High-efficiency second generation ethanol from the hemicellulosic fraction of softwood chips mixed with construction and demolition residues.

Using lignocellulosic residues for bioethanol production could provide an alternative solution to current approaches at competitive costs once challenges related to substrate recalcitrance, process complexity and limited knowledge are overcome. Thus, the impact of different process variables on the ethanol production by Saccharomyces cerevisiae using the hemicellulosic fraction extracted through the steam-treatment of softwood chips mixed with construction and demolition residues was assessed. A statistical design of experiments approach was developed and implemented in order to identify the influencing factors (various nutrient addition sources as well as yeast inoculum growth conditions and inoculation strategies) relevant for enhancing the ethanol production potential and substrate uptake. Ethanol yields of 74.12% and monomeric sugar uptakes of 82.12 g/L were predicted and experimentally confirmed in bench and bioreactor systems. This innovative approach revealed the factors impacting the ethanol yields and carbohydrate consumption allowing powerful behavioral predictions spanning different process inputs and outputs.

A two-step optimization strategy for 2nd generation ethanol production using softwood hemicellulosic hydrolysate as fermentation substrate.

Ethanol production using waste biomass represents a very attractive approach. However, there are considerable challenges preventing a wide distribution of these novel technologies. Thus, a fractional-factorial screening of process variables and Saccharomyces cerevisiae yeast inoculum conditions was performed using a synthetic fermentation media. Subsequently, a response-surface methodology was developed for maximizing ethanol yields using a hemicellulosic solution generated through the chemical hydrolysis of steam treatment broth obtained from residual softwood biomass. In addition, nutrient supplementation using starch-based ethanol production by-products was investigated. An ethanol yield of 74.27% of the theoretical maximum was observed for an initial concentration of 65.17g/L total monomeric sugars. The two-step experimental strategy used in this work represents the first successful attempt to developed and use a model to make predictions regarding the optimal ethanol production using both softwood feedstock residues as well as 1st generation ethanol production by-products.

Variation in Parasite Communities in Spottail Shiners (Notropis hudsonius) Linked with Precipitation.

The Richelieu River, Quebec, is a highly-regulated waterway subject to numerous anthropogenic influences from municipal effluents and agricultural activities. Parasite communities in 234 spottail shiners (Notropis hudsonius) were examined from 4 localities in late spring 2003 and 2004. Component species richness varied between 15 and 18 species in 2003 but declined to 9 to 13 in 2004, while total parasite abundance was consistently lower in 2003. Parasite component community similarity among localities could not be directly linked to available upstream water quality measurements or anthropogenic activity and was best explained by precipitation. Total precipitation in May 2003 was approximately 40% more than in May 2004, presumably altering patterns of runoff, river flow rates, and water quality. This study suggests that fish parasite species composition and richness in the Richelieu River are influenced by environmental parameters which in turn ultimately are driven by a combination of climatic conditions and anthropogenic activities in the watershed.

Responses of freshwater mussel (Elliptio complanata) hemocytes exposed in vitro to crude extracts of Microcystis aeruginosa and Lyngbya wollei.

Lyngbya wollei is a benthic filamentous cyanobacterium that produces a toxin analogous to the neurotoxic saxitoxin known as lyngbyatoxin (LYNGTX). Microcystis aeruginosa form blooms in the pelagic area of eutrophic lakes and produce a series of potent hepatotoxins-microcystins (MCYST). The aim of this study in vitro study was to examine the difference between the crude extracts of either M. aeruginosa or L. wollei toward the immune system of Elliptio complanata mussels. Freshly isolated hemolymph was plated and exposed to the crude extract of each species at LYNGTX or MCYST equivalent concentrations of 5, 10 and 25 µg/L for 18 h. Immunocompetence was characterized by following changes in hemocyte numbers, metabolic activity (viability), and phagocytosis. Hemocyte counts were not affected, indicating no turnover of hemocytes. Hemocyte metabolic activity was higher in cells exposed to crude extracts of L. wollei. Exposure to L. wollei extracts led to decreased pro-inflammatory precursors such as reactive oxygen species (ROS) and cyclooxygenase (COX) activities. Phagocytosis increased at 25 µg/L for both types of crude extracts. However, hemocytes exposed to crude extracts of M. aeruginosa produced more ROS and COX compared to hemocytes exposed to crude extracts of L. wollei. In conclusion, the data suggest that the crude extract of M. aeruginosa was more toxic than crude extract of L. wollei to mussel hemocytes.

Temporal and seasonal variation in acetylcholinesterase activity and glutathione-S-transferase in amphipods collected in mats of Lyngbya wollei in the St-Lawrence River (Canada).

Mats of filamentous algae Lyngbya wollei cover large areas on top of the sediments along the St-Lawrence River. Like most cyanobacteria, L. wollei produce toxins, named lyngbyatoxin (LYNGTX), an analog of saxitoxin. In order to verify the neurotoxic potential of these filamentous cyanobacteria, we collected amphipods within local vegetation at three sites in summer months: a site rich in L. wollei (Lake St-Louis [LSL]) and two urban sites composed of macrophytes but devoid of L. wollei mats (Richelieu River [RICH] and L'Assomption River [ASSO] outlets). The body weight of each amphipod was measured, and the neurotoxicity was estimated with the measurement of acetylcholinesterase (AChE) and glutathione-s-transferase (GST) activities. Levels of protein-bound phosphates were also evaluated, to rule out contamination by Microcystis sp. The results showed that the activity of acetylcholinesterase measured in amphipods in ASSO and RICH was lower than the activity found in amphipods collected in L. wollei mats in LSL. In addition, amphipods found in L. wollei mats at the LSL site have significantly higher GST activity than those located at the two devoid sites, RICH and ASSO. No significant change was observed for protein-bound phosphate levels. This study suggests that the presence of L. wollei triggers potential toxicity.

Energy status and immune system alterations in Elliptio complanata after ingestion of cyanobacteria Anabaena flos-aquae.

Cyanobacteria have often been described as nutritionally poor for herbivorous organisms. To gain additional information on the potential impacts of invertebrates feeding on cyanobacteria, we fed Elliptio complanata mussels with two types of algae: Anabaena flos-aquae (cyanobacteria) and Pseudokirchneriella subcapitata (green algae). Physiological parameters were examined at the energy status, immune system and oxidative stress levels. Energy status was examined by following the rate of electron transport activity in mitochondria (a measure of cellular energy expense) and lipid/sugar stores in the visceral mass. The cyanobacteria were not actively producing toxins. Based on the digestive gland index, the mussels fed equally on either regime. However, the energy status in mussels fed A. flos-aquae revealed that the total sugar was lower in the digestive gland, whereas mitochondrial electron transport activity (MET), once corrected against the digestive gland somatic index, showed increased energy expenses. Acetylcholinesterase activity and lipid peroxidation (LPO) were also higher in mussels fed with A. flos-aquae compared with mussels fed with P. subcapitata. LPO was correlated by mitochondrial activity in both the digestive gland and gills, suggesting that oxidative stress resulted from metabolic respiration. Immunocompetence (phagocytic activity, natural killer cell-like activity, haemocyte count and viability) and humoral level of lysozyme were not affected in mussels by the algae or cyanobacteria regime. Moreover, the xenobiotic conjugating enzyme, glutathione S-transferase, hemoprotein oxidase and vitellogenin-like proteins were not affected in mussel organs via ingestion of A. flos-aquae. Our study suggests that ingestion of cyanobacteria leads to increased energy expenses, oxidative stress and increased acetylcholine turnover in mussels.

Early biochemical effects of Microcystis aeruginosa extracts on juvenile rainbow trout (Oncorhynchus mykiss).

Microcystins (MC) are usually the predominant cyanotoxins associated with cyanobacterial blooms in natural surface waters. These toxins are well-known hepatotoxic agents that proceed by inhibiting protein phosphatase in aquatic biota; recent studies have also reported oxidative stress and disruption of ion regulation in aquatic organisms. In the present study, young trout (Oncorhynchus mykiss) were exposed to crude extracts of Microsystis aeruginosa for four days at 15 °C. The level of microcystins was calculated to confirm the presence of toxins in these crude extracts: 0, 0.75, 1.8 and 5 µg/L. Protein phosphatase measured in the liver increased by at least 3-fold and is significantly as a result of exposure to these sublethal concentrations of crude extract, his indicates an early defense response against protein phosphatase inhibition from cyanotoxins. This was corroborated by the decreased phosphate content in proteins found in the liver and brain. No increase in glutathione-S transferase (GST) activity was observed and lipid peroxidation was unaffected in both liver and brain tissue exposed to the cyanobacterial extracts. The data revealed that the proportion of the reduced (metal-binding) form of metallothionein (MT) decreased by two-fold relative to the control group (with a concomitant increase in the proportion of the oxidized form). The level of phosphate associated with MT increased by 1.5-fold at the highest concentration of crude extract. Acetylcholinesterase (AChE) activity in brain tissue was decreased after exposure to the highest concentration of crude extract, suggesting a slowdown in neural activity. However, no biotransformation processes or detoxification of GST was triggered. Our findings show early sign of biochemical effects of MC-LR in young trout.

Detection and confirmation of saxitoxin analogues in freshwater benthic Lyngbya wollei algae collected in the St. Lawrence River (Canada) by liquid chromatography-tandem mass spectrometry.

The presence of cyanotoxins in benthic Lyngbya wollei algae samples collected in a fluvial lake along the St. Lawrence River, Canada, was investigated using a multi-toxins method. Hydrophilic interaction liquid chromatography (HILIC) and reverse phased liquid chromatography (RPLC) were coupled to triple quadrupole mass spectrometry (LC-QqQMS) for quantification and to quadrupole-time of flight mass spectrometry (LC-QqTOFMS) for screening and confirmation. The presence of two saxitoxin analogues, LWTX-1 and LWTX-6, was confirmed in benthic Lyngbya wollei algae samples. Concentration of LWTX-1 was between 209±5 and 279±9 µg g(-1). No other targeted cyanotoxin (such as anatoxin-a, nodularin, microcystin-LR, microcystins-RR and saxitoxin) was found in the samples. The presence of LWTX-6 was observed by using a screening approach based on an in-house database of cyanotoxins, an algorithm of identification and high resolution mass spectrometry measurements on the precursor and product ions. This work demonstrates the need for more research on the fate of benthic cyanotoxins in aquatic ecosystems such the St. Lawrence River.