia and Peru. The conservation status of the new species is herein categorized following IUCN criteria.

Revised checklist of endemic vascular plants of Kazakhstan.

We compiled a checklist of endemic vascular plants occurring in Kazakhstan, employing an exhaustive examination of literature sources, herbarium collections, databases and field observations. Our study reveals that 451 taxa can be considered endemic to Kazakhstan, constituting 7.97% of the total vascular plant diversity in the country. These endemic taxa, originating from 139 genera and 34 families, predominantly thrive in the southern regions of Kazakhstan, specifically in the mountain ridges of the Kazakh part of the Tian Shan, including Karatau (123 taxa), Dzungarian Alatau (80 taxa) and Trans-Ili and Kungey Alatau (50 taxa). Notably, 107 endemic species are granted legal protection. Detailed information regarding life form, life cycle, conservation status and geographical distribution across floristic regions was meticulously compiled for each endemic taxon. Of the six groups of life forms, herbs include the highest part of endemic taxa (367 taxa), followed by dwarf semishrubs (25 taxa), shrubs (23 taxa), subshrubs (20 taxa), undershrubs (13 taxa) and trees (3 taxa). The observed life cycles are perennials (408 taxa), annuals (33 taxa) and biennials (10 taxa). This paper serves as a fundamental groundwork for prospective investigations aimed at assessing population sizes and hotspots of plant endemism throughout Kazakhstan, crucial for determining conservation status of endemic plants.

Considerable genetic diversity and structure despite narrow endemism and limited ecological specialization in the Hayden's ringlet, Coenonympha haydenii.

Understanding the processes that underlie the development of population genetic structure is central to the study of evolution. Patterns of genetic structure, in turn, can reveal signatures of isolation by distance (IBD), barriers to gene flow, or even the genesis of speciation. However, it is unclear how severe range restriction might impact the processes that dominate the development of genetic structure. In narrow endemic species, is population structure likely to be adaptive in nature, or rather the result of genetic drift? In this study, we investigated patterns of genetic diversity and structure in the narrow endemic Hayden's ringlet butterfly. Specifically, we asked to what degree genetic structure in the Hayden's ringlet can be explained by IBD, isolation by resistance (IBR) (in the form of geographic or ecological barriers to migration between populations), and isolation by environment (in the form of differences in host plant availability and preference). We employed a genotyping-by-sequencing (GBS) approach coupled with host preference assays, Bayesian modelling, and population genomic analyses to answer these questions. Our results suggest that despite their restricted range, levels of genetic diversity in the Hayden's ringlet are comparable to those seen in more widespread butterfly species. Hayden's ringlets showed a strong preference for feeding on grasses relative to sedges, but neither larval preference nor potential host availability at sampling sites correlated with genetic structure. We conclude that geography, in the form of IBR and simple IBD, was the major driver of contemporary patterns of differentiation in this narrow endemic species.

Choice of molecular marker influences spatial patterns of phylogenetic diversity.

Spatial patterns of phylogenetic diversity (PD) are increasingly becoming relevant for conservation decisions. PD measures are based on phylogenies estimated from molecular data. This paper addresses the question of how different molecular markers impact PD spatial patterns. We first conducted a simple simulation to explore the effect of deep and shallow changes in topology (simulating variations in molecular markers), using ultrametric and non-ultrametric trees, and then used a dataset of Chilean flora with four sets of markers to assess potential differences in spatial patterns of PD ranks using different markers and types of trees. The simulation consistently showed that the difference in PD rank was lower for ultrametric trees than for phylograms. A similar trend was observed using the Chilean flora dataset, with among-markers variability in spatial patterns of the PD metrics lower for ultrametric than for non-ultrametric trees, depicted as top 2.5 and 5% hotspots. Frequency distribution of PD values differed among markers as well, with this variation less apparent for ultrametric trees. We conclude that the choice of markers impacts spatial patterns of PD, and these results vary more strongly for phylograms, suggesting that ultrametric trees are more robust to the choice of marker.

Halichoeres sanchezi n. sp., a new wrasse from the Revillagigedo Archipelago of Mexico, tropical eastern Pacific Ocean (Teleostei: Labridae).

A new labrid fish species, Halichoeres sanchezi n. sp., is described from eight specimens collected in the Revillagigedo Archipelago in the tropical eastern Pacific Ocean, off the coast of Mexico. The new species belongs to the Halichoeres melanotis species complex that is found throughout the region, differing by 2.4% in the mtDNA cytochrome c oxidase I sequence from its nearest relative, H. melanotis from Panama, and 2.9% from Halichoeres salmofasciatus from Cocos Island, off Costa Rica. The complex is distinguished from others in the region by having a black spot on the opercular flap and a prominent black area on the caudal fin of males. The juveniles and initial phase of the new species closely resemble those of H. salmofasciatus and Halichoeres malpelo from Malpelo Island of Colombia, differing in having an oblong black spot with a yellow dorsal margin on the mid-dorsal fin of initial-phase adults as well as on juveniles. In contrast, the terminal-phase male color pattern is distinct from other relatives, being vermilion to orangish brown with dark scale outlines, a white patch on the upper abdomen, and a prominent black band covering the posterior caudal peduncle and base of the caudal fin. The new species adds to the list of endemic fish species for the isolated archipelago and is an interesting case of island endemism in the region. The discovery was made during the joint 2022 collecting expedition to the archipelago, which featured a pioneering collaborative approach to an inventory of an island ichthyofauna, specifically including expert underwater photographers systematically documenting specimens in situ, before hand-collection, and then photographed fresh, tissue-sampled, and subsequently vouchered in museum collections.

Monitoring data on the effect of domestic livestock and rabbits on Androcymbiumeuropaeum (Lange) K. Richt. and its xerophytiques pastures for thirteen years.

Background: Dataset of annual monitoring of herbivory effects on the conservation status of the endangered species Androcymbiumeuropaeum (Lange) K. Richt and its associated plant communities is presented in this manuscript. This dataset encompasses the annual monitoring of herbivory effects on the conservation status of the endangered species Androcymbiumeuropaeum. Since 2010, the SERPAM Department (Service of Evaluation, Restoration and Protection of Mediterranean Agrosystems) at the Zaidin Experimental Station, belonging to the Spanish National Research Council (CSIC-EEZ), has conducted annual sampling to assess the impact of both domestic and wild livestock, specifically rabbits, on the pastures where A.europaeum lives. The study consisted of a randomised block design, implementing three distinct treatments to evaluate different management strategies: (1) rabbit and domestic herbivory, (2) exclusion of domestic livestock and (3) exclusion of rabbits and domestic livestock. Within each treatment, two types of monitoring were conducted. Firstly, the abundance of A.europaeum was estimated by counting individuals within 50 cm x 50 cm quadrats. Secondly, plant species diversity was assessed along 2-m long transects using the modified Point-Quadrat method. The research was conducted within the Cabo de Gata-Níjar Natural Park in southern Spain, specifically in the Amoladeras Nature Reserve in Almería. New information: The dataset contains information spanning from 2010 to 2023, providing valuable insights into the annual monitoring of herbivory effects on the conservation status of A.europaeum, contributing to our understanding of the species' interaction with domestic and wild animal in the studied area.

Checklist of the genus Ridsdalea (Rubiaceae, Gardenieae) in Vietnam with description of the new species R.backanensis.

The paper provides an identification key and an annotated list of all six species of the genus Ridsdalea J.T.Pereira & K.M.Wong (Rubiaceae, Gardenieae) recorded in the flora of Vietnam, along with data on their valid names, synonyms, types, and distribution. A new species, R.backanensis, discovered in the limestone area of Bac Kan Province (northern Vietnam), is described and illustrated. Detailed data on its characters, ecology, distribution, phenology, preliminary IUCN conservation status, and taxonomical notes are given. The newly discovered species is potentially interesting for cultivation as an ornamental tree that may be effectively used in garden and urban green architecture.

Seed germination and vegetative and in vitro propagation of Hieracium lucidum subsp. lucidum (Asteraceae), a critically endangered endemic taxon of the Sicilian flora.

Hieracium lucidum subsp. lucidum is a critically endangered endemic taxa of the Sicilian flora. It is a relict of the Tertiary period surviving on the cliffs of Monte Gallo (NW-Sicily). This research focused on finding the best protocols for seed germination and vegetative and in vitro propagation to contribute to ex situ conservation. Seed germination tests were carried out using constant temperatures of 15 °C, 20 °C and 25 °C in continuous darkness and an alternating temperature of 30/15 °C (16 h/8 h, light/dark). The seeds had no dormancy, and a high germination capacity (70-95%) was obtained at all tested thermoperiods. The possibility of vegetative propagation of the taxon was evaluated through the rooting capacity of stem cuttings treated or not treated with indole-3-butyric acid (IBA). All cuttings were treated with IBA rooted within 2 months, while only 50% of the untreated cuttings were rooted within a longer time. An efficient protocol for rapid in vitro propagation from leaf portions was developed. The response of explants was tested on hormone-free Murashige and Skoog (MS) basal medium and MS enriched with different types of cytokinins: 6-Benzylaminopurine (BAP) and meta-Topolin (mT) in combination with naphthaleneacetic acid (NAA) and 2,4-Dichlorophenoxyacetic acid (2,4-D) at the same concentration. The combination of mT (2 mg L-1) and 2,4-D (1 mg L-1) in the medium was the most effective and showed the highest percentage of callus induction and the mean number of regenerated shoots. The maximum rate of root regeneration and the maximum number and length of roots were obtained on hormone-free MS and MS enriched with IBA at concentrations of 1 mg L-1. From the results obtained, it can be concluded that H. lucidum subsp. lucidum can be successfully propagated using one of the tested techniques, subject to the availability of the material for reproduction.

Pseudoscorpions (Arachnida, Pseudoscorpiones) from French Polynesia with first species records and description of new species.

A new species Olpiumcaputisp. nov. from Tahiti is described here based on external characters. This is the first record of the family Olpiidae Banks, 1895 from French Polynesia. Additionally, the genus Paratemnoides Harvey, 1991 is recorded from French Polynesia for the first time with the full description of new-found specimens of Paratemnoidesassimilis (Beier, 1932). New localities of Geogarypuslongidigitatus (Rainbow, 1897) are added. An identification key to pseudoscorpions of French Polynesia is provided.

Isodonxiaoluzhiensis (Lamiaceae, Nepetoideae), a new species from Yunnan, southwest China.

Isodonxiaoluzhiensis, a new species of the tribe Ocimeae in family Lamiaceae, is described and illustrated. The new species is known only from the type locality, Xiaoluzhi village in Luzhijang dry-hot valley of Yimen County, central Yunnan, southwest China. It is characterized by having a procumbent habit, gracile stems and branches, relatively small leaves and flowers, and the phenology of flowering in winter. The morphological comparisons with its putative closest relatives (I.adenanthus and I.hsiwenii) are also presented.

Jump dispersal drives the relationship between micro- and macroevolutionary dynamics in the Sicydiinae (Gobiiformes: Oxudercidae) of Sundaland and Wallacea.

Insular biodiversity hotspots of Southeast Asia are remarkable for their biodiverse faunas. With a marine larval phase lasting up to several months, the freshwater fish subfamily Sicydiinae has colonized most islands of these hotspots. However, Sicydiinae diversity is still poorly understood in Southeast Asia. With the objective to estimate intraspecific genetic diversity and infer past demography, we conducted the molecular inventory of Sicydiinae species in Sundaland and Wallacea using 652 bp of the mitochondrial cytochrome oxidase I gene, species delimitation methods and Bayesian Skyline plot reconstructions. In total, 24 Molecular Operational Taxonomic Units are delimited among the 603 sequences belonging to 27 species and five genera. Two cases of discordance between morphology and mitochondrial sequence are observed suggesting ongoing speciation and/or introgression in two genera. Multiple new occurrences are reported, either for a single biodiversity hotspot or both, some of which corresponding to observations of a few individuals far from the range distribution of their conspecifics. Among the ten species or species group whose intraspecific diversity was examined, high levels of genetic diversity and past population expansion are revealed by Tajima's D tests and Bayesian Skyline Plot reconstructions. Together these results indicate that long-distance dispersal is common and suggest that most endemic species originated through founder events followed by population expansion. Patterns of sexual dimorphism and males' coloration among diverging species pair seem to point to sexual selection as an important mechanism contributing to speciation in the Sicydiinae of Sundaland and Wallacea.

Patterns of Grewia (Malvaceae) diversity across geographic scales in Africa and Madagascar.

BACKGROUND AND AIMS: Quantifying spatial species richness is useful to describe biodiversity patterns across broad geographic areas, especially in large, poorly known plant groups. We explore patterns and predictors of species richness across Africa in one such group; the paleotropical genus Grewia L. (Malvaceae). METHODS: Grewia species richness was quantified by extracting herbarium records from GBIF and Tropicos and creating geographic grids at varying spatial scales. We assessed predictors of species richness using spatial regression models with 30 environmental variables. We explored species co-occurrence in Madagascar at finer resolutions using Schoener's index, and compared species' range sizes and IUCN status among ecoregions. Lastly, we derived a trait matrix for a subset of species found in Madagascar to characterize morphological diversity across space. KEY RESULTS: Grewia species occur in 50 countries in Africa, with the highest number of species in Madagascar (93, with 80 species endemic). Species richness is highest in Madagascar, with up to 23 Grewia species in a grid cell, followed by coastal Tanzania/Kenya (up to 13 species), and northern South Africa and central Angola (11 species each). Across Africa, higher species richness was predicted by variables related to aridity. In Madagascar, a greater range in environmental variables best predicted species richness, consistent with geographic grid cells of highest species richness occurring near biome/ecoregion transitions. In Madagascar we also observe increasing dissimilarity in species composition with increasing geographic distance. CONCLUSIONS: The spatial patterns and underlying environmental predictors that we uncover in Grewia represent an important step in our understanding of plant distribution and diversity patterns across Africa. Madagascar boasts nearly twice the Grewia species richness, compared to the second most species-rich country in Africa, which might be explained by complex topography and environmental conditions across small spatial scales.

The distribution and conservation of areas with microendemic species in a biodiversity hotspot: a multi-taxa approach.

Background: Microendemic species are species with very small geographic distributions (ranges). Their presence delimitates areas with microendemic species (AMs), denoting a spatial unit comprising at least one population of at least one microendemic species. AMs are assumed to be distributed distinctively and associated with specific ecological, historical, and anthropogenic attributes. However, the level of influence of these factors remains unclear. Thus, we studied the distribution patterns of microendemic species within the Brazilian Atlantic Forest to (a) identify the region's AMs; (b) evaluate whether ecological (latitude, altitude, distance from the coastline), historical (climate stability), and anthropogenic (ecological integrity) attributes distinguish AMs from non-AMs; and (c) assess the conservation status of the Atlantic Forest's AMs. Methods: We mapped the ranges of 1,362 microendemic species of angiosperms, freshwater fishes, and terrestrial vertebrates (snakes, passerine birds, and small mammals) to identify the region's AMs. Further, spatial autoregressive logit regression models were used to evaluate whether latitude, altitude, distance from the coastline, Climate Stability Index, and ecological integrity can be used to discern AMs from non-AMs. Moreover, the AMs' conservation status was assessed by evaluating the region's ecological integrity and conservation efforts (measured as the proportion of AMs in protected areas). Results: We identified 261 AMs for angiosperm, 205 AMs for freshwater fishes, and 102 AMs for terrestrial vertebrates in the Brazilian Atlantic Forest, totaling 474 AMs covering 23.8% of the region. The Brazilian Atlantic Forest is a large and complex biogeographic mosaic where AMs represent islands or archipelagoes surrounded by transition areas with no microendemic species. All local attributes help to distinguish AMs from non-AMs, but their impacts vary across taxonomic groups. Around 69% of AMs have low ecological integrity and poor conservation efforts, indicating that most microendemic species are under threat. This study provides insights into the biogeography of one of the most important global biodiversity hotspots, creating a foundation for comparative studies using other tropical forest regions.

Genetic approaches reveal a healthy population and an unexpectedly recent origin for an isolated desert spring fish.

Foskett Spring in Oregon's desert harbors a historically threatened population of Western Speckled Dace (Rhinichthys klamathensis). Though recently delisted, the dace's recruitment depends upon regular removal of encroaching vegetation. Previous studies assumed that Foskett Dace separated from others in the Warner Valley about 10,000 years ago, thereby framing an enigma about the population's surprising ability to persist for so long in a tiny habitat easily overrun by plants. To investigate that persistence and the effectiveness of interventions to augment population size, we assessed genetic diversity among daces inhabiting Foskett Spring, a refuge at Dace Spring, and three nearby streams. Analysis revealed a robust effective population size (Ne) of nearly 5000 within Foskett Spring, though Ne in the Dace Spring refuge is just 10% of that value. Heterozygosity is slightly lower than expected based on random mating at all five sites, indicating mild inbreeding, but not at a level of concern. These results confirm the genetic health of Foskett Dace. Unexpectedly, genetic differentiation reveals closer similarity between Foskett Dace and a newly discovered population from Nevada's Coleman Creek than between Foskett Dace and dace elsewhere in Oregon. Demographic modeling inferred Coleman Creek as the ancestral source of Foskett Dace fewer than 1000 years ago, much more recently than previously suspected and possibly coincident with the arrival of large herbivores whose grazing may have maintained open water suitable for reproduction. These results solve the enigma of persistence by greatly shortening the duration over which Foskett Dace have inhabited their isolated spring.