An overview of algae biofuel production and potential environmental impact.

Algae are among the most potentially significant sources of sustainable biofuels in the future of renewable energy. A feedstock with virtually unlimited applicability, algae can metabolize various waste streams (e.g., municipal wastewater, carbon dioxide from industrial flue gas) and produce products with a wide variety of compositions and uses. These products include lipids, which can be processed into biodiesel; carbohydrates, which can be processed into ethanol; and proteins, which can be used for human and animal consumption. Algae are commonly genetically engineered to allow for advantageous process modification or optimization. However, issues remain regarding human exposure to algae-derived toxins, allergens, and carcinogens from both existing and genetically modified organisms (GMOs), as well as the overall environmental impact of GMOs. A literature review was performed to highlight issues related to the growth and use of algal products for generating biofuels. Human exposure and environmental impact issues are identified and discussed, as well as current research and development activities of academic, commercial, and governmental groups. It is hoped that the ideas contained in this paper will increase environmental awareness of issues surrounding the production of algae and will help the algae industry develop to its full potential.

The potential environmental impact of waste from cellulosic ethanol production.

The increasing production of ethanol has been established as an important contributor to future energy independence. Although ethanol demand is increasing, a growing economic trend in decreased profitability and resource conflicts have called into question the future of grain-based ethanol production. Growing emphasis is being placed on utilizing cellulosic feedstocks to produce ethanol, and the need for renewable resources has made the development of cellulosic ethanol a national priority. Cellulosic ethanol production plants are being built in many areas of the United States to evaluate various feedstocks and processes. The waste streams from many varying processes that are being developed contain a variety of components. Differences in ethanol generation processes and feedstocks are producing waste streams unique to biofuel production, which could be potentially harmful to the environment if adequate care is not taken to manage those risks. Waste stream management and utilization of the cellulosic ethanol process are equally important components of the development of this industry.

Bacteriophages reduce experimental contamination of hard surfaces, tomato, spinach, broccoli, and ground beef by Escherichia coli O157:H7.

A bacteriophage cocktail (designated ECP-100) containing three Myoviridae phages lytic for Escherichia coli O157:H7 was examined for its ability to reduce experimental contamination of hard surfaces (glass coverslips and gypsum boards), tomato, spinach, broccoli, and ground beef by three virulent strains of the bacterium. The hard surfaces and foods contaminated by a mixture of three E. coli O157:H7 strains were treated with ECP-100 (test samples) or sterile phosphate-buffered saline buffer (control samples), and the efficacy of phage treatment was evaluated by comparing the number of viable E. coli organisms recovered from the test and control samples. Treatments (5 min) with the ECP-100 preparation containing three different concentrations of phages (10(10), 10(9), and 10(8) PFU/ml) resulted in statistically significant reductions (P = <0.05) of 99.99%, 98%, and 94%, respectively, in the number of E. coli O157:H7 organisms recovered from the glass coverslips. Similar treatments resulted in reductions of 100%, 95%, and 85%, respectively, in the number of E. coli O157:H7 organisms recovered from the gypsum board surfaces; the reductions caused by the two most concentrated phage preparations were statistically significant. Treatment with the least concentrated preparation that elicited significantly less contamination of the hard surfaces (i.e., 10(9) PFU/ml) also significantly reduced the number of viable E. coli O157:H7 organisms on the four food samples. The observed reductions ranged from 94% (at 120 +/- 4 h posttreatment of tomato samples) to 100% (at 24 +/- 4 h posttreatment of spinach samples). The data suggest that naturally occurring bacteriophages may be useful for reducing contamination of various hard surfaces, fruits, vegetables, and ground beef by E. coli O157:H7.

Testing antimicrobial cleaner efficacy on gypsum wallboard contaminated with Stachybotrys chartarum.

GOAL, SCOPE AND BACKGROUND: Reducing occupant exposure to indoor mold is the goal of this research, through the efficacy testing of antimicrobial cleaners. Often mold contaminated building materials are not properly removed, but instead surface cleaners are applied in an attempt to alleviate the problem. The efficacy of antimicrobial cleaners to remove, eliminate or control mold growth on surfaces can easily be tested on non-porous surfaces. However, the testing of antimicrobial cleaner efficacy on porous surfaces, such as those found in the indoor environment such as gypsum board can be more complicated and prone to incorrect conclusions regarding residual organisms. The mold Stachybotrys chartarum has been found to be associated with idiopathic pulmonary hemorrhage in infants and has been studied for toxin production and its occurrence in water damaged buildings. Growth of S. chartarum on building materials such as gypsum wallboard has been frequently documented. METHODS: Research to control S. chartarum growth using 13 separate antimicrobial cleaners on contaminated gypsum wallboard has been performed in laboratory testing. Popular brands of cleaning products were tested by following directions printed on the product packaging. RESULTS: A variety of gypsum wallboard surfaces were used to test these cleaning products at high relative humidity. The results indicate differences in antimicrobial efficacy for the six month period of testing. DISCUSSION: Results for the six types of GWB surfaces varied extensively. However, three cleaning products exhibited significantly better results than others. Lysol All-Purpose Cleaner-Orange Breeze (full strength) demonstrated results which ranked among the best in five of the six surfaces tested. Both Borax and Orange Glo Multipurpose Degreaser demonstrated results which ranked among the best in four of the six surfaces tested. CONCLUSIONS: The best antimicrobial cleaner to choose is often dependent on the type of surface to be cleaned of S. chartarum contamination. For Plain GWB, no paint, the best cleaners were Borax, Lysol All-Purpose Cleaner-Orange Breeze (full strength), Orange Glo Multipurpose Degreaser, and Fantastik Orange Action. RECOMMENDATIONS AND PERSPECTIVES: These results are not meant to endorse the incomplete removal of mold contaminated building materials. However, it is recognized that complete removal may not always be possible and solutions to control mold regrowth may contribute to reduced occupant exposure. Current recommendations of removal and replacement of porous building materials should be followed. It is not the intension of this discussion to endorse any product. Reporting on the performance of these products under the stated conditions was and remains the only purpose.

A simple polymerase chain reaction-sequencing analysis capable of identifying multiple medically relevant filamentous fungal species.

Due to the accumulating evidence that suggests that numerous unhealthy conditions in the indoor environment are the result of abnormal growth of the filamentous fungi (mold) in and on building surfaces it is necessary to accurately determine the organisms responsible for these maladies and to identify them in an accurate and timely manner. Historically, identification of filamentous fungal (mold) species has been based on morphological characteristics, both macroscopic and microscopic. These methods may often be time consuming and inaccurate, necessitating the development of identification protocols that are rapid, sensitive, and precise. To this end, we have devised a simple PAN-PCR approach which when coupled to cloning and sequencing of the clones allows for the unambiguous identification of multiple fungal organisms. Universal primers are used to amplify ribosomal DNA sequences which are then cloned and transformed into Escherichia coli. Individual clones are then sequenced and individual sequences analyzed and organisms identified. Using this method we were capable of identifying Stachybotrys chartarum, Penicillium purpurogenum, Aspergillus sydowii, and Cladosporium cladosporioides from a mixed culture. This method was found to be rapid, highly specific, easy to perform, and cost effective.

A simple polymerase chain reaction/restriction fragment length polymorphism assay capable of identifying medically relevant filamentous fungi.

Because of the accumulating evidence that suggests that numerous unhealthy conditions in the indoor environment are the result of abnormal growth of the filamentous fungi (mold) in and on building surfaces, it is necessary to accurately reflect the organisms responsible for these maladies and to identify them in precise and timely manner. To this end, we have developed a method that is cost effective, easy to perform, and accurate. We performed a simple polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis on multiple members of species known to negatively influence the indoor environment. The genera analyzed were Stachybotrys, Penicillium, Aspergillus, and Cladosporium. Each organism underwent PCR with universal primers that amplified ribosomal sequences generating products from 550 to 600 bp followed by enzymatic digestion with EcoRI, HaeIII, MspI, and HinfI. Our results show that using this combination of restriction enzymes enables the identification of these fungal organisms at the species level.

A simple multiplex polymerase chain reaction assay for the identification of four environmentally relevant fungal contaminants.

Historically, identification of filamentous fungal (mold) species has been based on morphological characteristics, both macroscopic and microscopic. These methods may often be time-consuming and inaccurate, necessitating the development of identification protocols that are rapid, sensitive, and precise. The polymerase chain reaction (PCR) has shown great promise in its ability to identify and quantify individual organisms from a mixed culture environment; however, the cost effectiveness of single organism PCR reactions is quickly becoming an issue. Our laboratory has developed a simple method to identify multiple fungal species, Stachybotrys chartarum, Aspergillus versicolor, Penicillium purpurogenum, and Cladosporium spp. by performing multiplex PCR and distinguishing the different reaction products by their mobility during agarose gel electrophoresis. The amplified genes include the beta-Tubulin gene from A. versicolor, the Tri5 gene from S. chartarum, and ribosomal sequences from both P. purpurogenum and Cladosporium spp. This method was found to be both rapid and easy to perform, while maintaining high sensitivity and specificity for characterizing isolates, even from a mixed culture.

A rapid DNA extraction method for PCR identification of fungal indoor air contaminants.

Following air sampling fungal DNA needs to be extracted and purified to a state suitable for laboratory use. Our laboratory has developed a simple method of extraction and purification of fungal DNA appropriate for enzymatic manipulation and Polymerase Chain Reaction (PCR) applications. The methodology described is both rapid and cost effective for use with multiple fungal organisms.