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USGS2014

Genomic characterization of H14 subtype influenza A viruses in New World waterfowl and experimental infectivity in mallards Anas platyrhynchos

Recent repeated isolation of H14 hemagglutinin subtype influenza A viruses (IAVs) in the New World waterfowl provides evidence to suggest that host and/or geographic ranges for viruses of this subtype may be expanding. In this study, we used genomic analyses to gain inference on the origin and evolution of H14 viruses in New World waterfowl and conducted an experimental challenge study in mallards ( Anas platyrhynchos ) to evaluate pathogenicity, viral replication, and transmissibility of a representative viral strain in a natural host species. Genomic characterization of H14 subtype IAVs isolated from New World waterfowl, including three isolates sequenced specifically for this study, revealed high nucleotide identity among individual gene segments (e.g. ≥95% shared identity among H14 HA gene segments). In contrast, lower shared identity was observed among internal gene segments. Furthermore, multiple neuraminidase subtypes were observed for H14 IAVs isolated in the New World. Gene segments of H14 viruses isolated after 2010 shared ancestral genetic lineages with IAVs isolated from wild birds throughout North America. Thus, genomic characterization provided evidence for viral evolution in New World waterfowl through genetic drift and genetic shift since purported introduction from Eurasia. In the challenge study, no clinical disease or lesions were observed among mallards experimentally inoculated with A/blue-winged teal/Texas/AI13-1028/2013(H14N5) or exposed via contact with infected birds. Titers of viral shedding for mallards challenged with the H14N5 IAV were highest at two days post-inoculation (DPI); however shedding was detected up to nine DPI using cloacal swabs. The distribution of viral antigen among mallards infected with H14N5 IAV was largely restricted to enterocytes lining the villi in the lower intestinal tract and in the epithelium of the bursa of Fabricius. Characterization of the infectivity of A/blue-winged teal/Texas/AI13-1028/2013(H14N5) in mallards provides support for similarities in viral replication and shedding as compared to previously described waterfowl-adapted, low pathogenic IAV strains in ducks.

PLoS ONE↗

USGS2004

Lack of spatial genetic structure among nesting and wintering King Eiders

The King Eider (Somateria spectabilis) has been delineated into two broadly distributed breeding populations in North America (the western and eastern Arctic) on the basis of banding data and their use of widely separated Pacific and Atlantic wintering areas. Little is known about the level of gene flow between these two populations. Also unknown is whether behavioral patterns common among migratory waterfowl, such as site fidelity to wintering areas and pair formation at these sites, have existed for sufficient time to create a population structure defined by philopatry to wintering rather than to nesting locations. We used six nuclear microsatellite DNA loci and cytochrome b mitochondrial DNA sequence data to estimate the extent of spatial genetic differentiation among nesting and wintering areas of King Eiders across North America and adjacent regions. Estimates of interpopulation variance in microsatellite allele and mtDNA haplotype frequency were both low and nonsignificant based on samples from three wintering and four nesting areas. Results from nested clade analysis, mismatch distributions, and coalescent-based analyses suggest historical population growth and gene flow that collectively may have homogenized gene frequencies. The presence of several unique mtDNA haplotypes among birds wintering near Greenland suggests that gene flow may now be more limited between the western and eastern Arctic, which is consistent with banding data.

Condor↗

USGS2023

A plea for Red Wolf conservation throughout Its recent distribution

Canis rufus (Red Wolf) is one of the most endangered mammals in North America. However, genes of the Red Wolf persist across much of the species' original range, carried predominantly within C. latrans (Coyote) populations. It is now known that such genes are distributed from extreme north-central Texas through most of eastern Texas to southern Louisiana. Publicizing of the most recent findings of Red Wolf genes in Coyotes of southern Louisiana emphasized that area for intensive conservation efforts. Such efforts could be applied throughout the entire known distribution of those rare genes, not just in the small area of southern Louisiana recently publicized. Because conservation efforts might be hindered by local conditions and circumstances, expanding geographic options for their application could make the difference in their success.

Southesatern Naturalist↗

USGS2006

Genetic variation among subspecies of Least Tern (Sterna antillarum): Implications for conservation

DNA sequence variation from two nuclear introns and part of the mitochondrial cytochrome-b gene were used to Evaluate population structure among three subspecies of Least Term that nest in the United States (California [Sterna antillarum browni], Interior [S. a. athalassos], Eastern [S. a. antillarum]). Sequence variation was highest for nuclear intron XI (Gadp) within the glyceraldehyde-3-phosphate dehydrogenase gene. The second nuclear intron was fixed for the same allele in all subspecies. Fixation indices, FST and MST, for Gadp indicated genetic divergence between California and Interior subspecies. Estimates of nuclear gene flow were <4 individuals/generation, except between the Interior and Eastern subspecies (4 individuals/generation). Genetic indices for mitochondrial DNA did not differ among subspecies, and gene flows (reflecting female dispersal) ranged from 10 to 83 individuals/generation. Reservations are expressed about the validity of the current subspecific divisions and further research is required, including their taxonomic relationship to the Little Tern (Sterna albifrons).

Waterbirds↗

USGS2008

Landscape Features Shape Genetic Structure in Threatened Northern Spotted Owls

Several recent studies have shown that landscape features can strongly affect spatial patterns of gene flow and genetic variation. Understanding landscape effects on genetic variation is important in conservation for defining management units and understanding movement patterns. The landscape may have little effect on gene flow, however, in highly mobile species such as birds. We tested for genetic breaks associated with landscape features in the northern spotted owl (Strix occidentalis caurina), a threatened subspecies associated with old forests in the U.S. Pacific Northwest and extreme southwestern Canada. We found little evidence for distinct genetic breaks in northern spotted owls using a large microsatellite dataset (352 individuals from across the subspecies' range genotyped at 10 loci). Nonetheless, dry low-elevation valleys and the Cascade and Olympic Mountains restrict gene flow, while the Oregon Coast Range facilitates it. The wide Columbia River is not a barrier to gene flow. In addition, inter-individual genetic distance and latitude were negatively related, likely reflecting northward colonization following Pleistocene glacial recession. Our study shows that landscape features may play an important role in shaping patterns of genetic variation in highly vagile taxa such as birds.

Open-File Report↗

USGS2014

2013 update on sea otter studies to assess recovery from the 1989 Exxon Valdez oil spill, Prince William Sound, Alaska

On March 24, 1989, the tanker vessel Exxon Valdez ran aground in Prince William Sound, Alaska, spilling an estimated 42 million liters of Prudhoe Bay crude oil. Oil spread in a southwesterly direction and was deposited on shores and waters in western Prince William Sound (WPWS). The sea otter (Enhydra lutris) was one of more than 20 nearshore species considered to have been injured by the spill. Since 1989, the U.S. Geological Survey has led a research program to evaluate effects of the spill on sea otters and assess progress toward recovery, as defined by demographic and biochemical indicators. Here, we provide an update on the status of sea otter populations in WPWS, presenting findings through 2013. To assess recovery based on demographic indicators, we used aerial surveys to estimate abundance and annual collections of sea otter carcasses to evaluate patterns in ages-at-death. To assess recovery based on biochemical indicators, we quantified transcription rates for a suite of genes selected as potential indicators of oil exposure in sea otters based on laboratory studies of a related species, the mink (Mustela vison). In our most recent assessment of sea otter recovery, which incorporated results from a subset of studies through 2009, we concluded that recovery of sea otters in WPWS was underway. This conclusion was based on increasing abundance throughout WPWS, including increasing numbers at northern Knight Island, an area that was heavily oiled in 1989 and where the local sea otter population had previously shown protracted injury and lack of recovery. However, we did not conclude that the WPWS sea otter population had fully recovered, due to indications of continuing reduced survival and exposure to lingering oil in sea otters at Knight Island, at least through 2009. Based on data available through 2013, we now conclude that the status of sea otters—at all spatial scales within WPWS—is consistent with the designation of recovery from the spill as defined by the Exxon Valdez Oil Spill Trustee Council. The support for this conclusion is based primarily on demographic data, including (1) a return to estimated pre-spill abundance of sea otters at northern Knight Island, and (2) a return to pre-spill mortality patterns. Gene transcription rates in 2012 were similar in sea otters from oiled, moderately oiled and unoiled areas, suggesting abatement of exposure effects in 2012. However, because 2012 gene transcription rates generally were low for sea otters from all areas relative to 2008, we cannot fully interpret these observations without data from a wider panel of genes. This slight uncertainty with respect to the data from the biochemical indicator is outweighed by the strength of the data for the demographic indicators. The return to pre-spill numbers and mortality patterns suggests a gradual dissipation of lingering oil over the past two decades, to the point where continuing exposure is no longer of biological significance to the WPWS sea otter population.

Alaska↗

USGS2019

Sampling across 20 years (1996–2017) reveals loss of diversity and genetic connectivity in the Coachella Valley fringe-toed lizard ( Uma inornata )

The Coachella Valley fringe-toed lizard ( Uma inornata ) is a federally threatened, aeolian sand dune obligate, endemic to the Coachella Valley, California. Historically, U. inornata is thought to have formed a large interconnected metapopulation across the valley, with local dune habitat and population size fluctuations linked to stochastic droughts and flooding. Since the 1950s, aeolian habitat in Coachella Valley has declined by 91–95 percent. What remains is highly fragmented by highways and development in the urban communities of the Coachella Valley, raising concerns that fringe-toed lizard movement and gene flow among remaining habitat fragments is limited or nonexistent. We examined population genetic structure across three sample periods (1996, 2008, and 2017). Over that time, this species has shifted from a panmictic condition (1996) with little or no genetic structure between sites to the current (2017) condition where there are now genetically distinct populations. Two severe droughts (2000–04 and 2012–16) may have accelerated this shift through drought-related population declines and subsequent genetic bottlenecks. Using a combination of microsatellite loci and single nucleotide polymorphisms, we found patterns of decreasing genetic connectivity and diversity over time. These patterns are consistent with reduced fringe-toed lizard movement and gene flow among isolated sand dune systems. Low effective population sizes were recovered in some sites, suggesting genetic drift in smaller and fluctuating populations is likely responsible for loss of genetic diversity. A U.S. Fish and Wildlife Service recovery objective for this species is to maintain genetic diversity; however, evidence of fragmentation suggests that genetic cohesiveness has been altered and that the diversity maintained in individual fragments is lower than in the total metapopulation. Management actions that increase genetic diversity could be implemented, including translocation. We modeled increasing gene flow between 1–10 percent, which showed that allelic richness could increase rapidly if translocated individuals can survive and reproduce. Establishing translocation protocols could help to avoid the high mortality that has occurred with other reptile translocations. Successful translocations could be a useful strategy to replenish lost genetic diversity after bottlenecks and could mitigate the loss of natural gene flow among populations.

California↗

USGS2005

Preliminary survey of antibiotic-resistant fecal indicator bacteria and pathogenic Escherichia coli from river-water samples collected in Oakland County, Michigan, 2003

A preliminary study was done in Oakland County, Michigan, to determine the concentration of fecal indicator bacteria (fecal coliform bacteria and enterococci), antibiotic resistance patterns of these two groups, and the presence of potentially pathogenic Escherichia coli ( E. coli ). For selected sites, specific members of these groups [ E. coli , Enterococcus faecium ( E. faecium ) and Enterococcus faecalis ( E. faecalis )] were isolated and tested for levels of resistance to specific antibiotics used to treat human infections by pathogens in these groups and for their potential to transfer these resistances. In addition, water samples from all sites were tested for indicators of potentially pathogenic E. coli by three assays: a growth-based assay for sorbitol-negative E. coli , an immunological assay for E. coli O157, and a molecular assay for three virulence and two serotype genes. Samples were also collected from two non-urbanized sites outside of Oakland County. Results from the urbanized Oakland County area were compared to those from these two non-urbanized sites. Fecal indicator bacteria concentrations exceeded State of Michigan recreational water-quality standards and (or) recommended U.S. Environmental Protection Agency (USEPA) standards in samples from all but two Oakland County sites. Multiple-antibiotic-resistant fecal coliform bacteria were found at all sites, including two reference sites from outside the county. Two sites (Stony Creek and Paint Creek) yielded fecal coliform isolates resistant to all tested antibiotics. Patterns indicative of extended-spectrum-β-lactamase (ESBL)- producing fecal coliform bacteria were found at eight sites in Oakland County and E. coli resistant to clinically significant antibiotics were recovered from the River Rouge, Clinton River, and Paint Creek. Vancomycin-resistant presumptive enterococci were found at six sites in Oakland County and were not found at the reference sites. Evidence of acquired antibiotic resistances was detected in bacteria from multiple sites in Oakland County but not detected in bacteria from the reference sites. Integrons capable of transferring resistance were detected in isolates from the River Rouge and Clinton River. E. faecium and E. faecalis identified in samples collected from Kearsley Creek and Evans Ditch were resistant to high levels of vancomycin and carried transferable genes responsible for resistance. Several sites in Oakland County had indicators of pathogenic E. coli in August and (or) September 2003. Two samples from the Clinton River in August tested positive for all three E. coli O157 tests. Both the August and September samples from one River Rouge site were positive for the immunological and molecular assay for E. coli O157. A combination of virulence genes commonly associated with human illness was detected at five sites in August and seven sites in September. Antibiotic-resistance profiles of clinical concern along with genes capable of transferring the resistance were found at several sites throughout Oakland County; samples from many of these sites also contained potentially pathogenic E. coli .

Michigan↗

USGS2014

Chemical and biological quality of water in Grand Lake St. Marys, Ohio, 2011-12, with emphasis on cyanobacteria

Grand Lake St. Marys (GLSM) is a shallow lake in northwest Ohio, which is about 9 miles long and 3 miles wide with depths averaging less than 8 feet. Cyanobacteria blooms are common in GLSM, and high concentrations of microcystins—toxins produced by cyanobacteria—have been documented therein. During 2011–12, the U.S. Geological Survey collected 11 sets of water samples at 6 locations in the lake. The water samples were analyzed for concentrations of nutrients, chlorophyll, and microcystin and to determine plankton community structure and abundance. Analysis by quantitative polymerase chain reaction (qPCR) and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to identify the relations between microcystin concentrations and Planktothrix and Microcystis genotypes (toxic versus non-toxic). The qPCR analysis targets deoxyribonucleic acid (DNA) genes and quantifies the potential for toxin production, whereas the qRT-PCR analysis targets ribonucleic acid (RNA) transcripts and quantifies the expression of the toxin gene. Water samples were collected six times at one site for analyses of major ions and trace elements. In addition, field measurements were made to determine transparency, temperature, dissolved oxygen, pH, and specific conductance of the water. GLSM is shallow with a long fetch, which contributes to the warm and turbid water conditions. Secchi-disk measurements generally ranged from 0.2 to 0.3 meters, and summer water temperatures in GLSM frequently exceed 25 degrees Celsius (°C), with peak temperatures greater than 30 °C. Dissolved oxygen readings below 0.5 milligrams per liter (mg/L) occurred at the lake bottom, which can lead to the internal recycling of phosphorus in the lake. Phytoplankton analyses indicated that GLSM is dominated by cyanobacteria with Planktothrix , the dominant genera during 2011–12. Nitrate ranged from 0.19 to 3.23 mg/L, although concentrations in most samples were less than 1 mg/L. Total nitrogen concentrations ranged from 1.86 to 5.42 mg/L. Orthophosphate (as P) concentrations ranged from less than 0.004 to 0.067 mg/L, although concentrations of most samples were less than 0.004 mg/L. Total phosphorus (as P) concentrations ranged from 0.12 to 0.43 mg/L. Microcystin concentrations ranged from 7.3 to 83 micrograms per liter. Microcystin concentrations were correlated to cyanobacteria biovolumes, and to concentrations of one ion (sodium) and three trace elements (molybdenum, antimony, and lithium). Concentrations of toxin genes ( mcyE ) determined by qPCR were consistently low for Microcystis and consistently high for Planktothrix throughout both sampling years. Concentrations of cyanobacteria found by qPCR were correlated to microcystin concentrations, cyanobacteria biovolumes, selected nutrient concentrations, and other parameters. Results from qRT-PCR assays showed that toxin gene expression was predominantly from the genus Planktothrix , and concentrations of the RNA transcript varied throughout the two sampling years. A number of conditions that may play a role in the dominance of Planktothrix and the production of microcystin were identified including water temperature; low-light transmission; low concentrations of silica and manganese; and relatively high concentrations of sodium, sulfate, and the trace elements of strontium, vanadium, and boron.

Ohio↗

USGS2015

Water Quality, Cyanobacteria, and Environmental Factors and Their Relations to Microcystin Concentrations for Use in Predictive Models at Ohio Lake Erie and Inland Lake Recreational Sites, 2013-14

Harmful cyanobacterial “algal” blooms (cyanoHABs) and associated toxins, such as microcystin, are a major water-quality issue for Lake Erie and inland lakes in Ohio. Predicting when and where a bloom may occur is important to protect the public that uses and consumes a water resource; however, predictions are complicated and likely site specific because of the many factors affecting toxin production. Monitoring for a variety of environmental and water-quality factors, for concentrations of cyanobacteria by molecular methods, and for algal pigments such as chlorophyll and phycocyanin by using optical sensors may provide data that can be used to predict the occurrence of cyanoHABs. To test these monitoring approaches, water-quality samples were collected at Ohio recreational sites during May–November in 2013 and 2014. In 2013, samples were collected monthly at eight sites at eight lakes to facilitate an initial assessment and select sites for more intensive sampling during 2014. In 2014, samples were collected approximately weekly at five sites at three lakes. Physical water-quality parameters were measured at the time of sampling. Composite samples were preserved and analyzed for dissolved and total nutrients, toxins, phytoplankton abundance and biovolume, and cyanobacterial genes by molecular methods. Molecular assays were done to enumerate (1) general cyanobacteria, (2) general Microcystis and Dolichospermum ( Anabaena ), (3) mcyE genes for Microcystis , Dolichospermum ( Anabaena ), and Planktothrix targeting deoxyribonucleic acid (DNA), and (4) mcyE transcripts for Microcystis , Dolichospermum ( Anabaena ), and Planktothrix targeting ribonucleic acid (RNA).The DNA assays for the mcyE gene provide data on cyanobacteria that have the potential to produce microcystin, whereas the RNA assays provide data on cyanobacteria that are actively transcribing the toxin gene. Environmental data were obtained from available online sources. Quality-control (QC) samples were collected and analyzed for all constituents to characterize bias and variability; however, QC data for molecular assays were examined in more detail than for the other constituents. The QC data for molecular assays suggested that sampling variability and qPCR variability were small in comparison with the combined variability associated with sample filtering, extraction and purification, and the matrix itself. A total of 46 water-quality samples were collected during 2013 at 8 beach sites—Buck Creek, Buckeye Crystal, Deer Creek, Harsha Main, Maumee Bay State Park (MBSP) Inland (negative control site), MBSP Lake Erie, Port Clinton, and Sandusky Bay. Microcystin was detected in 67–100 percent of samples at all sites except for MBSP Inland, where microcystin was detected in only 20 percent of samples. Microcystin concentrations ranged from <0.10 to 48 micrograms per liter (µ/L), with the widest range found at MBSP Lake Erie and the highest concentrations found at Buckeye Crystal. Saxitoxin was detected in five samples, and cylindrospermopsin was not detected in any samples. A total of 65 water-quality samples were collected during 2014 at 5 sites on 3 lakes—Buckeye Fairfield and Onion Island, Harsha Main and Campers, and MBSP Lake Erie beach. Four of the sites were bathing beaches and one site, Onion Island, was an offshore boater swim area. Concentrations of microcystin ranged from <0.10 to 240 µ/L and, as in 2013, the widest range was found at MBSP Lake Erie. At Buckeye Lake, microcystin concentrations were consistently high (greater than 20 µ/L), ranging from 23 to 81 µ/L. At Harsha Main and Campers, microcystin concentrations ranged from <0.10 to 15 µ/L. Saxitoxin was detected in four samples collected at MBSP Lake Erie. Throughout the 2014 season, the cyanobacterial community, as determined by molecular and microscopy methods, and the dominance associated with the highest microcystin concentrations were unique to individual lakes. At Buckeye Lake, Planktothrix dominated the cyanobacterial community throughout the season and Planktothrix DNA and RNA were found in 100 percent of samples; Microcystis mcyE DNA was found in low concentrations. At Harsha Lake, Dolichospermum and Microcystis were a substantial percentage of the community from late May through August, and the highest microcystin concentrations occurred in June and July. At MBSP Lake Erie, Microcystis generally dominated from mid-July through early November, and the highest microcystin concentrations occurred in August. Spearman’s correlation coefficient (rho) was computed to determine the relations between environmental and water-quality factors and microcystin concentrations at four sites—Buckeye Fairfield, Buckeye Onion Island, Harsha Main, and MBSP Lake Erie. Factors were evaluated for use as potential independent variables in two types of predictive models—daily and long-term models. Easily or continuously measured water-quality factors and available environmental data are used for daily predictions that do not require a site visit. Data from factors used in daily predictions and results from samples collected and analyzed in a laboratory are used for long-term predictions (a few days to several weeks). A few statistically significant correlations (p ≤ 0.05) between microcystin concentrations and factors for both daily and long-term predictions were found at Buckeye Onion Island, and many were found at Harsha Main and MBSP Lake Erie. There were only a few statistically significant factors for daily predictions at Buckeye Fairfield, likely because of the lack of variability in microcystin concentrations. Among factors for daily predictions, phycocyanin had the highest Spearman’s correlation to microcystin concentrations (rho = 0.79 to 0.93) at all sites except for Buckeye Fairfield. Turbidity, pH, algae category, and Secchi depth were significantly correlated to microcystin concentrations at Harsha Main and MBSP Lake Erie. Algae categories were observational categories from 0 (none) to 4 (extreme). Several discharge variables (Maumee River at Waterville, river mouth is approximately 3.5 miles from the beach) at MBSP Lake Erie were promising environmental factors for daily predictions. In addition to discrete water-quality measurements recorded at Harsha Main at the time of sampling, many manipulated measurements (factors derived from mathematical manipulation of time-series data) available from a nearby continuous monitor were strongly correlated to microcystin concentrations; the highest correlation was found for the relation between microcystin concentrations and the antecedent 7-day average phycocyanin (rho = 0.98). For long-term predictions, the most highly correlated molecular assays were Planktothrix mcyE DNA at Buckeye Onion Island and Microcystis mcyE DNA at Harsha Main and MBSP Lake Erie. Concentrations of several nutrient constituents were significantly correlated to microcystin concentrations including total nitrogen at Buckeye Onion Island, ammonia and nitrate plus nitrite (both negatively correlated) at Harsha Main and MBSP Lake Erie, and total phosphorus at MBSP Lake Erie. The results of this study showed that water-quality and environmental variables are promising for use in site-specific daily or long-term predictive models. In order to develop more accurate models to predict toxin concentrations at freshwater lake sites, data need to be collected more frequently and for consecutive days in future studies.

Ohio↗

USGS2018

Preliminary investigation of groundwater quality near a Michigan cemetery, 2016–17

The potential effect of cemetery leachate on groundwater quality in the United States has rarely been studied. Nutrients and other constituents associated with decomposition and burial processes (such as embalming) have the potential to reach shallow groundwater and could affect nearby drinking-water sources. The objective of this preliminary investigation was to evaluate the potential effect of cemetery leachate on shallow groundwater quality near Mt. Hope Cemetery in Ingham County, Lansing, Michigan, which is within the Well-head Protection Area for the City of Lansing. The constituents measured in this study include nutrients, trace metals, formaldehyde, fecal indicator bacteria, bacterial pathogen genes, contaminants of emerging concern (including pharmaceuticals, personal care products, and wastewater indicator compounds), and age-dating compounds. Three monitoring wells were installed 7 to 12 feet below land surface downgradient from the cemetery and sampled quarterly for 1 year. A fourth well (Fenner) was sampled to determine groundwater conditions outside the potential effects of cemetery leachate; samples from this well were collected near the water table. Nitrogen and phosphorus compounds were present at higher concentrations in two of the three monitoring wells (wells C1 and C3) than in the Fenner well. Formaldehyde and pharmaceuticals were not detected in any of the wells; however, several trace metals, including arsenic, manganese, and aluminum, were present in high concentrations, with arsenic concentrations typically exceeding the U.S. Environmental Protection Agency (EPA) drinking-water standard. Several wastewater indicator compounds, including atrazine, phenol, p-cresol, camphor, and skatole, were detected in the monitoring wells. Microbial data indicate the presence of staphylococci, enterococci, and Escherichia coli (E. coli) , with the highest concentrations being measured in the same two monitoring wells that exhibited elevated concentrations of nutrients in the groundwater (wells C1 and C3). Several bacterial pathogen genes were detected, including several Enterococcus species (spp.)— vanB (vancomycin-resistant enterococci), shiga-toxin-producing E. coli genes (including eaeA [attachment virulence trait] and stx1 [moderate toxin]), and the E. coli 16s ribosomal RNA (rDNA) gene ( E. coli species marker). These results were similar to results of studies conducted in Canada, Australia, and the United Kingdom, in which concentrations of bacteria, metals, and nutrients were elevated in groundwater near cemeteries.

Michigan↗

USGS2009

Genetic and serological typing of European infectious haematopoietic necrosis virus (IHNV) isolates

Infectious haematopoietic necrosis virus (IHNV) causes the lethal disease infectious haematopoietic necrosis (IHN) in juvenile salmon and trout. The nucleocapsid (N) protein gene and partial glycoprotein (G) gene (nucleotides 457 to 1061) of the European isolates IT-217A, FR-32/87, DE-DF 13/98 11621, DE-DF 4/99-8/99, AU-9695338 and RU-FR1 were sequenced and compared with IHNV isolates from the North American genogroups U, M and L. In phylogenetic studies the N gene of the Italian, French, German and Austrian isolates clustered in the M genogroup, though in a different subgroup than the isolates from the USA. Analyses of the partial G gene of these European isolates clustered them in the M genogroup close to the root while the Russian isolate clustered in the U genogroup. The European isolates together with US-WRAC and US-Col-80 were also tested in an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (MAbs) against the N protein. MAbs 136-1 and 136-3 reacted equally at all concentrations with the isolates tested, indicating that these antibodies identify a common epitope. MAb 34D3 separated the M and L genogroup isolates from the U genogroup isolate. MAb 1DW14D divided the European isolates into 2 groups. MAb 1DW14D reacted more strongly with DE-DF 13/98 11621 and RU-FR1 than with IT-217A, FR- 32/87, DE-DF 4/99-8/99 and AU-9695338. In the phylogenetic studies, the Italian, French, German and Austrian isolates clustered in the M genogroup, whereas in the serological studies using MAbs, the European M genogroup isolates could not be placed in the same specific group. These results indicate that genotypic and serotypic classification do not correlate. ?? 2009 Inter-Research.

Diseases of Aquatic Organisms↗

USGS2023

Coupling large-spatial scale larval dispersal modelling with barcoding to refine the amphi-Atlantic connectivity hypothesis in deep-sea seep mussels

In highly fragmented and relatively stable cold-seep ecosystems, species are expected to exhibit high migration rates and long-distance dispersal of long-lived pelagic larvae to maintain genetic integrity over their range. Accordingly, several species inhabiting cold seeps are widely distributed across the whole Atlantic Ocean, with low genetic divergence between metapopulations on both sides of the Atlantic Equatorial Belt (AEB, i.e. Barbados and African/European margins). Two hypotheses may explain such patterns: (i) the occurrence of present-day gene flow or (ii) incomplete lineage sorting due to large population sizes and low mutation rates. Here, we evaluated the first hypothesis using the cold seep mussels Gigantidas childressi, G. mauritanicus, Bathymodiolus heckerae and B. boomerang . We combined COI barcoding of 763 individuals with VIKING20X larval dispersal modelling at a large spatial scale not previously investigated. Population genetics supported the parallel evolution of Gigantidas and Bathymodiolus genera in the Atlantic Ocean and the occurrence of a 1-3 Million-year-old vicariance effect that isolated populations across the Caribbean Sea. Both population genetics and larval dispersal modelling suggested that contemporary gene flow and larval exchanges are possible across the AEB and the Caribbean Sea, although probably rare. When occurring, larval flow was eastward (AEB - only for B. boomerang ) or northward (Caribbean Sea - only for G. mauritanicus ). Caution is nevertheless required since we focused on only one mitochondrial gene, which may underestimate gene flow if a genetic barrier exists. Non-negligible genetic differentiation occurred between Barbados and African populations, so we could not discount the incomplete lineage sorting hypothesis. Larval dispersal modelling simulations supported the genetic findings along the American coast with high amounts of larval flow between the Gulf of Mexico (GoM) and the US Atlantic Margin, although the Blake Ridge population of B. heckerae appeared genetically differentiated. Overall, our results suggest that additional studies using nuclear genetic markers and population genomics approaches are needed to clarify the evolutionary history of the Atlantic bathymodioline mussels and to distinguish between ongoing and past processes.

Frontiers in Marine Science↗

USGS2018

Chronic physical disturbance substantially alters the response of biological soil crusts to a wetting pulse, as characterized by metatranscriptomic sequencing

Biological soil crusts (biocrusts) are microbial communities that are a feature of arid surface soils worldwide. In drylands where precipitation is pulsed and ephemeral, the ability of biocrust microbiota to rapidly initiate metabolic activity is critical to their survival. Community gene expression was compared after a short duration (1 hour) wetting pulse in both intact and soils disturbed by chronic foot trampling. Across the metatranscriptomes the majority of transcripts were cyanobacterial in origin, suggesting that cyanobacteria accounted for the bulk of the transcriptionally active cells. Chronic trampling substantially altered the functional profile of the metatranscriptomes, specifically resulting in a significant decrease in transcripts for nitrogen fixation. Soil depth (biocrust and below crust) was a relatively small factor in differentiating the metatranscriptomes, suggesting that the metabolically active bacteria were similar between shallow soil horizons. The dry samples were consistently enriched for hydrogenase genes, indicating that molecular hydrogen may serve as an energy source for the desiccated soil communities. The water pulse was associated with a restructuring of the metatranscriptome, particularly for the biocrusts. Biocrusts increased transcripts for photosynthesis and carbon fixation, suggesting a rapid resuscitation upon wetting. In contrast, the trampled surface soils showed a much smaller response to wetting, indicating that trampling altered the metabolic response of the community. Finally, several biogeochemical cycling genes in carbon and nitrogen cycling were assessed for their change in abundance due to wetting in the biocrusts. Different transcripts encoding the same gene product did not show a consensus response, with some more abundant in dry or wet biocrusts, highlighting the challenges in relating transcript abundance to biogeochemical cycling rates. These observations demonstrate that metatranscriptome sequencing was able to distinguish alterations in the function of arid soil microbial communities at two varying temporal scales, a long-term ecosystems disturbance through foot trampling, and a short term wetting pulse. Thus, community metatranscriptomes have the potential to inform studies on the response and resilience of biocrusts to various environmental perturbations.

Frontiers in Microbiology↗

USGS2025

Bacterial community diversity and potential eco-physiological roles in toxigenic blooms composed of Microcystis, Aphanizomenon or Planktothrix

Cyanobacterial toxicity, cyanotoxins, and their impact on aquatic ecosystems and human health are well documented. In comparison, less is known about bloom-associated bacterial communities. Co-occurring bacteria can influence bloom development, physiology and collapse, and may also provide a niche for pathogenic bacteria. Existing research focuses on the cyanosphere of Microcystis -dominated blooms, despite the increasing prevalence of filamentous genera ( Aphanizomenon and Planktothrix ). This pilot study aimed to broaden our understanding of the bacterial consortia attached to morphologically distinct cyanobacteria (coccoid and filamentous) dominating phytoplankton communities and to explore their potential roles in amplifying the impacts of cyanobacterial blooms. We investigated four shallow freshwater bodies across three continents and two climate zones: an urban pond in the USA, a dammed reservoir and a natural lake in Poland, and an urban water body in Singapore. Amplicon sequencing (16S rRNA gene) was used to characterize bacterial communities, while shotgun metagenomics identified nitrogen- and phosphorus-cycling genes to infer potential eco-physiological functions. Cyanobacteria dominated bacterioplankton assemblages at all sites (>35.6%), with bloom composition influencing toxigenic profiles. A mixed bloom of Microcystis , Snowella , and Aphanizomenon had the broadest range of cyanotoxin synthetase genes ( mcy E, cyr J, ana F and sxt A). Microcystis blooms correlated with increased Roseomonas , while Planktothrix co-occurred with Flavobacterium – both bacteria likely contribute to nutrient-cycling within blooms and represent potential opportunistic pathogens for aquatic organisms and humans. The Microcystis cyanosphere exhibited the highest number of significant positive correlations with bacteria (19 relations), compared to Planktothrix and Aphanizomenon (11 and 2 relations, respectively). Non-diazotrophic blooms of Microcystis and Planktothrix showed greater abundances of nitrogen – ( ure B, gln A, nar B, and nar HZ) and phosphorus-cycling genes ( pho BHPR and ppk 1), indicating a strong dependence on associated bacteria for nutrient acquisition compared to diazotrophic Aphanizomenon . These findings suggest that Aphanizomenon -dominated blooms may be sustained by simpler microbiomes. Our results provide preliminary evidence of cyanosphere heterogeneity potentially shaped by the dominance or coexistence of three morphologically and eco-physiologically distinct genera of cyanobacteria. A comprehensive knowledge of the taxonomy and functional roles of bloom-associated microbiomes is therefore essential to understand bloom activity, evaluate the environmental threat, and develop effective strategies for prevention and mitigation.

Frontiers in Microbiology↗

USGS2026

Comparative genomics reveals potential mechanisms of invasion in Phragmites australis (common reed)

Biological invasions are transforming ecosystems worldwide, yet the genomic bases enabling certain species to dominate new environments remain poorly understood. Phragmites australis , a widespread wetland grass with invasive and native subspecies co-occurring in North America, provides a powerful system to investigate genomic mechanisms of invasiveness. We generated independent chromosome-scale genome assemblies for invasive P. australis ssp. australis and co-occurring native ssp. americanus and used comparative genomic and transcriptomic analyses to identify lineage-specific innovations associated with invasive success. The invasive subspecies exhibits genomic novelties through functionally-biased single-copy orthologs, intronless genes, and subgenome expression asymmetry, along with a stress-ready basal transcriptome relative to the native subspecies. Following the removal of aboveground shoots (“cutback”), which measures the ability to recover from damage, the invasive subspecies undergoes stronger transcriptional reprogramming, increased shoot production, and higher biomass accumulation compared to the native. It also displays expansion of gene families and coordinately expressed gene modules that support resource mobilization, growth responses to light, and stress tolerance. Beyond Phragmites , comparative analyses across multiple grass genomes, including eight invasive species with related non-invasive species, revealed repeated expansion of gene families associated with abiotic stress tolerance and developmental regulation, suggesting convergent adaptive strategies in the grass family for invasive success. Together, these results demonstrate genomic architecture linked to invasion success and highlight potential targets for managing invasive grasses.

BioRxiv↗

USGS2018

Development and validation of quantitative PCR assays to measure cytokine transcript levels in the Florida manatee (Trichechus manatus latirostris)

Cytokines have important roles in the mammalian response to viral and bacterial infections, trauma, and wound healing. Because of early cytokine production after physiologic stresses, the regulation of messenger RNA (mRNA) transcripts can be used to assess immunologic responses before changes in protein production. To detect and assess early immune changes in endangered Florida manatees ( Trichechus manatus latirostris ), we developed and validated a panel of quantitative PCR assays to measure mRNA transcription levels for the cytokines interferon (IFN)-γ; interleukin (IL)-2, -6, and -10; tumor necrosis factor-α, and the housekeeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin (reference genes). Assays were successfully validated using blood samples from free-ranging, apparently healthy manatees from the east and west coasts of central Florida. No cytokine or housekeeping gene transcription levels were significantly different among age classes or sexes. However, the transcription levels for GAPDH, IL-2, IL-6, and IFN-γ were significantly higher ( P <0.05) in manatees from the east coast of Florida than they were from those from the west coast. We found IL-10 and β-actin to be consistent between sites and identified β-actin as a good candidate for use as a reference gene in future studies. Our assays can aid in the investigation of manatee immune response to physical trauma and novel or ongoing environmental stressors.

Florida↗

USGS2020

Effects of early life stage exposure of largemouth bass to atrazine or a model estrogen (17α-ethinylestradiol)

Endocrine disrupting contaminants are of continuing concern for potentially contributing to reproductive dysfunction in largemouth and smallmouth bass in the Chesapeake Bay watershed (CBW) and elsewhere. Exposures to atrazine (ATR) have been hypothesized to have estrogenic effects on vertebrate endocrine systems. The incidence of intersex in male smallmouth bass from some regions of CBW has been correlated with ATR concentrations in water. Fish early life stages may be particularly vulnerable to ATR exposure in agricultural areas, as a spring influx of pesticides coincides with spawning and early development. Our objectives were to investigate the effects of early life stage exposure to ATR or the model estrogen 17α-ethinylestradiol (EE2) on sexual differentiation and gene expression in gonad tissue. We exposed newly hatched largemouth bass (LMB, Micropterus salmoides ) from 7 to 80 days post-spawn to nominal concentrations of 1, 10, or 100 µg ATR/L or 1 or 10 ng EE2/L and monitored histological development and transcriptomic changes in gonad tissue. We observed a nearly 100% female sex ratio in LMB exposed to EE2 at 10 ng/L, presumably due to sex reversal of males. Many gonad genes were differentially expressed between sexes. Multidimensional scaling revealed clustering by gene expression of the 1 ng EE2/L and 100 µg ATR/L-treated male fish. Some pathways responsive to EE2 exposure were not sex-specific. We observed differential expression in male gonad in LMB exposed to EE2 at 1 ng/L of several genes involved in reproductive development and function, including star , cyp11a2 , ddx4 (previously vasa ), wnt5b , cyp1a and samhd1 . Expression of star , cyp11a2 and cyp1a in males was also responsive to ATR exposure. Overall, our results confirm that early development is a sensitive window for estrogenic endocrine disruption in LMB and are consistent with the hypothesis that ATR exposure induces some estrogenic responses in the developing gonad. However, ATR-specific and EE2-specific responses were also observed.

New York, Pennsylvania, Maryland, West Virginia, V↗