Food preference and food type innovation of surface- vs. cave-dwelling waterlouse (Asellus aquaticus) after 60 000 years of isolation

Behavioural innovativeness is important for colonising new habitats; however, it is also costly. Along the colonisation event of a simple, stable and isolated habitat offering only new food sources, one could hypothesize that the colonising individuals are more innovative than the average in their source population, showing preference to the new resource, while after colonisation, the adapted population will lose its innovativeness and become specialised to the new resource. To test this hypothesis, we compared food preference and food type innovation of a cave-dwelling waterlouse (Asellus aquaticus) population (genetically isolated for at least 60 000 years) to three surface-dwelling populations, also sampling individuals that have recently entered the cave (‘colonists’). In the cave, the only food sources are endogenous bacterial mats, while surface populations feed on various living and dead plant material together with their fungal and bacterial overgrow. We assayed all populations with the familiar and unfamiliar food types from the natural habitats and two novel food types not occurring in the natural habitats of the species. We found that all populations preferred surface to cave food and consumed the unnatural novel food types. Surface populations avoided cave food and colonists spent the most time with feeding on surface food. We conclude that the cave population maintained its preference for surface food and did not lose its food type innovativeness. We suggest that adapting to the special cave food was a major challenge in colonising the cave. Significance statement Behavioural innovativeness is a key trait for adapting to environmental changes or to colonise new habitats. However, it has developmental and maintenance costs due to the high energy need of the necessary sensory and neural organs. Therefore, we asked whether behavioural innovativeness decreases after colonising an isolated, stable and highly specialised habitat. By comparing food type innovativeness of surface-dwelling populations of waterlouse (Asellus aquaticus) to a population that has colonised a cave at least 60 000 years ago, we found that the high innovativeness towards unnatural food was retained in the cave population. Further, all populations preferred surface food (decaying leaves), with surface populations almost completely avoiding cave food (endogenous bacteria mats). We suggest that (i) food type innovativeness is evolutionary rigid in our system and (ii) the cave food was rather an obstacle against than a trigger of cave colonisation.

The Pennsylvania grotto sculpin: population genetics

The Pennsylvania grotto sculpin is known from just two caves of the Nippenose Valley in central Pennsylvania, USA. They exhibit emergent troglobitic morphological traits and are the second northern-most cave adapted fish in the world. Two mitochondrial (16S rRNA and D-loop gene) and one nuclear (S7 ribosomal protein gene intron) gene in both cave and epigean populations were sequenced. For the three markers, a large proportion of cave specimens possess unique haplotypes not found in their local surface counterparts, suggesting a vicariance in their evolutionary history. The cave population also has haplotypes from two separate lineages of surface sculpins of the Cottus cognatus/bairdii species complex. Since morphology, nuclear, and mitochondrial markers are not correlated among cave individuals, hybridization with introgression is suggested.

Contrasting Gene Decay in Subterranean Vertebrates: Insights from Cavefishes and Fossorial Mammals

Evolution sometimes proceeds by loss, especially when structures and genes become dispensable after an environmental shift relaxes functional constraints. Subterranean vertebrates are outstanding models to analyze this process, and gene decay can serve as a readout. We sought to understand some general principles on the extent and tempo of the decay of genes involved in vision, circadian clock, and pigmentation in cavefishes. The analysis of the genomes of two Cuban species belonging to the genus Lucifuga provided evidence for the largest loss of eye-specific genes and nonvisual opsin genes reported so far in cavefishes. Comparisons with a recently evolved cave population of Astyanax mexicanus and three species belonging to the Chinese tetraploid genus Sinocyclocheilus revealed the combined effects of the level of eye regression, time, and genome ploidy on eye-specific gene pseudogenization. The limited extent of gene decay in all these cavefishes and the very small number of loss-of-function mutations per pseudogene suggest that their eye degeneration may not be very ancient, ranging from early to late Pleistocene. This is in sharp contrast with the identification of several vision genes carrying many loss-of-function mutations in ancient fossorial mammals, further suggesting that blind fishes cannot thrive more than a few million years in cave ecosystems.

Contrasted gene decay in subterranean vertebrates: insights from cavefishes and fossorial mammals

Evolution sometimes proceeds by loss, especially when structures and genes become dispensable after an environmental shift relaxing functional constraints. Gene decay can serve as a read-out of this evolutionary process. Animals living in the dark are outstanding models, in particular cavefishes as hundreds of species evolved independently during very different periods of time in absence of light. Here, we sought to understand some general principals on the extent and tempo of decay of several gene sets in cavefishes. The analysis of the genomes of two Cuban species belonging to the genus Lucifuga provides evidence for the most massive loss of eye genes reported so far in cavefishes. Comparisons with a recently-evolved cave population of Astyanax mexicanus and three species belonging to the tetraploid Chinese genus Sinocyclocheilus revealed the combined effects of the level of eye regression, time and genome ploidy on the number of eye pseudogenes. In sharp contrast, most circadian clock and pigmentation genes appeared under strong selection. In cavefishes for which complete genomes are available, the limited extent of eye gene decay and the very small number of loss of function (LoF) mutations per pseudogene suggest that eye degeneration is never very ancient, ranging from early to late Pleistocene. This is in sharp contrast with the identification of several eye pseudogenes carrying many LoF mutations in ancient fossorial mammals. Our analyses support the hypothesis that blind fishes cannot thrive more than a few millions of years in cave ecosystems.

Evidence for rapid phenotypic and behavioural shifts in a recently established cavefish population

Cave colonization offers a natural laboratory to study an extreme environmental shift, and diverse cave species from around the world often have converged on robust morphological, physiological and behavioural traits. The Mexican tetra (Astyanax mexicanus) has repeatedly colonized caves in the Sierra de El Abra and Sierra de Guatemala regions of north-east Mexico ~0.20–1 Mya, indicating an ability to adapt to the cave environment. The time frame for the evolution of these traits in any cave animal, however, is poorly understood. Astyanax mexicanus from the Río Grande in South Texas were brought to Central Texas beginning in the early 1900s and colonized underground environments. Here, we investigate whether phenotypic and behavioural differences have occurred rapidly between a surface population and a geographically proximate cave population, probably of recent origin. Fish from the cave and surface populations differ significantly in morphological traits, including coloration, lateral line expansion and dorsal fin placement. Striking behavioural shifts in aggression, feeding and wall-following have also occurred. Together, our results suggest that morphological and behavioural changes accompanying cave colonization can be established rapidly, and this system offers an exciting and unique opportunity for isolating the genetic and environmental contributions to colonization of extreme environments.

Evidence for rapid phenotypic and behavioral change in a recently established cavefish population

Substantial morphological and behavioral shifts often accompany rapid environmental change, yet, little is known about the early stages of cave colonization. Relative to surface streams, caves are extreme environments with perpetual darkness and low nutrient availability. The Mexican tetra (Astyanax mexicanus), has repeatedly colonized caves throughout Mexico, suggesting an ability to adapt to these conditions. Here, we survey for phenotypic and behavioral differences between a surface population and a cave population of A. mexicanus that has recently colonized Honey Creek Cave, Comal County, Texas, likely within the last century. We found that fish from Honey Creek Cave and fish from Honey Creek surface populations differ significantly in morphological traits including length, coloration, body condition, eye size, and dorsal fin placement. Cavefish also exhibit an increased number of superficial neuromasts relative to surface fish. Behaviorally, cavefish consume fewer worms when trials are performed in both lighted and darkened conditions. Cavefish are more aggressive than surface fish and exhibit fewer behaviors associated with stress. Further in contrast to surface fish, cavefish prefer the edges to the center of an arena and are qualitatively more likely to investigate a novel object placed in the tank. While cavefish and surface fish were wild-caught and developmental environment likely play a role in shaping these differences, our work demonstrates morphological and behavioral shifts for Texas cavefish and offers an exciting opportunity for future work to explore the genetic and environmental contributions to early cave colonization.

A re-description of Cyrtodactylus chrysopylos Bauer (Squamata: Gekkonidae) with comments on the adaptive significance of orange coloration in hatchlings and descriptions of two new species from eastern Myanmar (Burma)

The karstic foothills and hilly western edge of the Shan Plateau of eastern Myanmar continue to be sources of discovery for new species of the gekkonid genus Cyrtodactylus. Two new karst-associated species within this unique landscape from Mandalay Region—C. aunglini sp. nov. from the Kyauk Nagar Cave and C. myaleiktaung sp. nov. from Mya Leik Taung Mountain—are described on the basis of color pattern, morphology, and genetics. A molecular phylogeny of the C. gansi group (defined herein) based on 1481 base pairs of the mitochondrial gene ND2 and its flanking tRNAs place C. aunglini sp. nov. as the sister species to C. gansi and C. myaleiktaung sp. nov. as the sister species of C. chrysopylos. A new population of C. chrysopylos from a boulder-strewn, karstic ridge near the village of Yane, Shan State at an elevation 951 m is reported here. Genetic and geographic variation as well as differences in natural history between it and the lowland cave population of C. chrysopylos from the type locality in the Panlaung-Pyadalin Cave Wildlife Sanctuary, Shan State at 303 m are discussed. Hatchlings of the new population from Yane are nearly uniformly bright-orange in color. We provide evidence in support of a hypothesis claiming this color functions as a mechanism for crypsis during low levels of illumination and is not involved in intraspecific communication.

Evidence for Late Pleistocene origin of Astyanax mexicanus cavefish

BackgroundCavefish populations belonging to the Mexican tetra species Astyanax mexicanus are outstanding models to study the tempo and mode of adaptation to a radical environmental change. They share similar phenotypic changes such as blindness and depigmentation resulting from independent and convergent evolution. As such they allow examining whether their evolution involved the fixation of preexisting standing genetic variations and/or de novo mutations. Cavefish populations are currently assigned to two main groups, the so-called "old" and "new" lineages, which would have populated several caves independently and at different times. However, we do not have yet accurate estimations of the time frames of evolution of these populations.ResultsFirst, we reanalyzed the geographic distribution of mitochondrial and nuclear DNA polymorphisms and we found that these data do not support the existence of two cavefish lineages, neither the ancient origin of the “old” lineage. Using IMa2, a program based on a method that does not assume that populations are at mutation/migration/drift equilibrium and thus allows dating population divergence in addition to demographic parameters, we found that microsatellite polymorphism strongly supports a very recent origin of cave populations (i.e. less than 20,000 years). Second, we identified a large number of single-nucleotide polymorphisms (SNPs) in transcript sequences of pools of embryos (Pool-seq) belonging to the “old” Pachón cave population and a surface population from Texas. Pachón cave population has accumulated more neutral substitutions than the surface population and we showed that it could be another signature of its recent origin. Based on summary statistics that can be computed with this SNP data set together with simulations of evolution of SNP polymorphisms in two recently isolated populations, we looked for sets of demographic parameters that allow the computation of summary statistics with simulated populations that are similar to the ones with the sampled populations. In most simulations for which we could find a good fit between the summary statistics of observed and simulated data, the best fit occurred when the divergence between simulated populations was less than 30,000 years.ConclusionsAlthough it is often assumed that some cave populations such as Pachón cavefish have a very ancient origin, within the range of the late Miocene to the middle Pleistocene, a recent origin of these populations is strongly supported by our analyses of two independent sets of nuclear DNA polymorphism using two very different methods of analysis. Moreover, the observation of two divergent haplogroups of mitochondrial and nuclear genes with different geographic distributions support a recent admixture of two divergent surface populations before the isolation of cave populations. If cave populations are indeed only several thousand years old, many phenotypic changes observed in cavefish would thus have mainly involved the fixation of genetic variants present in surface fish populations and within a very short period of time.