Specimen collection is essential for modern science.

Natural history museums are vital repositories of specimens, samples and data that inform about the natural world; this Formal Comment revisits a Perspective that advocated for the adoption of compassionate collection practices, querying whether it will ever be possible to completely do away with whole animal specimen collection.

Sex, not social behavior, predicts fecal glucocorticoid metabolite concentrations in a facultatively social rodent, the highland tuco-tuco (Ctenomys opimus).

Social relationships may influence circulating glucocorticoid levels, particularly in group-living species in which individuals regularly engage in interactions with conspecifics. The effects of such interactions appear to vary, with greater social contact being associated with increased glucocorticoid concentrations in some species but decreased concentrations in others. These distinct responses raise intriguing questions regarding relationships among social behavior, individual phenotypes, and glucocorticoid physiology. To explore such relationships in a free-living mammal with a dynamic social organization, we quantified variation in baseline glucocorticoids in a population of highland tuco-tucos (Ctenomys opimus) from Jujuy Province, Argentina. These subterranean rodents are facultatively social, with lone and group-living individuals regularly occurring within the same population. To assess potential endocrine correlates of this behavioral variability, we examined differences in baseline fecal glucocorticoid metabolite (fGCm) concentrations as a function of social group size and composition as well as several metrics of social behavior derived from social network analyses. Despite marked variability in social relationships among the 37 (12 male, 25 female) free-living tuco-tucos sampled, none of the measures of social behavior examined were significant predictors of variation in fGCm concentrations. In contrast, individual variation in glucocorticoid metabolites was best explained by sex, with males having higher fGCm concentrations than females. These analyses provide the first characterization of the glucocorticoid physiology of highland tuco-tucos and underscore the potential importance of intrinsic phenotypic factors (e.g., sex) in shaping glucocorticoid variation in free-living mammals.

Mating system is correlated with immunogenetic diversity in sympatric species of Peromyscine mice.

The number of reproductive partners per individual varies markedly across animal mating systems. This variation may be an important determinant of patterns of immunogenetic diversity, particularly at Major Histocompatibility Complex (MHC) Class I and II loci. To compare immunogenetic variation in taxa with markedly different mating systems, we used RNAseq-generated data to quantify genotypic diversity in three species of Peromyscine rodents: the monogamous California mouse (Peromyscus californicus) and the polygynandrous deer mouse (P. maniculatus) and brush mouse (P. boylii). By sampling populations of these species from multiple localities in California, we were able to conduct replicated analyses of the relationship between mating system and immunogenetic variation. Across the four localities sampled, diversity at MHC Class I and II genes was consistently higher in the two polygynandrous species. We found no evidence that sampling location (i.e., variation in habitat conditions) contributed to observed differences in MHC variation among populations or species. Collectively, our data indicate that immunogenetic variation in Peromyscine mice is associated with reproductive behavior, rather than geographic locality or habitat type. The consistently greater variability detected in the polygynandrous species examined suggests that balancing selection imposed by behaviorally-mediated pathogen exposure is important in maintaining variation at MHC genes in these animals.

Gut microbial diversity across a contact zone for California voles: Implications for lineage divergence of hosts and mitonuclear mismatch in the assembly of the mammalian gut microbiome.

Gut microbial diversity is thought to reflect the co-evolution of microbes and their hosts as well as current host-specific attributes such as genetic background and environmental setting. To explore interactions among these parameters, we characterized variation in gut microbiome composition of California voles (Microtus californicus) across a contact zone between two recently diverged lineages of this species. Because this contact zone contains individuals with mismatched mitochondrial-nuclear genomes (cybrids), it provides an important opportunity to explore how different components of the genotype contribute to gut microbial diversity. Analyses of bacterial 16S rRNA sequences and joint species distribution modelling revealed that host genotypes and genetic differentiation among host populations together explained more than 50% of microbial community variation across our sampling transect. The ranked importance (most to least) of factors contributing to gut microbial diversity in our study populations were: genome-wide population differentiation, local environmental conditions, and host genotypes. However, differences in microbial communities among vole populations (ß-diversity) did not follow patterns of lineage divergence (i.e., phylosymbiosis). Instead, among-population variation was best explained by the spatial distribution of hosts, as expected if the environment is a primary source of gut microbial diversity (i.e., dispersal limitation hypothesis). Across the contact zone, several bacterial taxa differed in relative abundance between the two parental lineages as well as among individuals with mismatched mitochondrial and nuclear genomes. Thus, genetic divergence among host lineages and mitonuclear genomic mismatches may also contribute to microbial diversity by altering interactions between host genomes and gut microbiota (i.e., hologenome speciation hypothesis).

Differential gene expression in relation to mating system in Peromyscine rodents.

Behaviors that increase an individual's exposure to pathogens are expected to have important effects on immunoactivity. Because sexual reproduction typically requires close contact among conspecifics, mating systems provide an ideal opportunity to study the immunogenetic correlates of behaviors with high versus low risks of pathogen exposure. Despite logical links between polygynandrous mating behavior, increased pathogen exposure, and greater immunoactivity, these relationships have seldom been examined in nonhuman vertebrates. To explore interactions among these variables in a different lineage of mammals, we used RNAseq to study the gene expression profiles of liver tissue-a highly immunoactive organ-from sympatric populations of the monogamous California mouse (Peromyscus californicus) and two polygynandrous congeners (P. maniculatus and P. boylii). Differential expression and co-expression analyses revealed distinct patterns of gene activity among species, with much of this variation associated with differences in mating system. This tendency was particularly pronounced for MHC genes, with multiple MHC Class I genes being upregulated in the two polygynandrous species, as expected if exposure to sexually transmitted pathogens varies with mating system. Our results underscore the role of mating behavior in influencing patterns of gene expression and highlight the use of emerging transcriptomic tools in behavioral studies of free-living animals.

Ecological and demographic impacts of a recent volcanic eruption on two endemic patagonian rodents.

Catastrophic events can significantly impact the demographic processes that shape natural populations of organisms. However, linking the outcomes of such events to specific demographic parameters is often challenging due to a lack of detailed pre-event data. The eruption of the Puyehue-Cordon Caulle volcanic complex on 4 June 2011 had profound consequences for the biota of southwestern Argentina. Our long-term behavioral, ecological, and demographic studies of two species of tuco-tucos (Ctenomys sociabilis and C. haigi) that occur in the region most heavily impacted by ash fall from the eruption provided an unusual opportunity to assess the effects of this event on natural populations of mammals. The post-eruption density of the study population for each species was markedly reduced compared to pre-eruption values, with the relative magnitude of this reduction being greater for the group-living C. sociabilis. The more extensive data set for this species indicated that ash fall from the eruption altered the food resources available to these animals; differences in pre- and post-eruption stable isotope signatures for fur samples from C. sociabilis were consistent with observed changes in vegetation. Per capita female reproductive success was also reduced in this species during the first breeding season following the eruption. Based on our detailed demographic records for C. sociabilis, neither survival of yearling females from 2010 to 2011 nor the percentage of unmarked females in the study population in 2011 differed from pre-eruption values. Instead, the post-eruption decrease in population density for C. sociabilis appeared to reflect reduced within-population recruitment of juvenile females to the 2011 breeding population. Although the eruption did not result in the local extinction of either study population, the demographic consequences detected are likely to have impacted the effective sizes of these populations, creating important opportunities to link specific demographic parameters to previously reported decreases in genetic variability detected after this significant natural event.

Genomic analysis of MHC-based mate choice in the monogamous California mouse.

Variation at Major Histocompatibility Complex (MHC) genes is thought to be an important mechanism underlying mate choice in vertebrates, with individuals typically predicted to prefer MHC-dissimilar reproductive partners. However, analyses based on individual MHC loci have generated contradictory results regarding the role of these genes in mate-choice decisions. To provide a more comprehensive assessment of relationships between MHC variation and mating behavior, we used an exome capture strategy to characterize variability at 13 MHC loci, 312 innate immune system genes, and 1044 nonimmune genes in 25 obligate monogamous pairs of California mice (Peromyscus californicus) from 2 free-living populations of this species in Monterey County, California. Pairwise genotypic comparisons and analyses of SNP-based allelic differences failed to detect disassortative mating based on MHC variability; reproductive partners were not more dissimilar than randomly generated male-female pairs at MHC, innate or nonimmune loci. Within populations, individuals tended to be more closely related at MHC genes than at innate or nonimmune genes. Consistent with the functional role of immunogenes, the 2 study populations were highly differentiated at MHC and innate genes but not at nonimmune loci. Collectively, our results suggest that MHC genetic variation in California mice reflects local differences in pathogen exposure rather than disassortative mating based on variability at MHC Class I and II genes.

Glucocorticoid-environment relationships align with responses to environmental change in two co-occurring congeners.

As more species undergo range shifts in response to climate change, it is increasingly important to understand the factors that determine an organism's realized niche. Physiological limits imposed by abiotic factors constrain the distributions of many species. Because glucocorticoids are essential to the maintenance of physiological homeostasis, identifying glucocorticoid-environment relationships may generate critical insights into both limits on species distributions and potential responses to environmental change. We explored relationships between variability in baseline glucocorticoids and sensitivity to environmental conditions in two chipmunk species characterized by divergent patterns of spatial, genetic, and morphological change over the past century. Specifically, we investigated whether the alpine chipmunk (Tamias alpinus), which has undergone pronounced changes, displays greater glucocorticoid sensitivity to environmental parameters than the lodgepole chipmunk (T. speciosus), which has exhibited little change over the same interval. From 2013 to 2015, we collected environmental data and fecal glucocorticoid metabolite (FGM) samples from these species. Using generalized linear mixed models and a model averaging approach, we examined the impacts of environmental and individual phenotypic parameters on FGMs. We found pronounced interspecific differences, with environmental parameters being better predictors of FGMs in T. alpinus. FGMs in this species were particularly elevated in less climatically suitable habitats and in areas with higher maximum daily temperatures. Individual phenotypic traits were not predictive of FGMs in T. alpinus, although they were highly predictive for T. speciosus. Collectively, these findings support the hypothesis that T. alpinus is more sensitive to environmental change. More generally, our results suggest that both phenotypic attributes and environmental conditions contribute to FGM responses but that the relative contributions of these factors differ among taxa, including among closely related species. Finally, our analyses underscore the value of glucocorticoids as bioindicators of sensitivity to environmental change in species for which the factors affecting stress physiology have been assessed.

Mito-nuclear discordance across a recent contact zone for California voles.

To examine the processes that maintain genetic diversity among closely related taxa, we investigated the dynamics of introgression across a contact zone between two lineages of California voles (Microtus californicus). We tested the prediction that introgression of nuclear loci would be greater than that for mitochondrial loci, assuming ongoing gene flow across the contact zone. We also predicted that genomic markers would show a mosaic pattern of differentiation across this zone, consistent with genomes that are semi-permeable. Using mitochondrial cytochrome b sequences and genome-wide loci developed via ddRAD-seq, we analyzed genetic variation for 10 vole populations distributed along the central California coast; this transect included populations from within the distributions of both parental lineages as well as the putative contact zone. Our analyses revealed that (1) the two lineages examined are relatively young, having diverged ca. 8.5-54 kya, (2) voles from the contact zone in Santa Barbara County did not include F1 or early generation backcrossed individuals, and (3) there appeared to be little to no recurrent gene flow across the contact zone. Introgression patterns for mitochondrial and nuclear markers were not concordant; only mitochondrial markers revealed evidence of introgression, putatively due to historical hybridization. These differences in genetic signatures are intriguing given that the contact zone occurs in a region of continuous vole habitat, with no evidence of past or present physical barriers. Future studies that examine specific isolating mechanisms, such as microhabitat use and mate choice, will facilitate our understanding of how genetic boundaries are maintained in this system.

Ecological specialization, variability in activity patterns and response to environmental change.

Differences in temporal patterns of activity can modulate the ambient conditions to which organisms are exposed, providing an important mechanism for responding to environmental change. Such differences may be particularly relevant to ecological generalists, which are expected to encounter a wider range of environmental conditions. Here, we compare temporal patterns of activity for partially sympatric populations of a generalist (the lodgepole chipmunk, Tamias speciosus) and a more specialized congener (the alpine chipmunk, Tamias alpinus) that have displayed divergent responses to the past century of environmental change. Although mean activity budgets were similar between species, analyses of individual-level variation in locomotion revealed that T. alpinus exhibited a narrower range of activity patterns than Tspeciosus Further analyses revealed that T. alpinus was more active earlier in the day, when temperatures were cooler, and that activity patterns for both species changed with increased interspecific co-occurrence. These results are consistent with the greater responsiveness of T. alpinus to changes in environmental conditions. In addition to highlighting the utility of accelerometers for collecting behavioural data, our findings add to a growing body of evidence, suggesting that the greater phenotypic variability displayed by ecological generalists may be critical to in situ responses to environmental change.

Dispersal limitation promotes the diversification of the mammalian gut microbiota.

The gut bacterial communities of mammals have profound effects on host fitness, but the processes that generate and maintain gut bacterial diversity remain poorly understood. We mapped compositional variation (i.e., ß-diversity) in the gut microbiotas of 136 pairs of wild mammalian species living throughout the Americas to assess how the distribution of mammals across geographic space influences the diversification of their gut bacteria. Comparing the gut microbiotas of sympatric and allopatric mammalian populations provided insights into the flow of gut bacteria within and between mammalian communities, revealing that spatial limits on bacterial dispersal promote ß-diversity between the gut microbiotas of mammalian species. Each geographic locale displayed a unique gut-microbiota composition that could not be fully explained by the diets and phylogenetic histories of the resident mammalian hosts, indicating that some gut bacteria are geographically restricted. Across the western hemisphere, the compositional overlap between the gut microbiotas of allopatric mammalian populations decayed exponentially with the geographic distance separating the hosts. The relationship between geographic distances among hosts and compositional differences among their gut microbiotas was independent of dietary and phylogenetic divergence among hosts. Within mammalian communities, we observed widespread sharing of gut bacteria between predator-prey host-species pairs, indicating horizontal transfer of gut bacteria through mammalian food chains. Collectively, these results indicate that compositional differences between the gut microbiotas of mammalian taxa are generated and maintained by limits to bacterial dispersal imposed by physical distance between hosts.

Genomic data reveal a loss of diversity in two species of tuco-tucos (genus Ctenomys) following a volcanic eruption.

Marked reductions in population size can trigger corresponding declines in genetic variation. Understanding the precise genetic consequences of such reductions, however, is often challenging due to the absence of robust pre- and post-reduction datasets. Here, we use heterochronous genomic data from samples obtained before and immediately after the 2011 eruption of the Puyehue-Cordón Caulle volcanic complex in Patagonia to explore the genetic impacts of this event on two parapatric species of rodents, the colonial tuco-tuco (Ctenomys sociabilis) and the Patagonian tuco-tuco (C. haigi). Previous analyses using microsatellites revealed no post-eruption changes in genetic variation in C. haigi, but an unexpected increase in variation in C. sociabilis. To explore this outcome further, we used targeted gene capture to sequence over 2,000 putatively neutral regions for both species. Our data revealed that, contrary to the microsatellite analyses, the eruption was associated with a small but significant decrease in genetic variation in both species. We suggest that genome-level analyses provide greater power than traditional molecular markers to detect the genetic consequences of population size changes, particularly changes that are recent, short-term, or modest in size. Consequently, genomic analyses promise to generate important new insights into the effects of specific environmental events on demography and genetic variation.

Mutualism in museums: A model for engaging undergraduates in biodiversity science.

Museums have an untapped potential to engage students in hands-on learning. Here, we describe the development of a tiered museum-based program at the University of California, Berkeley as a model for engaging undergraduates in biodiversity science. This decade-long effort to increase student participation in collections demonstrates the mutual benefits of undergraduate involvement. Museums benefit from critical help in collections care and an increased intellectual vitality, while students simultaneously gain essential research skills and an unparalleled exposure to biodiversity. Five first steps to creating a program are: dedicate a coordinator, offer credit, diversify participation, create a tiered structure, and build community.

Discrete but variable structure of animal societies leads to the false perception of a social continuum.

Animal societies are typically divided into those in which reproduction within a group is monopolized by a single female versus those in which it is shared among multiple females. It remains controversial, however, whether these two forms of social structure represent distinct evolutionary outcomes or endpoints along a continuum of reproductive options. To address this issue and to determine whether vertebrates and insects exhibit the same patterns of variation in social structure, we examined the demographic and reproductive structures of 293 species of wasps, ants, birds and mammals. Using phylogenetically informed comparative analyses, we found strong evidence indicating that not all reproductive arrangements within social groups are viable in nature and that in societies with multiple reproductives, selection favours instead taxon-specific patterns of decrease in the proportion of breeders as a function of group size. These outcomes suggest that the selective routes to sociality differ depending upon whether monopolization of reproduction by one individual is possible and that variation within and among taxonomic groups may lead to the false perception of a continuum of social structures. Thus, the occurrence of very large societies may require either complete reproductive monopolization (monogyny/singular breeding) or the maintenance of a taxon-specific range of values for the proportional decrease in the number of breeders within a group (polygyny/plural breeding), both of which may reduce reproductive conflict among females.

Genetic, spatial, and social relationships among adults in a group of howler monkeys (Alouatta palliata) from Barro Colorado Island, Panama.

Kinship plays an important role in the social behavior of many primate species, including patterns of intra-group affiliation and cooperation. Within social groups, kinship is strongly affected by dispersal patterns, with the degree of relatedness among group-mates expected to decrease as the tendency to disperse increases. In primate species characterized by bisexual dispersal, relatedness among adult group-mates is predicted to be low, with social interactions shaped largely by factors other than kinship. To date, however, few studies have examined the role of kinship in social interactions in bisexually dispersing species. Accordingly, we collected genetic, spatial and behavioral data on all adult members (three males, six females) in a group of free-ranging mantled howler monkeys (Alouatta palliata)--a bisexually dispersing species of atelid primate--from Barro Colorado Island (BCI), Panama. Analyses of microsatellite variation revealed that relatedness was greater among adult males in this group (mean pairwise relatedness = 0.32 for males versus 0.09 for females). Relatedness among individuals, however, was not associated with either spatial proximity or frequency of social interactions. Instead, sex was a better predictor of both of these aspects of social behavior. While relatedness among adults had no discernible effect on the intra-group social interactions documented in this study, we postulate that kinship may facilitate affiliative and cooperative behaviors among male group-mates when interacting competitively with neighboring howler groups over access to food or potential mates.

Morphological and dietary responses of chipmunks to a century of climate change.

Predicting how individual taxa will respond to climatic change is challenging, in part because the impacts of environmental conditions can vary markedly, even among closely related species. Studies of chipmunks (Tamias spp.) in Yosemite National Park provide an important opportunity to explore the reasons for this variation in response. While the alpine chipmunk (T. alpinus) has undergone a significant elevational range contraction over the past century, the congeneric and partially sympatric lodgepole chipmunk (T. speciosus) has not experienced an elevational range shift during this period. As a first step toward identifying the factors underlying this difference in response, we examined evidence for dietary changes and changes in cranial morphology in these species over the past century. Stable isotope analyses of fur samples from modern and historical museum specimens of these species collected at the same localities indicated that signatures of dietary change were more pronounced in T. alpinus, although diet breadth did not differ consistently between the study species. Morphometric analyses of crania from these specimens revealed significant changes in cranial shape for T. alpinus, with less pronounced changes in shape for T. speciosus; evidence of selection on skull morphology was detected for T. alpinus, but not for T. speciosus. These results are consistent with growing evidence that T. alpinus is generally more responsive to environmental change than T. speciosus, but emphasize the complex and often geographically variable nature of such responses. Accordingly, future studies that make use of the taxonomically and spatially integrative approach employed here may prove particularly informative regarding relationships between environmental conditions, range changes, and patterns of phenotypic variation.

Genetic Variation at Exon 2 of the MHC Class II DQB Locus in Blue Whale (Balaenoptera musculus) from the Gulf of California.

The genes of the Major Histocompatibility Complex (MHC) play an important role in the vertebrate immune response and are among the most polymorphic genes known in vertebrates. In some marine mammals, MHC genes have been shown to be characterized by low levels of polymorphism compared to terrestrial taxa; this reduction in variation is often explained as a result of lower pathogen pressures in marine habitats. To determine if this same reduction in variation applies to the migratory population of blue whales (Balaenoptera musculus) that occurs in the Gulf of California, we genotyped a 172 bp fragment of exon 2 of the MHC Class II DQB locus for 80 members of this population. Twenty-two putatively functional DQB allotypes were identified, all of which were homologous with DQB sequences from other cetacean species. Up to 5 putative alleles per individual were identified, suggesting that gene duplication has occurred at this locus. Rates of non-synonymous to synonymous substitutions (ω) and maximum likelihood analyses of models of nucleotide variation provided potential evidence of ongoing positive selection at this exon. Phylogenetic analyses of DQB alleles from B. musculus and 16 other species of cetaceans revealed trans-specific conservation of MHC variants, suggesting that selection has acted on this locus over prolonged periods of time. Collectively our findings reveal that immunogenic variation in blue whales is comparable to that in terrestrial mammals, thereby providing no evidence that marine taxa are subject to reduced pathogen-induced selective pressures.

Cranial morphological variation in Peromyscus maniculatus over nearly a century of environmental change in three areas of California.

Determining how species respond to prolonged environmental change is critical to understanding both their evolutionary biology and their conservation needs. In general, organisms can respond to changing environmental conditions by moving, by adapting in situ, or by going locally or globally extinct. Morphological changes, whether plastic or adaptive, are one way that species may respond in situ to local environmental change. Because cranial morphology is influenced by selective pressures arising from an organism's abiotic and biotic environments, including aspects of thermal physiology, diet, and sensory ecology, studies of cranial morphology may generate important insights into how species are responding to environmental change. To assess potential response of deer mice (Peromyscus maniculatus) to changing conditions in the Sierra Nevada Mountains of California, we quantified cranial variation in museum specimens of this species collected approximately 100 years apart. Specifically, we examined how cranial morphology varies in three populations of this geographically widespread, ecological generalist over elevation and time. Our analyses indicate that cranial morphology does not differ with elevation within either modern or historical samples but does vary between time periods, suggesting that in situ responses to environmental change have occurred. Contrary to predictions based on Bergmann's rule, we found no consistent relationship between body size and either elevation or time, suggesting that morphological differences detected between historic and modern specimens are specific to factors influencing cranial structure. Collectively, these analyses demonstrate the potential importance of in situ changes in morphology as a response to changing environmental conditions.

Taxon matters: promoting integrative studies of social behavior: NESCent Working Group on Integrative Models of Vertebrate Sociality: Evolution, Mechanisms, and Emergent Properties.

The neural and molecular mechanisms underlying social behavior - including their functional significance and evolution - can only be fully understood using data obtained under multiple social, environmental, and physiological conditions. Understanding the complexity of social behavior requires integration across levels of analysis in both laboratory and field settings. However, there is currently a disconnect between the systems studied in the laboratory versus the field. We argue that recent conceptual and technical advances provide exciting new opportunities to close this gap by making non-model organisms accessible to modern approaches in both laboratory and nature.

Contrasting stress responses of two co-occurring chipmunk species (Tamias alpinus and T. speciosus).

Glucocorticoid (GC) hormones are important mediators of responses to environmental conditions. Accordingly, differences in GC physiology may contribute to interspecific variation in response to anthropogenically-induced patterns of climate change. To begin exploring this possibility, we validated the use of fecal cortisol/corticosterone metabolites (FCM) to measure baseline glucocorticoid levels in two species of co-occurring chipmunks that have exhibited markedly different patterns of response to environmental change. In Yosemite National Park, the alpine chipmunk (Tamias alpinus) has undergone a significant upward contraction of its elevational range over the past century; in contrast, the lodgepole chipmunk (Tamiasspeciosus) has experienced no significant change in elevational distribution over this period. To determine if GC levels in these species vary in response to external stimuli and to assess whether these responses differ between species, we compared FCM levels for the same individuals (1) at the time of capture in the field, (2) after a short period of captivity, and (3) after adrenocorticotropic hormone (ACTH), (4) handling, and (5) trapping challenges conducted while these animals were held in captivity. Our analyses indicate that T. alpinus was more responsive to several of these changes in external conditions. Although both species displayed a significant FCM response to ACTH challenge, only T. alpinus showed a significant response to our handling challenge and to captive housing conditions. These findings underscore the importance of species-specific validation studies and support the potential for studies of GC physiology to generate insights into interspecific differences in response to environmental change.