Lizard thermoregulation revisited after two decades of global warming

CRUE-CSIC (Acuerdos Transformativos 2022)1.Although the effects of global warming on thermoregulation are usually explored using predictions of climate envelop modelling, such effects should best be analysed empirically, studying the same population with the same methods after a long enough period of temperature rise. 2. We used a 30-year long database about body temperatures (Tbs) of field-active Psammodromus algirus lizards inhabiting a well-conserved temperate open forest, and we focused on the summers of 1997 and 2017 to compare Tbs, environmental operative temperatures (Tes), their proximity to the selected thermal range (Tsel), and the selection of sunlit and shaded patches all along the day. From these data, we estimated the precision (standard deviation of Tbs), accuracy (average distance between Tbs and Tsel) and effectiveness (extent to which Tbs are closer to Tsel than Tes) of thermoregulation. 3. Of the highest 5% of all Tbs in the database, 95% were recorded in 2017, when the adjustment to Tsel was much better for Tbs selected in a laboratory thermogradient than for field Tbs (percentages of Tbs above Tsel of 2% and 52% respectively). 4. In 2017, especially after 12:00 h, the selection of shaded patches (87% of lizards in full shade vs. <1% in full sun) was more intense than in 1997, contributed more to overall thermoregulation, and produced a larger difference between Tes and Tbs. In spite of this, Tbs were lower—and closer to Tsel—in 1997 (when most shaded patches offered favourable thermal opportunities, with Tes within or below Tsel) than in 2017 (when only 33% of full shade Tes, and 8% of all Tes, were within or below Tsel). As a consequence, estimates of the accuracy and effectiveness of thermoregulation decreased over the 20-year period examined. 5. We conclude that given the low availability of Tes within or below Tsel, lizards cannot longer prevent the rise of their Tbs above Tsel, at least in hot summer days. Thus, the effects of global warming are already hindering the ability of lizards to buffer environmental change by behavioural means, even in temperate forests with a fine-grained mosaic of sun and shade patches.Depto. de Biodiversidad, Ecología y EvoluciónFac. de Ciencias BiológicasTRUEMinisterio de Ciencia e Innovación (MICINN)pu

Transgenerational inheritance of environmental information in Caenorhabditis elegans

Motivation: In 1906 Luther Burbank, an American botanist, said “Heridity is only the sum of all past environment” (Burbank, 1906). This idea has always been present in spite of being left behind after sustaining that the germline was isolated of the rest of the organism and, therefore, of the environment. With the rediscovery of epigenetics as a regulatory system which controls gene expression without affecting the composition of genes themselves, this idea has regained importance. Not only a great number of cases of environmental changes that influenced the epigenetics in a great variety of species began to be observed; but also the effects of these changes could last for three or more generations. This phenomenon was known as transgenerational epigenetic inheritance (Heard and Martienssen, 2014). This project aims to study the phenomenon of transgenerational epigenetic inheritance in the Caenorhabditis elegans model animal. The idea is to set the parameters where we can identify the phenomenon and reveal the molecular mechanism involved in the process.Methods: We set two different experiments, resistance to pathogen and behavioral respond to the pathogen. For the resistance experiment, plates with Escherichia coli (OP50, main source of nematode's food) and plates with Pseudomonas aeruginosa (PA14, a pathogen) were used to grow mothers and then the percentage of descendants that reach adulthood were scored. For the behavioral experiment, plates with OP50 and plates with both bacteria (PA14 and OP50) were used to grow mothers. Then their eggs were placed on plates containing OP50 and PA14. We counted the percent of worms that went to OP50 and to PA14.Results: It has been observed that individuals whose mothers have been exposed to Pseudomonas aeruginosa are more resistant to it than those whose mothers have never been in contact with the pathogen. On the other hand, it has been observed that this inheritance not only affects the defense mechanisms of the organism, but also influences its behavior. At this time we are studying the genetic pathways involved in this process by using mutants affected in epigenetic and in other candidate pathways.Conclusions: Our results suggest that there is not only an inheritance to stress resistance but also an acquired behavioral inheritance. Genes involved in epigenetic seem to be involved in the process

There is more to the picture than meets the eye: adaptation for crypsis blurs phylogeographical structure in a lizard

Aim: We examined dorsal coloration in and genetic relationships among Iberian populations of the lizard Psammodromus algirus to determine the extent to which the current distribution of phenotypic variation is correlated with phylogeographical history or local environmental conditions. Location: Iberian Peninsula, western Palaearctic.Methods: We sequenced mitochondrial DNA (ND4 and adjacent tRNAs genes) in 36 populations, and seven microsatellite loci in eight representative populations. In 23 populations, lizards were classified according to the presence and intensity of a dorsal striped pattern, the heritability of which was estimated by means of mother–offspring regressions. To determine whether colour pattern is an adaptation for crypsis, we compared the time taken by humans to detect striped and unstriped lizards in different environments.Results: The analysis of mtDNA revealed an ancient split between a western clade, subdivided into south- and north-western haplogroups, and an eastern clade with central, south-eastern and eastern haplogroups. In contrast, nuclear markers showed a post-glacial admixture of central and western haplogroups, with the central haplogroup apparently isolated from the rest of its clade. This was consistent with variation in the dorsal striped pattern, a heritable phenotypic trait: central and western lizards were unstriped, whereas eastern lizards were striped. We then suggest that dorsal coloration promotes crypsis: in eastern locations detection times were longer for striped than for unstriped lizards, whereas the opposite was true in western and central locations.Main conclusions: Our results indicate that natural selection for crypsis may promote not only divergence within clades, as suggested by the apparent isolation between unstriped central lizards and striped members of eastern haplogroups, but also admixture between them. We conclude that ecologically driven selection is crucial for understanding the phylogeographical background of phenotypic variation, because recent adaptation to the environment can blur the effects of ancestral isolation.This study is a contribution to projects CGL2010-17928/BOS and CGL2013-41642-P, funded by the Spanish Ministry of Education and SciencePeer reviewe

Infection of laboratory-colonized Anopheles darlingi mosquitoes by Plasmodium vivax.

Anopheles darlingi Root is the most important malaria vector in the Amazonia region of South America. However, continuous propagation of An. darlingi in the laboratory has been elusive, limiting entomological, genetic/genomic, and vector-pathogen interaction studies of this mosquito species. Here, we report the establishment of an An. darlingi colony derived from wild-caught mosquitoes obtained in the northeastern Peruvian Amazon region of Iquitos in the Loreto Department. We show that the numbers of eggs, larvae, pupae, and adults continue to rise at least to the F6 generation. Comparison of feeding Plasmodium vivax ex vivo of F4 and F5 to F1 generation mosquitoes showed the comparable presence of oocysts and sporozoites, with numbers that corresponded to blood-stage asexual parasitemia and gametocytemia, confirming P. vivax vectorial capacity in the colonized mosquitoes. These results provide new avenues for research on An. darlingi biology and study of An. darlingi-Plasmodium interactions

Adaptive divergence under gene flow along an environmental gradient in two coexisting stickleback species

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The combined use of raw and phylogenetically independent methods of outlier detection uncovers genome‐wide dynamics of local adaptation in a lizard

Local adaptation is a dynamic process by which different allele combinations are se-lected in different populations at different times, and whose genetic signature can be inferred by genome-wide outlier analyses. We combined gene flow estimates with two methods of outlier detection, one of them independent of population coances-try (CIOA) and the other one not (ROA), to identify genetic variants favored when ecology promotes phenotypic convergence. We analyzed genotyping-by-sequencing data from five populations of a lizard distributed over an environmentally heteroge-neous range that has been changing since the split of eastern and western lineages ca. 3 mya. Overall, western lizards inhabit forest habitat and are unstriped, whereas eastern ones inhabit shrublands and are striped. However, one population (Lerma) has unstriped phenotype despite its eastern ancestry. The analysis of 73,291 SNPs confirmed the east–west division and identified nonoverlapping sets of outliers (12 identified by ROA and 9 by CIOA). ROA revealed ancestral adaptive variation in the uncovered outliers that were subject to divergent selection and differently fixed for eastern and western populations at the extremes of the environmental gradient. Interestingly, such variation was maintained in Lerma, where we found high levels of heterozygosity for ROA outliers, whereas CIOA uncovered innovative variants that were selected only there. Overall, it seems that both the maintenance of ancestral variation and asymmetric migration have counterbalanced adaptive lineage split-ting in our model species. This scenario, which is likely promoted by a changing and heterogeneous environment, could hamper ecological speciation of locally adapted populations despite strong genetic structure between lineages

Low genome‐wide divergence between two lizard populations with high adaptive phenotypic differentiation

Usually, adaptive phenotypic differentiation is paralleled by genetic divergence between locally adapted populations. However, adaptation can also happen in a scenario of nonsignificant genetic divergence due to intense gene flow and/or recent differentiation. While this phenomenon is rarely published, findings on incipient ecologically driven divergence or isolation by adaptation are relatively common, which could confound our understanding about the frequency at which they actually occur in nature. Here, we explore genome-wide traces of divergence between two populations of the lacertid lizard Psammodromus algirus separated by a 600 m elevational gradient. These populations seem to be differentially adapted to their environments despite showing low levels of genetic differentiation (according to previously studies of mtDNA and microsatellite data). We performed a search for outliers (i.e., loci subject to selection) trying to identify specific loci with FST statistics significantly higher than those expected on the basis of overall, genome-wide estimates of genetic divergence. We find that local phenotypic adaptation (in terms of a wide diversity of characters) was not accompanied by genome-wide differentiation, even when we maximized the chances of unveiling such differentiation at particular loci with FSTbased outlier detection tests. Instead, our analyses confirmed the lack of genomewide differentiation on the basis of more than 70,000 SNPs, which is concordant with a scenario of local adaptation without isolation by environment. Our results add evidence to previous studies in which local adaptation does not lead to any kind of isolation (or early stages of ecological speciation), but maintains phenotypic divergence despite the lack of a differentiated genomic background

Environmental association modelling with loci under divergent selection predicts the distribution range of a lizard

During the historical building of a species range, individual colonizers have to confront different ecological challenges, and the capacity of the species to broaden its range may depend on the total amount of adaptive genetic variation supplied by evolution. We set out to increase our understanding of what defines a range and the role of underlying genetics by trying to predict an entire species’ range from the geographical distribution of its genetic diversity under selection. We sampled five populations of the western Mediterranean lizard Psammodromus algirus that inhabit a noticeable environmental gradient of temperature and precipitation. We correlated the genotypes of 95 individuals (18–20 individuals per population) for 21 SNPs putatively under selection with environmental scores on a bioclimatic gradient, using 1 × 1 km2 grid cells as sampling units. By extrapolating the resulting model to all possible combinations of alleles, we inferred all the geographic cells that were theoretically suitable for a given amount of genetic variance under selection. The inferred distribution range overlapped to a large extent with the realized range of the species (77.46% of overlap), including an accurate prediction of internal gaps and range borders. Our results suggest an adaptability threshold determined by the amount of genetic variation available that would be required to warrant adaptation beyond a certain limit of environmental variation. These results support the idea that the expansion of a species’ range can be ultimately linked to the arising of new variants under selection (either newly selected variants from standing genetic variation or innovative mutations under selection)

Intestinal P-glycoprotein exports endocannabinoids to prevent inflammation and maintain homeostasis

Neutrophil influx into the intestinal lumen is a critical response to infectious agents, but is also associated with severe intestinal damage observed in idiopathic inflammatory bowel disease. The chemoattractant hepoxilin A3, an eicosanoid secreted from intestinal epithelial cells by the apically restricted efflux pump multidrug resistance protein 2 (MRP2), mediates this neutrophil influx. Information about a possible counterbalance pathway that could signal the lack of or resolution of an apical inflammatory signal, however, has yet to be described. We now report a system with such hallmarks. Specifically, we identify endocannabinoids as the first known endogenous substrates of the apically restricted multidrug resistance transporter P-glycoprotein (P-gp) and reveal a mechanism, which we believe is novel, for endocannabinoid secretion into the intestinal lumen. Knockdown or inhibition of P-gp reduced luminal secretion levels of N-acyl ethanolamine-type endocannabinoids, which correlated with increased neutrophil transmigration in vitro and in vivo. Additionally, loss of CB2, the peripheral cannabinoid receptor, led to increased pathology and neutrophil influx in models of acute intestinal inflammation. These results define a key role for epithelial cells in balancing the constitutive secretion of antiinflammatory lipids with the stimulated secretion of proinflammatory lipids via surface efflux pumps in order to control neutrophil infiltration into the intestinal lumen and maintain homeostasis in the healthy intestine

Whole genome sequencing analysis of Plasmodium vivax using whole genome capture

Background: Malaria caused by Plasmodium vivax is an experimentally neglected severe disease with a substantial burden on human health. Because of technical limitations, little is known about the biology of this important human pathogen. Whole genome analysis methods on patient-derived material are thus likely to have a substantial impact on our understanding of P. vivax pathogenesis and epidemiology. For example, it will allow study of the evolution and population biology of the parasite, allow parasite transmission patterns to be characterized, and may facilitate the identification of new drug resistance genes. Because parasitemias are typically low and the parasite cannot be readily cultured, on-site leukocyte depletion of blood samples is typically needed to remove human DNA that may be 1000X more abundant than parasite DNA. These features have precluded the analysis of archived blood samples and require the presence of laboratories in close proximity to the collection of field samples for optimal pre-cryopreservation sample preparation. Results: Here we show that in-solution hybridization capture can be used to extract P. vivax DNA from human contaminating DNA in the laboratory without the need for on-site leukocyte filtration. Using a whole genome capture method, we were able to enrich P. vivax DNA from bulk genomic DNA from less than 0.5% to a median of 55% (range 20%-80%). This level of enrichment allows for efficient analysis of the samples by whole genome sequencing and does not introduce any gross biases into the data. With this method, we obtained greater than 5X coverage across 93% of the P. vivax genome for four P. vivax strains from Iquitos, Peru, which is similar to our results using leukocyte filtration (greater than 5X coverage across 96% of the genome). Conclusion: The whole genome capture technique will enable more efficient whole genome analysis of P. vivax from a larger geographic region and from valuable archived sample collections.National Institutes of Health [R21-AI085374-01A1, U19AI089681, 1K24AI068903, D43TW007120, R01AI067727]UCSD Genetics Training Program through an institutional training grant from the National Institute of General Medical Sciences [T32 GM008666]NIH/NCRR Grant [UL1 RR025774