Master of Science / Food Science Institute / Curtis Kastner / Harmful Algal Blooms (HABs) including Cyanobacteria and other toxic marine biota are responsible for similar harmful effects on human health, food safety, ecosystem maintenance, economic losses and liability issues for aquaculture farms as well as the food industry. Detection, monitoring and mitigation are all key factors in decreasing the deleterious effects of these toxic algal blooms. Harmful algal blooms can manifest toxic effects on a number of facets of animal physiology, elicit noxious taste and odor events and cause mass fish as well as animal kills. Such blooms can adversely impact the perception of the efficacy and safety of the food industry, water utilities, the quality of aquaculture and land farming products, as well as cause ripple effects experienced by coastal communities. HABs can adversely impact coastal areas and other areas reliant on local aquatic ecosystems through the loss of revenues experienced by local restaurants, food manufacturers as well as seafood harvesting/processing plants; loss of tourism revenue, decreased property values and a fundamental shift in the lives of those that are reliant upon those industries for their quality of life. This paper discusses Cyanobacteria, macroalgae, HABs, Cyanobacteria toxins, mitigation of HAB populations and their products as well as the ramifications this burgeoning threat to aquatic/ landlocked communities including challenges these toxic algae pose to the field of food science and the economy.

A crescente eutrofização dos ambientes aquáticos é consequência das atividades humanas que desequilibram os ecossistemas. A descarga de esgotos urbanos, a utilização de adubos químicos, os efluentes das agroindústrias e dos diversos setores industriais promovem a entrada de quantidades significativas de nutrientes e matéria orgânica em corpos d´água, favorecendo o aumento das florações de algas, entre elas as cianobactérias. O excesso de matéria orgânica e os despejos de efluentes agroindustriais nos mananciais hídricos representam os principais fatores do excesso de floração das cianobactérias, destacando-se a Microcystis aeruginosa e da sua cianotoxina microcistina. Nesta tese foi aplicado o Processo Oxidativo Avançado (POA) de Fotoeletroxidação (FEO) para a degradação de Microcystis aeruginosa e da cianotoxina microcistina. Investigou-se a otimização de parâmetros como tempo de tratamento e intensidade de corrente elétrica aplicada. Os resultados obtidos nas condições aplicadas demonstram valores de 99% para a degradação de Microcystis aeruginosa e cianotoxina Microcistina com o tempo de 10min e densidade de corrente de 2mA.cm-2. Os testes de toxicidade aguda com ajuste de pH de 6,3 para 7,3 não apresentaram toxidade para a espécie teste Pimephales promelas. / The increasing eutrophication of aquatic environments is a consequence of human activities that disrupt ecosystems. The discharge of urban sewage, the use of chemical fertilizers, effluents from agro-industries and other industrial sectors promote the input of significant amounts of nutrients and organic matter in water, favoring increased blooms of algae, including cyanobacteria. Excessive organic matter and the discharge of agroindustrial effluents in the water bodies, represent the main factors for the excessive bloom of cyanobacteria, especially Microcystis aeruginosa and microcystin cianotoxin. In this study, an Advanced Oxidation Process named photoelectrooxidation, was used to the degradation of Microcystis aeruginosa and cianotoxin microcystin. The optimization of parameters such as, treatment time and intensity of electrical current applied in the process of PEO were evaluated. The results achieved under the conditions applied, show 99% of degradation with experimental times at 10 minutes and a current density of 2mA.cm-2. The acute toxicity tests, with pH adjustment from 6.3 to 7.3, did not show toxicity to the species Pimephales promelas.

A crescente eutrofização dos ambientes aquáticos é consequência das atividades humanas que desequilibram os ecossistemas. A descarga de esgotos urbanos, a utilização de adubos químicos, os efluentes das agroindústrias e dos diversos setores industriais promovem a entrada de quantidades significativas de nutrientes e matéria orgânica em corpos d´água, favorecendo o aumento das florações de algas, entre elas as cianobactérias. O excesso de matéria orgânica e os despejos de efluentes agroindustriais nos mananciais hídricos representam os principais fatores do excesso de floração das cianobactérias, destacando-se a Microcystis aeruginosa e da sua cianotoxina microcistina. Nesta tese foi aplicado o Processo Oxidativo Avançado (POA) de Fotoeletroxidação (FEO) para a degradação de Microcystis aeruginosa e da cianotoxina microcistina. Investigou-se a otimização de parâmetros como tempo de tratamento e intensidade de corrente elétrica aplicada. Os resultados obtidos nas condições aplicadas demonstram valores de 99% para a degradação de Microcystis aeruginosa e cianotoxina Microcistina com o tempo de 10min e densidade de corrente de 2mA.cm-2. Os testes de toxicidade aguda com ajuste de pH de 6,3 para 7,3 não apresentaram toxidade para a espécie teste Pimephales promelas. / The increasing eutrophication of aquatic environments is a consequence of human activities that disrupt ecosystems. The discharge of urban sewage, the use of chemical fertilizers, effluents from agro-industries and other industrial sectors promote the input of significant amounts of nutrients and organic matter in water, favoring increased blooms of algae, including cyanobacteria. Excessive organic matter and the discharge of agroindustrial effluents in the water bodies, represent the main factors for the excessive bloom of cyanobacteria, especially Microcystis aeruginosa and microcystin cianotoxin. In this study, an Advanced Oxidation Process named photoelectrooxidation, was used to the degradation of Microcystis aeruginosa and cianotoxin microcystin. The optimization of parameters such as, treatment time and intensity of electrical current applied in the process of PEO were evaluated. The results achieved under the conditions applied, show 99% of degradation with experimental times at 10 minutes and a current density of 2mA.cm-2. The acute toxicity tests, with pH adjustment from 6.3 to 7.3, did not show toxicity to the species Pimephales promelas.

A crescente eutrofização dos ambientes aquáticos é consequência das atividades humanas que desequilibram os ecossistemas. A descarga de esgotos urbanos, a utilização de adubos químicos, os efluentes das agroindústrias e dos diversos setores industriais promovem a entrada de quantidades significativas de nutrientes e matéria orgânica em corpos d´água, favorecendo o aumento das florações de algas, entre elas as cianobactérias. O excesso de matéria orgânica e os despejos de efluentes agroindustriais nos mananciais hídricos representam os principais fatores do excesso de floração das cianobactérias, destacando-se a Microcystis aeruginosa e da sua cianotoxina microcistina. Nesta tese foi aplicado o Processo Oxidativo Avançado (POA) de Fotoeletroxidação (FEO) para a degradação de Microcystis aeruginosa e da cianotoxina microcistina. Investigou-se a otimização de parâmetros como tempo de tratamento e intensidade de corrente elétrica aplicada. Os resultados obtidos nas condições aplicadas demonstram valores de 99% para a degradação de Microcystis aeruginosa e cianotoxina Microcistina com o tempo de 10min e densidade de corrente de 2mA.cm-2. Os testes de toxicidade aguda com ajuste de pH de 6,3 para 7,3 não apresentaram toxidade para a espécie teste Pimephales promelas. / The increasing eutrophication of aquatic environments is a consequence of human activities that disrupt ecosystems. The discharge of urban sewage, the use of chemical fertilizers, effluents from agro-industries and other industrial sectors promote the input of significant amounts of nutrients and organic matter in water, favoring increased blooms of algae, including cyanobacteria. Excessive organic matter and the discharge of agroindustrial effluents in the water bodies, represent the main factors for the excessive bloom of cyanobacteria, especially Microcystis aeruginosa and microcystin cianotoxin. In this study, an Advanced Oxidation Process named photoelectrooxidation, was used to the degradation of Microcystis aeruginosa and cianotoxin microcystin. The optimization of parameters such as, treatment time and intensity of electrical current applied in the process of PEO were evaluated. The results achieved under the conditions applied, show 99% of degradation with experimental times at 10 minutes and a current density of 2mA.cm-2. The acute toxicity tests, with pH adjustment from 6.3 to 7.3, did not show toxicity to the species Pimephales promelas.

Algal blooms caused by toxigenic cyanobacterial species are an increasing economic burden globally, as high anthropogenic inputs of nitrogen and phosphorous, coupled with rising levels of atmospheric CO2, promote eutrophication and enhance bloom proliferation. Of the freshwater bloom forming species, Microcystis aeruginosa has garnered the most attention due to the production of toxic secondary metabolites known as microcystins. These cyclic peptides are potent eukaryotic protein phosphatase 1 and 2A inhibitors, and can induce hepatic damage if concentration levels reach above the World Health Organization level of 1 µg/L. Current mitigation strategies of water column disruption or by use of broad acting chemicals, are limited in their range and may cause unwanted off target effects to the surrounding biota. Antisense oligonucleotides are short single-stranded DNA polymers that hybridize with transcribed mRNA, and suppress translation of protein products through steric hindrance of ribosomes, or by RNAse H degradation of the DNA/RNA bound complex. While antisense oligonucleotide applications have proven successful in the pharmaceutical industry, their potential remains largely unexplored in environmental contexts. For this reason, we investigated the knockdown of microcystin synthetase gene cluster mcyE in M. aeruginosa. We found that ionic charge neutralization coupled with heat shock were effective chemical competence based methods for delivery, mcyE transcript abundance in cells treated with phosphodiester linked antisense oligonucleotides significantly decreased in RT-qPCR analysis, and production of intracellular microcystin significantly decreased over a 24 hour period (-1.9 fg/cell). This work demonstrates a novel proof of concept for the potential use of exogenous antisense oligonucleotides to target M. aeruginosa in harmful algal bloom occurrences.

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Cyanobacterial blooms are of great concern to the drinking water treatment industry due to their capacity to produce microcystins and other cyanotoxins that are deadly to humans, livestock, pets, and aquatic life at low doses. Unfortunately, the strategies currently employed for cyanotoxin detection involve laborious analyses requiring significant expertise or bioassay kits that are subject to numerous false positives and negatives. These methods are incapable of providing rapid, inexpensive, and robust information to differentiate between the >80 cyanotoxin variants potentially present in an aqueous sample. The use of Raman spectroscopy for identification and quantification of the ubiquitous cyanotoxin microcystin-LR (MC-LR) was examined. Raman spectra readily reflect minute changes in molecular structure, spectra can be collected through water or glass, portable Raman spectrometers are increasingly available, and through surface enhanced Raman spectroscopy (SERS) it is possible to achieve femto or picomolar detection limits for a variety of target species. Drop coating deposition Raman (DCDR) was successfully implemented for quantitation of 2-100 ng of MC-LR deposited in 2 ?L of aqueous sample, even without the use of a specifically designed DCDR substrate or Raman signal enhancements. Reproducible MC-LR Raman spectra were observed for both fresh and aged DCDR samples, and the MC-LR Raman spectrum remained identifiable through a matrix of >80% DOM by mass. DCDR methods show tremendous potential for the rapid, simple, and economical detection of cyanotoxins in environmental matricies at environmentally relevant concentrations. / Master of Science