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Biomedical subjects

J R Puleo

Publications and source records attributed to J R Puleo.

At least 19 recordsLinked to original sources
PubMed1979

Wipe-rinse technique for quantitating microbial contamination on large surfaces.

The evaluation of an improved wipe-rinse technique for the bioassay of large areas was undertaken due to inherent inadequacies in the cotton swab-rinse technique to which assay of spacecraft is currently restricted. Four types of contamination control cloths were initially tested. A polyester-bonded cloth (PBC) was selected for further evaluation because of its superior efficiency and handling characteristics. Results from comparative tests with PBC and cotton swabs on simulated spacecraft surfaces indicated a significantly higher recovery efficiency for the PBC than for the cotton (90.4 versus 75.2%). Of the sampling areas sites studied, PBC was found to be most effective on surface areas not exceeding 0.74 m2 (8.0 feet 2).

Aircraft↗
PubMed1978

Thermal resistance of naturally occurring airborne bacterial spores.

Simulation of a heat process used in the terminal dry-heat decontamination of the Viking spacecraft is reported. Naturally occurring airborne bacterial spores were collected on Teflon ribbons in selected spacecraft assembly areas and subsequently subjected to dry heat. Thermal inactivation experiments were conducted at 105, 111.7, 120, 125, 130, and 135 degrees C with a moisture level of 1.2 mg of water per liter. Heat survivors were recovered at temperatures of 135 degrees C when a 30-h heating cycle was employed. Survivors were recovered from all cycles studied and randomly selected for identification. The naturally occurring spore population was reduced an average of 2.2 to 4.4 log cycles from 105 to 135 degrees C. Heating cycles of 5 and 15 h at temperature were compared with the standard 30-h cycle at 111.7, 120, and 125 degrees C. No significant differences in inactivation (alpha = 0.05) were observed between 111.7 and 120 degrees C. The 30-h cycle differs from the 5-and 15-h cycles at 125 degrees C. Thus, the heating cycle can be reduced if a small fraction (about 10-3 to 10-4) of very resistant spores can be tolerated.

Air Microbiology↗
PubMed1977

Microbiological profiles of the Viking spacecraft.

Planetary quarantine requirements associated with the launch of two Viking spacecraft necessitated microbiological assessment during assembly and testing at Cape Canaveral and the Kennedy Space Center. Samples were collected from selected surface of the Viking Lander Capsules (VLC), Orbiters, (VO), and Shrouds at predetermined intervals during assembly and testing. Approximately 7,000 samples were assayed. Levels of bacterial spores per square meter on the VLC-1 and VLC-2 were 1.6 x 10(2) and 9.7 x 10(1), respectively, prior to dry-heat sterilization. The ranges of aerobic mesophilic microorganisms detected on the VO-1 and VO-2 at various sampling events were 4.2 x 10(2) to 4.3 x 10(3) and 2.3 x 10(2) to 8.9 x 10(3)/m2, respectively. Approximately 1,300 colonies were picked from culture plates, identified, lypholipized, and stored for future reference. About 75% of all isolates were microorganisms considered indigenous to humans; the remaining isolates were associated with soil and dust in the environment. The percentage of microorganisms of human origin was consistent with results obtained with previous automated spacecraft but slightly lower than those observed for manned (Apollo) spacecraft.

Actinomycetales↗
PubMed1977

Pyrolysis gas-liquid chromatography of the genus Bacillus: effect of growth media on pyrochromatogram reproducibility.

Pyrolysis gas-liquid chromatography was performed on dried Bacillus microorganisms to evaluate the effects of growth media. Six cultures of Bacillus and six lot numbers of Trypticase soy agar (BBL) were used to test the hypothesis that a microorganism grown on various lot numbers of the same chromatogram. Also tested was the effect of three different media on chromatogram reproduction using the same six cultures. Results show little or no differences observed between the chromatograms of the individual Bacillus spp. grown on the six lot numbers of Trypticase soy agar. When chromatograms of the three different media were compared, several differences were observed, particularly in the areas most characteristic of individual species. Pryolysis gas-liquid chromatography can be a useful tool for the characterization or identification of the genus Bacillus if the chromatographic and cultural conditions are maintained.

Bacillus↗
PubMed1975

Method for collecting naturally occurring airborne bacterial spores for determining their thermal resistance.

The ability to determine the thermal resistance of naturally occurring airborne bacterial spores associated with spacecraft and their assembly areas has been hindered by lack of an effective collecting system. Efforts to collect and concentrate spores with air samplers or from air filters have not been successful. A fallout method was developed for this purpose and tested. Sterile Teflon ribbons (7.6 by 183 cm) were exposed in pertinent spacecraft assembly areas and subsequently treated with dry heat. Thermal inactivation experiments were conducted at 125 and 113 C. Heating intervals ranged from 1 to 12 h at 125 C and 6, 12, 18, and 24 h at 113 C. Eight hours was the longest heating time yielding survivors at 125 C, whereas survivors were recovered at all of the heating intervals at 113 C. D125C values were calculated using the fractional-replicate-unit-negative technique of Pflug and Schmidt (1968) and ranged from 25 to 126 min. This variation indicated that the most probable number of survivors at each heating interval did not fall on a straight line passing through the initial spore population. However, the most-probable-number values taken alone formed a straight line suggesting logarithmic thermal destruction of a subpopulation of spores with a D125C value of 6.3 h.

Air Microbiology↗
PubMed1975

Quantitative relationship between airborne viable and total particles.

The numbers of viable and total particles in a microbiological laboratory and in a class 100,000 clean room were examined for a predictable relationship to aid the monitoring of airborne microbial contamination. Six-stage Andersen samplers were used to enumerate viable particles and a modified Royco particle counter was used to enumerate total particles. Over 99% of the total particles present in both areas were less than 1 micron in size; however, only 1 in 10,000 of the particles this size were viable. At the other end of the particle size scale, it was found that less than 0.1% of the total particles were greater than 5.4 micron in size, but only 4.5% of these particles were viable. Viable particles make up only a very small portion of the total particles making any correlation undetectable. An analysis of the combined data from both areas using only total and viable particles over 5.4 micron showed a positive correlation. An analysis of the data from each area individually showed no correlation. Estimation of viable particulates from enumeration of total particulates in the large particle ranges may be possible only in areas where a high degree of environmental control is exercised.

Air Microbiology↗
PubMed1974

Evaluation fo membrane filter field monitors for microbiological air sampling.

Due to area constraints encountered in assembly and testing areas of spacecraft, the membrane filter field monitor (MF) and the National Aeronautics and Space Administration-accepted Reyniers slit air sampler were compared for recovery of airborne microbial contamination. The intramural air in a microbiological laboratory area and a clean room environment used for the assembly and testing of the Apollo spacecraft was studied. A significantly higher number of microorganisms was recovered by the Reyniers sampler. A high degree of consistency between the two sampling methods was shown by a regression analysis, with a correlation coefficient of 0.93. The MF samplers detected 79% of the concentration measured by the Reyniers slit samplers. The types of microorganisms identified from both sampling methods were similar. Variables in the MF samplers, such as pore size, relative humidity, and flow rates, have been studied, but no effect was noted on recovery. The results show that the MF method could be used to estimate the number and types of microorganisms found in the air.

Actinomycetales↗
PubMed1974

Comparison of media for detection of fungi on spacecraft.

Five media, including Trypticase soy agar (TSA; BBL) pour plates, spread plates of TSA, Mycophil agar with chloromycetin, Mycophil agar with chloromycetin and Actidione, and cornmeal agar with chloromycetin were quantitatively and qualitatively compared for the detection of fungi on spacecraft. Cornmeal agar with chloromycetin yielded the highest number of fungal colonies, although not always significantly higher than Mycophil agar with chloromycetin or TSA spread plates. Cornmeal agar with chloromycetin also gave the best qualitative representation of fungi on the spacecraft, recovering 68% of the genera found from all media. This medium yielded 10 times the number of fungal colonies and 3 times the number of genera found on TSA pour plates as currently used for spacecraft assay.

Agar↗
PubMed1974

Mathematical estimation on the level of microbial contamination on spacecraft surfaces by volumetric air sampling.

Microbiological sampling methods presently used for enumeration of microorganisms on spacecraft surfaces require contact with easily damaged components. Estimation of viable particles on surfaces using air sampling methods in conjunction with a mathematical model would be desirable. Parameters necessary for the mathematical model are the effect of angled surfaces on viable particle collection and the number of viable cells per viable particle. Deposition of viable particles on angled surfaces closely followed a cosine function, and the number of viable cells per viable particle was consistent with a Poisson distribution. Other parameters considered by the mathematical model included deposition rate and fractional removal per unit time. A close nonlinear correlation between volumetric air sampling and airborne fallout on surfaces was established with all fallout data points falling within the 95% confidence limits as determined by the mathematical model.

Actinomycetales↗
PubMed1973

Microbiological profiles of four Apollo spacecraft.

Selected surfaces from the Command Module, Lunar Module (ascent and descent stages), Instrument Unit, Saturn S-4B engine, and Spacecraft Lunar Module Adapter comprised the various components of four Apollo spacecraft which were assayed quantitatively and qualitatively for microorganisms. In addition, the first Lunar Roving Vehicle was assayed. Average levels of microbial contamination (10(4) per square foot of surface) on the Command Module, Instrument Unit, and Saturn S-4B engine were relatively consistent among spacecraft. The first postflight sampling of interior surfaces of the Command Module was possible due to elimination of the 21-day back-contamination quarantine period. Results of the pre- and postflight samples revealed increases in the postflight samples of 3 logs/inch(2). A total of 5,862 microbial isolates was identified; 183 and 327 were obtained from the Command Module at preflight and postflight sampling periods, respectively. Although the results showed that the majority of microorganisms isolated were those considered to be indigenous to humans, an increase in organisms associated with soil and dust was noted with each successive Apollo spacecraft.

Actinomycetales↗
PubMed1970

Quantitative and qualitative microbiological profiles of the Apollo 10 and 11 spacecraft.

Microbiological profiles were determined for surfaces of the command module, lunar module (ascent and descent stages), instrument unit, Saturn S-4B stage, and the spacecraft lunar module adapter of the Apollo 10 and 11 spacecraft. Average levels of contamination of the command module were 2.1 x 10(4) and 2.7 x 10(4) microorganisms per ft(2) for Apollo 10 and 11, respectively. With the exception of the exterior surfaces of the ascent stage of the lunar module and the interior surfaces of the command module, average levels of microbial contamination on all components of the Apollo 11 were found to be lower than those observed on Apollo 10. For each Apollo mission, approximately 2,000 colonies were picked from a variety of media and identified. The results showed that approximately 95% of all isolates were those considered indigenous to humans; the remaining were associated with soil and dust in the environment. However, the ratio of these two general groups varied depending on the degrees of personnel density and environmental control associated with each module.

Bacteria↗
PubMed1968

Comparison of microbial contamination levels among hospital operating rooms and industrial clean rooms.

Microbial contamination in industrial clean rooms was compared quantitatively and qualitatively with that of hospital operating rooms. The number of aerobic mesophilic microorganisms which accumulated on stainless-steel strips exposed for periods up to 21 weeks to the intramural air of four operating rooms was at least 1 log higher than the accumulation on strips exposed in four clean rooms, and was essentially the same as that found in two factory areas. Volumetric air samplings showed that there were significantly higher numbers of airborne viable particles per cubic foot of air in operating rooms than in industrial clean rooms. In contrast to clean rooms, where most of the airborne contaminants were those associated with human hair, skin, and respiratory tract, the hospital operating rooms showed a very high level of microorganisms associated with dust and soil.

Air Microbiology↗
PubMed1967

Use of ultrasonic energy in assessing microbial contamination on surfaces.

Ultrasonic tanks were evaluated for their ability to remove viable microorganisms from various surfaces for subsequent enumeration. Test surfaces were polished stainless steel, smooth glass, frosted glass, and electronic components. The position of contaminated surfaces in relation to the ultrasonic energy source, distance of the ultrasonic source from the test surfaces, and temperature of the rinse fluid were some of the factors which influenced recovery. Experimental systems included both naturally occurring microbial contamination and artificial contamination with spores of Bacillus subtilis var. niger. The results showed that ultrasonic energy was more reliable and efficient than mechanical agitation for recovering surface contaminants. Conditions which increased the number and percentage of microorganisms recovered by ultrasonic energy were: using a cold rinse fluid, placing the sample bottle on the bottom of the ultrasonic tank, and facing the contaminated surfaces toward the energy source. It was also demonstrated that ultrasonic energy could be effectively used for eluting microorganisms from cotton swabs.

Journal Article↗