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At least 307 records · Page 17Linked to original sources

USGS2017

Screening of duplicated loci reveals hidden divergence patterns in a complex salmonid genome

A whole-genome duplication (WGD) doubles the entire genomic content of a species and is thought to have catalysed adaptive radiation in some polyploid-origin lineages. However, little is known about general consequences of a WGD because gene duplicates (i.e., paralogs) are commonly filtered in genomic studies; such filtering may remove substantial portions of the genome in data sets from polyploid-origin species. We demonstrate a new method that enables genome-wide scans for signatures of selection at both nonduplicated and duplicated loci by taking locus-specific copy number into account. We apply this method to RAD sequence data from different ecotypes of a polyploid-origin salmonid ( Oncorhynchus nerka ) and reveal signatures of divergent selection that would have been missed if duplicated loci were filtered. We also find conserved signatures of elevated divergence at pairs of homeologous chromosomes with residual tetrasomic inheritance, suggesting that joint evolution of some nondiverged gene duplicates may affect the adaptive potential of these genes. These findings illustrate that including duplicated loci in genomic analyses enables novel insights into the evolutionary consequences of WGDs and local segmental gene duplications.

Molecular Ecology↗

USGS2006

Dissimilatory arsenate and sulfate reduction in sediments of two hypersaline, arsenic-rich soda lakes: Mono and Searles Lakes, California

A radioisotope method was devised to study bacterial respiratory reduction of arsenate in sediments. The following two arsenic-rich soda lakes in California were chosen for comparison on the basis of their different salinities: Mono Lake (∼90 g/liter) and Searles Lake (∼340 g/liter). Profiles of arsenate reduction and sulfate reduction were constructed for both lakes. Reduction of [ 73 As]arsenate occurred at all depth intervals in the cores from Mono Lake (rate constant [ k ] = 0.103 to 0.04 h −1 ) and Searles Lake ( k = 0.012 to 0.002 h −1 ), and the highest activities occurred in the top sections of each core. In contrast, [ 35 S]sulfate reduction was measurable in Mono Lake ( k = 7.6 ×10 4 to 3.2 × 10 −6 h −1 ) but not in Searles Lake. Sediment DNA was extracted, PCR amplified, and separated by denaturing gradient gel electrophoresis (DGGE) to obtain phylogenetic markers (i.e., 16S rRNA genes) and a partial functional gene for dissimilatory arsenate reduction ( arrA ). The amplified arrA gene product showed a similar trend in both lakes; the signal was strongest in surface sediments and decreased to undetectable levels deeper in the sediments. More arrA gene signal was observed in Mono Lake and was detectable at a greater depth, despite the higher arsenate reduction activity observed in Searles Lake. A partial sequence (about 900 bp) was obtained for a clone (SLAS-3) that matched the dominant DGGE band found in deeper parts of the Searles Lake sample (below 3 cm), and this clone was found to be closely related to SLAS-1, a novel extremophilic arsenate respirer previously cultivated from Searles Lake.

Applied and Environmental Microbiology↗

USGS1994

Use of molecular techniques to evaluate the survival of a microorganism injected into an aquifer

A PCR primer set and an internal probe that are specific for Pseudomonas sp. strain B13, a 3-chlorobenzoate-metabolizing strain, were developed. Using this primer set and probe, we were able to detect Pseudomonas sp. strain B13 DNA sequences in DNA extracted from aquifer samples 14.5 months after Pseudomonas sp. strain B13 had been injected into a sand and gravel aquifer. This primer set and probe were also used to analyze isolates from 3-chlorobenzoate enrichments of the aquifer samples by Southern blot analysis. Hybridization of Southern blots with the Pseudomonas sp. strain B13-specific probe and a catabolic probe in conjunction with restriction fragment length polymorphism (RFLP) analysis of ribosome genes was used to determine that viable Pseudomonas sp. strain B13 persisted in this environment. We isolated a new 3-chlorobenzoate-degrading strain from one of these enrichment cultures. The B13-specific probe does not hybridize to DNA from this isolate. The new strain could be the result of gene exchange between Pseudomonas sp. strain B13 and an indigenous bacterium. This speculation is based on an RFLP pattern of ribosome genes that differs from that of Pseudomonas sp. strain B13, the fact that identically sized restriction fragments hybridized to the catabolic gene probe, and the absence of any enrichable 3-chlorobenzoate-degrading strains in the aquifer prior to inoculation.

Massachusetts↗

USGS2002

A single Alal 39-to-Glu substitution in the Renibacterium salmoninarum virulence-associated protein p57 results in antigenic variation and is associated with enhanced p57 binding to Chinook salmon leukocytes

The gram-positive bacterium Renibacterium salmoninarum produces relatively large amounts of a 57-kDa protein (p57) implicated in the pathogenesis of salmonid bacterial kidney disease. Antigenic variation in p57 was identified by using monoclonal antibody 4C11, which exhibited severely decreased binding to R. salmoninarum strain 684 p57 and bound robustly to the p57 proteins of seven other R. salmoninarum strains. This difference in binding was not due to alterations in p57 synthesis, secretion, or bacterial cell association. The molecular basis of the 4C11 epitope loss was determined by amplifying and sequencing the two identical genes encoding p57, msa1 and msa2 . The 5′ and coding sequences of the 684 msa1 and msa2 genes were identical to those of the ATCC 33209 msa1 and msa2 genes except for a single C-to-A nucleotide mutation. This mutation was identified in both the msa1 and msa2 genes of strain 684 and resulted in an Ala 139 -to-Glu substitution in the amino-terminal region of p57. We examined whether this mutation in p57 altered salmonid leukocyte and rabbit erythrocyte binding activities. R. salmoninarum strain 684 extracellular protein exhibited a twofold increase in agglutinating activity for chinook salmon leukocytes and rabbit erythrocytes compared to the activity of the ATCC 33209 extracellular protein. A specific and quantitative p57 binding assay confirmed the increased binding activity of 684 p57. Monoclonal antibody 4C11 blocked the agglutinating activity of the ATCC 33209 extracellular protein but not the agglutinating activity of the 684 extracellular protein. These results indicate that the Ala 139 -to-Glu substitution altered immune recognition and was associated with enhanced biological activity of R. salmoninarum 684 p57.

Applied and Environmental Microbiology↗

USGS2004

In situ expression of nifD in Geobacteraceae in subsurface sediments

In order to determine whether the metabolic state of Geobacteraceae involved in bioremediation of subsurface sediments might be inferred from levels of mRNA for key genes, in situ expression of nifD , a highly conserved gene involved in nitrogen fixation, was investigated. When Geobacter sulfurreducens was grown without a source of fixed nitrogen in chemostats with acetate provided as the limiting electron donor and Fe(III) as the electron acceptor, levels of nifD transcripts were 4 to 5 orders of magnitude higher than in chemostat cultures provided with ammonium. In contrast, the number of transcripts of recA and the 16S rRNA gene were slightly lower in the absence of ammonium. The addition of acetate to organic- and nitrogen-poor subsurface sediments stimulated the growth of Geobacteraceae and Fe(III) reduction, as well as the expression of nifD in Geobacteraceae . Levels of nifD transcripts in Geobacteraceae decreased more than 100-fold within 2 days after the addition of 100 μM ammonium, while levels of recA and total bacterial 16S rRNA in Geobacteraceae remained relatively constant. Ammonium amendments had no effect on rates of Fe(III) reduction in acetate-amended sediments or toluene degradation in petroleum-contaminated sediments, suggesting that other factors, such as the rate that Geobacteraceae could access Fe(III) oxides, limited Fe(III) reduction. These results demonstrate that it is possible to monitor one aspect of the in situ metabolic state of Geobacteraceae species in subsurface sediments via analysis of mRNA levels, which is the first step toward a more global analysis of in situ gene expression related to nutrient status and stress response during bioremediation by Geobacteraceae .

Applied and Environmental Microbiology↗

USGS2009

Clostridium botulinum type E occurs and grows in the alga Cladophora glomerata

In recent years, massive avian die-offs from Clostridium botulinum type E infection have occurred in the Sleeping Bear Dunes National Lakeshore (SLBE) area of Lake Michigan. These outbreaks have been coincidental with massive blooms of the green algae Cladophora , mostly Cladophora glomerata . We tested the hypothesis that Clostridium botulinum type E can grow under suitable conditions in these algal mats. In a lab mesocosm study, Cladophora from four outbreak-impacted beaches from SLBE were compared with four unimpacted beaches in the Milwaukee–Racine area for bontE gene of Clostridium botulinum . Frequency of the bontE gene was higher after incubation (25 °C for up to 6 weeks) of Cladophora from impacted vs. the unimpacted area. Since no type E gene was detected initially in Cladophora from any of the eight locations, we infer that the increased occurrence of type E gene arose from spore germination or vegetative Clostridium growth within the existing algal mats of SLBE. Moreover, we found that the congener Clostridium perfringens readily grows in mesocosms containing Cladophora .

Lake Michigan;Sleeping Bear Dunes National Lakesho↗

USGS2016

Genetic and phenotypic variation along an ecological gradient in lake trout Salvelinus namaycush

Background Adaptive radiation involving a colonizing phenotype that rapidly evolves into at least one other ecological variant, or ecotype, has been observed in a variety of freshwater fishes in post-glacial environments. However, few studies consider how phenotypic traits vary with regard to neutral genetic partitioning along ecological gradients. Here, we present the first detailed investigation of lake trout Salvelinus namaycush that considers variation as a cline rather than discriminatory among ecotypes. Genetic and phenotypic traits organized along common ecological gradients of water depth and geographic distance provide important insights into diversification processes in a lake with high levels of human disturbance from over-fishing. Results Four putative lake trout ecotypes could not be distinguished using population genetic methods, despite morphological differences. Neutral genetic partitioning in lake trout was stronger along a gradient of water depth, than by locality or ecotype. Contemporary genetic migration patterns were consistent with isolation-by-depth. Historical gene flow patterns indicated colonization from shallow to deep water. Comparison of phenotypic ( Pst ) and neutral genetic variation ( Fst ) revealed that morphological traits related to swimming performance (e.g., buoyancy, pelvic fin length) departed more strongly from neutral expectations along a depth gradient than craniofacial feeding traits. Elevated phenotypic variance with increasing water depth in pelvic fin length indicated possible ongoing character release and diversification. Finally, differences in early growth rate and asymptotic fish length across depth strata may be associated with limiting factors attributable to cold deep-water environments. Conclusion We provide evidence of reductions in gene flow and divergent natural selection associated with water depth in Lake Superior. Such information is relevant for documenting intraspecific biodiversity in the largest freshwater lake in the world for a species that recently lost considerable genetic diversity and is now in recovery. Unknown is whether observed patterns are a result of an early stage of incipient speciation, gene flow-selection equilibrium, or reverse speciation causing formerly divergent ecotypes to collapse into a single gene pool.

Lake Superior, Isle Royale↗

USGS2018

New insights into the phylogenetics and population structure of the prairie falcon (Falco mexicanus)

Background Management requires a robust understanding of between- and within-species genetic variability, however such data are still lacking in many species. For example, although multiple population genetics studies of the peregrine falcon ( Falco peregrinus ) have been conducted, no similar studies have been done of the closely-related prairie falcon ( F. mexicanus ) and it is unclear how much genetic variation and population structure exists across the species’ range. Furthermore, the phylogenetic relationship of F. mexicanus relative to other falcon species is contested. We utilized a genomics approach (i.e., genome sequencing and assembly followed by single nucleotide polymorphism genotyping) to rapidly address these gaps in knowledge. Results We sequenced the genome of a single female prairie falcon and generated a 1.17 Gb (gigabases) draft genome assembly. We generated maximum likelihood phylogenetic trees using complete mitochondrial genomes as well as nuclear protein-coding genes. This process provided evidence that F. mexicanus is an outgroup to the clade that includes the peregrine falcon and members of the subgenus Hierofalco. We annotated > 16,000 genes and almost 600,000 high-quality single nucleotide polymorphisms (SNPs) in the nuclear genome, providing the raw material for a SNP assay design featuring > 140 gene-associated markers and a molecular-sexing marker. We subsequently genotyped ~ 100 individuals from California (including the San Francisco East Bay Area, Pinnacles National Park and the Mojave Desert) and Idaho (Snake River Birds of Prey National Conservation Area). We tested for population structure and found evidence that individuals sampled in California and Idaho represent a single panmictic population. Conclusions Our study illustrates how genomic resources can rapidly shed light on genetic variability in understudied species and resolve phylogenetic relationships. Furthermore, we found evidence of a single, randomly mating population of prairie falcons across our sampling locations. Prairie falcons are highly mobile and relatively rare long-distance dispersal events may promote gene flow throughout the range. As such, California’s prairie falcons might be managed as a single population, indicating that management actions undertaken to benefit the species at the local level have the potential to influence the species as a whole.

BMC Genomics↗

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.

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