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At least 1,351 records · Page 75Linked to original sources

Ecological status of aquatic communities in selected streams in the Milwaukee Metropolitan Sewerage District planning area of Wisconsin, 2004–13

A total of 14 wadable streams in urban or urbanizing watersheds near Milwaukee, Wisconsin, were sampled in 2004, 2007, 2010, and 2013 to assess the ecological status of aquatic communities (biota), including benthic algae and invertebrates, and fish. To assess temporal variation, additional community sampling was also done at a subset of three sites in 2011 and 2012. Relative abundances of each type of organism were used to calculate biological metrics, such as richness and diversity, percentages of intolerant and tolerant organisms, and indexes of biotic integrity for invertebrates and fish. Selected environmental (physical and chemical) data in the streams were collected to evaluate potential relations to the biota and the ecological health of the stream. Physical and chemical data included land use/land cover, stream discharge from U.S. Geological Survey (USGS) streamgages (except at 2 creeks that were not gaged), stream habitat, microhabitat at invertebrate collection locations, water quality (except at 2 creeks that were not gaged), field measurements of several water-quality constituents, measures of benthic algal biomass, and toxicity and chemical tests on extracts from passive samplers deployed at a subset of 6 sites. Relative abundances of organisms and biological metrics were compared among sampling years and with environmental metrics to evaluate the ecological status of these streams and determine primary stressors on the aquatic communities, with the aim of helping resource managers understand and work toward improving the ecological health of these and other urban and urbanizing rivers in the study area. Biological metrics for most sites indicated some level of diminished ecological status when compared across all sampled sites and when compared with rating scales for selected metrics. The least degraded sites among all those sampled—indicated by aggregate bioassessments for algae, invertebrates, and fish metrics and in order starting with the best overall condition—were the Milwaukee River near Cedarburg, Menomonee River at Menomonee Falls, Jewel Creek, and Milwaukee River at Milwaukee. The most degraded sites were Menomonee River at Wauwatosa, Root River at Greenfield, Lincoln Creek, and the Kinnickinnic River. Differences in aggregate bioassessments indicate that aquatic communities at the Menomonee River at Wauwatosa site and the Root River at Greenfield site were worse in 2013 than in 2004; however, Oak Creek and Honey Creek sites were better. In 2013, several sites had less than 30-percent pollution-sensitive diatoms indicating degraded algal assemblages. Invertebrate metrics for most of the 14 sites in 2013 were lower than in 2004 and indicate that invertebrate assemblages at most sampled sites were more degraded in 2013. Tolerant fish taxa made up more than 40 percent of assemblages at most sites and nearly 100 percent of assemblages at four sites. At times, in some smaller streams, too few fish were captured to compute an Index of Biotic Integrity with confidence, and invertebrates provided a better means for assessing the ecological status and water quality. With these few exceptions, the use of all three groups of biota provided the most robust assessments at the 14 sites in 2004–13. Physical and chemical stressors were correlated to adverse effects on aquatic biota at the sampled streams. Passive samplers were deployed at a subset of six sites in 2013. Microtox results indicated there was little or no toxicity at the Milwaukee River near Cedarburg site and at the Oak Creek site, slight toxicity at the Lincoln Creek and Honey Creek sites, and moderate toxicity at the Milwaukee River at Milwaukee site and the Little Menomonee River site; however, based on cytochrome-P450 reporter gene system toxicity tests, potential toxicity from hydrophobic organic contaminants was measured at all six sites. For all 14 sites, physical and chemical stressors related to urbanization correlated with biological metrics for algae, invertebrates, and fish. Most stressors for aquatic biota reflected an urban signature. Stressors related to ecological condition in our study were chemical and physical, such as developed land, impervious surface in the watershed, urban land in a buffer area around the stream (a 100-foot [30-meter]-wide area on each side of the stream, and maximum instantaneous discharge normalized by drainage area (a measure of flood and scour effects). Chemical stressors included low waterborne concentrations of dissolved oxygen and high concentrations of chloride, zinc and other metals, nutrients (nitrite and phosphorus), and fecal coliform bacteria. Although algae, invertebrates, and fish did not always demonstrate a significant response to the same stressors, higher abundances of high total phosphorus-indicator diatoms, lower ratings for invertebrate biotic integrity indexes and percentages of mayflies-stoneflies-caddisflies, and lower values for fish biotic integrity indexes underscored possible adverse effects of even low levels of developed land. Developed land is typically associated with more rapid runoff, which washes chemicals from impervious surfaces into area waterways and degrades stream habitat for aquatic communities. However, with respect to at least chloride from road salt, diatoms tolerant to dissolved salts were significantly lower with as little as 1-percent mixed forest in the watershed. Lower percentages of urban land in the stream buffer correlated with healthier aquatic assemblages of algae, invertebrates, and fish. The assessment of algal, invertebrate, and fish assemblages coupled with physical and chemical data were highly useful in evaluating the ecological status of aquatic communities at the 14 sites and for determining environmental stressors that may be contributing to reduced stream condition. Some of the stressors could potentially be removed or lessened with stream rehabilitation or changes in watershed management.

Wisconsin↗

Identification of bacteria in groundwater used for domestic supply in the southeast San Joaquin Valley, California, 2014

Groundwater is an important source of drinking water in California. Water-borne diseases caused by microbial contamination are a growing concern. The MI test, a membrane filtration method for the chromogenic/fluorogenic detection of total coliforms and Escherichia coli , was used for samples collected January to April 2014 from 42 domestic wells in the southeastern San Joaquin Valley. The wells were sampled as part of the Groundwater Ambient Monitoring and Assessment Program Priority Basin Project (GAMA-PBP), a cooperative study between the U.S. Geological Survey and the California State Water Resources Control Board. Polymerase chain reaction analysis and sequencing of deoxyribonucleic acid (DNA) were used for 34 target and nontarget colonies that grew on the MI media from samples collected from 13 of the domestic wells to identify what genera of bacteria could exist in groundwater used by domestic wells. Gene sequences obtained using the Sanger method were entered into the basic local alignment search tool (BLAST) database, and 17 genera of bacteria were identified. Of these, 13 genera contain species that are human pathogens or opportunistic human pathogens. All the genera that include human pathogens are naturally present in soil, plants, or water; one of the pathogens also can be found in fecal matter. Six of the human pathogens were from non-target colony growth on the MI media. Target and non-target microbial growth on MI media are indicators of the possible presence of pathogenic bacteria even if the bacteria naturally are from soil rather than from a fecal source.

California↗

Isolation and identification of microcystin-degrading bacteria in Lake Erie source waters and drinking-water plant sand filters

The increasing prevalence of cyanobacterial harmful algal blooms and the toxins they produce is a global water-quality issue. In the Western Basin of Lake Erie, high microcystin concentrations have led to water-quality advisories, process adjustments for treating drinking water, and increased water-quality monitoring. Biodegradation is an environmentally friendly and cost-effective way to reduce concentrations of microcystins in drinking water; however, few studies have been done to determine biodegradation potential of bacteria indigenous to the Lake Erie watershed. As part of a cooperative program between the U.S. Geological Survey and the U.S. Environmental Protection Agency, this study aimed to identify naturally occurring microcystin-degrading bacteria in source waters and in the sand filters of drinking-water treatment plants in the Western Basin of Lake Erie. Biodegradation of microcystin-LR was found to occur in microcosms developed with three different Lake Erie-area sources—Lake Erie water, water from storage reservoirs supplied by inland streams, and water or solid medium from sand/anthracite filters at drinking-water plants. In microplates with microcystin-LR as the sole carbon source, 10 isolates exhibited cellular respiration and were, therefore, identified as promising microcystin biodegraders; 4 of those isolates subsequently were found to have potential to form biofilms. The 10 promising isolates along with 14 additional isolates from the microcosms were identified by 16S ribosomal RNA sequencing: 15 isolates were γ-proteobacteria, 6 isolates were β-proteobacteria, 1 isolate was an α-proteobacterium, 1 isolate was a flavobacterium in the phylum Bacteroidetes , and 1 isolate was in the phylum Actinobacteria . Isolates were screened for possession of the mlrA gene (found to encode for the protein responsible for cleaving the cyclic structure of microcystin), and results indicate that, for Lake Erie source waters and elsewhere, more work would be required to identify microcystin-biodegradation pathways and products and to confirm biodegradation rates in pure culture isolates.

Lake Erie↗

Colorimetric determination of nitrate plus nitrite in water by enzymatic reduction, automated discrete analyzer methods

This report documents work at the U.S. Geological Survey (USGS) National Water Quality Laboratory (NWQL) to validate enzymatic reduction, colorimetric determinative methods for nitrate + nitrite in filtered water by automated discrete analysis. In these standard- and low-level methods (USGS I-2547-11 and I-2548-11), nitrate is reduced to nitrite with nontoxic, soluble nitrate reductase rather than toxic, granular, copperized cadmium used in the longstanding USGS automated continuous-flow analyzer methods I-2545-90 (NWQL laboratory code 1975) and I-2546-91 (NWQL laboratory code 1979). Colorimetric reagents used to determine resulting nitrite in aforementioned enzymatic- and cadmium-reduction methods are identical. The enzyme used in these discrete analyzer methods, designated AtNaR2 by its manufacturer, is produced by recombinant expression of the nitrate reductase gene from wall cress (Arabidopsis thaliana) in the yeast Pichia pastoris. Unlike other commercially available nitrate reductases we evaluated, AtNaR2 maintains high activity at 37&deg;C and is not inhibited by high-phenolic-content humic acids at reaction temperatures in the range of 20&deg;C to 37&deg;C. These previously unrecognized AtNaR2 characteristics are essential for successful performance of discrete analyzer nitrate + nitrite assays (henceforth, DA-AtNaR2) described here. Method detection levels (or limits; MDL) estimated for standard- and low-level DA-AtNaR2 nitrate + nitrite methods were 0.02 milligrams nitrogen per liter (mg-N/L) and 0.002 mg-N/L, respectively, which are comparable to 2010 NWQL long-term MDLs of the continuous-flow analyzer, cadmium-reduction methods (henceforth, CFA-CdR) they replace. Typically, reagent-water blanks for standard- and low-level DAAtNaR2 nitrate + nitrite methods are one half MDL or less. Nitrate + nitrite concentration differences for between-day replicates were 3 percent or less at or above 5 times the MDL and were as great as 35 percent near the MDL. Typically, nitrate spike recoveries from reagent water, surface water, groundwater, and high-phenolic-content, humic-acid-amended reagent water were 100&plusmn;20 percent. In addition to operational details and performance benchmarks for these new DA-AtNaR2 nitrate + nitrite assays, this report also provides results of interference studies for common inorganic and organic matrix constituents at 1, 10, and 100 times their median concentrations in surface-water and groundwater samples submitted annually to the NWQL for nitrate + nitrite analyses. Paired t-test and Wilcoxon signed-rank statistical analyses of results determined by CFA-CdR methods and DA-AtNaR2 methods indicate that nitrate concentration differences between population means or sign ranks were either statistically equivalent to zero at the 95 percent confidence level (p &ge; 0.05) or analytically equivalent to zero-that is, when p < 0.05, concentration differences between population means or medians were less than MDLs.

Techniques and Methods↗

Comparison of genome size and synthesis of structural proteins of Hirame Rhabdovirus, infectious hematopoietic necrosis virus, and viral hemorrhagic Septicemia virus

Genomic RNA was extracted from purified virions of hirame rhabdovirus (HRV), infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV). The full-length RNA was analyzed using formaldehyde agarose gel electrophoresis followed by ethidium bromide staining. Compared with an internal RNA size standard, all three viral genomic RNAs appeared to have identical relative mobilities and were estimated to be approximately 10.7 kilobases in length or about 3.7 megadaltons in molecular mass. Structural protein synthesis of HRV, IHNV, and VHSV was studied using cell cultures treated with actinomycin D. At 2 h intervals, proteins were labeled with 35 S-methionine, extracted, and analyzed by SDS-polyacrylamide gel electrophoresis and autoradiography. The five structural proteins of each of the three viruses appeared in the following order : nucleoprotein (N), matrix protein 1 (M1), matrix protein 2 (M2), glycoprotein (G), and polymerase (L) reflecting both the approximate relative abundance of each protein within infected cells and the gene order within the viral genome.

Fish Pathology↗

A novel picorna-like virus in a Wabash Pigtoe (Fusconaia flava) from the upper Mississippi River, USA

Unionid mussels are threatened by multiple environmental stressors and have experienced mass mortality events over the last several decades, but the role of infectious disease in unionid health and population declines remains poorly understood. Although several microbial agents have been found in unionids, to date only one virus has been documented—Lea plague virus ( Arenaviridae ) in propagated Triangle Shell mussels ( Hyriopsis cumingii ) in China. We used next-generation DNA sequencing to screen hemolymph of seven individuals of five unionid species from the Upper Mississippi River basin, USA for viruses. We identified the complete polyprotein gene of a novel picornalike virus in one individual of the Wabash Pigtoe ( Fusconaia flava ). The virus is a member of the Nora virus clade of picornalike viruses and is most closely related to viruses from arthropods in China. We did not detect viruses in another Wabash Pigtoe or in animals of the other four species. It is premature to make inferences about the role of this virus in the health of Wabash Pigtoes or other unionid species or the origin or transmission of this virus. Nevertheless, to our knowledge, our results represent the first report of a virus in wild North American unionids. Technologies based on next-generation DNA sequencing should prove useful for identifying new viruses and investigating their role in unionid health and disease.

Minnesota, Wisconsin↗

Detection of group 1 coronaviruses in bats in North America

The epidemic of severe acute respiratory syndrome (SARS) was caused by a newly emerged coronavirus (SARS-CoV). Bats of several species in southern People's Republic of China harbor SARS-like CoVs and may be reservoir hosts for them. To determine whether bats in North America also harbor coronaviruses, we used reverse transcription-PCR to detect coronavirus RNA in bats. We found coronavirus RNA in 6 of 28 fecal specimens from bats of 2 of 7 species tested. The prevalence of viral RNA shedding was high: 17% in Eptesicus fuscus and 50% in Myotis occultus. Sequence analysis of a 440-bp amplicon in gene 1b showed that these Rocky Mountain bat coronaviruses formed 3 clusters in phylogenetic group 1 that were distinct from group 1 coronaviruses of Asian bats. Because of the potential for bat coronaviruses to cause disease in humans and animals, further surveillance and characterization of bat coronaviruses in North America are needed.

Emerging Infectious Diseases↗

Gnathostoma spinigerum in live Asian swamp eels (Monopterus spp.) from food markets and wild populations, United States

In Southeast Asia, swamp eels (Synbranchidae: Monopterus spp.) are a common source of human gnathostomiasis, a foodborne zoonosis caused by advanced third-stage larvae (AL3) of Gnathostoma spp. nematodes. Live Asian swamp eels are imported to US ethnic food markets, and wild populations exist in several states. To determine whether these eels are infected, we examined 47 eels from markets and 67 wild-caught specimens. Nematodes were identified by morphologic features and ribosomal intergenic transcribed spacer&ndash;2 gene sequencing. Thirteen (27.7%) M. cuchia eels from markets were infected with 36 live G. spinigerum AL3: 21 (58.3%) in liver; 7 (19.4%) in muscle; 5 (13.8%) in gastrointestinal tract, and 3 (8.3%) in kidneys. Three (4.5%) wild-caught M. albus eels were infected with 5 G. turgidum AL3 in muscle, and 1 G. lamothei AL3 was found in a kidney (both North American spp.). Imported live eels are a potential source of human gnathostomiasis in the United States.

Emerging Infectious Diseases↗

Evidence for free-living Bacteroides in Cladophora along the shores of the Great Lakes

Bacteroides is assumed to be restricted to the alimentary canal of animals and humans and is considered to be non-viable in ambient environments. We hypothesized that Bacteroides could persist and replicate within beach-stranded Cladophora glomerata mats in southern Lake Michigan, USA. Mean Bacteroides concentration (per GenBac3 Taqman quantitative PCR assay) during summer 2012 at Jeorse Park Beach was 5.2 log calibrator cell equivalents (CCE) g -1 dry weight (dw), ranging from 3.7 to 6.7. We monitored a single beach-stranded mat for 3 wk; bacterial concentrations increased by 1.6 log CCE g -1 dw and correlated significantly with ambient temperature (p = 0.003). Clonal growth was evident, as observed by >99% nucleotide sequence similarity among clones. In in vitro studies, Bacteroides concentrations increased by 5.5 log CCE g -1 after 7 d (27&deg;C) in fresh Cladophora collected from rocks. Partial sequencing of the 16S rRNA gene of 36 clones from the incubation experiment showed highly similar genotypes (&ge;97% sequence overlap). The closest enteric Bacteroides spp. from the National Center for Biotechnology Information database were only 87 to 91% similar. Genomic similarity, clonality, growth, and persistence collectively suggest that putative, free-living Bacteroides inhabit Cladophora mats of southern Lake Michigan. These findings may have important biological, medical, regulatory, microbial source tracking, and public health implications.

southern Lake Michigan↗

Covariance of bacterioplankton composition and environmental variables in a temperate delta system

We examined seasonal and spatial variation in bacterioplankton composition in the Sacramento-San Joaquin River Delta (CA) using terminal restriction fragment length polymorphism (T-RFLP) analysis. Cloned 16S rRNA genes from this system were used for putative identification of taxa dominating the T-RFLP profiles. Both cloning and T-RFLP analysis indicated that Actinobacteria , Verrucomicrobia , Cytophaga-Flavobacterium and Proteobacteria were the most abundant bacterioplankton groups in the Delta. Despite the broad variety of sampled habitats (deep water channels, lakes, marshes, agricultural drains, freshwater and brackish areas), and the spatial and temporal differences in hydrology, temperature and water chemistry among the sampling campaigns, T-RFLP electropherograms from all samples were similar, indicating that the same bacterioplankton phylotypes dominated in the various habitats of the Delta throughout the year. However, principal component analysis (PCA) and partial least-squares regression (PLS) of T-RFLP profiles revealed consistent grouping of samples on a seasonal, but not a spatial, basis. b- Proteobacteria related to Ralstonia , Actinobacteria related to Microthrix , and b- Proteobacteria identical to the environmental Clone LD12 had the highest relative abundance in summer/fall T-RFLP profiles and were associated with low river flow, high pH, and a number of optical and chemical characteristics of dissolved organic carbon (DOC) indicative of an increased proportion of phytoplankton-produced organic material as opposed to allochthonous, terrestrially derived organic material. On the other hand, Geobacter -related b- Proteobacteria showed a relative increase in abundance in T-RFLP analysis during winter/spring, and probably were washed out from watershed soils or sediment. Various phylotypes associated with the same phylogenetic division, based on tentative identification of T-RFLP fragments, exhibited diverse seasonal patterns, suggesting that ecological roles of Delta bacterioplankton were partitioned at the genus or species level.

Aquatic Microbial Ecology↗

Sequence analysis of the internal transcribed spacer (ITS) region reveals a novel clade of Ichthyophonus sp. from rainbow trout

The mesomycetozoean parasite Ichthyophonus hoferi is most commonly associated with marine fish hosts but also occurs in some components of the freshwater rainbow trout Oncorhynchus mykiss aquaculture industry in Idaho, USA. It is not certain how the parasite was introduced into rainbow trout culture, but it might have been associated with the historical practice of feeding raw, ground common carp Cyprinus carpio that were caught by commercial fisherman. Here, we report a major genetic division between west coast freshwater and marine isolates of Ichthyophonus hoferi. Sequence differences were not detected in 2 regions of the highly conserved small subunit (18S) rDNA gene; however, nucleotide variation was seen in internal transcribed spacer loci (ITS1 and ITS2), both within and among the isolates. Intra-isolate variation ranged from 2.4 to 7.6 nucleotides over a region consisting of ~740 bp. Majority consensus sequences from marine/anadromous hosts differed in only 0 to 3 nucleotides (99.6 to 100% nucleotide identity), while those derived from freshwater rainbow trout had no nucleotide substitutions relative to each other. However, the consensus sequences between isolates from freshwater rainbow trout and those from marine/anadromous hosts differed in 13 to 16 nucleotides (97.8 to 98.2% nucleotide identity).

Diseases of Aquatic Organisms↗

Emergence of viral hemorrhagic septicemia virus in the North American Great Lakes region is associated with low viral genetic diversity

Viral hemorrhagic septicemia virus (VHSV) is a fish rhabdovirus that causes disease in a broad range of marine and freshwater hosts. The known geographic range includes the Northern Atlantic and Pacific Oceans, and recently it has invaded the Great Lakes region of North Ame­rica. The goal of this work was to characterize genetic diversity of Great Lakes VHSV isolates at the early stage of this viral emergence by comparing a partial glycoprotein (G) gene sequence (669 nt) of 108 isolates collected from 2003 to 2009 from 31 species and at 37 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVb within the major VHSV genetic group IV. Among these 108 isolates, genetic diversity was low, with a maximum of 1.05% within the 669 nt region. There were 11 unique sequences, designated vcG001 to vcG011. Two dominant sequence types, vcG001 and vcG002, accounted for 90% (97 of 108) of the isolates. The vcG001 isolates were most widespread. We saw no apparent association of sequence type with host or year of isolation, but we did note a spatial pattern, in which vcG002 isolates were more prevalent in the easternmost sub-regions, including inland New York state and the St. Lawrence Seaway. Different sequence types were found among isolates from single disease outbreaks, and mixtures of types were evident within 2 isolates from ­individual fish. Overall, the genetic diversity of VHSV in the Great Lakes region was found to be extremely low, consistent with an introduction of a new virus into a geographic region with ­previously naïve host populations.

Diseases of Aquatic Organisms↗

Genetic basis of differences in myxospore count between whirling disease-resistant and -susceptible strains of rainbow trout

We used a quantitative genetics approach and estimated broad sense heritability (h 2 b ) of myxospore count and the number of genes involved in myxospore formation to gain a better understanding of how resistance to Myxobolus cerebralis, the parasite responsible for whirling disease, is inherited in rainbow trout Oncorhynchus mykiss. An M. cerebralis-resistant strain of rainbow trout, the German Rainbow (GR), and a wild, susceptible strain of rainbow trout, the Colorado River Rainbow (CRR), were spawned to create 3 intermediate crossed populations (an F1 cross, F2 intercross, and a B2 backcross between the F1 and the CRR). Within each strain or cross, h 2 b was estimated from the between-family variance of myxospore counts using full-sibling families. Estimates of h 2 b and average myxospore counts were lowest in the GR strain, F1 cross, and F2 intercross (h 2 b = 0.34, 0.42, and 0.34; myxospores fish −1 = 275, 9566, and 45780, respectively), and highest in the B2 backcross and CRR strain (h 2 b = 0.93 and 0.89; myxospores fish −1 = 97865 and 187595, respectively). Comparison of means and a joint-scaling test suggest that resistance alleles arising from the GR strain are dominant to susceptible alleles from the CRR strain. Resistance was retained in the intermediate crosses but decreased as filial generation number increased (F2) or backcrossing occurred (B2). The estimated number of segregating loci responsible for differences in myxospore count in the parental strains was 9 ± 5. Our results indicate that resistance to M. cerebralis is a heritable trait within these populations and would respond to either artificial selection in hatcheries or natural selection in the wild.

Diseases of Aquatic Organisms↗

Disease surveillance of Atlantic herring: molecular characterization of hepatic coccidiosis and a morphological report of a novel intestinal coccidian

Surveillance for pathogens of Atlantic herring, including viral hemorrhagic septicemia virus (VHSV), Ichthyophonus hoferi , and hepatic and intestinal coccidians, was conducted from 2012 to 2016 in the NW Atlantic Ocean, New Jersey, USA. Neither VHSV nor I. hoferi was detected in any sample. Goussia clupearum was found in the livers of 40 to 78% of adult herring in varying parasite loads; however, associated pathological changes were negligible. Phylogenetic analysis based on small subunit 18S rRNA gene sequences placed G. clupearum most closely with other extraintestinal liver coccidia from the genus Calyptospora , though the G. clupearum isolates had a unique nucleotide insertion between 604 and 729 bp that did not occur in any other coccidian species. G. clupearum oocysts from Atlantic and Pacific herring were morphologically similar, though differences occurred in oocyst dimensions. Comparison of G. clupearum genetic sequences from Atlantic and Pacific herring revealed 4 nucleotide substitutions and 2 gaps in a 1749 bp region, indicating some divergence in the geographically separate populations. Pacific G. clupearum oocysts were not directly infective, suggesting that a heteroxenous life cycle is likely. Intestinal coccidiosis was described for the first time from juvenile and adult Atlantic herring. A novel intestinal coccidian species was detected based on morphological characteristics of exogenously sporulated oocysts. A unique feature in these oocysts was the presence of 3 long (15.1 &plusmn; 5.1 &micro;m, mean &plusmn;SD) spiny projections on both ends of the oocyst. The novel morphology of this coccidian led us to tentatively name this parasite G. echinata n. sp.

New Jersey↗

Complete sequences of 4 viral hemorrhagic septicemia virus IVb isolates and their virulence in northern pike fry

Four viral hemorrhagic septicemia virus (VHSV) genotype IVb isolates were sequenced, their genetic variation explored, and comparative virulence assayed with experimental infections of northern pike Esox lucius fry. In addition to the type strain MI03, the complete 11183 bp genome of the first round goby Neogobius melanostomus isolate from the St. Lawrence River, and the 2013 and 2014 isolates from gizzard shad Dorosoma cepedianum die-offs in Irondequoit Bay, Lake Ontario and Dunkirk Harbor, Lake Erie were all deep sequenced on an Illumina platform. Mutations documented in the 11 yr since the MI03 index case from Lake St. Clair muskellunge Esox masquinongy showed 87 polymorphisms among the 4 isolates. Twenty-six mutations were non-synonymous and located at 18 different positions within the matrix protein, glycoprotein, non-virion protein, and RNA polymerase genes. The same 4 isolates were used to infect northern pike fry by a single 1 h bath exposure. Cumulative percent mortality varied from 42.5 to 62.5%. VHSV was detected in 57% (41/72) of the survivors at the end of the 21-d trial, suggesting that the virus was not rapidly cleared. Lesions were observed in many of the moribund and dead northern pike, such as hemorrhaging in the skin and fins, as well as hydrocephalus. Mean viral load measured from the trunk and visceral tissues of MI03-infected pike was significantly higher than the quantities detected in fish infected with the most recent isolates of genotype IVb, but there were no differences in cumulative mortality observed.

Diseases of Aquatic Organisms↗

Hyperpigmented melanistic skin lesions of smallmouth bass Micropterus dolomieu from the Chesapeake Bay watershed

Hyperpigmented melanistic skin lesions (HPMLs) of smallmouth bass Micropterus dolomieu are observed in the Potomac and Susquehanna rivers, Chesapeake Bay watershed, USA. Routine, nonlethal population surveys were conducted at 8 sites on the mainstem Susquehanna River and 9 on the Juniata River, a tributary of the Susquehanna River, between 2012 and 2018, and the prevalence of HPMLs was documented. A total of 4078 smallmouth bass were collected from the mainstem Susquehanna River and 6478 from the Juniata River. Lesions were primarily seen in bass greater than 200 mm, and prevalence in the Susquehanna River (8%) was higher (p < 0.001) than in the Juniata River (2%). As part of ongoing fish health monitoring projects, smallmouth bass were collected at additional sites, primarily tributaries of the Susquehanna (n = 758) and Potomac (n = 545) rivers between 2013 and 2018. Prevalence in the Susquehanna River (13%) was higher (p < 0.001) than the Potomac (3%). Microscopically, HPMLs were characterized by an increased number of melanocytes in the epidermis or within the dermis and epidermis. RNAseq analyses of normal and melanistic skin identified 3 unique sequences in HPMLs. Two were unidentified and the third was a viral helicase (E1). Transcript abundance in 16 normal skin samples and 16 HPMLs showed upregulation of genes associated with melanogenesis and cell proliferation in HPMLs. The E1 transcript was detected in 12 of the 16 melanistic areas but in no samples from normal skin. Further research will be necessary to identify the putative new virus and determine its role in melanocyte proliferation.

Delaware, Maryland, Pennsylvania, Virginia, West V↗

Evaluation of the protective immunogencity of the N, P, M, NV and G proteins of infectious hematopoietic necrosis virus in rainbow trout Oncorhynchus mykiss using DNA vaccines

The protective immunogenicity of the nucleoprotein (N), phosphoprotein (P), matrix protein (M), non-virion protein (NV) and glycoprotein (G) of the rhabdovirus infectious hematopoietic necrosis virus (IHNV) was assessed in rainbow trout using DNA vaccine technology. DNA vaccines were produced by amplifying and cloning the viral genes in the plasmid pCDNA 3.1. The protective immunity elicited by each vaccine was evaluated through survival of immunized fry after challenge with live virus. Neutralizing antibody titers were also determined in vaccinated rainbow trout Oncorhynchus mykiss fry (mean weight 2 g) and 150 g sockeye salmon Oncorhynchus nerka . The serum from the 150 g fish was also used in passive immunization studies with naïve fry. Our results showed that neither the internal structural proteins (N, P and M) nor the NV protein of IHNV induced protective immunity in fry or neutralizing antibodies in fry and 150 g fish when expressed by a DNA vaccine construct. The G protein, however, did confer significant protection in fry up to 80 d post-immunization and induced protective neutralizing antibodies. We are currently investigating the role of different arms of the fish immune system that contribute to the high level of protection against IHNV seen in vaccinated fish.

Diseases of Aquatic Organisms↗