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Tony Goldberg

Publications and source records attributed to Tony Goldberg.

15 recordsLinked to original sources

Bacteriological analysis of unionid hemolymph collected from freshwater mussel populations in the Pacific northwestern United States

Native freshwater mussel (Unionidae) mortality events have been occurring with increased frequency in recent decades, with few investigations into potential etiological agents. In the western United States, no surveys have been published regarding the bacteria associated with unionid mussels. Herein, we examine locations of known mussel mortality events in the Chehalis River (Washington), in the Crooked River (Oregon), and Owyhee River (Oregon). Mussel populations considered healthy were sampled in the Skookumchuck River (Washington) for comparison. A variety of bacteria were isolated from these populations, and most notably, Acinetobacter spp. were identified from 82% of moribund individuals of Gonidea angulata in the Owyhee River. Future work evaluating whether Acinetobacter spp. are pathogenic to freshwater mussels could be valuable in unraveling the factors associated with these enigmatic mortality events.

Oregon, Washington

Mussel mass mortality in the Clinch River, USA: Metabolomics detects affected pathways and biomarkers of stress

Biologists monitoring freshwater mussel (order Unionida) populations rely on behavioral, often subjective, signs to identify moribund (“sick”) or stressed mussels, such as gaping valves and slow response to probing, and they lack clinical indicators to support a diagnosis. As part of a multi-year study to investigate causes of reoccurring mortality of pheasantshell ( Ortmanniana pectorosa; synonym Actinonaias pectorosa ) in the Clinch River, Virginia and Tennessee, USA, we analyzed the hemolymph metabolome of a subset of mussels from the 2018 sampling period. Mussels at the mortality sites were diagnosed in the field as affected (case) or unaffected (control) based on behavioral and physical signs. Hemolymph was collected in the field by non-lethal methods from the anterior adductor muscle for analysis. We used ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectroscopy to detect targeted and untargeted metabolites in hemolymph and compared metabolomic profiles by field assessment of clinical status. Targeted biomarker analysis found 13 metabolites associated with field assessments of clinical status. Of these, increased gamma-linolenic acid and N -methyl- L -alanine were most indicative of case mussels, while adenine and inosine were the best indicators of control mussels. Five pathways in the targeted analysis differed by clinical status; two of these, purine metabolism and glycerophospholipid metabolism, were also indicated in the untargeted analysis. In the untargeted nalysis, 22 metabolic pathways were associated with clinical status. Many of the impacted pathways in the case group were catabolic processes, such as degradation of amino acids and fatty acids. Hierarchical clustering analysis matched clinical status in 72% (18 of 25) of mussels, with control mussels more frequently (5 of 16) not matching clinical status. Our study demonstrated that metabolomic analysis of hemolymph is suitable for assessing mussel condition and complements field-based indicators of health.

Tennessee, Virginia

Long-term assessment of relationships between changing environmental conditions and the physiology of southern Beaufort Sea polar bears (Ursus maritimus)

Climate change is influencing polar bear ( Ursus maritimus ) habitat, diet, and behavior but the effects of these changes on their physiology is not well understood. Blood-based biomarkers are used to assess the physiologic health of individuals but their usefulness for evaluating population health, especially as it relates to changing environmental conditions, has rarely been explored. We describe links between environmental conditions and physiologic functions of southern Beaufort Sea polar bears using data from blood samples collected from 1984 to 2018, a period marked by extensive environmental change. We evaluated associations between 13 physiologic biomarkers and circumpolar (Arctic oscillation index) and regional (wind patterns and ice-free days) environmental metrics and seasonal and demographic co-variates (age, sex, season, and year) known to affect polar bear ecology. We observed signs of dysregulation of water balance in polar bears following years with a lower annual Arctic oscillation index. In addition, liver enzyme values increased over time, which is suggestive of potential hepatocyte damage as the Arctic has warmed. Biomarkers of immune function increased with regional-scale wind patterns and the number of ice-free days over the Beaufort Sea continental shelf and were lower in years with a lower winter Arctic oscillation index, suggesting an increased allocation of energetic resources for immune processes under these conditions. We propose that the variation in polar bear immune and metabolic function is likely indicative of physiologic plasticity, a response that allows polar bears to remain in homeostasis even as they experience changes in nutrition and habitat in response to changing environments.

Alaska

Serum virome of southern Beaufort Sea polar bears (Ursus maritimus) during a period of rapid climate change

Climate change affects the behavior, physiology and life history of many Arctic wildlife species. It can also influence the distribution and ecology of infectious agents. The southern Beaufort Sea (SB) subpopulation of polar bears ( Ursus maritimus ) has experienced dramatic behavioral changes due to retreating sea ice and other climate-related factors, but the effects of these changes on physiology and infection remain poorly understood. Using serum from polar bears sampled between 2004 and 2015 and metagenomic DNA sequencing, we identified 48 viruses, all of the family Anelloviridae . Anelloviruses are small, ubiquitous infectious agents with circular single-stranded DNA genomes that are not known to cause disease but, in humans, covary in diversity and load with immunological compromise. We therefore examined the usefulness of anelloviruses as biomarkers of polar bear physiological stress related to climate and habitat use. Polar bear anelloviruses sorted into two distinct clades on a phylogenetic tree, both of which also contained anelloviruses of giant pandas ( Ailuropoda melanoleuca ), another ursid. Neither anellovirus diversity nor load were associated with any demographic variables, behavioral factors or direct physiological measures. However, pairwise genetic distances between anelloviruses were positively correlated with pairwise differences in sampling date, suggesting that the polar bear “anellome”is evolving over time. These findings suggest that anelloviruses are not a sensitive indicator of polar physiological stress, but they do provide a baseline for evaluating future changes to polar bear viromes.

Alaska

A bacteriological comparison of the hemolymph from healthy and moribund unionid mussel populations in the upper Midwestern U.S.A. prompts the development of diagnostic assays to detect Yokenella regensburgei

Recent bacteriological investigations of freshwater mussel mortality events in the southeastern United States have identified a variety of bacteria and differences in bacterial communities between sick and healthy mussels. In particular, Yokenella regensburgei and Aeromonas spp. have been shown to be associated with moribund mussels, although it remains unclear whether these bacteria are causes or consequences of disease. To further understand the role of bacteria in mussel epizootics, we investigated mortality events that occurred in the upper Midwest in the Embarrass River (Wisconsin) and the Huron River (Michigan). For comparison, we also studied mussels from an unaffected population in the St. Croix River (Wisconsin). Diverse bacterial genera were identified from these sites, including Y. regensburgei from moribund mussels in the Embarrass River (Wisconsin). This bacterium has also been consistently isolated during ongoing mortality events in the Clinch River (Virginia). Subsequently, we developed and validated molecular assays for the detection of Yokenella to use in future investigations of mussel mortality events and to identify environmental reservoirs of this bacterium.

Michigan, Minnesota, Wisconsin

Further bacteriological analysis of annual Pheasantshell (Actinonaias pectorosa) mussel mortality events in the Clinch River (Virginia/Tennessee), USA, reveals a consistent association with Yokenella Regensburgei

Pheasantshell ( Actinonaias pectorosa ) mussels in the Clinch River (Tennessee/Virginia, USA) have declined dramatically in recent years. The bacterium Yokenella regensburgei was first isolated with high prevalence from Pheasantshells during the peak of a 2017 mortality event, but it was not identified after mortality subsided a few months later. Since 2017, Pheasantshell mortality in the Clinch River has occurred each autumn. We extended the investigation of culturable bacterial communities in the Clinch River during mussel mortality events in 2018, 2019, and 2020 and examined the spatial and temporal distribution of bacterial genera among Pheasantshells, as well as among other unionid mussels. We identified Y. regensburgei each year, almost exclusively during active mortality events. The significance of Y. regensburgei remains unclear, but the continued association of this bacterium with mussel mortality events warrants further study.

Tennessee, Virginia

Freshwater mussels show elevated viral richness and intensity during a mortality event

Freshwater mussels (Unionida) are among the world’s most imperiled taxa, but the relationship between freshwater mussel mortality events and infectious disease is largely unstudied. We surveyed viromes of a widespread and abundant species (mucket, Actinonaias ligamentina ; syn: Ortmanniana ligamentina ) experiencing a mortality event of unknown etiology in the Huron River, Michigan, in 2019–2020 and compared them to viromes from mucket in a healthy population in the St. Croix River, Wisconsin and a population from the Clinch River, Virginia and Tennessee, where a mortality event was affecting the congeneric pheasantshell ( Actinonaias pectorosa ; syn: Ortmanniana pectorosa ) population. We identified 38 viruses, most of which were associated with mussels collected during the Huron River mortality event. Viral richness and cumulative viral read depths were significantly higher in moribund mussels from the Huron River than in healthy controls from each of the three populations. Our results demonstrate significant increases in the number and intensity of viral infections for freshwater mussels experiencing mortality events, whereas individuals from healthy populations have a substantially reduced virome comprising a limited number of species at low viral read depths.

Michigan, Tennessee, Virginia, Wisconsin

Mussel mass mortality and the microbiome: Evidence for shifts in the bacterial microbiome of a declining freshwater bivalve

Freshwater mussels (Unionida) are suffering mass mortality events worldwide, but the causes remain enigmatic. Here, we describe an analysis of bacterial loads, community structure, and inferred metabolic pathways in the hemolymph of pheasantshells ( Actinonaias pectorosa ) from the Clinch River, USA, during a multi-year mass mortality event. Bacterial loads were approximately 2 logs higher in moribund mussels (cases) than in apparently healthy mussels (controls). Bacterial communities also differed between cases and controls, with fewer sequence variants (SVs) and higher relative abundances of the proteobacteria Yokenella regensburgei and Aeromonas salmonicida in cases than in controls. Inferred bacterial metabolic pathways demonstrated a predominance of degradation, utilization, and assimilation pathways in cases and a predominance of biosynthesis pathways in controls. Only two SVs correlated with Clinch densovirus 1, a virus previously shown to be strongly associated with mortality in this system: Deinococcota and Actinobacteriota, which were associated with densovirus-positive and densovirus-negative mussels, respectively. Overall, our results suggest that bacterial invasion and shifts in the bacterial microbiome during unionid mass mortality events may result from primary insults such as viral infection or environmental stressors. If so, bacterial communities in mussel hemolymph may be sensitive, if generalized, indicators of declining mussel health.

Tennessee, Virginia

Mass mortality in freshwater mussels (Actinonaias pectorosa) in the Clinch River, USA, linked to a novel densovirus

Freshwater mussels (order Unionida) are among the world’s most biodiverse but imperiled taxa. Recent unionid mass mortality events around the world threaten ecosystem services such as water filtration, nutrient cycling, habitat stabilization, and food web enhancement, but causes have remained elusive. To examine potential infectious causes of these declines, we studied mussels in Clinch River, Virginia and Tennessee, USA, where the endemic and once-predominant pheasantshell ( Actinonaias pectorosa ) has suffered precipitous declines since approximately 2016. Using metagenomics, we identified 17 novel viruses in Clinch River pheasantshells. However, only one virus, a novel densovirus ( Parvoviridae ; Densovirinae ), was epidemiologically linked to morbidity. Clinch densovirus 1 was 11.2 times more likely to be found in cases (moribund mussels) than controls (apparently healthy mussels from the same or matched sites), and cases had 2.7 (log 10 ) times higher viral loads than controls. Densoviruses cause lethal epidemic disease in invertebrates, including shrimp, cockroaches, crickets, moths, crayfish, and sea stars. Viral infection warrants consideration as a factor in unionid mass mortality events either as a direct cause, an indirect consequence of physiological compromise, or a factor interacting with other biological and ecological stressors to precipitate mortality.

Virginia, Tennessee

Reference intervals for blood-based biochemical analytes of southern Beaufort Sea polar bears

Accurate reference intervals (RI) for commonly measured blood-based analytes are essential for health monitoring programs. Baseline values for a panel of analytes can be used to monitor physiologic and pathophysiologic processes such as organ function, electrolyte balance, and protein catabolism. Our reference population includes 651 serum samples from polar bears (Ursus maritimus) from the southern Beaufort Sea subpopulation sampled in Alaska, USA, between 1983 - 2016. To establish RI for 13 biochemical analytes, we defined specific criteria for characterizing the reference population and relevant subgroups. To account for differences in seasonal life history characteristics, we determined separate RI for the spring and fall seasons, when prey availability and energetic requirements of bears differ. We established RI for five subgroups in spring based on sex, age class, and denning status, and three subgroups in fall based on sex and age class in females. Alkaline phosphatase activities were twice as high in subadult as in adult polar bears in spring (zmales = 4.08, Pmales < 0.001, zfemales = 3.90, Pfemales < 0.001), and did not differ between seasons. Denning females had significantly higher glucose concentrations than non-denning females (z = 4.94, P < 0.001), possibly reflecting differences in energy expenditure during lactation. Ten of the 13 analytes differed significantly between seasons in either males or females; however, the physiologic importance of these differences may be minimal. Establishing these RI allows for temporal monitoring of polar bear health in the southern Beaufort Sea and may prove useful for assessing and monitoring additional polar bear subpopulations in a changing Arctic environment.

Alaska

A comparison of bacteria cultured from unionid mussel hemolymph between stable populations in the upper Mississippi River and a mortality event in the Clinch River

The diagnosis of bacterial disease in freshwater unionid mussels has been hindered by a lack of baseline information regarding the microbial communities associated with these animals. In this study, we cultured and identified bacteria from the hemolymph of stable mussel populations from the upper Mississippi River basin and compared results to those from mussel populations experiencing a mortality event in the Clinch River, VA and TN. Several bacterial genera were consistently identified across mussel species and locations, appearing to be part of the natural bacterial flora. One noteworthy bacterial species was identified from the Clinch River: Yokenella regensbergei. Y.regensbergei occurred in relatively high prevalence during the mortality event but was absent from samples acquired after the mortality event. Its role, if any, in the mortality event is unknown but deserves further investigation. We suggest that future studies of freshwater mussel health incorporate hemolymph as a sample type due to its relative separation from the aquatic environment, its role in the circulatory system and the fact that it can be collected nonlethally.

Mississippi River basin

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

Building a response network to investigate potential pathogens associated with unionid mortality events

Unexplained mortality events have confounded the mussel conservation community for over 30 years. While the effects of chemical pollutants and environmental factors have been examined, few investigations have focused on the identification of potential mussel pathogens. Consequently, very little is known regarding the impact that pathogens have on freshwater mussel health, especially as they pertain to propagation efforts. There are several barriers that have hindered progress in the study of freshwater mussel disease, one of which is lack of appropriate diagnostic tools, and another is lack of guidance for field personnel regarding sampling procedures and laboratory contacts.

Ellipsaria

Temporal variation in viral hemorrhagic septicemia virus antibodies in freshwater drum ( Aplodinotus grunniens ) indicates cyclic transmission in Lake Winnebago, Wisconsin

Viral hemorrhagic septicemia virus (VHSV) is an emerging pathogen that causes mass mortality in multiple fish species. In 2007, the Great Lakes freshwater strain, type IVb, caused a large die-off of freshwater drum ( Aplodinotus grunniens ) in Lake Winnebago, Wisconsin, USA. To evaluate the persistence and transmission of VHSV, freshwater drum from Lake Winnebago were tested for antibodies to the virus using recently developed virus neutralization (VN) and enzyme-linked immunosorbent (ELISA) assays. Samples were also tested by real-time reverse transcription-PCR (rRT-PCR) to detect viral RNA. Of 548 serum samples tested, 44 (8.03%) were positive by VN (titers ranging from 1:16 to 1:1,024) and 45 (8.21%) were positive by ELISA, including 7 fish positive by both assays. Antibody prevalence increased with age and was higher in one northwestern area of Lake Winnebago than in other areas. Of 3,864 tissues sampled from 551 fish, 1 spleen and 1 kidney sample from a single adult female fish collected in the spring of 2012 tested positive for VHSV by rRT-PCR, and serum from the same fish tested positive by VN and ELISA. These results suggest that VHSV persists and viral transmission may be active in Lake Winnebago even in years following outbreaks and that wild fish may survive VHSV infection and maintain detectable antibody titers while harboring viral RNA. Influxes of immunologically naive juvenile fish through recruitment may reduce herd immunity, allow VHSV to persist, and drive superannual cycles of transmission that may sporadically manifest as fish kills.

Wisconsin

Development and evaluation of a blocking enzyme-linked immunosorbent assay and virus neutralization assay to detect antibodies to viral hemorrhagic septicemia virus

Viral hemorrhagic septicemia virus (VHSV) is a target of surveillance by many state and federal agencies in the United States. Currently, the detection of VHSV relies on virus isolation, which is lethal to fish and indicates only the current infection status. A serological method is required to ascertain prior exposure. Here, we report two serologic tests for VHSV that are nonlethal, rapid, and species independent, a virus neutralization (VN) assay and a blocking enzyme-linked immunosorbent assay (ELISA). The results show that the VN assay had a specificity of 100% and sensitivity of 42.9%; the anti-nucleocapsid-blocking ELISA detected nonneutralizing VHSV antibodies at a specificity of 88.2% and a sensitivity of 96.4%. The VN assay and ELISA are valuable tools for assessing exposure to VHSV.

Clinical and Vaccine Immunology