Migratory singers dynamically overlap the signal space of a breeding warbler community.

Migratory species inhabit many communities along their migratory routes. Across taxa, these species repeatedly move into and out of communities, interacting with each other and locally breeding species and competing for resources and niche space. However, their influence is rarely considered in analyses of ecological processes within the communities they temporarily occupy. Here, we explore the impact of migratory species on a breeding community using the framework of acoustic signal space, a limited resource in which sounds of species within communities co-exist. Migrating New World warblers (Parulidae, hereafter referred to as migrant species) often sing during refueling stops in areas and at times during which locally breeding warbler species (hereafter breeding species) are singing to establish territories and attract mates. We used eBird data to determine the co-occurrence of 19 migrant and 11 breeding warbler species across spring migration in SW Michigan, generated a signal space from song recordings of these species, and examined patterns of signaling overlap experienced by breeding species as migrants moved through the community. Migrant species were present for two-thirds of the breeding season of local species, including periods when breeding species established territories and attracted mates. Signaling niche overlap experienced by individual breeding species was idiosyncratic and varied over time, yet niche overlap between migrant and breeding species occurred more commonly than between breeding species or between migrant species. Nevertheless, the proportion of niche overlap between migrant and breeding warblers was similar to overlap among breeding species. Our findings showed that singing by migrant species overlapped the signals of many breeding species, suggesting that migrants could have unexplored impacts on communication in breeding species, potentially affecting song detection and song evolution. Our study contributes to a growing body of research documenting the impacts of migratory species on communities and ecosystems.

Diet composition and diversity does not explain fewer, smaller urban nestlings.

The reproductive success of animals breeding in cities is often lower compared to counterparts that inhabit rural, suburban, and peri-urban areas. Urban dwelling may be especially costly for offspring development and survival. Diet composition and diversity may underlie factors that lead to lower fitness, particularly if prey abundance and quality decline in modified environments. Moreover, breeding success may change over the course of a season, an effect that may be augmented in urban areas. In this study, we tested the hypothesis that habitat and date affected nestling house wren (Troglodytes aedon) body condition and survival, and examined whether diet explained differences in nestling success. We monitored urban and rural populations of house wrens breeding in nest boxes, and tested whether clutch size, nestling survivorship, and nestling body condition varied by habitat or by date, and then characterized the diet of a subset of nestlings with DNA metabarcoding of fecal samples. Urbanization had clear impacts on house wren nestling fitness: urban broods contained fewer, smaller nestlings. Early nestling survival decreased as the breeding season progressed, and this effect was more pronounced in the urban population. However, the diets of urban and rural nestlings were similar and did not explain differences in body condition. Instead, across populations, diet changed with date, becoming less diverse, with fewer Lepidoptera and more Orthoptera. Regardless of habitat, adult house wrens provide nestlings with similar types of foods, but other factors, such as quantity or quality of prey delivered, may lead to fitness disparities between urban and rural nestlings.

Host gene expression in wildlife disease: making sense of species-level responses.

Emerging infectious diseases are significant threats to wildlife conservation, yet the impacts of pathogen exposure and infection can vary widely among host species. As such, conservation biologists and disease ecologists have increasingly aimed to understand species-specific host susceptibility using molecular methods. In particular, comparative gene expression assays have been used to contrast the transcriptomic responses of disease-resistant and disease-susceptible hosts to pathogen exposure. This work usually assumes that the gene expression responses of disease-resistant species will reveal the activation of molecular pathways contributing to host defence. However, results often show that disease-resistant hosts undergo little gene expression change following pathogen challenge. Here, we discuss the mechanistic implications of these "null" findings and offer methodological suggestions for future molecular studies of wildlife disease. First, we highlight that muted transcriptomic responses with minimal immune system recruitment may indeed be protective for nonsusceptible hosts if they limit immunopathology and promote pathogen tolerance in systems where susceptible hosts suffer from genetic dysregulation. Second, we argue that overly narrow investigation of responses to pathogen exposure may overlook important, constitutively active molecular pathways that underlie species-specific defences. Finally, we outline alternative study designs and approaches that complement interspecific transcriptomic comparisons, including intraspecific gene expression studies and genomic methods to detect signatures of selection. Collectively, these insights will help ecologists extract maximal information from conservation-relevant transcriptomic data sets, leading to a deeper understanding of host defences and, ultimately, the implementation of successful conservation interventions.

Host association influences variation at salivary protein genes in the bat ectoparasite Cimex adjunctus.

Parasite-host relationships create strong selection pressures that can lead to adaptation and increasing specialization of parasites to their hosts. Even in relatively loose host-parasite relationships, such as between generalist ectoparasites and their hosts, we may observe some degree of specialization of parasite populations to one of the multiple potential hosts. Salivary proteins are used by blood-feeding ectoparasites to prevent hemostasis in the host and maximize energy intake. We investigated the influence of association with specific host species on allele frequencies of salivary protein genes in Cimex adjunctus, a generalist blood-feeding ectoparasite of bats in North America. We analysed two salivary protein genes: an apyrase, which hydrolyses ATP at the feeding site and thus inhibits platelet aggregation, and a nitrophorin, which brings nitrous oxide to the feeding site, inhibiting platelet aggregation and vasoconstriction. We observed more variation at both salivary protein genes among parasite populations associated with different host species than among populations from different spatial locations associated with the same host species. The variation in salivary protein genes among populations on different host species was also greater than expected under a neutral scenario of genetic drift and gene flow. Finally, host species was an important predictor of allelic divergence in genotypes of individual C. adjunctus at both salivary protein genes. Our results suggest differing selection pressures on these two salivary protein genes in C. adjunctus depending on the host species.

Nestedness in assemblages of helminth parasites of bats: a function of geography, environment, or host nestedness?

Nested subsets occur in ecological communities when species-poor communities are subsets of larger, species-rich communities. Understanding this pattern can help elucidate species colonization abilities, extinction risks, and general structuring of biological communities. Here, we evaluate nestedness in a poorly studied host-parasite system, bats and their helminths, across the Japanese archipelago and within its different bioclimatic regions. We hypothesized that (1) if helminth communities are nested across geographic sites at the level of the archipelago, then broad-scale processes, like colonization-extinction dynamics, mainly structure parasite assemblages; (2) if helminth communities are nested across geographic sites at the level of the bioclimatic region, then fine-scale environmental variation plays a significant role in species nestedness; (3) if helminth community nestedness mirrors host species nestedness, then communities are nested because the habitats they occupy are nested; and (4) if nestedness does not occur or if it is not correlated with any geographical or host data, then passive sampling could be responsible for the patterns of parasite assemblage in our sample. We found that helminth communities were nested across host species throughout the archipelago but, when considering each bioclimatic region, helminths in only one region were significantly more nested than the null model. Helminth communities were also nested across sites within all four bioclimatic regions. These results suggest that helminths form nested subsets across the archipelago due to broad-scale processes that reflect the overall lineages of their mammalian hosts; however, at the regional scale, environmental processes related to nestedness of their habitats drive parasite community nestedness.

From individual heterogeneity to population-level overdispersion: quantifying the relative roles of host exposure and parasite establishment in driving aggregated helminth distributions.

In most host-parasite systems, variation in parasite burden among hosts drives transmission dynamics. Heavily infected individuals introduce disproportionate numbers of infective stages into host populations or surrounding environments, causing sharp increases in frequency of infection. Parasite aggregation within host populations may result from variation among hosts in exposure to infective propagules and probability of subsequent establishment of parasites in the host. This is because individual host heterogeneities contribute to a pattern of parasite overdispersion that emerges at the population level. We quantified relative roles of host exposure and parasite establishment in producing variation in parasite burdens, to predict which hosts are more likely to bear heavy burdens, using big brown bats (Eptesicus fuscus) and their helminths as a model system. We captured bats from seven colonies in Michigan and Indiana, USA, assessed their helminth burdens, and collected data on intrinsic and extrinsic variables related to exposure, establishment, or both. Digenetic trematodes had the highest prevalence and mean abundance while cestodes and nematodes had much lower prevalence and mean abundance. Structural equation modeling revealed that best-fitting models to explain variations in parasite burden included genetic heterozygosity and immunocompetence as well as distance to the nearest water source and the year of host capture. Thus, both differential host exposure and differential parasite establishment significantly influence heterogeneous helminth burdens, thus driving population-level patterns of parasite aggregation.

Prevalence and diversity of avian Haemosporida infecting songbirds in southwest Michigan.

Avian blood parasites from the genera Plasmodium, Haemoproteus, and Leucocytozoon (Haemosporida) affect hosts in numerous ways. They influence species interactions, host behavior, reproductive success, and cause pathology and mortality in birds. The Great Lakes region of North America has extensive aquatic and wetland habitat and supports a diverse vector community. Here we describe the community of bird-infecting Haemosporida in southwest Michigan and their host associations by measuring parasite prevalence, diversity, and host breadth across a diverse community of avian hosts. Over 700 songbirds of 55 species were screened for Haemosporida infection across southwest Michigan, including 11 species that were targeted for larger sample sizes. In total, 71 parasite lineages infected over 40% of birds. Of these, 42 were novel, yet richness estimates suggest that approximately half of the actual parasite diversity in the host community was observed despite intensive sampling of multiple host species. Parasite prevalence varied among parasite genera (7-24%) and target host species (0-85%), and parasite diversity was consistently high across most target species. Host breadth varied widely across the most prevalent parasite lineages, and we detected around 60% of host species richness for these parasite lineages. We report many new lineages and novel host-parasite associations, but substantial parasite diversity remains undiscovered in the Midwest.

Antibiotic resistant bacteria are widespread in songbirds across rural and urban environments.

The widespread use of antibiotics in human and veterinary medicine to treat pathogenic bacteria has resulted in the rapid emergence of antibiotic-resistant bacteria (ARB). Wild animals may enable the spread of pathogenic and non-pathogenic ARB when they are exposed to reservoirs (e.g., contaminated soil, water, or crops) and carry ARB in and on their bodies to other environments. We tested for the presence of ARB in four songbird species in southwest Michigan across a gradient of land use. Our specific objectives were to: 1) quantify the prevalence of ARB found in the gut microbiome of birds; 2) identify the specific bacteria exhibiting resistance; 3) assess whether ARB prevalence and identity varied among bird species; and 4) assess whether anthropogenic land use influenced the prevalence and identity of ARB found on birds. We sampled birds across a land use gradient consisting of urban, agricultural, and natural land covers using a randomized, spatially-balanced sampling design and cultured bacteria from fecal samples in the presence of three different antibiotics (amoxicillin, tetracycline, and ciprofloxacin). Overall prevalence of ARB was high, with 88% of total birds carrying ARB resistant to one of three antibiotics that we tested. Resistance to amoxicillin was more common (83% of sampled birds) than resistance to tetracycline (15%) or ciprofloxacin (1%). Identified ARB were diverse, and included 135 isolates representing 5 bacterial phyla and 22 genera. There was no effect of land use on ARB prevalence, with 90% of sampled birds captured in rural sites and 85% of sampled birds in urban sites carrying ARB. We provide the first analysis of ARB prevalence across multiple bird species and land uses utilizing a spatially-balanced, randomized study design. Our results demonstrate that nearly all sampled birds carried at least some ARB, and that they may serve as important dispersal agents of ARB across large spatial scales.

Comparative analysis of landscape effects on spatial genetic structure of the big brown bat and one of its cimicid ectoparasites.

Identification of landscape features that correlate with genetic structure permits understanding of factors that may influence gene flow in a species. Comparing effects of the landscape on a parasite and host provides potential insights into parasite-host ecology. We compared fine-scale spatial genetic structure between big brown bats (Eptesicus fuscus) and their cimicid ectoparasite (Cimex adjunctus; class Insecta) in the lower Great Lakes region of the United States, in an area of about 160,000 km2. We genotyped 142 big brown bat and 55 C. adjunctus samples at eight and seven microsatellite loci, respectively, and inferred effects of various types of land cover on the genetic structure of each species. We found significant associations between several land cover types and genetic distance in both species, although different land cover types were influential in each. Our results suggest that even in a parasite that is almost entirely reliant on its hosts for dispersal, land cover can affect gene flow differently than in the hosts, depending on key ecological aspects of both species.

Host association and selection on salivary protein genes in bed bugs and related blood-feeding ectoparasites.

Reciprocal selective pressures can drive coevolutionary changes in parasites and hosts, and result in parasites that are highly specialized to their hosts. Selection and host co-adaptation are better understood in endoparasites than in ectoparasites, whose life cycles may be more loosely linked to that of their hosts. Blood-feeding ectoparasites use salivary proteins to prevent haemostasis in the host, and maximize energy intake. Here we looked for signals of selection in salivary protein genes of ectoparasite species from a single genus (Cimex) that associate with a range of hosts including mammals (bats and humans) and birds (swallows). We analysed two genes that code for salivary proteins that inhibit platelet aggregation and vasoconstriction and may directly affect the efficiency of blood feeding in these species. Significant positive selection was detected at five codons in one gene in all bat-associated species groups. Our results suggest association with bats, versus humans or swallows, has posed a selective pressure on the salivary apyrase gene in species of Cimex.

Decomposing an urban soundscape to reveal patterns and drivers of variation in anthropogenic noise.

Continuous and intermittent noise may have different effects on humans and wildlife, therefore distinguishing temporal patterns of noise and their drivers is important for policy regarding both public health and wildlife management. We visualized patterns and explored land-use drivers of continuous and high-amplitude intermittent sound pressure levels (SPLs) on an urban campus in Michigan, U.S.A. To visualize patterns of SPLs, we introduce decibel duration curves (DDCs), which show the cumulative frequency distribution of SPLs and aid in the interpretation of statistical SPLs (Ln values) that reflect continuous versus intermittent sounds. DDCs and Ln values reveal that our 24 recording locations varied in the intensity of both continuous and intermittent noise, with intermittent high-amplitude sound events in particular contributing to variability in SPLs over the study site. Time of day influenced both continuous and intermittent SPLs, as locations relatively close to manmade structures (buildings, roads and parking lots) experienced higher SPLs as the day progressed. Continuous SPLs increased with decreasing distance to manmade structures, whereas intermittent SPLs increased with decreasing distance to roads and increasing distance to buildings. Thus, different land-use factors influenced patterns of continuous and intermittent noise, which suggests that different policy and strategies may be needed to ameliorate their effects on the public and wildlife.

Range-wide genetic structure and demographic history in the bat ectoparasite Cimex adjunctus.

BACKGROUND: Evolutionary histories of parasite and host populations are intimately linked such that their spatial genetic structures may be correlated. While these processes have been relatively well studied in specialist parasites and their hosts, less is known about the ecological and evolutionary consequences of relationships between generalist ectoparasites and their hosts. The aim of this study was to investigate the genetic structure and demographic history of a bat ectoparasite, Cimex adjunctus, whose host affinity is weak but the biology of the potential hosts have been well studied. This ectoparasite has been hypothesized to rely on its hosts for dispersal due to its low inherent dispersal potential. Here we describe genetic diversity and demographic history in C. adjunctus through most of its range in North America. We investigated variation at the cytochrome c oxidase 1 mitochondrial gene and nine microsatellite markers, and tested the prediction that genetic diversity in C. adjunctus is spatially structured. We also tested the prediction that demographic history in C. adjunctus is characterized by range and demographic expansion as a consequence of post-Pleistocene climate warming. RESULTS: We found stronger spatial structuring of genetic diversity in C. adjunctus than has been quantified in two of its hosts, but contrast in amount of variation explained by host association with different genetic markers (i.e., nuclear vs mitochondrial DNA). Also, C. adjunctus' history is not primarily characterized by demographic and range expansion, as is the case with two of its key hosts. CONCLUSIONS: Our study shows different patterns of genetic structure and demographic history in C. adjunctus than have been detected in two of its key hosts. Our results suggest an effect of a loose parasite-host relationship and anti-parasitism strategies on genetic structure and post-Pleistocene recovery of population size.

Relationships between host body condition and immunocompetence, not host sex, best predict parasite burden in a bat-helminth system.

Sex-biased parasitism highlights potentially divergent approaches to parasite resistance resulting in differing energetic trade-offs for males and females; however, trade-offs between immunity and self-maintenance could also depend on host body condition. We investigated these relationships in the big brown bat, Eptesicus fuscus, to determine if host sex or body condition better predicted parasite resistance, if testosterone levels predicted male parasite burdens, and if immune parameters could predict male testosterone levels. We found that male and female hosts had similar parasite burdens and female bats scored higher than males in only one immunological measure. Top models of helminth burden revealed interactions between body condition index and agglutination score as well as between agglutination score and host sex. Additionally, the strength of the relationships between sex, agglutination, and helminth burden is affected by body condition. Models of male parasite burden provided no support for testosterone predicting helminthiasis. Models that best predicted testosterone levels did not include parasite burden but instead consistently included month of capture and agglutination score. Thus, in our system, body condition was a more important predictor of immunity and worm burden than host sex.

Quantifying Global Drivers of Zoonotic Bat Viruses: A Process-Based Perspective.

Emerging infectious diseases (EIDs), particularly zoonoses, represent a significant threat to global health. Emergence is often driven by anthropogenic activity (e.g., travel, land use change). Although disease emergence frameworks suggest multiple steps from initial zoonotic transmission to human-to-human spread, there have been few attempts to empirically model specific steps. We create a process-based framework to separate out components of individual emergence steps. We focus on early emergence and expand the first step, zoonotic transmission, into processes of generation of pathogen richness, transmission opportunity, and establishment, each with its own hypothesized drivers. Using this structure, we build a spatial empirical model of these drivers, taking bat viruses shared with humans as a case study. We show that drivers of both viral richness (host diversity and climatic variability) and transmission opportunity (human population density, bushmeat hunting, and livestock production) are associated with virus sharing between humans and bats. We also show spatial heterogeneity between the global patterns of these two processes, suggesting that high-priority locations for pathogen discovery and surveillance in wildlife may not necessarily coincide with those for public health intervention. Finally, we offer direction for future studies of zoonotic EIDs by highlighting the importance of the processes underlying their emergence.

Range-Wide Genetic Analysis of Little Brown Bat (Myotis lucifugus) Populations: Estimating the Risk of Spread of White-Nose Syndrome.

The little brown bat (Myotis lucifugus) is one of the most widespread bat species in North America and is experiencing severe population declines because of an emerging fungal disease, white-nose syndrome (WNS). To manage and conserve this species effectively it is important to understand patterns of gene flow and population connectivity to identify possible barriers to disease transmission. However, little is known about the population genetic structure of little brown bats, and to date, no studies have investigated population structure across their entire range. We examined mitochondrial DNA and nuclear microsatellites in 637 little brown bats (including all currently recognized subspecific lineages) from 29 locations across North America, to assess levels of genetic variation and population differentiation across the range of the species, including areas affected by WNS and those currently unaffected. We identified considerable spatial variation in patterns of female dispersal and significant genetic variation between populations in eastern versus western portions of the range. Overall levels of nuclear genetic differentiation were low, and there is no evidence for any major barriers to gene flow across their range. However, patterns of mtDNA differentiation are highly variable, with high ΦST values between most sample pairs (including between all western samples, between western and eastern samples, and between some eastern samples), while low mitochondrial differentiation was observed within two groups of samples found in central and eastern regions of North America. Furthermore, the Alaskan population was highly differentiated from all others, and western populations were characterized by isolation by distance while eastern populations were not. These data raise the possibility that the current patterns of spread of WNS observed in eastern North America may not apply to the entire range and that there may be broad-scale spatial variation in the risk of WNS transmission and occurrence if the disease continues to spread west.

Two Tickets to Paradise: Multiple Dispersal Events in the Founding of Hoary Bat Populations in Hawai'i.

The Hawaiian islands are an extremely isolated oceanic archipelago, and their fauna has long served as models of dispersal in island biogeography. While molecular data have recently been applied to investigate the timing and origin of dispersal events for several animal groups including birds, insects, and snails, these questions have been largely unaddressed in Hawai'i's only native terrestrial mammal, the Hawaiian hoary bat, Lasiurus cinereus semotus. Here, we use molecular data to test the hypotheses that (1) Hawaiian L. c. semotus originated via dispersal from North American populations of L. c. cinereus rather than from South American L. c. villosissimus, and (2) modern Hawaiian populations were founded from a single dispersal event. Contrary to the latter hypothesis, our mitochondrial data support a biogeographic history of multiple, relatively recent dispersals of hoary bats from North America to the Hawaiian islands. Coalescent demographic analyses of multilocus data suggest that modern populations of Hawaiian hoary bats were founded no more than 10 kya. Our finding of multiple evolutionarily significant units in Hawai'i highlights information that should be useful for re-evaluation of the conservation status of hoary bats in Hawai'i.

Genetic approaches to the conservation of migratory bats: a study of the eastern red bat (Lasiurus borealis).

Documented fatalities of bats at wind turbines have raised serious concerns about the future impacts of increased wind power development on populations of migratory bat species. However, for most bat species we have no knowledge of the size of populations and their demographic trends, the degree of structuring into discrete subpopulations, and whether different subpopulations use spatially segregated migratory routes. Here, we utilize genetic data from eastern red bats (Lasiurus borealis), one of the species most highly affected by wind power development in North America, to (1) evaluate patterns of population structure across the landscape, (2) estimate effective population size (Ne ), and (3) assess signals of growth or decline in population size. Using data on both nuclear and mitochondrial DNA variation, we demonstrate that this species forms a single, panmictic population across their range with no evidence for the historical use of divergent migratory pathways by any portion of the population. Further, using coalescent estimates we estimate that the effective size of this population is in the hundreds of thousands to millions of individuals. The high levels of gene flow and connectivity across the population of eastern red bats indicate that monitoring and management of eastern red bats must integrate information across the range of this species.

Genetic structure of little brown bats (Myotis lucifugus) corresponds with spread of white-nose syndrome among hibernacula.

Until recently, the little brown bat (Myotis lucifugus) was one of the most common bat species in North America. However, this species currently faces a significant threat from the emerging fungal disease white-nose syndrome (WNS). The aims of this study were to examine the population genetic structure of M. lucifugus hibernating colonies in Pennsylvania (PA) and West Virginia (WV), and to determine whether that population structure may have influenced the pattern of spread of WNS. Samples were obtained from 198 individuals from both uninfected and recently infected colonies located at the crest of the disease front. Both mitochondrial (636bp of cytochrome oxidase I) and nuclear (8 microsatellites) loci were examined. Although no substructure was evident from nuclear DNA, female-mediated gene flow was restricted between hibernacula in western PA and the remaining colonies in eastern and central PA and WV. This mitochondrial genetic structure mirrors topographic variation across the region: 3 hibernating colonies located on the western Appalachian plateau were significantly differentiated from colonies located in the central mountainous and eastern lowland regions, suggesting reduced gene flow between these clusters of colonies. Consistent with the hypothesis that WNS is transmitted primarily through bat-to-bat contact, these same 3 hibernating colonies in westernmost PA remained WNS-free for 1-2 years after the disease had swept through the rest of the state, suggesting that female migration patterns may influence the spread of WNS across the landscape.

Host phylogeny constrains cross-species emergence and establishment of rabies virus in bats.

For RNA viruses, rapid viral evolution and the biological similarity of closely related host species have been proposed as key determinants of the occurrence and long-term outcome of cross-species transmission. Using a data set of hundreds of rabies viruses sampled from 23 North American bat species, we present a general framework to quantify per capita rates of cross-species transmission and reconstruct historical patterns of viral establishment in new host species using molecular sequence data. These estimates demonstrate diminishing frequencies of both cross-species transmission and host shifts with increasing phylogenetic distance between bat species. Evolutionary constraints on viral host range indicate that host species barriers may trump the intrinsic mutability of RNA viruses in determining the fate of emerging host-virus interactions.

Social calls used by a leaf-roosting bat to signal location.

Social calls in bats have many functions, including mate attraction and maintaining contact during flight. Research suggests that social calls may also be used to transfer information about roosts, but no studies have yet demonstrated that calls are used to actively attract conspecifics to roosting locations. We document the social calls used by Spix's disc-winged bat (Thyroptera tricolor) to actively recruit group members to roosts. In acoustic trials, we recorded two sets of calls; one from flying individuals termed 'inquiry calls', and another from roosting bats termed 'response calls'. Inquiry calls were emitted by flying bats immediately upon release, and quickly (i.e. 178 ms) elicited production of response calls from roosting individuals. Most flying bats entered the roost when roosting individuals responded, while few bats entered the roost in the absence of a response. We argue that information transfer concerning roost location may facilitate sociality in T. tricolor, given the ephemeral nature of roosting structures used by this species.