Development and Application of a Monoclonal-Antibody Technique for Counting Aureococcus anophagefferens, an Alga Causing Recurrent Brown Tides in the Mid-Atlantic United States

ABSTRACT A method was developed for the rapid detection and enumeration of Aureococcus anophagefferens, the cause of harmful algal blooms called “brown tides” in estuaries of the Mid-Atlantic United States. The method employs a monoclonal antibody (MAb) and a colorimetric, enzyme-linked immunosorbent assay format. The MAb obtained exhibits high reactivity with A. anophagefferens and very low cross-reactivities with a phylogenetically diverse array of other protists and bacteria. Standard curves are constructed for each 96-well microtiter plate by using known amounts of a preserved culture of A. anophagefferens. This approach allows estimation of the abundance of the alga in natural samples. The MAb method was compared to an existing method that employs polyclonal antibodies and epifluorescence microscopy and to direct microscopic counts of A. anophagefferens in samples with high abundances of the alga. The MAb method provided increased quantitative accuracy and greatly reduced sample processing time. A spatial survey of several Long Island estuaries in May 2000 using this new approach documented a range of abundances of A. anophagefferens in these bays spanning nearly 3 orders of magnitude.

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Marine harmful algal blooms (HABs) in the United States: History, current status and future trends

Harmful Algae ◽

10.1016/j.hal.2021.101975 ◽

2021 ◽

pp. 101975

Author(s):

Donald M. Anderson ◽

Elizabeth Fensin ◽

Christopher J. Gobler ◽

Alicia E. Hoeglund ◽

Katherine A. Hubbard ◽

...

Keyword(s):

United States ◽

Harmful Algal Blooms ◽

Algal Blooms ◽

United States History ◽

The United States ◽

Current Status ◽

Future Trends ◽

States History

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Mapping the Distribution of Cysts from the Toxic Dinoflagellate Cochlodinium polykrikoides in Bloom-Prone Estuaries by a Novel FluorescenceIn SituHybridization Assay

Applied and Environmental Microbiology ◽

10.1128/aem.03457-15 ◽

2015 ◽

Vol 82 (4) ◽

pp. 1114-1125 ◽

Cited By ~ 10

Author(s):

Theresa K. Hattenrath-Lehmann ◽

Yu Zhen ◽

Ryan B. Wallace ◽

Ying-Zhong Tang ◽

Christopher J. Gobler

Keyword(s):

Harmful Algal Blooms ◽

Algal Blooms ◽

Large Subunit ◽

Cross Reactivity ◽

Epifluorescence Microscopy ◽

Lsu Rdna ◽

Cochlodinium Polykrikoides ◽

Content Type ◽

Ecosystem Studies ◽

Resting Cysts

ABSTRACTCochlodinium polykrikoidesis a cosmopolitan dinoflagellate that is notorious for causing fish-killing harmful algal blooms (HABs) across North America and Asia. While recent laboratory and ecosystem studies have definitively demonstrated thatCochlodiniumforms resting cysts that may play a key role in the dynamics of its HABs, uncertainties regarding cyst morphology and detection have prohibited even a rudimentary understanding of the distribution ofC. polykrikoidescysts in coastal ecosystems. Here, we report on the development of a fluorescencein situhybridization (FISH) assay using oligonucleotide probes specific for the large subunit (LSU) ribosomal DNA (rDNA) ofC. polykrikoides. The LSU rDNA-targeted FISH assay was used with epifluorescence microscopy and was iteratively refined to maximize the fluorescent reaction withC. polykrikoidesand minimize cross-reactivity. The final LSU rDNA-targeted FISH assay was found to quantitatively recover cysts made by North American isolates ofC. polykrikoidesbut not cysts formed by other common cyst-forming dinoflagellates. The method was then applied to identify and mapC. polykrikoidescysts across bloom-prone estuaries. Annual cyst and vegetative cell surveys revealed that elevated densities ofC. polykrikoidescysts (>100 cm−3) during the spring of a given year were spatially consistent with regions of dense blooms the prior summer. The identity of cysts in sediments was confirmed via independent amplification ofC. polykrikoidesrDNA. This study mappedC. polykrikoidescysts in a natural marine setting and indicates that the excystment of cysts formed by this harmful alga may play a key role in the development of HABs of this species.

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Harmful algal blooms and eutrophication: Examining linkages from selected coastal regions of the United States

Harmful Algae ◽

10.1016/j.hal.2008.08.017 ◽

2008 ◽

Vol 8 (1) ◽

pp. 39-53 ◽

Cited By ~ 346

Author(s):

Donald M. Anderson ◽

Joann M. Burkholder ◽

William P. Cochlan ◽

Patricia M. Glibert ◽

Christopher J. Gobler ◽

...

Keyword(s):

United States ◽

Harmful Algal Blooms ◽

Algal Blooms ◽

The United States ◽

Coastal Regions

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Natural Infection of Verbena and Phlox by a Recently Described Member of the Carmovirus Genus

Plant Disease ◽

10.1094/pd-90-1115b ◽

2006 ◽

Vol 90 (8) ◽

pp. 1115-1115 ◽

Cited By ~ 6

Author(s):

F. M. Assis Filho ◽

A. Harness ◽

M. Tiffany ◽

A. Gera ◽

S. Spiegel ◽

...

Keyword(s):

United States ◽

Online Publication ◽

Polyclonal Antibodies ◽

Natural Infection ◽

The United States ◽

Enzyme Linked Immunosorbent Assay ◽

Virus Species ◽

Virus Elimination ◽

Das Elisa ◽

Virus Testing

A novel carmovirus infecting angelonia (Angelonia angustifolia) was recently described independently by researchers in the United States, Israel, and Germany (1,2,4). Angelonia flower break virus (AnFBV) and Angelonia flower mottle virus were proposed as appropriate names for this carmovirus. The virus, causing stunting, mild leaf mottle, flower mottling, and flower breaking symptoms has been detected in naturally infected angelonia in the United States, Israel, and Germany (2,4). Here we report the first detection of natural infection of verbena (in the United States and Israel) and phlox (in the United States) by using a recently developed double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA; Agdia, Elkhart, IN). Prior to this report, verbena was considered insusceptible to carmovirus infection (3) and phlox was known as an experimental host for AnFBV (2). A comparative serological study including 27 virus species, demonstrated that DAS-ELISA did not cross-react with any viruses that commonly infect ornamentals or are related to carmoviruses, showing that the polyclonal antibodies are specific to AnFBV. Antibody specificity was confirmed by the carmovirus group PCR test (Agdia). Furthermore, reverse transcription-polymerase chain reaction with AnFBV specific primers (2) produced the expected 1172-bp band from all ELISA-positive samples tested. Between November 2005 and March 2006, AnFBV was detected in 181 of 567 verbena, 26 of 143 phlox, and 193 of 267 angelonia samples submitted to Agdia Testing Services by commercial ornamental propagators for virus testing. Most samples were asymptomatic, although a few exhibited mild leaf mottle. It should be noted that the number of AnFBV-infected samples might not accurately reflect the actual number of commercially produced plants infected with AnFBV because most of the samples analyzed originated from virus elimination programs. The detection of natural AnFBV infection of verbena, phlox, and angelonia suggests that AnFBV may be more widespread in the ornamental industry than previously thought. References: (1) S. Adkins et al. Phytopathology (Abstr.) 95(suppl.):S2, 2005. (2) S. Adkins et al. Phytopathology 96:460, 2006. (3) G. P. Martelli and M. Russo. Online publication. ICTVdB-The Universal Virus Database. 00.074.0.02, 2004. (4) S. Winter et al. New Disease Reports. Vol 12. Brit. Soc. Plant Pathol. Online publication, 2005.

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Characterisation of M2e Antigenicity using anti-M2 Monoclonal Antibody and anti-M2e Polyclonal Antibodies

Jurnal Ilmu Ternak dan Veteriner ◽

10.14334/jitv.v24i3.1987 ◽

2019 ◽

Vol 24 (3) ◽

pp. 122

Author(s):

Sumarningsih Sumarningsih ◽

Simson Tarigan ◽

H Farhid ◽

Jagoda Ignjatovic

Keyword(s):

Monoclonal Antibody ◽

Influenza A ◽

Polyclonal Antibodies ◽

Enzyme Linked Immunosorbent Assay ◽

Discriminating Power ◽

Antigenic Sites ◽

Rabbit Polyclonal Antibody ◽

Adjacent Position ◽

Important Position ◽

M2e Protein

Matrix 2 ectodomain (M2e) protein is a potential antigen for detection of influenza A virus infection in vaccinated poultry (DIVA test). However the M2e antigenicity and immune response it induces in either humans or animals are poorly understood. Seventeen M2e peptides and sixteen recombinant M2e (rM2e) proteins with amino acid (aa) changes introduced at position 10, 11, 12, 13 14, 16, 18 and 20 were compared by western blot (WB) and enzyme-linked immunosorbent assay (ELISA) using mouse anti-M2 monoclonal antibody (mAb) 14C2, and anti-M2e peptide chicken and rabbit polyclonal antibody (pAb). The mAb 14C had the best discriminating power and indicated that all six positions contributed to the M2e antigenicity. Position 11 was the important immunodominant and affected Mab14C binding to a greatest degree. Changes in the adjacent position 14, 16 and 18 also influenced the binding, and it detected regardless of the method (WB or ELISA), or the antigen used (M2e peptide or rM2e). For chicken pAb and rabbit pAb, the immunodominant aa was position 10 and the antibody reaction was not affected by aa change at 11. The binding of rabbit pAb was also affected by changes at 14 and 16, which confirm the contribution of these positions to the M2e antigenicity. Position 10 was the only important position for the binding of chicken pAb to M2e. Overall, the study showed that the M2e antigenic sites are located between residues 10 – 18 and that aa changes at position 10, 11, 12, 14, 16 and 18 may all affect the antibody binding within the M2e protein.

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Immunochemical Analysis of UMP Kinase fromEscherichia coli

Journal of Bacteriology ◽

10.1128/jb.181.3.833-840.1999 ◽

1999 ◽

Vol 181 (3) ◽

pp. 833-840 ◽

Cited By ~ 17

Author(s):

Stéphanie Landais ◽

Pierre Gounon ◽

Christine Laurent-Winter ◽

Jean-Claude Mazié ◽

Antoine Danchin ◽

...

Keyword(s):

Monoclonal Antibody ◽

Polyclonal Antibodies ◽

Intact Protein ◽

Enzyme Linked Immunosorbent Assay ◽

Bacterial Membranes ◽

E Coli ◽

Specific Localization ◽

Ump Kinase ◽

Carboxy Terminal

ABSTRACT Mono- and polyclonal antibodies directed against UMP kinase fromEscherichia coli were tested with the intact protein or with fragments obtained by deletion mutagenesis. As detected in enzyme-linked immunosorbent assay tests, the carboxy-terminal quarter of UMP kinase is immunodominant. Polyclonal antibodies inhibited the enzyme activity with partial or total loss of allosteric effects exerted by UTP and GTP, respectively. These data indicate that the UTP and GTP binding sites in UMP kinase are only partially overlapping. One monoclonal antibody (44-2) recognized a linear epitope in UMP kinase between residues 171 and 180. A single substitution (D174N) in this segment of the enzyme abolished its interaction with the monoclonal antibody (44-2). Polyclonal antisera were used to identify UMP kinase in the bacterial proteome. The enzyme appears as a single spot on two-dimensional electrophoresis at a pI of 7.24 and an apparent molecular mass of 26 kDa. Immunogold labeling of UMP kinase in wholeE. coli cells shows a localization of the protein near the bacterial membranes. Because the protein does not contain sequences usually required for compartmentalization, the aggregation properties of UMP kinase observed in vitro might play a role in this phenomenon. The specific localization of UMP kinase might also be related to its putative role in cell division.

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The brown tide algae, Aureococcus anophagefferens and Aureoumbra lagunensis (Pelagophyceae), allelopathically inhibit the growth of competing microalgae during harmful algal blooms

Limnology and Oceanography ◽

10.1002/lno.10714 ◽

2017 ◽

Vol 63 (2) ◽

pp. 985-1003 ◽

Cited By ~ 6

Author(s):

Yoonja Kang ◽

Christopher J. Gobler

Keyword(s):

Harmful Algal Blooms ◽

Algal Blooms ◽

Brown Tide ◽

Aureococcus Anophagefferens ◽

Aureoumbra Lagunensis

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The rise in climate change-induced federal fishery disasters in the United States

PeerJ ◽

10.7717/peerj.11186 ◽

2021 ◽

Vol 9 ◽

pp. e11186

Author(s):

Lyall Bellquist ◽

Vienna Saccomanno ◽

Brice X. Semmens ◽

Mary Gleason ◽

Jono Wilson

Keyword(s):

Climate Change ◽

United States ◽

Harmful Algal Blooms ◽

Algal Blooms ◽

Ad Hoc ◽

The United States ◽

Societal Benefit ◽

Disaster Assistance ◽

Environmental Events ◽

The Impact

Commercial, recreational, and indigenous fisheries are critical to coastal economies and communities in the United States. For over three decades, the federal government has formally recognized the impact of fishery disasters via federal declarations. Despite these impacts, national syntheses of the dynamics, impacts, and causes of fishery disasters are lacking. We developed a nationwide Federal Fishery Disaster database using National Oceanic and Atmospheric Administration (NOAA) fishery disaster declarations and fishery revenue data. From 1989-2020, there were 71 federally approved fishery disasters (eleven are pending), which spanned every federal fisheries management region and coastal state in the country. To date, we estimate fishery disasters resulted in $2B (2019 USD) in Congressional allocations, and an additional, conservative estimate of $3.2B (2019 USD) in direct revenue loss. Despite this scale of impact, the disaster assistance process is largely ad hoc and lacks sufficient detail to properly assess allocation fairness and benefit. Nonetheless, fishery disasters increased in frequency over time, and the causes of disasters included a broad range of anthropogenic and environmental factors, with a recent shift to disasters now almost exclusively caused by extreme environmental events (e.g., marine heatwaves, hurricanes, and harmful algal blooms). Nationwide, 84.5% of fishery disasters were either partially or entirely attributed to extreme environmental events. As climate change drives higher rates of such extreme events, and as natural disaster assistance requests reach an all-time high, the federal system for fisheries disaster declaration and mitigation must evolve in order to effectively protect both fisheries sustainability and societal benefit.

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