George L. Marchin

Nanoscale Powders and Formulations with Biocidal Activity Toward Spores and Vegetative Cells of Bacillus Species, Viruses, and Toxins

Certain formulations of nanoscale powders possess antimicrobial properties. These formulations are made of simple, nontoxic metal oxides such as magnesium oxide (MgO) and calcium oxide (CaO, lime) in nanocrystalline form, carrying active forms of halogens, for example, MgO · Cl2 and MgO · Br2. When these ultrafine powders contact vegetative cells of Escherichia coli, Bacillus cereus, or Bacillus globigii, over 90% are killed within a few minutes. Likewise, spore forms of the Bacillus species are decontaminated within several hours. Dry contact with aflatoxins and contact with MS2 bacteriophage (surrogate of human enterovirus) in water also causes decontamination in minutes.

A multifunctional biocide/sporocide and photocatalyst based on titanium dioxide (TiO2) codoped with silver, carbon, and sulfur.

Composite nanostructured samples of Ag (0.5-20%)/(C, S)-TiO(2) were synthesized and characterized by EDX, XRD, FT-IR, UV-vis, BET, XPS, and zeta potential measurements. Photocatalytic and biocidal tests revealed that the amount of the codoped silver (Ag(+)) in (C, S)-TiO(2) played a crucial, distinctive role in the photodegradation of gas-phase acetaldehyde as well as in the inactivation of Escherichia coli cells and Bacillus subtilis spores. Very interestingly, Ag/(C, S)-TiO(2) nanoparticles (crystallite size <10 nm) have shown very strong antimicrobial properties without light activation against both E. coli (log kill >8) and B. subtilis spores (log kill >5) for 30 min exposures, compared with P25-TiO(2). Thus, for the first time, we have demonstrated that titanium dioxide (an environmentally friendly photocatalyst) codoped with silver, carbon, and sulfur can serve as a multifunctional generic biocide as well as a visible light activated photocatalyst.

Contact and demand‐release disinfectants

A new class of solid‐phase disinfectants has emerged in recent years. Certain types of these disinfectants, the iodinated strong base resins, release iodine upon demand to microorganisms in water. These triiodide and pentaiodide resins have proven effective against a large number of bacterial genera, viruses, and representative parasites such as Giardia lamblia. Although federal regulatory policies have prevented their use for large scale water purification, they have been selectively applied in the U.S. Space Program and in situations for short‐term water disinfection. They appear to hold promise as components of hybrid devices utilizing filtration and halogen scavenging to produce high quality, potable water.

Effect of microgravity on Escherichia coli and MS-2 bacteriophage disinfection by iodinated resins☆

Experiments on chemical disinfection by iodinated resins were conducted on STS 50 (USML-1), which flew a 13 day mission during 1992. Fluid processing apparatus containing microorganisms and iodinated resins was assembled in either Manhattan, Kansas, or Boulder, Colorado, and loaded on-board the Space Shuttle for the mission. Pentaiodide resin was more effective than the triiodide resin against Escherichia coli. Both resins were more effective bactericides at unit gravity than microgravity because of cosedimentation of bacteria and iodinated resin beads. In bacteriophage experiments, the triiodide resin reduced the viable titer of MS-2 by nine logs. The few viable phage surviving chemical disinfection were associated with precipitant formation in the fluid processing apparatus.

Cryopreservation of Giardia lamblia with dimethyl sulfoxide using a dewar flask

This study examined the effect of varying freezing conditions on the human intestinal parasite Giardia lamblia (Portland-1 strain) using a constant vacuum in a Dewar flask and an ethanol bath to regulate the cooling rate. The cryopreservation of the trophozoite stage was investigated. Dimethyl sulfoxide (Me2SO), the cryoprotective agent of choice, was added directly to Giardia growth medium. Me2SO toxicity assays were conducted on those concentrations used in the freezing protocol. The results of this study indicated a 6.5% (v/v) Me2SO concentration yields a 90% survival based upon organism motility. A 30.9% cell viability was obtained by freezing in medium without a cryoprotective agent. Recommendations are offered concerning alternate viability criteria.

Efficacy of a pentaiodide resin disinfectant on Cryptosporidium parvum (Apicomplexa: Cryptosporidiidae) oocysts in vitro

The resin-I5 column developed at Kansas State University was tested for efficacy against oocysts of Cryptosporidium parvum (Apicomplexa: Cryptosporidiidae). Cesium chloride gradient-purified oocysts were passed through 1.0-cm-diameter columns with lengths of 2.5, 5.0, and 10.0 cm at 23 C. Following column passage, oocyst viability was determined both in vitro by excystation and in vivo by the ability to establish infections in suckling mice. Oocysts were found to be retained by the pentaiodide resin in a linear fashion, probably by electrostatic interactions. Linear regression analysis revealed 100% of the oocysts should be removed in such a manner using a column length of greater than or equal to 25.7 cm. When compared to untreated control oocysts, less than 12% of the oocysts that passed through the columns appeared to be affected by the resin, as assessed by excystation. Inoculation of suckling mice with these column-treated oocysts supported the excystation data and revealed the coccidian to be viable. These results indicate that oocysts of C. parvum are retained on the pentaiodide column in a 1-hit manner and that, although killing of parasites may occur within the column, the greatest effect that the column may have on the parasite is as an electrostatic retention device.

Resistance to chemical disinfection under conditions of microgravity

In unit gravity, bacteria and disinfecting resin beads co-sediment to the septum in a fluid processing apparatus (FPA) resulting in effective chemical disinfection. In microgravity bacteria in suspension have access to a larger volume of the FPA because of a lack of sedimentation. Further, when disinfecting resin beads are added to the FPA they also remain in suspension reducing their effective concentration. Typically, therefore, disinfection experiments in microgravity return larger numbers of viable bacteria than ground-based controls. Preliminary experiments aboard the MIR Space Station with Pseudomonas aeruginosa additionally suggest that the longer bacteria are retained in microgravity the more resistant they become to chemical disinfection. This phenomenon is probably due to additional time to develop resistant biofilms on the interior of the FPA. To partially solve these problems we have developed additional disinfecting materials to use in conjunction with polyiodide containing resin beads. One o...

Chemical disinfection under conditions of microgravity

There is enormous potential for point-of-use water purifiers where central water treatment does not exist or distribution systems are faulty and allow incursion of pathogenic organisms after primary treatment. Manned space missions on the Space Shuttle and planned missions on the Space Station also employ point-of-use water purifiers termed microbial check valves (MCVs). Polyiodide resin materials in use on the Space Shuttle within the MCV and in terrestrial water purifiers, silver and copper chelex resins, zirconium peroxide chelex resin, and a quaternary ammonium compound-Dow Corning 5700-polymerized to carbon and polystyrene beads, were compared for disinfection ability. Experiments were conducted in fluid processing apparatus (FPAs) at unit gravity and in microgravity conditions aboard seven STS missions. These new materials may have applications in both space and terrestrial water treatment devices.

Development of resistance to chemical disinfection by Pseudomonas aeruginosa during long-term space flight

Two long-term experiments have been conducted aboard the Mir Space Station to evaluate the development of resistance by Pseudomonas aeruginosa to chemical disinfection by polyiodide quaternary ammonium strong base resin disinfectants. The first preliminary experiment was launched aboard STS 79 and a second more extensive experiment aboard STS 86. During both experiments, after two months in a microgravity environment, aqueous suspensions of P. aeruginosa contained viable bacteria after having the iodinated resin added to them. In the second experiment identical ground based controls did not exhibit a similar phenomenon. Also in the second experiment, individual colonies from the surviving bacteria were evaluated for resistance to aqueous iodine disinfection. Compared to individual colonies from the original inoculum no resistance was observed. The data are consistent with slow development of a resistant biofilm in the bacterial suspensions flown aboard the Mir Space Station.

A Cloning Strategy for the Bacteriophage T4 vs Gene

The product of the bacteriophage T4 vs gene, the tau peptide, has been shown to thermally stabilize temperature-sensitive valyl-tRNA synthetase (EC 6.1.1.9). To clone the bacteriophage T4 vs gene, recombinant pBR322-T4 DNA molecules were used to transform Escherichia coli CP 790302 (valSts). Transformants that grew at 43.5 degrees C on selective medium exhibited some properties indicative of a phage-modified valyl-tRNA synthetase. The data are consistent with successful cloning of the vs gene and atypical modification of the valyl-tRNA synthetase in CP 790302.