Mold growth on gypsum wallboard--a summary of three techniques.

Reducing occupant exposure to mold growing on damp gypsum wallboard and controlling mold contamination in the indoor environment was studied through 1) delineation of environmental conditions required to promote and avoid mold growth and 2) efficacy testing of antimicrobial products, specifically cleaners and paints, on gypsum wallboard (GWB) surfaces. The effects of moisture and relative humidity (RH) on mold growth and transport are important in avoiding and eliminating problems. These effects have been demonstrated on GWB and are discussed in this article for use as control guidance. The authors discuss the efficacy of antimicrobial cleaners and paints to remove, eliminate, or control mold growth on GWB. Research to control Stachybotrys chartarum growth using 13 separate antimicrobial cleaners and nine varieties of antimicrobial paint on contaminated GWB was performed in laboratory testing. GWB surfaces were subjected to high RH. GWB control measures are summarized and combined, and the antimicrobial product results are explained.

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

The goal of this research was to reduce occupant exposure to indoor mold through the efficacy testing of antimicrobial paints. An accepted method for handling Stachybotrys chartarum-contaminated gypsum wallboard (GWB) is removal and replacement. This practice is also recommended for water-damaged or mold-contaminated GWB but is not always followed completely. The efficacy of antimicrobial paints to eliminate or control mold regrowth on surfaces can be tested easily on nonporous surfaces. The testing of antimicrobial efficacy on porous surfaces found in the indoor environment, such as gypsum wallboard, can be more complicated and prone to incorrect conclusions regarding residual organisms. The mold S. chartarum has been studied for toxin production and its occurrence in water-damaged buildings. Research to control its growth using seven different antimicrobial paints and two commonly used paints on contaminated, common gypsum wallboard was performed in laboratory testing at high relative humidity. The results indicate differences in antimicrobial efficacy for the period of testing, and that proper cleaning and resurfacing of GWB with an antimicrobial paint can be an option in those unique circumstances when removal may not be possible.

Comparison of four different methods for extraction of Stachybotrys chartarum spore DNA and verification by real-time PCR.

A comparison of four different methods for the extraction of spore DNA from Stachybotrys chartarum was conducted. Spore DNA was extracted and purified using either one of three different commercial kits or water. All preparations utilized bead milling. Genomic DNA extracted from 10(1) to 10(7) spores was assessed by both agarose gel electrophoresis and real-time quantitative polymerase chain reaction (qPCR) performed against multi-copy (rRNA) and single-(tubulin) gene targets. The spore isolation technique we employed was verified to be pure by light microscopy. Although all preparatory methods led to successful detection by qPCR, S. chartarum spore DNA prepared using the Qiagen Plant kit was notably better over the extraction range.

Solvent comparison in the isolation, solubilization, and toxicity of Stachybotrys chartarum spore trichothecene mycotoxins in an established in vitro luminescence protein translation inhibition assay.

It is well known that non-viable mold contaminants such as macrocyclic trichothecene mycotoxins of Stachybotrys chartarum are highly toxinigenic to humans. However, the method of recovering native mycotoxin has been without consensus. Inconsistencies occur in the methods of isolation, suspension, preparation, and quantitation of the mycotoxin from the spores. The purpose of this study was to provide quantitatively comparative data on three concurrent preparations of 10(6)S. chartarum spores. The experiments were designed to specifically evaluate a novel method of mycotoxin extraction, solubilization, and the subsequent inhibitory effect in an established in vitro luminescence protein translation assay from 30 day-old spores. The mycotoxin-containing spores swabbed from wallboard cultures were milled with and without glass beads in 100% methanol, 95% ethanol, or water. Milled spore lysates were cleared of cell debris by filter centrifugation followed by a second centrifugation through a 5000 MWCO filter to remove interfering proteins and RNases. Cleared lysate was concentrated by centrivap and suspended in either alcohol or water as described. The suspensions were used immediately in the in vitro luminescence protein translation assay with the trichothecene, T-2 toxin, as a control. Although, mycotoxin is reported to be alcohol soluble, the level of translation inhibition was not reliably satisfactory for either the methanol or ethanol preparations. In fact, the methanol and ethanol control reactions were not significantly different than the alcohol prepared spore samples. In addition, we observed that increasing amounts of either alcohol inhibited the reaction in a dose dependent manner. This suggests that although alcohol isolation of mycotoxin is desirable in terms of time and labor, the presence of alcohol in the luminescence protein translation reaction was not acceptable. Conversely, water extraction of mycotoxin demonstrated a dose dependent response, and there was significant difference between the water controls and the water extracted mycotoxin reactions. In our hands, water was the best extraction agent for mycotoxin when using this specific luminescence protein translation assay kit.

An analytical method for the measurement of nonviable bioaerosols.

Exposures from indoor environments are a major issue for evaluating total long-term personal exposures to the fine fraction (<2.5 microm in aerodynamic diameter) of particulate matter (PM). It is widely accepted in the indoor air quality (IAQ) research community that biocontamination is one of the important indoor air pollutants. Major indoor air biocontaminants include mold, bacteria, dust mites, and other antigens. Once the biocontaminants or their metabolites become airborne, IAQ could be significantly deteriorated. The airborne biocontaminants or their metabolites can induce irritational, allergic, infectious, and chemical responses in exposed individuals. Biocontaminants, such as some mold spores or pollen grains, because of their size and mass, settle rapidly within the indoor environment. Over time they may become nonviable and fragmented by the process of desiccation. Desiccated nonviable fragments of organisms are common and can be toxic or allergenic, depending upon the specific organism or organism component. Once these smaller and lighter fragments of biological PM become suspended in air, they have a greater tendency to stay suspended. Although some bioaerosols have been identified, few have been quantitatively studied for their prevalence within the total indoor PM with time, or for their affinity to penetrate indoors. This paper describes a preliminary research effort to develop a methodology for the measurement of nonviable biologically based PM, analyzing for mold and ragweed antigens and endotoxins. The research objectives include the development of a set of analytical methods and the comparison of impactor media and sample size, and the quantification of the relationship between outdoor and indoor levels of bioaerosols. Indoor and outdoor air samples were passed through an Andersen nonviable cascade impactor in which particles from 0.2 to 9.0 microm were collected and analyzed. The presence of mold, ragweed, and endotoxin was found in all eight size ranges. The presence of respirable particles of mold and pollen found in the fine particle size range from 0.2 to 5.25 microm is evidence of fragmentation of larger source particles that are known allergens.

Investigation of the potential antimicrobial efficacy of sealants used in HVAC systems.

Recent experiments confirm field experience that duct cleaning alone may not provide adequate protection from regrowth of fungal contamination on fiberglass duct liner (FGDL). Current recommendations for remediation of fungally contaminated fiberglass duct materials specify complete removal of the materials. But removal of contaminated materials can be extremely expensive. Therefore, a common practice in the duct-cleaning industry is the postcleaning use of antimicrobial surface coatings with the implication that they may contain or limit regrowth. Little information is available on the efficacy of these treatments. This paper describes a study to evaluate whether three commercially available antimicrobial coatings, placed on a cleaned surface that 1 year previously had been actively growing microorganisms, would be able to prevent regrowth. The three coatings contained different active antimicrobial compounds. All three of the coatings were designed for use on heating, ventilation, and air conditioning (HVAC) system components or interior surfaces of lined and unlined duct systems. Coating I was a polyacrylate copolymer containing zinc oxide and borates. Coating II was an acrylic coating containing decabromodiphenyl oxide and antimony trioxide. Coating III was an acrylic primer containing a phosphated quaternary amine complex. The study included field and laboratory assessments. The three treatments were evaluated in an uncontrolled field setting in an actual duct system. The laboratory study broadened the field study to include a range of humidities under controlled conditions. Both static and dynamic chamber laboratory experiments were performed. The results showed that two of the three antimicrobial coatings limited the regrowth of fungal contamination, at least in the short term (the 3-month time span of the study); the third did not. Before use in the field, testing of the efficacy of antimicrobial coatings under realistic use conditions is recommended because antimicrobials have different baseline activities and interact differently with the substrate that contains them and their local environment.

Legislation, policy, and tobacco use among youth: implications for health care providers.

This paper addresses implications of recent tobacco legislation, policy, and tobacco use among youth in the context of health care policy and services. Tobacco use prevalence and definitions and diagnoses of nicotine addiction and dependence are described. Assessment of smoking prevalence in Texas provides a case study of the problem and potential solutions for tobacco use among youth. The case study highlights specific implications to be considered when providing health care focused on prevention and risk reduction for youth. The paper concludes with implications and critical Internet resources for health care providers engaging in youth tobacco control.

The distribution of gram negative bacteria and endotoxin on raw cotton components.

The botanical composition of representative raw cottons from seven different growing regions was determined by manual removal and identification of all trash components greater than 50 micron in size. The number of gram negative bacteria (GNB) and the amount of endotoxin present in each of the separated raw cotton components were quantified. Low middling cotton contained significantly more bract-leaf trash than that found in higher quality cottons such as those in the middling grade division. Significantly more GNB and endotoxin were found in botanical trash components as well as lint of raw cotton derived from the southwest and southeast growing regions as compared to similar botanical components from far west cottons. For representative raw cottons from the 1980 USA crop we determined that 67% of the GNB and 89% of the endotoxin resided on white lint itself, from which all particulate larger than 50 micron in size had been removed manually.