Adaptive Evolution of Nearctic Deepwater Fish Vision: Implications for Assessing Functional Variation for Conservation.

Intraspecific functional variation is critical for adaptation to rapidly changing environments. For visual opsins, functional variation can be characterized in vitro and often reflects a species' ecological niche but is rarely considered in the context of intraspecific variation or the impact of recent environmental changes on species of cultural or commercial significance. Investigation of adaptation in postglacial lakes can provide key insight into how rapid environmental changes impact functional evolution. Here, we report evidence for molecular adaptation in vision in 2 lineages of Nearctic fishes that are deep lake specialists: ciscoes and deepwater sculpin. We found depth-related variation in the dim-light visual pigment rhodopsin that evolved convergently in these 2 lineages. In vitro characterization of spectral sensitivity of the convergent deepwater rhodopsin alleles revealed blue-shifts compared with other more widely distributed alleles. These blue-shifted rhodopsin alleles were only observed in deep clear postglacial lakes with underwater visual environments enriched in blue light. This provides evidence of remarkably rapid and convergent visual adaptation and intraspecific functional variation in rhodopsin. Intraspecific functional variation has important implications for conservation, and these fishes are of conservation concern and great cultural, commercial, and nutritional importance to Indigenous communities. We collaborated with the Saugeen Ojibway Nation to develop and test a metabarcoding approach that we show is efficient and accurate in recovering the ecological distribution of functionally relevant variation in rhodopsin. Our approach bridges experimental analyses of protein function and genetics-based tools used in large-scale surveys to better understand the ecological extent of adaptive functional variation.

First phylogenetic analysis of the Nearctic madicolous midges of the genus Androprosopa Mik (Diptera: Thaumaleidae).

Molecular phylogenetic analyses were conducted to infer relationships between the eastern and western Nearctic Androprosopa Mik and amongst the considerably more diverse western Nearctic species. Fresh, molecular-grade material was obtained for all Nearctic Androprosopa species except two Mexican species, An. sonorensis (Arnaud & Boussy) and An. zempoala Sinclair & Huerta, that eluded capture. Molecular sequences from two nuclear protein-coding genes, big zinc finger (BZF) and molybdenum cofactor sulfurase (MCS), were sampled from representatives of several outgroup and ingroup taxa and analyzed phylogenetically using maximum likelihood criteria to confirm identifications of females and immatures using a barcoding approach, test species boundaries among morphologically similar species, and infer relationships among more morphologically disparate groups. Resulting phylogenies suggest the following with significant node (bootstrap) support: (1) the eastern Nearctic Androprosopa species form the sister group to the lineage comprised of all sampled Palearctic thaumaleids, i.e., An. larvata (Mik), An. striata (Okada), and Thaumalea testacea Ruthe; (2) the aforementioned lineage is the sister group to the clade comprised of western Nearctic Androprosopa species; (3) the western Nearctic Androprosopa species form three multispecies lineages, two of which can be further divided into three or more well founded species groups. Our results suggest that Androprosopa as currently defined is paraphyletic. Additionally, we propose several new species groups within the western Nearctic Androprosopa based on molecular and morphological data.

Formation of the Holarctic Fauna: Dated molecular phylogenetic and biogeographic insights from the Quedius-lineage of Ground-Dwelling Rove Beetles (Coleoptera, Staphylinidae).

Although the Holarctic fauna has been explored for centuries, many questions on its formation are still unanswered. For example, i) what was the impact of the uplift of the Himalaya and Tibetan Plateau?, ii) what were the timings and climate of the faunal bridges connecting the Nearctic and Palearctic regions?, and iii) how did insect lineages respond to the late Paleogene global cooling and regional aridification? To answer these, we developed a phylogenetic dataset of 1229 nuclear loci for a total of 222 species of rove beetles (Staphylinidae) with emphasis in the tribe Quediini, especially Quedius-lineage and its subclade Quedius sensu stricto. Using eight fossils for calibrating molecular clock, we estimated divergence times and then analysed in BioGeoBEARS paleodistributions of the most recent common ancestor for each target lineage. For each species we generated climatic envelopes of the temperature and precipitation and mapped them across the phylogeny to explore evolutionary shifts. Our results suggest that the warm and humid Himalaya and Tibetan Plateau acted as an evolutionary cradle for the Quedius-lineage originating during the Oligocene from where, in the Early Miocene, the ancestor of the Quedius s. str. dispersed into the West Palearctic. With the climate cooling from the Mid Miocene onwards, new lineages within Quedius s. str. emerged and gradually expanded distributions across the Palearctic. In Late Miocene, a member of the group dispersed to the Nearctic region via Beringia before the closure of this land bridge 5.3 Ma. Paleogene global cooling and regional aridification largely shaped the current biogeographic pattern for Quedius s. str. species, many of them originating during the Pliocene and shifting or contracting their ranges during Pleistocene.

Tapping into natural history collections to assess latitudinal gradients of parasite diversity.

Parasites are key components of the biosphere not only due to their huge diversity, but also because they exert important influences on ecological processes. Nevertheless, we lack an understanding of the biogeographical patterns of parasite diversity. Here, we tap into the potential of biodiversity collections for understanding parasite biogeography. We assess species richness of supracommunities of helminth parasites infecting mammal assemblages in the Nearctic, and describe its relation to latitude, climate, host diversity, and land area. We compiled data from parasitology collections and assessed parasite diversity in Nearctic ecoregions for the entire parasite supracommunity of mammals in each ecoregion, as well as separately from carnivores and rodents to explore the effect of host taxonomic resolution on observed patterns. For carnivores, we found evidence of a negative latitudinal gradient, while parasites of rodents displayed no clear pattern. We found that parasite diversity was positively correlated with mean annual temperature and negatively correlated with seasonal precipitation. Parasite richness shows a diversity peak at intermediate host richness values and in carnivores correlates with temperature and seasonal precipitation. Rodent parasite diversity did not correlate with explored factors. Other researchers are encouraged to use parasitology collections to continue exploring patterns of parasite biogeography and macroecology.

First report of Orthostrongylus sp. (Nematoda: Protostrongylidae) in wild reindeer (Rangifer tarandus) from the Taimyr, Russia: Nearctic parasites in a Palearctic host.

First stage larvae of an unknown lungworm (Protostrongylidae) were isolated in the feces of a wild reindeer (Rangifer tarandus) from Taimyr, Russia. Larvae were 365-366 µm long and had a tail spike lacking a dorsal spine. DNA analyses using BLAST showed that nuclear sequences obtained (LSU rDNA, 825 bp and ITS2 rDNA, 395 bp) were highly similar (99.50% and 98.88% identity, respectively) to an isolate of Orthostrongylus macrotis (GenBank: EU595592.1) from North America. It cannot be confirmed whether these larvae represent an uncharacterized species of Orthostrongylus or can be referred to O. macrotis, a species that has historically only been reported from the Nearctic. This is the first report of lungworms attributable to Protostrongylinae in R. tarandus across its vast geographic in the Holarctic.

Community modeling reveals the importance of elevation and land cover in shaping migratory bird abundance in the Andes.

The tropical Andes are characterized by extreme topographic and climatic complexity, which has likely contributed to their outstanding current species diversity, composed of many range-restricted species. However, little is known about how the distribution and abundance of highly mobile organisms, like long-distance migratory birds, varies across different land covers, elevations, and climatic conditions within the Andes. We conducted 1,606 distance-sampling point counts across the Colombian Andes, spanning elevations from 253 to 3,708 m, a range of precipitation regimes and representative land covers. We then employed a novel application of a multispecies hierarchical modeling approach to evaluate how elevation, local land cover, aboveground woody biomass, cloud cover, precipitation, and seasonality in precipitation shape the abundance of the migratory land bird community in the Andes. We detected 1,824 individuals of 29 species of migratory land birds, six of which were considered incidental in our study region. We modeled the abundance of the remaining 23 species, while considering observer and time of day effects on detectability. We found that both elevation and land cover had an overriding influence on the abundance of migratory species across the Andes, with strong evidence for a mid-elevation peak in abundance, and species-specific responses to both variables. As a community, migratory birds had the highest mean abundance in shade coffee plantations, secondary forest, and mature forest. Aboveground woody biomass did not affect the abundance of all species as a group, but a few showed strong responses to this variable. Contrary to predictions of a positive correlation between abundance and precipitation, we found no evidence for community-level responses to precipitation, aside for a weak tendency for birds to select areas with intermediate levels of precipitation. This novel use of a multispecies model sheds new light on the mechanisms shaping the winter distribution of migratory birds and highlights the importance of elevation and land cover types over climatic variables in the context of the Colombian Andes.

Migration tactics and connectivity of a Nearctic-Neotropical migratory shorebird.

During long-distance spring migrations, birds may rest and refuel at numerous stopover sites while minimizing the time to reach the breeding grounds. If habitat is limited along the migration route, pre-breeding birds optimize flight range by having longer stopovers at higher quality sites compared to poorer quality sites. Stopover duration also depends on distance remaining to breeding grounds, ecological barriers and individual characteristics. We assessed spring migration tactics and connectivity of a Nearctic-Neotropical migratory shorebird, the semipalmated sandpiper Calidris pusilla, at two sites with known relative habitat quality on the northern Gulf of Mexico (NGOM) coast, the first land encountered after crossing the Gulf of Mexico (GOM). We used automated radio telemetry (Motus) to estimate stopover duration and probability of departure. Migration speed was estimated for individuals detected at subsequent receivers on the Motus network. To measure migratory connectivity, we used morphometrics and the Motus network to assign general breeding regions. Additionally, feather stable isotope ratios of C and N provided coarse information about overwintering regions. Stopover duration declined with higher fuel loads at capture as expected under a time-minimizing strategy. After accounting for fuel load, stopover duration was approximately 40% longer at the higher quality site. We found no detectable effect of age, sex or breeding location on stopover behaviour. The probability of departure was strongly affected by humidity and also by tailwind and weather conditions. Birds stopping at the higher quality site had earlier apparent arrival to the breeding grounds. The Louisiana coast is an apparent stopover hub for this species, since the individuals were departing to range-wide breeding regions and isotope values suggested birds were also using widespread wintering regions. Our study shows how high-quality, coastal wetlands along the NGOM coast serve a critical role in the annual cycle of a migratory shorebird. Stopover behaviour indicated that high-quality habitat may be limited for this species during spring migration. As threats to the GOM coast increase, protection of these already limited wetlands is vitally important.

Assessing the genetic diversity of the metacercariae of Posthodiplostomum minimum (Trematoda: Diplostomidae) in Middle American freshwater fishes: one species or more?

Trematode taxonomy is mainly based on the morphological traits of adults. The identification of metacercariae is challenging because such traits are not developed in larval forms, and they even may show some level of morphological variability. Studies testing the potential correspondence between morphological differences and genetic variation of parasites are still lacking. The metacercariae of Posthodiplostomum minimum are probably the diplostomids more widely distributed in North and Middle American freshwater fish, and their intraspecific morphological variability has been attributed to the effect exerted by the host. Here, we tested the hypothesis whether they represent a single species, or a species complex by assessing the genetic divergence and phylogenetic relationships of metacercariae sampled from several host species in a wide geographical range across Middle America. The internal transcribed spacers (ITS1-5.8S-ITS2), and the mitochondrial COI gene were sequenced for 124 and 55 metacercariae, respectively. Phylogenetic analysis inferred from ITS sequences uncovered six well-supported monophyletic lineages. The six lineages show no correspondence to any Posthodiplostomum species for which sequences are available thus far in GenBank. Lineages exhibit some degree of host specificity; Lineages I, II, IV and V are primarily parasites of cyprinodontiforms of the families Poeciliidae, Goodeidae, Profundulidae and Fundulidae. In poeciliids there are at least four candidate species of Posthodiplostomum, some of them occurring in sympatry; instead, Lineages II and VI are exclusively parasites of cichlids. This study contributes to our understanding of the diversity of larval forms of diplostomids and provides an opportunity to further study their life cycles.

A place to land: spatiotemporal drivers of stopover habitat use by migrating birds.

Migrating birds require en route habitats to rest and refuel. Yet, habitat use has never been integrated with passage to understand the factors that determine where and when birds stopover during spring and autumn migration. Here, we introduce the stopover-to-passage ratio (SPR), the percentage of passage migrants that stop in an area, and use 8 years of data from 12 weather surveillance radars to estimate over 50% SPR during spring and autumn through the Gulf of Mexico and Atlantic coasts of the south-eastern US, the most prominent corridor for North America's migratory birds. During stopovers, birds concentrated close to the coast during spring and inland in forested landscapes during autumn, suggesting seasonal differences in habitat function and highlighting the vital role of stopover habitats in sustaining migratory communities. Beyond advancing understanding of migration ecology, SPR will facilitate conservation through identification of sites that are disproportionally selected for stopover by migrating birds.

Contrasting migratory journeys and changes in hippocampal astrocyte morphology in shorebirds.

Semipalmated sandpiper (Calidris pusilla) migration to the Southern Hemisphere includes a 5-day non-stop flight over the Atlantic Ocean, whereas semipalmated plover (Charadrius semipalmatus) migration, to the same area, is largely over land, with stopovers for feeding and rest. We compared the number and 3D morphology of hippocampal astrocytes of Ch. semipalmatus before and after autumnal migration with those of C. pusilla to test the hypothesis that the contrasting migratory flights of these species could differentially shape hippocampal astrocyte number and morphology. We captured individuals from both species in the Bay of Fundy (Canada) and in the coastal region of Bragança (Brazil) and processed their brains for selective GFAP immunolabeling of astrocytes. Hierarchical cluster analysis of astrocyte morphological features distinguished two families of morphological phenotypes, named type I and type II, which were differentially affected after migratory flights. Stereological counts of hippocampal astrocytes demonstrated that the number of astrocytes decreased significantly in C. pusilla, but did not change in Ch. semipalmatus. In addition, C. pusilla and Ch. semipalmatus hippocampal astrocyte morphological features were differentially affected after autumnal migration. We evaluated whether astrocyte morphometric variables were influenced by phylogenetic differences between C. pusilla and Ch. semipalmatus, using phylogenetically independent contrast approach, and phylogenetic trees generated by nuclear and mitochondrial markers. Our findings suggest that phylogenetic differences do not explain the results and that contrasting long-distance migratory flights shape plasticity of type I and type II astrocytes in different ways, which may imply distinct physiological roles for these cells.

Repeated sampling of individuals reveals impact of tropical and temperate habitats on microbiota of a migratory bird.

Migratory animals experiencing substantial change in diet and habitat across the annual cycle may have corresponding shifts in host-associated microbial diversity. Using automated telemetry and radio tags to recapture birds, we examined gut microbiota structure in the same population and often same individual of Kirtland's Warblers (Setophaga kirtlandii) initially sampled on their wintering grounds in The Bahamas and subsequently resampled within their breeding territories in Michigan, USA. Initial sampling occurred in March and April and resampling occurred in May, June and early July. The composition of the most abundant phyla and classes of the warblers' microbiota is similar to that of other migratory birds. However, we detected notable variation in abundance and diversity of numerous bacterial taxa, including a decrease in microbial richness and significant differences in microbial communities when comparing the microbiota of birds first captured in The Bahamas to that of birds recaptured in Michigan. This is observed at the individual and population level. Furthermore, we found that 22 bacterial genera exhibit heightened abundance within specific sampling periods and are probably associated with diet and environmental change. Finally, we described a small, species-specific shared microbial profile that spans multiple time periods and environments within the migratory cycle. Our research highlights that the avian gut microbiota is dynamic over time, most significantly impacted by changing environments associated with migration. These results support the need for full annual cycle monitoring of migratory bird microbiota to improve understanding of seasonal host movement ecologies and response to recurrent physiological stressors.

Molecular evidence of three closely related species of Biacetabulum Hunter, 1927 (Cestoda: Caryophyllidea): a case of recent speciation in different fish hosts (Catostomidae)?

Monozoic tapeworms (Caryophyllidea) are dominant components of parasite communities of suckers (Catostomidae) in North America, with Biacetabulum Hunter, 1927 representing one of the more species-rich genera. Molecular (28S rDNA) and morphological (including scanning electron microscopy and histology) evaluation of newly collected tapeworms from different fish hosts revealed the existence of four similar (and three closely related) species of Biacetabulum. These four species differ from their congeners by having a long body (up to 48 mm long) with a very long, slender neck (its length represents ≥30% of total body length), a large, globular scolex with a prominent central acetabulum-like loculus on the dorsal and ventral sides, two pairs of shallow lateral loculi and a distinct, slightly convex apical disc, and a cirrus-sac that is situated between the anterior arms of the ovarian wings. Taken together, the morphological and molecular data and the host associations of these species provide evidence of their host specificity. Biacetabulum isaureae n. sp. occurs in notch clip redhorse, Moxostoma collapsum, in South Carolina (USA), B. longicollum n. sp. in silver redhorse, Moxostoma anisurum (type host), and golden redhorse, M. erythrurum, in Manitoba (Canada) and West Virginia (USA), B. overstreeti n. sp. in a spotted sucker, Minytrema melanops, in Mississippi, and B. hypentelii n. sp. in northern hogsucker, Hypentelium nigricans, in Tennessee (USA). The new species differ from each other in the number of postovarian vitelline follicles, the posterior extent of preovarian vitelline follicles and relative size of the cirrus sac.

Disentangling taxonomy of Biacetabulum (Cestoda, Caryophyllidea), parasites of catostomid fishes in North America: proposal of Megancestus gen. n. to accommodate B. carpiodi.

A new genus, Megancestus n. gen., is proposed to accommodate the caryophyllidean tapeworm Biacetabulum carpiodi Mackiewicz, 1969 from carpsuckers and quillback (Carpiodes spp.) in North America. This species is not closely related to other species of Biacetabulum Hunter, 1927 and is transferred to a newly erected genus. This new genus is typified by the possession of a small body (total length of 3.1-7.5 mm) with a scolex that bears a pair of large acetabulum-like loculi, two pairs of shallow lateral loculi, and a slightly convex apical disc, testes arranged in one or two layers, oval, thick-walled cirrus-sac, well-developed external seminal vesicle, separate gonopores, H-shaped ovary, few median vitelline follicles, and the uterus extending by a single loop anterior to the cirrus-sac. Megancestus differs from all Nearctic caryophyllidean genera (family Capingentidae), including Biacetabulum, by vitelline follicles dorsal to the ovary that connect the preovarian and postovarian vitelline fields. The most closely related Hunterella Mackiewicz et McCrae, 1962 differs by shape of the scolex (tholate, i.e., devoid of any loculi), dumbbell-shaped ovary and the uterus not extending anterior to the cirrus-sac. Megancestus carpiodi (Mackiewicz, 1969) n. comb. is the only species of the genus and it is a stenoxenous parasite, which has been found only in the river carpsucker (Carpiodes carpio-type host), quillback (Carpiodes cyprinus) and highfin carpsucker (Carpiodes velifer) (Catostomidae: Ictiobinae) in the lower and middle Mississippi basin.

Pattern of climate connectivity and equivalent niche of Triatominae species of the Phyllosoma complex.

The Phyllosoma complex is a Triatominae (Hemiptera: Reduviidae) group of medical importance involved in Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) transmission. Most of the members of this group are endemic and sympatric species with distribution in Mexico and the southern U.S.A. We employed MaxEnt to construct ecological niche models of nine species of Triatominae to test three hypothesis: (a) whether species with a broad climatic niche breadth occupy a broader geographical range than species with a narrow climatic breadth, (b) whether species with broad distribution present high degree of climatic fragmentation/isolation, which was tested through landscape metrics; and (c) whether the species share the same climatic niche space (niche conservatism) considered through an equivalence test implemented in ENMtools. Overall, our results suggest that the geographical distribution of this complex is influenced mainly by temperature seasonality where all suitable areas are places of current and potential transmission of T. cruzi. Niche breadth in the Phyllosoma complex is associated with the geographical distribution range, and the geographical range affects the climatic connectivity. We found no strong evidence of niche climatic divergence in members of this complex. We discuss the epidemiological implications of these results.

Patterns of blood mercury variation in two long-distance migratory thrushes on Mount Mansfield, Vermont.

We investigated mercury (Hg) blood concentrations in Bicknell's thrush (Catharus bicknelli) and Swainson's thrush (C. ustulatus), congeneric long-distance migratory songbirds, from 2000-2017 at a montane forest site in north-central Vermont. We analyzed variation in blood Hg of both species using mixed-effects models, incorporating atmospheric wet Hg deposition data from a nearby sampling location. Although Hg deposition varied among years and seasonally, we detected no temporal trend in either atmospheric deposition or blood Hg, nor evidence of a relationship between the two. Sampling date had the strongest effect on blood Hg concentration, which declined seasonally, followed by age and sex of the individual. The data did not support an effect of species. We believe that the absence of a clear relationship between local atmospheric deposition and thrush blood Hg concentrations suggests that Hg cycling dynamics, mechanisms of transfer, and timing of uptake by montane forest biota are complex and poorly understood. The blood Hg concentrations of ~0.07-0.1 µg/g we documented in Bicknell's and Swainson's thrush are below those found to negatively impact physiological or reproductive endpoints in other invertivorous terrestrial passerines. To better evaluate the validity of Bicknell's thrush as a bioindicator of MeHg availability in montane forest ecosystems, we recommend (1) effects-based investigations, (2) a more robust understanding of Hg and MeHg cycling, (3) more clear geospatial and temporal links between Hg deposition and biotic uptake, and (4) more thorough documentation of Hg burdens across the species' annual cycle.

Phylogeography of higher Diptera in glacial and postglacial grasslands in western North America.

BACKGROUND: Pleistocene glaciations have had an important impact on the species distribution and community composition of the North American biota. Species survived these glacial cycles south of the ice sheets and/or in other refugia, such as Beringia. In this study, we assessed, using mitochondrial DNA from three Diptera species, whether flies currently found in Beringian grasslands (1) survived glaciation as disjunct populations in Beringia and in the southern refugium; (2) dispersed northward postglacially from the southern refugium; or (3) arose by a combination of the two. Samples were collected in grasslands in western Canada: Prairies in Alberta and Manitoba; the Peace River region (Alberta); and the southern Yukon Territory. We sequenced two gene regions (658 bp of cytochrome c oxidase subunit I, 510 bp of cytochrome b) from three species of higher Diptera: one with a continuous distribution across grassland regions, and two with disjunct populations between the regions. We used a Bayesian approach to determine population groupings without a priori assumptions and performed analysis of molecular variance (AMOVA) and exact tests of population differentiation (ETPD) to examine their validity. Molecular dating was used to establish divergence times. RESULTS: Two geographically structured populations were found for all species: a southern Prairie and Peace River population, and a Yukon population. Although AMOVA did not show significant differentiation between populations, ETPD did. Divergence time between Yukon and southern populations predated the Holocene for two species; the species with an ambiguous divergence time had high haplotype diversity, which could suggest survival in a Beringian refugium. CONCLUSIONS: Populations of Diptera in Yukon grasslands could have persisted in steppe habitats in Beringia through Pleistocene glaciations. Current populations in the region appear to be a mix of Beringian relict populations and, to a lesser extent, postglacial dispersal northward from southern prairie grasslands.

Up high and down low: Molecular systematics and insight into the diversification of the ground beetle genus Rhadine LeConte.

Rhadine LeConte is a Nearctic genus of flightless ground beetles that is poorly studied despite its relevance to evolutionary studies of subterranean fauna. Adults are notable for their slender and leggy habitus and the wide variety of habitat preferences among species, with several known only from mountaintops while others are restricted to caves or more general subterranean habitats. In central Texas, USA there are several cave endemics relevant to conservation. Here we present the first phylogenetic hypothesis for the overall structure of the genus with an emphasis on the troglobites in central Texas. We infer the phylogeny of Rhadine from ∼2.4-kb of aligned nucleotide sites from the nuclear genes, 28S rDNA and CAD, and the mitochondrial gene COI. These data were obtained for 30 species of Rhadine as well as from members of their putative sister group, Tanystoma Motschulsky. Results reveal that Rhadine is polyphyletic, and morphological characters that have been traditionally used to classify the genus into species groups are shown to be convergent in many cases. Rhadine aside from two species of uncertain placement is composed of two major clades, Clades I and II that both include epigean and subterranean species in very unequal proportions. Clade I is primarily composed of subterranean species, and Clade II includes many epigean species and high altitude montane endemics. A clade of troglobitic, cave-restricted species in Texas includes several species of large-eyed cave Rhadine. The slender habitus typical of some species [e.g., R. exilis (Barr and Lawrence), R. subterranea (Van Dyke), R. austinica Barr] evolved independently at least three times. Major biogeographic and evolutionary patterns based on these results include: troglobitic species north of the Colorado River in Texas (that also lack lateral pronotal setae) are found to comprise a monophyletic group, beetles in caves south of the Colorado River likely form another monophyletic group, and the "species pairs" of troglobitic Rhadine known to occur in the same caves are not resolved as each other's sister group, suggesting that these caves were colonized independently by more than one lineage of Rhadine. Our divergence time estimates support a Miocene age for the split between Clade I and II Rhadine and indicate that all subterranean Clade I Rhadine began diversifying in the late Miocene-early Pliocene, contemporary with cave formation in the Balcones Escarpment.

A synoptic review of Promonobothrium Mackiewicz, 1968 (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America, with description of two new species.

Monozoic cestodes of the recently amended genus Promonobothrium Mackiewicz, 1968 (Cestoda: Caryophyllidea), parasites of suckers (Cypriniformes: Catostomidae) in North America, are reviewed, with information on their host specificity, distribution and data on the scolex morphology of seven species studied for the first time using scanning electron microscopy (SEM). Evaluation of type and voucher specimens from museum collections and newly collected material of most species indicated the following valid nominal species: Promonobothrium minytremi Mackiewicz, 1968 (type species); P. ingens (Hunter, 1927); P. hunteri (Mackiewicz, 1963); P. ulmeri (Calentine et Mackiewicz, 1966); P. fossae (Williams, 1974) and P. mackiewiczi (Williams, 1974). Rogersus Williams, 1980 with its only species R. rogersi is transferred to Promonobothrium based on morphological and molecular data. Promonobothrium currani sp. n. and P. papiliovarium sp. n. are described from Ictiobus bubalus (Rafinesque) and Ictiobus niger (Rafinesque), and Erimyzon oblongus (Mitchill), respectively. The newly described species can be distinguished from the other congeners by the morphology of the scolex, the position of the anteriormost vitelline follicles and testes, the presence of postovarian vitelline follicles and the shape of the ovary. Molecular phylogenetic analyses of six species based on sequences of the small and large subunits of the nuclear ribosomal RNA genes (ssrDNA, lsrDNA) confirmed the monophyletic status of the genus and supported the validity of the species analysed. A key to identification of all species of Promonobothrium based on morphological characteristics is provided.

A New Species of Culicoides (Selfia) (Diptera: Ceratopogonidae) from Southeastern Utah.

A new species of biting midge, Culicoides (Selfia) moabensis, is described and illustrated from southeastern Utah. Its relationship to Culicoides (Selfia) multipunctatus Malloch and Culicoides (Selfia) brookmani Wirth is discussed, and modifications to existing keys to adult males and females of C. (Selfia) species are provided. Its abundance, seasonal distribution, and aspects of its reproductive and feeding biology and potential as an arbovirus vector are discussed.

Extensive gene flow characterizes the phylogeography of a North American migrant bird: Black-headed Grosbeak (Pheucticus melanocephalus).

We describe range-wide phylogeographic variation in the Black-headed Grosbeak (Pheucticus melanocephalus), a songbird that is widely distributed across North American scrublands and forests. Phylogenetic analysis of mitochondrial DNA (mtDNA, n=424) revealed three geographically structured clades. One widespread clade occurs throughout the Rocky Mountains, Great Basin, and Mexican Plateau, a second clade is found on the Pacific coast and in coastal ranges; and, a third in the Sierra Madre del Sur of Oaxaca and Guerrero. Some geographical structuring occurs in Mexican Plateau and Sierra Madre Oriental mtDNA clade, presumably because these populations have been more stable over time than northern populations. Multiple mitochondrial groups are found sympatrically in the Okanogan River Valley in Washington, the eastern Sierra Nevada, and the Transvolcanic Belt across central Mexico, indicating that there is a potential for introgression. Analyses of 12 nuclear loci did not recover the same geographically structured clades. Population analyses show high levels of gene flow in nucDNA from the Interior into the Sierra Madre del Sur and Pacific population groups, possibly indicating expansion of the Interior population at the expense of peripheral populations.