Evaluation of a one-way airflow system in an animal room based on counts of airborne dust particles and bacteria and measurements of ammonia levels

Air cleanliness in the working area of an animal room equipped with a conventional turbulent flow air distribution systems was compared with that in a similar room fitted with a one-way-flow air distribution system; in this, the supply air flowed from the working area through the racks of cages and was removed from the exhaust side. Before the introduction of animals, the air in the working and exhaust areas of both rooms was ascertained to be Class 100. With animals in situ, however, whereas in the turbulent airflow room both the workspace and exhaust air reached about Class 10,000 (with particle counts, bacterial counts and ammonia levels being almost the same) in the one-way-flow room, the air in the work space only went up to about Class 1000. With the addition of sliding doors or curtains in front of the rack in the one-way-flow room the work space air was maintained at Class 100 with almost no dust particles over 1 microns in size, airborne bacteria or ammonia being detectable. A comparison of all factors measured showed that whereas in the turbulent flow room the contamination of the work space air was 91% of that of the exhaust air, in the one-way-flow room it was only 47%, with curtains added this was reduced to 7% and with sliding doors to only 2%. In the latter case, contamination levels increased markedly on both sides during and immediately after cage changing, but recovered to the pre-cage changing levels within 30 min in the personnel working area and within 60 min on the exhaust side.     

Evaluation of laminar flow microbiological safety cabinets

The microbiological control efficiency of two class 100 laminar down-flow hoods was determined by using aerosols of Bacillus subtilis var. niger spores. The first unit challenged utilized a slanted eyelid to partially enclose the front work opening. This hood showed nearly perfect control of ambient organisms in the work area. It also gave a 10(6) or greater drop in the number of organisms passing out of the exhaust system. However, when the interior work area of the hood was challenged, significant numbers of spores penetrated the air barrier and escaped into the ambient air. A redesigned laminar flow hood was built incorporating a vertical eyelid and a reduced opening to the work area. This hood showed the same excellent characteristics for controlling ambient contamination. Exhaust system leakage was also extremely low. Air barrier efficiency for the newer hood was increased with lower amounts of spore penetration into the ambient air.     

Biohazard Hood to Prevent Infection During Microbiological Procedures

A microbiological hood was designed to reduce the danger of airborne infection of laboratory workers. The hood uses absolute filters to deliver sterile air in a laminar flow to the work area. An air curtain across the hood opening permits easy access but separates the worker from aerosols produced in the hood, and protects material inside the hood from contamination by room air. Tests with bacterial and viral aerosols showed that the air curtain is at least 99.96% effective in preventing airborne particles from entering the work area.     

屏障系统动物室单向流与乱流空气净化方式的细菌比较分析

目的探讨屏障系统动物室控制微生物污染,维持洁净的方法。方法将屏障系统动物室的乱流空气净化方式进行改进,在乱流室内增加能改变气流方式的无动力的设施,使室内气流改变为单向流动的方式。结果乱流动物室的沉降菌平均为47个,浮游菌平均为154.5个;单向气流动物室内的沉降菌平均为1.5个,浮游菌平均为7个。结论单向流优于乱流。这将为今后我国制定新的实验动物屏障系统环境标准提供技术数据。     

Evaluation of indoor passive pollen sedimentation over 1 year: a possible source of contamination?

The study aimed to evaluate the possible contamination by passive indoor pollen sedimentation in a laboratory where routine pollen analyses were conducted, but where no particular laboratory policies were adopted to limit contamination. Gravimetric pollen deposition was observed on traps (petri dishes soaked with glycerol) set in the palynology laboratory, under an extractor hood and on the bench beside it over 1 year (1995–1996), and in an air filtered room in a flow cabinet and on its roof over 1 month for comparison. Under the extractor hood, three types of airborne particles were deposited: pollen grains, spores and algae, representing 32.35, 67.28 and 0.37%, respectively, of the total sedimented particles over 1 year. The number of pollen grains deposited on the surface trap ranged from 0 (27 November to 4 December 1995) to 707 (10–18 April 1995). The highest number of taxa during a weekly collection was 23 (9–15 May 1995). The pollen flora represented by anemophilous pollen (>90% of the trapped pollen) was related to the vegetation next to the laboratory:Acer, Carpinus, Castanea, Corylus, Cupressaceae, Pinus, Quercus, Salix, Taxus for trees and shrubs andArtemisia, Brassicaceae, Plantago, Poaceae, andUrticaceae for grasses and weeds. Indoor pollen deposition corresponded to the period of the outdoor pollination (macroscopic field observation) which lasted from March to the beginning of August. However, some pollen were almost always present in the collection (Poaceae, Salix, Castanea, Betula), reflecting the occurrence of pollen grains in the atmosphere out of the pollination period. Moreover, about five times more entomophilous pollen was found under the extractor hood compared to the other area of the laboratory; even in the flow cabinet of the air filtered room, 237 particles were captured (versus only 15 on the roof). These data suggest a possible human contamination during operations under the extractor hood or in the flow cabinet. Although few airborne pollen were found, possible contamination has to be considered in investigations where even low pollen quantities are of interest.     

An Assessment of Air As a Source of DNA Contamination Encountered When Performing PCR

Sensitive molecular methods, such as the PCR, can detect low-level contamination, and careful technique is required to reduce the impact of contaminants. Yet, some assays that are designed to detect high copy-number target sequences appear to be impossible to perform without contamination, and frequently, personnel or laboratory environment are held responsible as the source. This complicates diagnostic and research analysis when using molecular methods. To investigate the air specifically as a source of contamination, which might occur during PCR setup, we exposed tubes of water to the air of a laboratory and clean hood for up to 24 h. To increase the chances of contamination, we also investigated a busy open-plan office in the same way. All of the experiments showed the presence of human and rodent DNA contamination. However, there was no accumulation of the contamination in any of the environments investigated, suggesting that the air was not the source of contamination. Even the air from a busy open-plan office was a poor source of contamination for all of the DNA sequences investigated (human, bacterial, fungal, and rodent). This demonstrates that the personnel and immediate laboratory environment are not necessarily to blame for the observed contamination.     

Reduction of skin bacteria in theatre air with comfortable,non-woven disposable clothing for operating-theatre staff.

Conventional loose-weave cotton operating garments were compared with clothing of a non-woven fabric to test their efficacy in reducing the dispersal of skin bacteria into theatre air. When men wore operating suits made of the non-woven fabric dispersal of skin bacteria was reduced by 72%. When all the operating-theatre staff wore suits and dresses of this fabric air bacterial counts during operating sessions were reduced by 55%; no reduction occurred when the fabric was worn by only the scrubbed team. The lowest levels of microbial contamination of the air in the operating theatre occurred when both the unscrubbed and scrubbed theatre staff wore clothes of non-woven fabric.     

Assessment of microbial parameters as indicators of viral contamination of aerosol from urban sewage treatment plants

In order to evaluate possible indicators of viral aerosol contamination in sewage treatment plants, a year-long study was carried out on the relationships between the presence of cytopathogenic viruses and the counts of total bacteria, faecal streptococci and somatic coliphages in samples collected at various distances from the aerosol source (aeration tank). The activated sludge plant studied proved to be a significant source of microbe-bearing aerosol with high levels of viral contamination. When the virus was found in sewage, it was also found in the air, at least in the sites closest to the aeration tank. With regard to the possibility of using the chosen parameters as markers of viral contamination, the total bacteria and faecal streptococci counts were generally positively correlated with viral presence, while coliphage counts yielded no analogous relationship.     

Quantitative analysis of bacterial aerosols in two different dental clinic environments.

Microbial aerosols are generated during dental treatments and may represent an important source of infection. This study was designed to quantify bacterial air contamination during dental treatments in both a closed dental operatory and a multichair dental clinic. Air was sampled by using a slit type of biological air sampler. Following air sampling, blood-supplemented Trypticase soy agar plates were incubated at 37 degrees C under anaerobic conditions for 7 days. The maximum levels of air contamination in the closed dental operatory were observed while dental treatments were being performed (four trials; 216 +/- 75 CFU/m3 for ultrasonic scaling treatments and 75 +/- 22 CFU/m3 for operative treatments). At 2 h after completion of the treatments, the bacterial counts were about the same as the pretreatment levels (12 to 14 CFU/m3). In the second part of the study, a multichair dental clinic was divided into four areas, and air contamination was monitored at each site. Three sites were located in active dental treatment areas, whereas no dental treatments were performed within an 11-m radius of the fourth site. At 3 h after the beginning of dental treatments, the highest bacterial counts were obtained in the three active dental treatment areas (76 to 114 CFU/m3). However, there was noticeable contamination in the inactive dental treatment area (42 CFU/m3). Thus, bacterial aerosols were able to spread into areas where there was no dental activity. My data show that dental treatments significantly increased the levels of bacterial air contamination in both a closed dental operatory and a multichair dental clinic.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative levels and types of microbial contamination detected in industrial clean rooms

The primary objective of this study was to determine quantitatively and qualitatively the predominant types of microbial contamination occurring in conventional and laminar flow clean rooms. One horizontal laminar flow, three conventional industrial clean rooms, and three open factory areas were selected for microbiological tests. The results showed that as the environment and personnel of a clean room were controlled in a more positive manner with respect to the reduction of particulate contamination, the levels of airborne and surface microbial contaminants were reduced accordingly. The chief sources of microbial contamination were associated with the density and activity of clean room personnel. In addition, the majority of microorganisms isolated from the intramural air by air samplers were those indigenous to humans. Studies on the fallout and accumulation of airborne microorganisms on stainless-steel surfaces showed that, although there were no significant differences in the levels of microbial contamination among the conventional clean rooms, the type of microorganism detected on stainless-steel surfaces was consistently and significantly different. In addition, the "plateau phenomenon" occurred in all environments studied. It was concluded that the stainless-steel strip method for detecting microbial accumulation on surfaces is efficient and sensitive in ultra-clean environments and is the most reliable and practical method for monitoring microbial contamination in future class 100 clean rooms to be used for the assembly of spacecraft which will be sterilized.     

The turbulent boundary layer over a flapping Populus leaf

Abstract The air-flow regime over a Populus leaf was investigated using a constant temperature hot-film sensor which could be glued on the abaxial surface. The leaf was exposed to a laminar or turbulent flow of air in a wind-tunnel, while being free to undergo normal motion. In the laminar air-flow the boundary layer remained laminar, despite the fluttering of the leaf, until a Reynolds number of 2.3 × 10⁴ was reached. When the air incident on the leaf was made turbulent, to resemble natural conditions, the boundary layer became turbulent at a Reynolds number of 0.4 × 10⁴. The experiments suggest that the boundary layers over leaves are always turbulent in the natural wind.     

Effect of flow regimes on the presence of Legionella within the biofilm of a model plumbing system

AIMS: Stagnation is widely believed to predispose water systems to colonization by Legionella. A model plumbing system was constructed to determine the effect of flow regimes on the presence of Legionella within microbial biofilms. METHODS AND RESULTS: The plumbing model contained three parallel pipes where turbulent, laminar and stagnant flow regimes were established. Four sets of experiments were carried out with Reynolds number from 10,000 to 40,000 and from 355 to 2,000 in turbulent and laminar pipes, respectively. Legionella counts recovered from biofilm and planktonic water samples of the three sampling pipes were compared with to determine the effect of flow regime on the presence of Legionella. Significantly higher colony counts of Legionella were recovered from the biofilm of the pipe with turbulent flow compared with the pipe with laminar flow. The lowest counts were in the pipe with stagnant flow. CONCLUSIONS: We were unable to demonstrate that stagnant conditions promoted Legionella colonization. SIGNIFICANCE AND IMPACT OF THE STUDY: Plumbing modifications to remove areas of stagnation including deadlegs are widely recommended, but these modifications are tedious and expensive to perform. Controlled studies in large buildings are needed to validate this unproved hypothesis.     

The effect of filters on aseptic pipetting lifetime of mechanical and electronic pipettors and carryover during pipetting

The aseptic pipetting lifetime of air displacement pipettors during serial pipetting was investigated. When a Micrococcus luteus ^T culture was pipetted using no filter, less than 100 pipettings were required to contaminate the interiors of both mechanical and electronic pipettors with more than 10³ cfu (n = 1890 pipettings). When a filter was placed in the pipettor tip cone, the sterility of the pipettor barrel was maintained for more than 500 pipettings (n = 2620) or 50–250 pipettings (n = 2520), depending on the filter type used. When a radioactive liquid or plasmid DNA was pipetted using no filter, contamination of the pipettor barrel occurred within less than 100 pipettings. A filter placed in the pipettor tip cone protected the interior of the pipettors against radioactive contamination for more than 250 pipettings and against DNA contamination for more than 500 pipettings. A pipettor barrel contaminated with M. luteus ^T caused carryover in 13% of the pipetting series. When a SCF filter was placed in the tip cone, no carryover was observed within 2620 pipettings. A tip cone filter replaced at intervals of 50–250 (polyethylene filter) or 500 (SCF filter) pipettings will protect the pipettor barrel from contamination and the samples from carryover.     

Mass Airflow Cabinet for Control of Airborne Infection of Laboratory Rodents

A mass airflow cabinet for handling and housing of laboratory rodents has been developed and tested. The unit consists of a high-efficiency particulate air filter and uniform distribution of air at a vertical velocity of 19 cm per s. Animals are maintained without bedding in mesh-bottomed cages that rest on rollers for rotation inside the cabinet. There is an air barrier of 90 cm per s separating the cabinet air from room air. Sampling for airborne bacteria yielded an average of 0.03 colony-forming units (CFU) per ft(3) of air inside the cabinet, whereas 28.8 CFU per ft(3) was simultaneously detected outside the cabinet during housekeeping, a reduction of almost three logs. The efficiency of the air barrier was tested by aerosolization of T3 phage. When phage was aerosolized 5 cm outside the cabinet, no phage could be detected 5 cm inside when the fans were operating; with the fans off an average of 1.6 x 10(4) plaque-forming units (PFU) per ft(3) was detected in six tests. Aerosolization of phage inside the cabinet yielded an average of 9 x 10 PFU per ft(3) outside; an average of 4.1 x 10(6) PFU per ft(3) were detected with the fans not in operation, a reduction of more than four logs. In-use studies on effectiveness showed that the cabinet significantly reduced the incidence of mice originally titer-free to Reo-3 virus. Hemagglutination inhibition antibodies to Reo-3 were detected in 9/22 (42%) mice housed in a conventionally ventilated animal laboratory while no seroconversion was detected in any of 22 mice housed in the mass air flow cabinet in the same laboratory.     

Reaerosolization of fluidized spores in ventilation systems

This project examined dry, fluidized spore reaerosolization in a heating, ventilating, and air conditioning duct system. Experiments using spores of Bacillus atrophaeus, a nonpathogenic surrogate for Bacillus anthracis, were conducted to delineate the extent of spore reaerosolization behavior under normal indoor airflow conditions. Short-term (five air-volume exchanges), long-term (up to 21,000 air-volume exchanges), and cycled (on-off) reaerosolization tests were conducted using two common duct materials. Spores were released into the test apparatus in turbulent airflow (Reynolds number, 26,000). After the initial pulse of spores (approximately 10(10) to 10(11) viable spores) was released, high-efficiency particulate air filters were added to the air intake. Airflow was again used to perturb the spores that had previously deposited onto the duct. Resuspension rates on both steel and plastic duct materials were between 10(-3) and 10(-5) per second, which decreased to 10 times less than initial rates within 30 min. Pulsed flow caused an initial spike in spore resuspension concentration that rapidly decreased. The resuspension rates were greater than those predicted by resuspension models for contamination in the environment, a result attributed to surface roughness differences. There was no difference between spore reaerosolization from metal and that from plastic duct surfaces over 5 hours of constant airflow. The spores that deposited onto the duct remained a persistent source of contamination over a period of several hours.     

Direct visualizations of air flow in the human upper airway using in-vitro models

A better understanding of airflow characteristics in the upper airway(UA) is crucial in investigating obstructive sleep apnea(OSA), particle sedimentation, drug delivery, and many biomedical problems. Direct visualization of air flow patterns in in-vitro models with realistic anatomical structures is a big challenge. In this study, we constructed unique half-side transparent physical models of normal UA based on realistic anatomical structures. A smoke-wire method was developed to visualize the air flow in UA models directly. The results revealed that the airflow through the pharynx was laminar but not turbulent under normal inspiration, which suggested that compared with turbulent models, a laminar model should be more suitable in numerical simulations. The flow predicted numerically using the laminar model was consistent with the observations in the physical models. The comparison of the velocity fields predicted numerically using the half-side and complete models confirmed that it was reasonable to investigate the flow behaviors in UA using the half-side model. Using the laminar model, we simulated the flow and evaluated the effects of UA narrowing caused by rostral fluid shift on pharyngeal resistance. The results suggested that fluid shift could play an important role in the formation of hypopnea or OSA during sleep.     

Cardiorespiratory effects of heliox using a model of upper airway obstruction]

Heliox is a mixture of Oxygen and Helium. The low density of Helium allows this mixture to flow in a laminar pattern where oxygen, nitrogen or air flow would be turbulent. Therefore the force necessary to move a volume of gas (e.g. Heliox) is greatly reduced in comparison to a turbulent gas flow. In a respiratory loading experiment we investigated the effects which Heliox exerts on hemodynamic as well respiratory variables. 10 volunteers were breathing spontaneously and through three different endotracheal (ET-) tubes (ID 4.0, 4.5, 5.0 mm).The subjects were switched from room air to Heliox and differences in the variables heart rate (HR), blood pressure (BP), stroke volume (SV), stroke index (SI), peripheral vascular resistance (TPRI) and left ventricular work index (LVWI) were measured. Furthermore the (PhAng) between abdomen and thorax was detected using respiratory inductance plethysmography (n=2) and the sense of dyspnoe under the different conditions was assessed by the use of a dyspnea score (DS). The means of BP, SV, SI, TPRI and LVWI did not significantly differ between the resting and the different loading conditions irrespective of the gas that was used. The variability of hemodynamic measures was significant larger during loaded vs. unloaded breathing. Heliox could significantly reduce this degree of variability. In two subjects Heliox could also significantly reduce the PhAng as well as DS. Heliox showed effects on hemodynamic as well as respiratory and subjective variables. These effects can be interpreted as a reduction of the extent of pressure variations in the intrapleural space leading to less impact on hemodynamic variables while breathing Heliox vs. room air in a resistive loading experiment. In the future the combined measurement of hemodynmic variables as well as non-invasive assessment of respiration might shed new light on cardio-respiratory interaction and effects of Heliox during airway obstruction.     

Aerodynamic investigation of the thermo-dependent flow structure in the wake of a cyclist

The main purpose of this study was to assess the influence of the environmental temperature on both the aerodynamic flow evolving around the bicycle and cycling power output. The CFD method was used to investigate the detailed flow field around the cyclist/bicycle system for a constant speed of 11.1 m/s (40 km/h) without wind. In complement, a mathematical model was used to determine the temperature-dependent power output in the range [−10; 40 °C]. The numerical investigation gives valuable information about the turbulent flow field in the cyclist's wake which evolves accordingly the surrounding temperature. A major result of this study is that the areas of overpressure upstream of the cyclist and of underpressure downstream of him are less extensive for a temperature of 40 °C compared to −10 °C. The results suggest that the aerodynamic braking effect of the bicycle is minimized when the air temperature is high, as a lower air density results in a reduction in drag on the cyclist. This study showed that the power required to maintain a constant speed is reduced when the temperature is high, the reason being a lower aerodynamic resistance.