Spatial analysis of toxic or otherwise bioactive cyanobacterial peptides in Green Bay, Lake Michigan.

Cyanobacterial harmful algal blooms (cyanoHABs) are a growing problem in freshwater systems worldwide. CyanoHABs are well documented in Green Bay, Lake Michigan but little is known about cyanoHAB toxicity. This study characterized the diversity and spatial distribution of toxic or otherwise bioactive cyanobacterial peptides (TBPs) in Green Bay. Samples were collected in 2014 and 2015 during three cruises at sites spanning the mouth of the Fox River north to Chambers Island. Nineteen TBPs were analyzed including 11 microcystin (MC) variants, nodularin, three anabaenopeptins, three cyanopeptolins and microginin-690. Of the 19 TBPs, 12 were detected in at least one sample, and 94% of samples had detectable TBPs. The most prevalent TBPs were MCRR and MCLR, present in 94% and 65% of samples. The mean concentration of all TBPs was highest in the Fox River and lower bay, however, the maximum concentration of all TBPs occurred in the same sample north of the lower bay. MCs were positively correlated with chlorophyll and negatively correlated with distance to the Fox River in all cruises along a well-established south-to-north trophic gradient in Green Bay. The mean concentration of MC in the lower bay across all cruises was 3.0 +/- 2.3 µg/L. Cyanopeptolins and anabaenopeptins did not trend with the south-north trophic gradient or varied by cruise suggesting their occurrence is driven by different environmental factors. Results from this study provides evidence that trends in TBP concentration differ by congener type over a trophic gradient.

Chemical speciation, reactivity, and long-term burial of sedimentary phosphorus in Green Bay, a seasonally hypoxia-influenced freshwater estuary.

Sediment cores were collected along a trophic gradient in Green Bay, a seasonally hypoxia-influenced freshwater estuary in Lake Michigan, to measure various phosphorus (P) species, including exchangeable-P (Ex-P), iron-bound-P (Fe-P), biogenic-apatite and/or CaCO3-associated-P (CFA-P), organic-P (Org-P) and detrital-apatite-P (Detr-P). Although total phosphorus (TP) decreased with increasing depth, different P species exhibited distinct vertical distribution patterns with different post-depositional behaviors. The Ex-P, Fe-P and CFA-P species were identified as potentially bioavailable-P (BAP). Little variation was observed for Org-P and Detr-P species, especially below the upper-active-layer, both serving as the primary sink for P in sediment. Detr-P% decreased consistently from the near river plume station to the open bay in the north. P accumulation rates were estimated at 25.1 mmol-P/m2/yr (779 mg-P/m2/yr) in the south, 10.9 mmol-P/m2/yr (338 mg-P/m2/yr) in the central region, and 8.1 mmol-P/m2/yr (252 mg-P/m2/yr) in the north of Green Bay, showing a decrease in the depth of the upper active layer for P regeneration along the south-north transect. The overall potential P regeneration back into the water column increased from 2.8 mmol-P/m2/yr (87 mg-P/m2/yr) in the south, and 3.3 mmol-P/m2/yr (101 mg-P/m2/yr) in the central region to 5.6 mmol-P/m2/yr (173 mg-P/m2/yr) in the north of the bay, corresponding to P burial efficiencies of ∼89 %, 70 % and 31 % along the trophic gradient. The recent decrease in Detr-P and thus the increase in BAP over the last 2-3 decades could be related to anthropogenic activities, such as damming and implementation of agricultural conservation practices. Conversely, a recent increase in TOC/TOP ratios may reflect the increased extent of trophic status and seasonal hypoxia in bottom waters and enhanced regeneration and recycling of particulate P in Green Bay since the 1960s. New results from this study provide an improved understanding of the linkage between sources, internal cycling, and long-term burial of P in the basin.

Distinct variations in fluorescent DOM components along a trophic gradient in the lower Fox River-Green Bay as characterized using one-sample PARAFAC approach.

Variations in molecular weight distributions of dissolved organic matter (DOM) and PARAFAC-derived fluorescent components were investigated along a transect in the seasonally hypereutrophic lower Fox River-Green Bay using the one-sample PARAFAC approach coupling flow field-flow fractionation for size-separation with fluorescence excitation-emission matrix (EEM) and PARAFAC analysis. Concentrations of dissolved organic carbon and nitrogen, chromophoric-DOM, specific UV absorbance at 254 nm, and humification index all decreased monotonically from river to open bay, showing a strong river-dominated DOM source and a dynamic change in DOM quality along the river-lake transect. The relative abundance of colloidal DOM (>1 kDa) derived from ultrafiltration exhibited minimal variation, averaging 71 ± 4 % of the bulk DOM, across the entire estuarine transect although the colloidal concentration decreased in general. Using the one-sample EEM-PARAFAC approach, the identified major fluorescent components were distinct between stations along the river-estuary-open bay continuum, with four components in river/upper-estuary but three components in open bay waters. Among the four common fluorescent components (C475, C410, C320 and C290), the most abundant and refractory humic-like component, C475, behaved conservatively and its relative abundance (%ΣFmax) remained fairly constant (50 ± 4 %) along the transect, while the semi-labile humic-like component, C410, consistently decreased from river to estuary and eventually vanished in open Green Bay. In contrast, the two autochthonous protein-like components (C320 and C290) increased from river to open bay along the trophic gradient. The new results presented here provide an improved understanding of the diverse and fluctuating characteristics in DOM composition, lability, and estuarine mixing behavior across the river-lake interface and demonstrate the efficacy of the one-sample PARAFAC approach.

Assessment of concentration and distribution of total mercury and polychlorinated biphenyls in Green Bay, Wisconsin, USA.

Under the 1987 Great Lakes Water Quality Agreement, the lower Green Bay and Fox River estuary have been labeled as areas of concern due to the contamination of mercury and polychlorinated biphenyls (PCBs) from industrialization. These pollutants pose substantial health and environmental hazards for the Green Bay region. The PCBs reported in this region, including Aroclor 1242, are known to trigger carcinogenic responses in animals and mercury targets the central nervous system and vital organs. Furthermore, these compounds are extremely difficult to remove from the environment once introduced. Extensive remedial actions have been implemented including dredging sediments in the Lower Fox River from DePere to Green Bay. The purpose of this study is to assess the concentration and distribution of Aroclor 1242 and total mercury in the Green Bay region sediments and pore waters and to assess the impact of interventions and the natural rates of change previously found.

Partitioning and transformation of organic and inorganic phosphorus among dissolved, colloidal and particulate phases in a hypereutrophic freshwater estuary.

Phosphorus (P) loadings to the Great Lakes have been regulated for decades, but re-eutrophication and seasonal hypoxia have recently been increasingly reported. It is of paramount importance to better understand the fate, transformation, and biogeochemical cycling processes of different P species across the river-lake interface. We report here results on chemical speciation of P in the seasonally hypoxic Fox River-Green Bay system and variations in sources and partitioning of P species along the aquatic continuum. During midsummer when productivity is generally high, phosphate and dissolved organic P (DOP) were the major species in river water while particulate-organic-P predominated in open bay waters, showing a dynamic change in the chemical speciation of P along the river-bay transect with active transformations between inorganic and organic P and between colloidal and particulate phases. Colloidal organic P (COP, >1 kDa) comprised 33‒65% of the bulk DOP, while colloidal inorganic P was generally insignificant and undetectable especially in open bay water. Sources of COP changed from mainly allochthonous in the Fox River, having mostly smaller sized colloids (1-3 kDa) and a lower organic carbon to phosphorus (C/P) ratio, to predominantly autochthonous in open bay waters with larger sized colloids (>10 kDa) and a higher organic C/P ratio. The observed high apparent distribution coefficients (Kd) of P between dissolved and particulate phases and high-abundant autochthonous colloidal and particulate organic P in the hypereutrophic environment suggest that, in addition to phosphate, colloidal/particulate organic P may play a critical role in the biogeochemical cycling of P and the development of seasonal hypoxia.

Variations in size and composition of colloidal organic matter in a negative freshwater estuary.

Dynamic variations in chemical composition and size distribution of dissolved organic matter (DOM) along the river-lake interface in the Fox River plume were investigated using ultrafiltration, flow field-flow fractionation, UV-Vis and fluorescence spectroscopy and parallel factor analysis. On average, ~67% of bulk dissolved organic carbon (DOC) were partitioned in the <1kDa (actual cutoff 2.5kDa) low molecular weight fraction, and the other 33% were in the 1kDa-0.7µm colloidal phase. Concentrations of DOC and chromophoric DOM in the bulk and size-fractionated samples decreased monotonously with decreasing conductivity from river to bay waters, demonstrating a dominant terrestrial source and quasi conservative mixing behavior. However, the percentages of colloidal fluorescent-DOM increased while those of carbohydrates decreased from river to bay waters, showing different mixing behavior in the river plume. Colloidal chromophores and humic-like fluorophores were mainly partitioned in the size range of 1-6nm, but a bimodal distribution (with peaks at 1-6 and 35-45nm) was observed for colloidal protein-like DOM. Along the river-lake transect, the peak locations of chromophores, humic-like and small-sized protein-like colloids remained almost constant, while the larger-sized protein-like colloids exhibited a slight peak shift from 38.3 to 40.4nm, showing a molecular size enhancement from high to low conductivity waters, with physical mixing, photochemical/microbial degradation, and disaggregation/repartitioning being the important processes affecting the variations of DOM size and composition. New results herein should enhance our understanding of the heterogeneity of DOM in size and composition and its fate, transport and transformation at the river-lake interface and along the aquatic continuum as a whole.

Spatiotemporal variations in the abundance and composition of bulk and chromophoric dissolved organic matter in seasonally hypoxia-influenced Green Bay, Lake Michigan, USA.

Green Bay, Lake Michigan, USA, is the largest freshwater estuary in the Laurentian Great Lakes and receives disproportional terrestrial inputs as a result of a high watershed to bay surface area ratio. While seasonal hypoxia and the formation of "dead zones" in Green Bay have received increasing attention, there are no systematic studies on the dynamics of dissolved organic matter (DOM) and its linkage to the development of hypoxia. During summer 2014, bulk dissolved organic carbon (DOC) analysis, UV-vis spectroscopy, and fluorescence excitation-emission matrices (EEMs) coupled with PARAFAC analysis were used to quantify the abundance, composition and source of DOM and their spatiotemporal variations in Green Bay, Lake Michigan. Concentrations of DOC ranged from 202 to 571µM-C (average=361±73µM-C) in June and from 279 to 610µM-C (average=349±64µM-C) in August. In both months, absorption coefficient at 254nm (a254) was strongly correlated to bulk DOC and was most abundant in the Fox River, attesting a dominant terrestrial input. Non-chromophoric DOC comprised, on average, ~32% of bulk DOC in June with higher terrestrial DOM and ~47% in August with higher aquagenic DOM, indicating that autochthonous and more degraded DOM is of lower optical activity. PARAFAC modeling on EEM data resulted in four major fluorescent DOM components, including two terrestrial humic-like, one aquagenic humic-like, and one protein-like component. Variations in the abundance of DOM components further supported changes in DOM sources. Mixing behavior of DOM components also indicated that while bulk DOM behaved quasi-conservatively, significant compositional changes occurred during transport from the Fox River to the open bay.

Dynamics of dissolved and particulate phosphorus influenced by seasonal hypoxia in Green Bay, Lake Michigan.

Despite major investments in point source reductions, portions of the Great Lakes, like Green Bay, remain hypereutrophic and are subject to persistent seasonal hypoxia. Phosphorus (P) is generally a limiting nutrient in the Great Lakes ecosystem, but not all P species are equally bioavailable, and the dynamics of nutrients and their correlation to algal bloom remain poorly characterized, in part, due to a lack of adequate quantification of P chemical speciation. During summer 2014, water samples were collected from seasonally hypoxic Green Bay for measurements of dissolved and particulate inorganic and organic P to examine P cycling dynamics along a steep nutrient gradient ranging from Fox River inflow dominated eutrophic waters in the southern bay to mesotrophic northern waters near the bay's connection with open Lake Michigan. River-derived dissolved and particulate P was quickly removed from the water column in southern Green Bay through biological uptake and sedimentation. Concentrations of phosphate or dissolved inorganic P (DIP) dramatically decreased from 828 ± 216 nM in the Fox River, comprising 57 ± 1% of the total dissolved P, to 24 ± 9 nM in northern Green Bay where dissolved organic P (DOP) became predominant (>80%). Generally low phosphate concentrations and extremely high dissolved organic C/P ratios (2090 ± 1160 in August 2014) suggested high DOP turnover rates and active transformation between DOP and DIP through organic degradation during P-limited conditions in Green Bay. Elevated DIP levels were accompanied by low dissolved oxygen in deeper waters (10-15m) of central Green Bay where hypoxia-development occurred, suggesting the release of DIP through particle regeneration under hypoxic conditions enhanced by lateral transport and sediment resuspension. High partition coefficients (Kd) of both inorganic and organic P and their significant negative correlation with suspended particulate matter concentrations indicated the particle-reactive nature of P in freshwater environments and may imply that DOP could also be bioavailable under P-limitation.