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Muruleedhara N. Byappanahalli

Publications and source records attributed to Muruleedhara N. Byappanahalli.

At least 19 recordsLinked to original sources

Evaluation of a eutrophication Beneficial Use Impairment in the Grand Calumet River Area of Concern in northwest Indiana, 2021–22

Eutrophication has been regularly documented in the Grand Calumet River and Indiana Harbor Canal in northwest Indiana. The area has undergone various remediation efforts since the development of a Remedial Action Plan for the area in response to a 1987 amendment to the Great Lakes Water Quality Agreement of 1978 between the United States and Canada and the designation of the Grand Calumet River Area of Concern by the U.S. Environmental Protection Agency. To address concerns of eutrophication and its effects, the U.S. Geological Survey, in cooperation with the U.S. Environmental Protection Agency and the Indiana Department of Environmental Management, collected data from the Grand Calumet River and Indiana Harbor Canal in 2021 and 2022 to document the effects of remediation and combined sewer overflow releases on water quality. This study used continuous monitors to collect real time data for dissolved oxygen, water temperature, chlorophyll fluorescence, specific conductance, and pH. Discrete sample analysis included biological oxygen demand, chemical oxygen demand, nutrients, nitrogen and oxygen isotopes in nitrate, nitrogen isotopes in ammonia, hydrogen and oxygen isotopes in water, and identification of algal communities (cyanobacteria and eukaryotic algae) by metabarcoding. Eutrophic conditions were found throughout the area. Isotopic results indicated the source of nitrate in samples was either from soil organic nitrogen or combined sewer overflows. Combined sewer overflows were shown to have considerable effects on the sites, and remediation status did not have a great effect. Algal community results identified several taxa capable of becoming nuisance species, including Microcystaceae (cyanobacteria) and Chrysophyceae, Cryptophyceae, and Bacillariophyceae (all eukaryotic algae). When sites with irregular flow patterns were excluded from datasets, minimum dissolved oxygen concentrations were often higher downstream from remediated sites than from unremediated sites. This study shows the potential for further and more targeted exploration into the unusual conditions found throughout the Grand Calumet River and Indiana Harbor Canal area.

Indiana

Bacterial community structure across a sand dune chronosequence at the Indiana Dunes National Park

The microbial role in dune succession along the Great Lakes freshwater sand dunes remains poorly understood. A chronosequence study was conducted to understand the relationships among soil bacterial communities, soil chemistry, and prescribed burning at the Indiana Dunes National Park. Soil bacterial communities and chemistry, as well as groundlayer vegetation were sampled during 2015 and 2017 from seven successional stages from the beach (contemporary) to the 14,000-year-old oak forest. Bacterial communities from unburned and burned sites among stages were determined by 16S rRNA gene amplicon sequencing. Soil pH and cations decreased from early (beach, foredune, secondary dune, and woodland transition) to late (oak savanna, woodland, and oak forest) successional stages, while organic matter and organic carbon concentrations increased in the late successional stages. Bacterial alpha diversity showed no significant differences among stages, but a significant interaction was found between stage and prescribed burning (H = 39.7, p < 0.001). Bacterial communities separated mainly along stage by all four beta diversity metrics used (Bray Curtis, Jaccard, and Weighted and Unweighted UniFrac), with the main difference observed along the primary axis (weighted UniFrac, 48 %). Bacterial phyla were differentially abundant in older soil stages compared to beach (ANCOM-BC, q < 0.05); likewise, differential abundances in genera were evident when burned and unburned sites were compared. A Mantel test indicated stronger congruency between the bacterial communities and soil chemistry than between bacterial communities and vegetation. Collectively, soil chemical and microbial parameters along with management practices contribute to dunal successional patterns in the Great Lakes.

Indiana

Bacteria common to rhizosphere communities of Asiatic bittersweet across a post-glacial landscape

Invasive plants such as Asiatic bittersweet ( Celastrus orbiculatus Thunb.) are a significant problem for land managers as they impact plant species composition, disrupt nutrient dynamics and structure of native ecosystems, and are difficult to eradicate. As a result of the increasing abundance of Asiatic bittersweet across the eastern U.S., we have been investigating underlying factors potentially contributing to the success of this plant. Recently, ecologists have been investigating the role of plant-soil-microbe interactions contributing to plant invasion. This work has led to question: are there certain microbes (e.g., bacteria, fungi) contributing to the success of some invasive plants? We hypothesize that despite differences in geologic age of soils where Asiatic bittersweet has established in the Indiana Dunes National Park there are sufficient common factors that led to common bacterial taxa in their rhizosphere. The objectives were to determine differences and commonalities in the soil chemistry, plant community and bacterial communities of Asiatic bittersweet plants. To achieve these objectives, bittersweet plants were collected at thirteen locations in the national park from soils ranging in geologic age from 150 to over 14,500 years. Surrounding soil chemistry, plant cover and the 16S rRNA gene amplicon sequences of rhizosphere soil bacterial communities of these Asiatic bittersweet were compared. Asiatic bittersweet coverage of sampling sites ranged from 2 to 77% averaging 52 ± 2%. There were statistically significant differences ( p < 0.05) in alpha diversity (Shannon, Faith’s PD and Pielou’s evenness) and beta diversity (Bray Curtis, Jaccard, unweighted Unifrac, weighted Unifrac) among the samples when grouped by soil age or habitat. Despite these differences in the bacterial communities from different soil ages and habitats, some bacterial taxa (e.g., Bacillus, Streptomyces, Sphingomonas and Rhizobiales) previously found in other studies to be beneficial to plant growth were found in every rhizosphere community sampled. These microbes provide insight into a possible contributing factor to the success of this invasive plant at the Indiana Dunes National Park, and a strategy for future work to reduce the impact of Asiatic bittersweet establishment and offer some new strategies to manage this nuisance species.

Frontiers of Ecology and Evolution

Microbial source tracking and evaluation of best management practices for restoring degraded beaches of Lake Michigan

Attempts to mitigate shoreline microbial contamination require a thorough understanding of pollutant sources, which often requires multiple years of data collection (e.g., point/nonpoint) and the interacting factors that influence water quality. Because restoration efforts can alter shoreline or beach morphology, revisiting source inputs is often necessary. Microbial source tracking (MST) using source-specific molecular markers, genomic community analyses, and physical modeling was used to identify contamination sources along three Lake Michigan beaches of the Laurentian Great Lakes with historically high fecal indicator bacteria (FIB, E. coli ) concentrations. Genetic markers for human (Bacteroides HF183) and mixed gull species ( Catellicoccus marimammalium ) fecal sources were tested from water and sediment. Gene sequencing (16S rRNA) was used to identify similarities in bacterial communities in nearshore water, river inputs, sand, sediment, and groundwater. Synoptic surveys of water exchange were conducted to determine nearshore-offshore interactions of FIB. In addition to these MST studies, best management practices to mitigate FIB, including gull deterrence, slope grading, wetland establishment, and shoreline plantings, were reviewed for their effectiveness at reducing FIB concentrations over time. Using multiple tools for MST helped identify primary and secondary sources of FIB (gulls, stormwater inputs) and the physical processes that exacerbate FIB concentrations (onshore currents, limited circulation). Management actions were successful in the short-term at reducing FIB, but scope of success was temporally limited, with FIB concentrations often rebounding. Results highlight the usefulness of MST to inform best management practices and the need for a sustained adaptive approach that adjusts for changes in the coastal system.

Illinois, Wisconsin

Gut microbiota associated with different sea lamprey (Petromyzon marinus) life stages

Sea lamprey (SL; Petromyzon marinus ), one of the oldest living vertebrates, have a complex metamorphic life cycle. Following hatching, SL transition into a microphagous, sediment burrowing larval stage, and after 2–10+ years, the larvae undergo a dramatic metamorphosis, transforming into parasitic juveniles that feed on blood and bodily fluids of fishes; adult lamprey cease feeding, spawn, and die. Since gut microbiota are critical for the overall health of all animals, we examined the microbiota associated with SLs in each life history stage. We show that there were significant differences in the gut bacterial communities associated with the larval, parasitic juvenile, and adult life stages. The transition from larval to the parasitic juvenile stage was marked with a significant shift in bacterial community structure and reduction in alpha diversity. The most abundant SL-associated phyla were Proteobacteria, Fusobacteria, Bacteroidetes, Verrucomicrobia, Actinobacteria, and Firmicutes, with their relative abundances varying among the stages. Moreover, while larval SL were enriched with unclassified Fusobacteriaceae, unclassified Verrucomicrobiales and Cetobacterium, members of the genera with fastidious nutritional requirements, such as Streptococcus , Haemophilus , Cutibacterium , Veillonella , and Massilia , were three to four orders of magnitude greater in juveniles than in larvae. In contrast, adult SLs were enriched with Aeromonas , Iodobacter , Shewanella , and Flavobacterium . Collectively, our findings show that bacterial communities in the SL gut are dramatically different among its life stages. Understanding how these communities change over time within and among SL life stages may shed more light on the role that these gut microbes play in host growth and fitness.

Michigan, Ontario

Influence of filter pore size on composition and relative abundance of bacterial communities and select host-specific MST markers in coastal waters of southern Lake Michigan

Water clarity is often the primary guiding factor in determining whether a prefiltration step is needed to increase volumes processed for a range of microbial endpoints. In this study, we evaluate the effect of filter pore size on the bacterial communities detected by 16S rRNA gene sequencing and incidence of two host-specific microbial source tracking (MST) markers in a range of coastal waters from southern Lake Michigan, using two independent data sets collected in 2015 (bacterial communities) and 2016–2017 (MST markers). Water samples were collected from river, shoreline, and offshore areas. For bacterial communities, each sample was filtered through a 5.0-μm filter, followed by filtration through a 0.22-μm filter, resulting in 70 and 143 filter pairs for bacterial communities and MST markers, respectively. Following DNA extraction, the bacterial communities were compared using 16S rRNA gene amplicons of the V3–V4 region sequenced on a MiSeq Illumina platform. Presence of human ( Bacteroides HF183) and gull (Gull2, Catellicoccus marimammalium ) host-specific MST markers were detected by qPCR. Actinobacteriota, Bacteroidota, and Proteobacteria, collectively represented 96.9% and 93.9% of the relative proportion of all phyla in the 0.22- and 5.0-μm pore size filters, respectively. There were more families detected in the 5.0-μm pore size filter (368) than the 0.22-μm (228). There were significant differences in the number of taxa between the two filter sizes at all levels of taxonomic classification according to linear discriminant analysis (LDA) effect size (LEfSe) with as many as 986 taxa from both filter sizes at LDA effect sizes greater than 2.0. Overall, the Gull2 marker was found in higher abundance on the 5.0-μm filter than 0.22 μm with the reverse pattern for the HF183 marker. This discrepancy could lead to problems with identifying microbial sources of contamination. Collectively, these results highlight the importance of analyzing pre- and final filters for a wide range of microbial endpoints, including host-specific MST markers routinely used in water quality monitoring programs. Analysis of both filters may increase costs but provides more complete genomic data via increased sample volume for characterizing microbial communities in coastal waters.

Illinois, Indiana, Wisconsin

Quantifying microplastics in Indiana’s Lake Michigan watershed and submerged aquatic vegetation

Environmental breakdown of plastics produces synthetic microfibers, a class of microplastics that are most prevalent in surface waters. A main source of these pollutants is wastewater treatment plants which discharge into surface waters, including those that flow into Lake Michigan. Microplastics can settle into aquatic sediment or exist for lengths of time in the swash zone of the lakes; they can be ingested by organisms or concentrated in algae or other lake plankton, among other possibilities. As part of a larger Great Lakes study of the growth and senescence of submerged aquatic vegetation (SAV), the loads of synthetic microfibers in the SAV have been determined from Lake Michigan samples collected during the summer months of 2018, 2019 and 2020, and from various samples collected in the watershed and along the shoreline in Northwest Indiana. In order to effectively address microplastics pollution, a complete understanding of the distribution and fate of these pollutants is needed; this project is working toward that goal.

Indiana

Virulence and biodegradation potential of dynamic microbial communities associated with decaying Cladophora in Great Lakes

Cladophora mats that accumulate and decompose along shorelines of the Great Lakes create potential threats to the health of humans and wildlife. The decaying algae create a low oxygen and redox potential environment favoring growth and persistence of anaerobic microbial populations, including Clostridium botulinum , the causal agent of botulism in humans, birds, and other wildlife. In addition to the diverse population of microbes, a dynamic chemical environment is generated, which involves production of numerous organic and inorganic substances, many of which are believed to be toxic to the sand and aquatic biotic communities. In this study, we used 16S-rDNA-based-amplicon sequencing and microfluidic-based quantitative PCR approaches to characterize the bacterial community structure and the abundances of human pathogens associated with Cladophora at different stages (up to 90 days) of algal decay in laboratory microcosms. Oxygen levels were largely depleted after a few hours of incubation. As Cladophora decayed, the algal microbial biodiversity decreased within 24 h, and the mat transitioned from an aerobic to anaerobic environment. There were increasing abundances of enteric and pathogenic bacteria during decomposition of Cladophora , including Acinetobacter , Enterobacter , Kluyvera , Cedecea , and others. In contrast, there were no or very few sequences (< 0.07%) assigned to such groups in fresh Cladophora samples. Principal coordinate analysis indicated that the bacterial community structure was dynamic and changed significantly with decay time. Knowledge of microbial communities and chemical composition of decaying algal mats is critical to our further understanding of the role that Cladophora plays in a beach ecosystem's structure and function, including the algal role in trophic interactions. Based on these findings, public and environmental health concerns should be considered when decaying Cladophora mats accumulate Great Lakes shorelines.

Great Lakes

Application of a microfluidic quantitative polymerase chain reaction technique to monitor bacterial pathogens in beach water and complex environmental matrices

Microfluidic quantitative polymerase chain reaction (MFQPCR) and conventional quantitative polymerase chain reaction methods were compared side by side in detecting and quantifying 19 genetic markers associated with Escherichia coli and select bacterial pathogens in algae, beach sand, and water from Lake Michigan. Enteropathogenic E. coli (EPEC), Shiga toxin-producing E. coli , Salmonella spp., Campylobacter jejuni , and Clostridium perfringens were among the pathogens tested. Of the pathogenic markers, eaeA that encodes intimin in EPEC was detected in all sample types: water (5%), detached/floating algae (42%), exposed/stranded algae (43%), sand below exposed algae (27%), and nearshore sand with no algae (22%). Other pathogenic markers, however, were detected sporadically. Despite comparable results from the two methods for the genetic markers tested in this study, the MFQPCR method may be superior, with the advantage of detecting and quantifying multiple pathogens simultaneously in environmental matrices.

Indiana

U.S. recreational water quality criteria: a vision for the future

This manuscript evaluates the U.S. Recreational Water Quality Criteria (RWQC) of 2012, based upon discussions during a conference held 11&ndash;13 March 2013, in Honolulu, Hawaii. The RWQC of 2012 did not meet expectations among the research community because key recommended studies were not completed, new data to assess risks to bathers exposed to non-point sources of fecal indicator bacteria (FIB) were not developed, and the 2012 RWQC did not show marked improvements in strategies for assessing health risks for bathers using all types of recreational waters. The development of the 2012 RWQC was limited in scope because the epidemiologic studies at beach sites were restricted to beaches with point sources of pollution and water samples were monitored for only enterococci. The vision for the future is development of effective RWQC guidelines based on epidemiologic and quantitative microbial risk assessment (QMRA) studies for sewage specific markers, as well as human enteric pathogens so that health risks for bathers at all recreational waters can be determined. The 2012 RWQC introduced a program for states and tribes to develop site-specific water quality criteria, and in theory this approach can be used to address the limitations associated with the measurements of the traditional FIB.

International Journal of Environmental Research an

Wildlife, urban inputs, and landscape configuration are responsible for degraded swimming water quality at an embayed beach

Jeorse Park Beach, on southern Lake Michigan, experiences frequent closures due to high Escherichia coli ( E. coli ) levels since regular monitoring was implemented in 2005. During the summer of 2010, contaminant source tracking techniques, such as the conventional microbial and physical surveys and hydrodynamic models, were used to determine the reasons for poor water quality at Jeorse Park. Fecal indicator bacteria ( E. coli , enterococci) were high throughout the season, with densities ranging from 12&ndash;2419 (culturable E. coli ) and 1&ndash;2550 and < 1&ndash;5831 (culturable and qPCR enterococci, respectively). Genetic markers for human ( Bacteroides HF183) and gull ( Catellicoccus marimammalium ) fecal contamination were found in 15% and 37% of the samples indicating multiple sources contributing to poor water quality. Nesting colonies of double-crested cormorants ( Phalacrocorax auritus ) have steadily increased since 2005, coinciding with high E. coli levels. A hydrodynamic model indicated that limited circulation allows bacteria entering the embayed area to be retained in nearshore areas; and bacterial resuspension from sand and stranded beach wrack during storm events compounds the problem. The integration of hydrodynamics, expanded use of chemical and biological markers, as well as more complex statistical multivariate techniques can improve microbial source tracking, informing management actions to improve recreational water quality. Alterations to embayed structures to improve circulation and reduce nuisance algae as well as growing native plants to retain sand to improve beach morphometry are among some of the restoration strategies under consideration in ongoing multi-agency collaborations.

Indiana

Prevalence of toxin-producing Clostridium botulinum associated with the macroalga Cladophora in three Great Lakes: Growth and management

The reemergence of avian botulism caused by Clostridium botulinum type E has been observed across the Great Lakes in recent years. Evidence suggests an association between the nuisance algae, Cladophora spp., and C. botulinum in nearshore areas of the Great Lakes. However, the nature of the association between Cladophora and C. botulinum is not fully understood due, in part, to the complex food web interactions in this disease etiology. In this study, we extensively evaluated their association by quantitatively examining population size and serotypes of C. botulinum in algal mats collected from wide geographic areas in lakes Michigan, Ontario, and Erie in 2011&ndash;2012 and comparing them with frequencies in other matrices such as sand and water. A high prevalence (96%) of C. botulinum type E was observed in Cladophora mats collected from shorelines of the Great Lakes in 2012. Among the algae samples containing detectable C. botulinum , the population size of C. Botulinum type E was 10 0 &ndash;10 4 MPN/g dried algae, which was much greater (up to 10 3 fold) than that found in sand or the water column, indicating that Cladophora mats are sources of this pathogen. Mouse toxinantitoxin bioassays confirmed that the putative C. botulinum belonged to the type E serotype. Steam treatment was effective in reducing or eliminating C. botulinum type E viable cells in Cladophora mats, thereby breaking the potential transmission route of toxin up to the food chain. Consequently, our data suggest that steam treatment incorporated with a beach cleaning machine may be an effective treatment of Cladophora -borne C. botulinum and may reduce bird mortality and human health risks.

Lake Erie, Lake Michigan, Lake Ontario

Detection limits and cost comparisons of human- and gull-associated conventional and quantitative PCR assays in artificial and environmental waters

Some molecular methods for tracking fecal pollution in environmental waters have both PCR and quantitative PCR (qPCR) assays available for use. To assist managers in deciding whether to implement newer qPCR techniques in routine monitoring programs, we compared detection limits (LODs) and costs of PCR and qPCR assays with identical targets that are relevant to beach water quality assessment. For human-associated assays targeting Bacteroidales HF183 genetic marker, qPCR LODs were 70 times lower and there was no effect of target matrix (artificial freshwater, environmental creek water, and environmental marine water) on PCR or qPCR LODs. The PCR startup and annual costs were the lowest, while the per reaction cost was 62% lower than the Taqman based qPCR and 180% higher than the SYBR based qPCR. For gull-associated assays, there was no significant difference between PCR and qPCR LODs, target matrix did not effect PCR or qPCR LODs, and PCR startup, annual, and per reaction costs were lower. Upgrading to qPCR involves greater startup and annual costs, but this increase may be justified in the case of the human-associated assays with lower detection limits and reduced cost per sample.

Journal of Environmental Management

Enterococcus phages as potential tool for identifying sewage inputs in the Great Lakes region

Bacteriophages are viruses living in bacteria that can be used as a tool to detect fecal contamination in surface waters around the world. However, the lack of a universal host strain makes them unsuitable for tracking fecal sources. We evaluated the suitability of two newly isolated Enterococcus host strains (ENT-49 and ENT-55) capable for identifying sewage contamination in impacted waters by targeting phages specific to these hosts. Both host strains were isolated from wastewater samples and identified as E. faecium by 16S rRNA gene sequencing. Occurrence of Enterococcus phages was evaluated in sewage samples ( n = 15) from five wastewater treatment plants and in fecal samples from twenty-two species of wild and domesticated animals (individual samples; n = 22). Levels of Enterococcus phages, F + coliphages, Escherichia coli and enterococci were examined from four rivers, four beaches, and three harbors. Enterococcus phages enumeration was at similar levels (Mean = 6.72 Log PFU/100 mL) to F + coliphages in all wastewater samples, but were absent from all non-human fecal sources tested. The phages infecting Enterococcus spp. and F + coliphages were not detected in the river samples (detection threshold < 10 PFU/100 mL), but were present in the beach and harbor samples (range = 1.83 to 2.86 Log PFU/100 mL). Slightly higher concentrations (range = 3.22 to 3.69 Log MPN/100 mL) of E. coli and enterococci when compared to F + coliphages and Enterococcus phages, were observed in the river, beach and harbor samples. Our findings suggest that the bacteriophages associated with these particular Enterococcus host strains offer potentially sensitive and human-source specific indicators of enteric pathogen risk.

Journal of Great Lakes Research

Probabilistic analysis showing that a combination of bacteroides and methanobrevibacter source tracking markers is effective for identifying waters contaminated by human fecal pollution

Microbial source tracking assays to identify sources of waterborne contamination typically target genetic markers of host-specific microorganisms. However, no bacterial marker has been shown to be 100% host-specific, and cross-reactivity has been noted in studies evaluating known source samples. Using 485 challenge samples from 20 different human and animal fecal sources, this study evaluated microbial source tracking markers including the Bacteroides HF183 16S rRNA, M. smithii nifH, and Enterococcus esp gene targets that have been proposed as potential indicators of human fecal contamination. Bayes' Theorem was used to calculate the conditional probability that these markers or a combination of markers can correctly identify human sources of fecal pollution. All three human-associated markers were detected in 100% of the sewage samples analyzed. Bacteroides HF183 was the most effective marker for determining whether contamination was specifically from a human source, and greater than 98% certainty that contamination was from a human source was shown when both Bacteroides HF183 and M. smithii nifH markers were present. A high degree of certainty was attained even in cases where the prior probability of human fecal contamination was as low as 8.5%. The combination of Bacteroides HF183 and M. smithii nifH source tracking markers can help identify surface waters impacted by human fecal contamination, information useful for prioritizing restoration activities or assessing health risks from exposure to contaminated waters.

Environmental Science & Technology

Choices in recreational water quality monitoring: new opportunities and health risk trade-offs

With the recent release of new recreational water quality monitoring criteria, there are more options for regulatory agencies seeking to protect beachgoers from waterborne pathogens. Included are methods that can reduce analytical time, providing timelier estimates of water quality, but the application of these methods has not been examined at most beaches for expectation of health risk and management decisions. In this analysis, we explore health and monitoring outcomes expected at Lake Michigan beaches using protocols for indicator bacteria including culturable Escherichia coli (E. coli; EC), culturable enterococci (ENT), and enterococci as analyzed by qPCR (QENT). Correlations between method results were generally high, except at beaches with historically high concentrations of EC. The “beach action value” was exceeded most often when using EC or ENT as the target indicator; QENT exceeded the limit far less frequently. Measured water quality between years was varied. Although methods with equivalent health expectation have been established, the lack of relationship among method outcomes and annual changes in mean indicator bacteria concentrations complicates the decision-making process. The monitoring approach selected by beach managers may be a combination of available tools that maximizes timely health protection, cost efficiency, and collaboration among beach jurisdictions.

Environmental Science & Technology

Association of toxin-producing Clostridium botulinum with the macroalga Cladophora in the Great Lakes

Avian botulism, a paralytic disease of birds, often occurs on a yearly cycle and is increasingly becoming more common in the Great Lakes. Outbreaks are caused by bird ingestion of neurotoxins produced by Clostridium botulinum, a spore-forming, gram-positive, anaerobe. The nuisance, macrophytic, green alga Cladophora (Chlorophyta; mostly Cladophora glomerata L.) is a potential habitat for the growth of C. botulinum. A high incidence of botulism in shoreline birds at Sleeping Bear Dunes National Lakeshore (SLBE) in Lake Michigan coincides with increasingly massive accumulations of Cladophora in nearshore waters. In this study, free-floating algal mats were collected from SLBE and other shorelines of the Great Lakes between June and October 2011. The abundance of C. botulinum in algal mats was quantified and the type of botulism neurotoxin (bont) genes associated with this organism were determined by using most-probable-number PCR (MPN-PCR) and five distinct bont gene-specific primers (A, B, C, E, and F). The MPN-PCR results showed that 16 of 22 (73%) algal mats from the SLBE and 23 of 31(74%) algal mats from other shorelines of the Great Lakes contained the bont type E (bont/E) gene. C. botulinum was present up to 15 000 MPN per gram dried algae based on gene copies of bont/E. In addition, genes for bont/A and bont/B, which are commonly associated with human diseases, were detected in a few algal samples. Moreover, C. botulinum was present as vegetative cells rather than as dormant spores in Cladophora mats. Mouse toxin assays done using supernatants from enrichment of Cladophora containing high densities of C. botulinum (>1000 MPN/g dried algae) showed that Cladophora-borne C. botulinum were toxin-producing species (BoNT/E). Our results indicate that Cladophora provides a habitat for C. botulinum, warranting additional studies to better understand the relationship between this bacterium and the alga, and how this interaction potentially contributes to botulism outbreaks in birds.

Great Lakes

Notable decomposition products of senescing Lake Michigan Cladophora glomerata

Massive accumulations of Cladophora , a ubiquitous, filamentous green alga, have been increasingly reported along Great Lakes shorelines, negatively affecting beach aesthetics, recreational activities, public health and beachfront property values. Previously, the decomposition byproducts of decaying algae have not been thoroughly examined. To better understand the negative consequences and potential merit of the stranded Cladophora , a three month mesocosm study of the dynamic chemical environment of the alga was conducted using fresh samples collected from southern Lake Michigan beaches. Typical fermentation products, such as organic acids, sulfide compounds, and alcohols were detected in the oxygen–deprived algae. Short chain carboxylic acids peaked on day seven, in correspondence with the lowest pH value. Most low molecular mass carbon compounds were eventually consumed, but 4-methylphenol, indole, and 3-methylindole were detected throughout the incubation period. Natural oils were detected in fresh and decomposing algae, indicating the stable nature of these compounds. The mesocosm experiment was validated by directly sampling the fluid within decomposing Cladophora mats in the field; many of the same compounds were found. This study suggests that the problematic Cladophora accumulations may be harvested for useful byproducts, thereby reducing the odiferous and potentially harmful mats stranded along the shorelines.

Lake Michigan