The contribution of incomplete lineage sorting and introgression to the evolutionary history of the fast-evolving genus Ctenomys (Rodentia, Ctenomyidae).

Incomplete lineage sorting (ILS) and introgression have been increasingly recognized as important processes involved in biological differentiation. Both ILS and introgression result in incongruences between gene trees and species trees, consequently causing difficulties in phylogenetic reconstruction. This is particularly the case for rapid radiations, as short internodal distances and incomplete reproductive isolation increase the likelihood of both ILS and introgression. Estimation of the relative frequency of these processes requires assessments across many genomic regions. We use transcriptomics to test for introgression and estimate the frequency of ILS in a set of three closely related and geographically adjacent South American tuco-tucos species (Ctenomys), a genus comprising 64 species resulting from recent, rapid radiation. After cleaning and filtering, 5764 orthologous genes strongly support paraphyly of C. pearsoni relative to C. brasiliensis (putatively represented by the population of Villa Serrana). In line with earlier phylogenetic work, the C. pearsoni - C. brasiliensis pair is closely related to C. torquatus, whereas C. rionegrensis is more distantly related to these three nominal species. Classical Patterson's d-statistic shows significant signals of introgression from C. torquatus into C. brasiliensis. However, a 5-taxon test shows no significant results. ILS was estimated to have involved about 9% of the loci, suggesting it represents an important process in the incipient diversification of tuco-tucos.

An association between differential expression and genetic divergence in the Patagonian olive mouse (Abrothrix olivacea).

Recent molecular studies have found striking differences between desert-adapted species and model mammals regarding water conservation. In particular, aquaporin 4, a classical gene involved in water regulation of model species, is absent or not expressed in the kidneys of desert-adapted species. To further understand the molecular response to water availability, we studied the Patagonian olive mouse Abrothrix olivacea, a species with an unusually broad ecological tolerance that exhibits a great urine concentration capability. The species is able to occupy both the arid Patagonian steppe and the Valdivian and Magellanic forests. We sampled 95 olive mouse specimens from four localities (two in the steppe and two in the forests) and analysed both phenotypic variables and transcriptomic data to investigate the response of this species to the contrasting environmental conditions. The relative size of the kidney and the ratio of urine to plasma concentrations were, as expected, negatively correlated with annual rainfall. Expression analyses uncovered nearly 3,000 genes that were differentially expressed between steppe and forest samples and indicated that this species resorts to the "classical" gene pathways for water regulation. Differential expression across biomes also involves genes that involved in immune and detoxification functions. Overall, genes that were differentially expressed showed a slight tendency to be more divergent and to display an excess of intermediate allele frequencies, relative to the remaining loci. Our results indicate that both differential expression in pathways involved in water conservation and geographical allelic variation are important in the occupation of contrasting habitats by the Patagonian olive mouse.

[Measurement of the carbon footprint at a regional hospital]. / Huella de carbono en el Hospital Base de Puerto Montt.

BACKGROUND: Measuring the carbon footprint (CF) makes it possible to estimate the contribution of clinical activity to global warming. AIM: To measure the emitter components of CO2 equivalents (CO2e) at the Hospital Base, Puerto Montt (HBPM). MATERIAL AND METHODS: Descriptive study with data collected retrospectively between January and December 2016 from the HBPM database. The data analyzes direct and indirect emissions as well as other indirect emissions beyond the organizational limits. RESULTS: Of the 9,660.3 tons of CO2e emitted by the HBPM in 2016, 46% were derived from consumption of electricity, 29% derived from the generation of residues, and 10% from clinical gas consumption, of which Sevoflurane was the greatest contributor. CONCLUSIONS: Clinical gases are a significant source of CO2e emissions. Sevoflurane alone is in fourth place in CO2e emissions at the HBPM. Estimating the CF produced by HBPM is the first step in the discussion of measures to reduce the environmental impact of our activity.

Testing for the Occurrence of Selective Episodes During the Divergence of Otophysan Fishes: Insights from Mitogenomics.

How natural selection shapes biodiversity constitutes a topic of renewed interest during the last few decades. The division Otophysi comprises approximately two-thirds of freshwater fish diversity and probably underwent an extensive adaptive radiation derived from a single invasion of the supercontinent Pangaea, giving place to the evolution of the main five Otophysan lineages during a short period of time. Little is known about the factors involved in the processes that lead to lineage diversification among this group of fishes and identifying directional selection acting over protein-coding genes could offer clues about the processes acting on species diversification. The main objective of this study was to explore the otophysan mitochondrial genome evolution, in order to account for the possible signatures of selective events in this lineage, and to explore for the functional connotations of these molecular substitutions. Mainly, three different approaches were used: the "ω-based" BS-REL and MEME methods, implemented in the DATAMONKEY web server, and analysis of selection on amino acid properties, implemented in the software TreeSAAP. We found evidence of selective episodes along several branches of the evolutionary history of othophysan fishes. Analyses carried out using the BS-REL algorithm suggest episodic diversifying selection at basal branches of the otophysan lineage, which was also supported by analyses implemented in MEME and TreeSAAP. These results suggest that throughout the Siluriformes radiation, an important number of adaptive changes occurred in their mitochondrial genome. The metabolic consequences and ecological correlates of these molecular substitutions should be addressed in future studies.

Sex beyond species: the first genetically analyzed case of intergeneric fertile hybridization in pinnipeds.

A species, according to the biological concept, is a natural group of potentially interbreeding individuals isolated by diverse mechanisms. Hybridization is considered the production of offspring resulting from the interbreeding of two genetically distinct taxa. It has been documented in over 10% of wild animals, and at least in 34 cases for Artic marine mammals. In Otariids, intergeneric hybridization has been reported though neither confirming it through genetic analyses nor presenting evidence of fertile offspring. In this study, we report the finding of a hybrid adult female between a South American fur seal (Arctocephalus australis) and a South American sea lion (Otaria byronia), and its offspring, a male pup, in Uruguay. Further based on morphological constraints and breeding seasons, sex-biased hybridization between the two species is hypothesized. Morphological and genetic (nuclear and mitochondrial) results confirm de hybrid nature of the female-pup pair. Here we discuss a genetic dilution effect, considering other hybridization events must be occurring, and how isolation mechanisms could be circumvented. Moreover, the results obtained from stable isotope analysis suggest feeding habits may be a trait transmitted maternally, leading to consider broader issues regarding hybridization as an evolutionary innovation phenomenon.

Characterization of the kidney transcriptome of the South American olive mouse Abrothrix olivacea.

BACKGROUND: The olive mouse Abrothrix olivacea is a cricetid rodent of the subfamily Sigmodontinae that inhabits a wide range of contrasting environments in southern South America, from aridlands to temperate rainforests. Along its distribution, it presents different geographic forms that make the olive mouse a good focal case for the study of geographical variation in response to environmental variation. We chose to characterize the kidney transcriptome because this organ has been shown to be associated with multiple physiological processes, including water reabsorption. RESULTS: Transcriptomes of thirteen kidneys from individuals from Argentina and Chile were sequenced using Illumina technology in order to obtain a kidney reference transcriptome. After combining the reads produced for each sample, we explored three assembly strategies to obtain the best reconstruction of transcripts, TrinityNorm and DigiNorm, which include its own normalization algorithms for redundant reads removal, and Multireads, which simply consist on the assembly of the joined reads. We found that Multireads strategy produces a less fragmented assembly than normalization algorithms but recovers fewer number of genes. In general, about 15000 genes were annotated, of which almost half had at least one coding sequence reconstructed at 99% of its length. We also built a list of highly expressed genes, of which several are involved in water conservation under laboratory conditions using mouse models. CONCLUSION: Based on our assembly results, Trinity's in silico normalization is the best algorithm in terms of cost-benefit returns; however, our results also indicate that normalization should be avoided if complete or nearly complete coding sequences of genes are desired. Given that this work is the first to characterize the transcriptome of any member of Sigmodontinae, a subfamily of cricetid rodents with about 400 living species, it will provide valuable resources for future ecological and evolutionary genomic analyses.

Differential expression analyses and detection of SNP loci associated with environmental variables: Are salinity and temperature factors involved in population differentiation and speciation in Odontesthes?

Environmental factors play a key role in individual adaptation to different local conditions. Because of this, studies about the physiological and genetic responses of individuals exposed to different natural environments offer clues about mechanisms involved in population differentiation, and as a subsequent result, speciation. Marine environments are especially suited to survey this kind of phenomena because they commonly harbor species adapted to different local conditions along a geographic continuum. Silversides belonging to Odontesthes are commonly distributed in tropical and temperate regions of South America and exhibit noticeable phenotypic plasticity, which allows them to adapt to contrasting environments. In this study, the genetic expression of O. argentinensis sampled along the Uruguayan Atlantic coast and estuarine adjacent areas was investigated. In addition, the correlation between individual genotypes and environmental variables was also analysed in O. argentinensis and O. bonariensis. Results obtained suggest a differential expression pattern of low magnitude among individuals from the different areas sampled and a correlation between several SNP loci and environmental variables. The analyses carried out did not show a clear differentiation among individuals sampled along different salinity regimens, but enriched GOTerms seem to be driven by water oxygen content. On the other hand, a total of 46 SNPs analysed in O. argentinensis and O. bonariensis showed a correlation with salinity and temperature. Although none of the correlated SNPs and corresponding genes from our both analyses were directly associated with hypoxia, genes related to the cardiovascular system and muscle cell differentiation were found. All these genes are interesting candidates for future studies since they are closely related to the differentially expressed genes. Although salinity was also mentioned as an important parameter limiting introgression between O. argentinensis and O. bonariensis, it was found that salinity does not drive differential expression in O. argentinensis, but rather oxygen levels. Moreover, salinity does not directly affect the structure and genetic divergence of the populations, they appear to be structured based on their degree of isolation and geographical distance between them. Further studies, like genome-wide analyses, could help to elucidate additional genes adapted to the different environments in these silverside species.

Senecio spp. transboundary introduction and expansion affecting cattle in Uruguay: Clinico-pathological, epidemiological and genetic survey, and experimental intoxication with Senecio oxyphyllus.

The genus Senecio is distributed worldwide, being responsible of poisoning in livestock and humans. Many species of Senecio have high invasion and expansion capacity, highly competitive with agricultural and native plant species, causing ecological damage. Particularly in Uruguay, poisoning by Senecio have grown exponentially to reach epidemic proportions. Herein we describe Seneciosis as a re-emerging and expanding epidemic disease affecting cattle, by describing clinico-pathological, epidemiological and genetic variation of species involved, as well as an experimental intoxication with Senecio oxyphyllus. For this, a study was carried out on 28 cattle farms in Eastern Uruguay, with history of seneciosis from 2010 to 2016. Plants of fifty populations of Senecio were sampled, in 2015 and 2016, for identification, analysis of alkaloids and study of genetic variation. In turn, post-mortem examination was performed in cattle of natural and an experimental case to confirm the intoxication, showing microscopic characteristic lesions (hepatomegalocytosis, diffuse fibrosis and ductal reaction). Four species of Senecio were identified: S. oxyphyllus, S. madagascariensis, S. selloi and S. brasiliensis. In the genetic study, 489 molecular markers of amplified sequence-related polymorphisms (SRAP), associated with species and pasture, were used for genetic variation analysis. There was no statistically significant association between genetic variation determined by molecular markers and population (specimens of same species collected from the same farm), botanically determined species, or geographical origin. The increase of seneciosis in cattle in the last years, the presence of species not identified to the moment with implication in the poisoning outbreaks and expansion of these plants shows that the disease is in an epidemic growing active stage. In turn, the experimental poisoning with S. oxyphyllus confirms its chronic hepatotoxic effect, being an emergent species for the region, of high distribution and toxic risk. This latter turned out the main Senecio species involved. This case of expansion of harmful plant for animal production and desirable plant species, can be useful as a model of ecopathological characterization, which is likely to occur with other toxic plants in different geographical ranges globally.

Characterization of the kidney transcriptome of the long-haired mouse Abrothrix hirta (Rodentia, Sigmodontinae) and comparison with that of the olive mouse A. olivacea.

To understand how small mammals cope with the challenge of water homeostasis is a matter of interest for ecologists and evolutionary biologists. Here we take a step towards the understanding of the transcriptomic functional response of kidney using as a model the long-haired mouse (Abrothrix hirta) a species that distributes across Patagonian steppes and Austral temperate rainforests in Argentina and Chile. Specifically, we i) characterize the renal transcriptome of A. hirta, and ii) compare it with that-already available-of the co-generic and co-distributed A. olivacea. Renal mRNA transcripts from 16 specimens of A. hirta from natural populations were analyzed. Over 500 million Illumina paired-end reads were assembled de novo under two approaches, an individual assembly for each specimen, and a single in-silico normalized joint assembly including all reads from all specimens. The total number of annotated genes was similar with both strategies: an average of 14,956 in individual assemblies and 14,410 in the joint assembly. Overall, 15,463 distinct genes express in the kidney of A. hirta. Transcriptomes of A. hirta and A. olivacea were similar in terms of gene abundance and composition: 95.6% of the genes of A. hirta were also found in A. olivacea making their functional profiles also similar. However, differences in the transcriptome of these two species were observed in the set of highly expressed genes, in terms of private genes for each species and the functional profiles of highly expressed genes. As part of the novel transcriptome characterization, we provide distinct gene lists with their functional annotation that would constitute the basis for further research on these or any other species of the subfamily Sigmodontinae, which includes about 400 living species distributed from Tierra del Fuego to southern United States.

Huella de carbono en el Hospital Base de Puerto Montt / Measurement of the carbon footprint at a regional hospital

Background: Measuring the carbon footprint (CF) makes it possible to estimate the contribution of clinical activity to global warming. Aim: To measure the emitter components of CO2 equivalents (CO2e) at the Hospital Base, Puerto Montt (HBPM). Material and methods: Descriptive study with data collected retrospectively between January and December 2016 from the HBPM database. The data analyzes direct and indirect emissions as well as other indirect emissions beyond the organizational limits. Results: Of the 9,660.3 tons of CO2e emitted by the HBPM in 2016, 46% were derived from consumption of electricity, 29% derived from the generation of residues, and 10% from clinical gas consumption, of which Sevoflurane was the greatest contributor. Conclusions: Clinical gases are a significant source of CO2e emissions. Sevoflurane alone is in fourth place in CO2e emissions at the HBPM. Estimating the CF produced by HBPM is the first step in the discussion of measures to reduce the environmental impact of our activity.