Strain differentiation of epidemic typhus rickettsiae (Rickettsia prowazekii) by DNA restriction endonuclease analysis

We have investigated the number of methanogenic bacteria in different samples of fresh feces and effluents from the anaerobic digestion of cattle and swine waste and municipal sludges. We have further compared conventional anaerobic techniques with the rigorous, oxygen-free, anaerobic, experimental conditions obtained in a controlled glove cabinet atmosphere. It was found that the total number of oxygen-intolerant methanogenic bacteria was about 10–100 times higher than that of anaerobic oxygen-tolerant methanogenic bacteria. We have also developed a simple technique for the growth of methanogenic bacteria that permits the realization of rigorous anaerobic conditions without the use of a glove cabinet.     

国内生物安全柜产品标准（JG170和YY0569）比较

国内生物安全柜执行的两份标准（ JG170-2005和YY0569-2011）存在一定差异,导致实际执行中存在分歧和疑惑。该文就两份标准中对生物安全柜的结构要求,性能要求和测试方法进行了比较,并对其逐一进行客观分析解释,对生物安全柜在实际维护和检验时提供了指导。     

Bacteriological testing of a modified laminar flow microbiological safety cabinet

A modified microbiological safety cabinet which can be used as a class II and a class III safety cabinet has been bacteriologically tested. This cabinet makes use of a high-speed down-flow air curtain in the front opening to minimize the amount of air escaping over the arms of the operator. By using artificial aerosols and a dummy or a test person placing his arms into the working opening of the cabinet, a transfer from the inside to the environment was detected only when the highest concentration of the test aerosol was used. Since the number of bacteria detected was very low, this is considered to be acceptable. when the cabinet was used as a class III type, with a glove panel mounted in the front opening, leakage from the environment occurred. This could be completely prevented by fixing tape over the hinge of the front panel.The conclusion is drawn that this type of biohazard hood can be safely used as a class II and a class III microbiological safety cabinet, provided the construction of the hinge of the front panel will be adapted to prevent transfer from the environment to the working area.     

Cryopreservation of somatic embryos from <Emphasis Type="Italic">Petiveria alliacea</Emphasis> L. by different techniques based on vitrification

Petiveria alliacea L. is a medicinal plant originating from the Amazon region. This study describes an efficient cryopreservation protocol for somatic embryos (SEs) produced from roots of P. alliacea based on the comparison of vitrification, encapsulation-dehydration, and D cryo-plate techniques. With the vitrification technique, SEs treated with PVS2 solution (0.4 M sucrose, 3.3 M glycerol, 2.4 M ethylene glycol, and 1.9 M DMSO) for 30 min displayed high viability (85%) and intermediate proliferation recovery (about 12 adventitious SEs produced from original SEs [SEs/SE] after 90 d of culture). With the encapsulation-dehydration technique, lower viability (70%) and very low proliferation recovery (about two SEs/SE) were achieved with cryopreserved SEs dehydrated for 10 min in a laminar air flow cabinet. The D cryo-plate technique led to high viability (85%) and proliferation recovery (19 SEs/SE) of cryopreserved SEs after 90 min dehydration. In the experimental conditions tested, the D cryo-plate method was the most efficient technique for cryopreservation of P. alliacea SEs.     

Containment of Microbial Aerosols in a Microbiological Safety Cabinet

A microbiological safety cabinet was evaluated to determine conditions under which microorganisms might escape. Tests were conducted under three cabinet-closure conditions, various airflow velocities, and different laboratory operations, with 10(5), 1.1 x 10(5), and 10(6) microorganisms per cubic foot of cabinet space released per min for 5 min. The data revealed that (i) escape of a human infectious dose is possible when the cabinet is used with the glove panel off; (ii) the number of organisms that escaped from the cabinet increased with a decrease in air velocity; and (iii) an increase in the number of laboratory operations resulted in an increase in the number of organisms that escaped. Thus, when the glove panel was off, the cabinet was only safe for operations that released a small number of microorganisms into the cabinet, whereas the cabinet was safe for operations of significantly greater hazard when used with the glove panel on but with the gloves unattached.     

Evaluation of a class III biological safety cabinet for enclosure of an ultracentrifuge

An evaluation of a special safety cabinet housing a high-speed centrifuge was made. The cabinet enclosed both the top access port and the drive and pumping machinery of the centrifuge. A titanium rotor was loaded with tubes containing a bacterial culture, weakened, and driven until rotor rupture occurred. There were several bent and broken components in the centrifuge, and bacteria leaked from the vacuum chamber. Although the forces were sufficient to displace the cabinet, none of the test bacteria were found outside the cabinet.     

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.     

Microbiological Studies on the Performance of a Laminar Airflow Biological Cabinet

Engineering and microbiological tests indicated that a typical, commercial laminar airflow cabinet was not effective in providing either product protection or agent containment. The cabinet was modified and tested through a series of alternate configurations to establish a set of design criteria. A mock-up cabinet was developed from these design criteria. The mock-up unit was evaluated for efficiency in providing both product protection and agent containment. In these evaluations, challenge methods were developed to simulate normal, in-use laboratory operations. Controlled bacterial or viral aerosol challenges were used at higher than normal levels to provide stringent test conditions. Test results indicated that the mock-up unit was considerably better in preventing agent penetration (0.1 to 0.2 particles per 100 ft(3) of air) than the commercial cabinet (5 to 6 particles per 100 ft(3) of air) during product protection tests. Similarly, agent containment was considerably better in the new cabinet (particle escape of 2 to 3 per 100 ft(3) of air at only one of the five test sites) than in the commercial cabinet (particle escape of 2 to 14 per 100 ft(3) of air at three of the five test sites).     

Electrochromatography of amino acid mixtures in fermentation liquors

The substitution of the phenolic chromatography dimension, by electrophoresis at 500 to 1000 volts, in a veronal buffer, pH 8.6, μ 0.02 followed by triple rechromatography in a completely miscible butanol, acetic acid, water solvent system, leads to clear, easily reproducible electrochromatography patterns. A reliable economical chromatography cabinet is described. The technique allows the analysis of many samples conveniently, and some important characteristics of unidentifiable components can be readily predicted.     

Influence of crossdrafts on the performance of a biological safety cabinet.

A biological safety cabinet was tested to determine the effect of crossdrafts (such as those created by normal laboratory activity or ventilation) upon the ability of the cabinet to protect both experiments and investigators. A simple crossdraft, controllable from 50 to 200 feet per min (fpm; 15.24 to 60.96 m/min), was created across the face of the unit. Modifications of standardized procedures involving controlled bacterial aerosol challenges provided stringent test conditions. Results indicated that, as the crossflow velocities exceeded 100 fpm, the ability of the cabinet to protect either experiments or investigators decreased logarithmically with increasing crossdraft speed. Because 100 fpm is an airspeed easily achieved by some air conditioning and heating vents (open windows and doorways may create velocities far in excess of 200 fpm), the proper placement of a biological safety cabinet within the laboratory--away from such disruptive air currents--is essential to satisfactory cabinet performance.     

Influence of crossdrafts on the performance of a biological safety cabinet.

A biological safety cabinet was tested to determine the effect of crossdrafts (such as those created by normal laboratory activity or ventilation) upon the ability of the cabinet to protect both experiments and investigators. A simple crossdraft, controllable from 50 to 200 feet per min (fpm; 15.24 to 60.96 m/min), was created across the face of the unit. Modifications of standardized procedures involving controlled bacterial aerosol challenges provided stringent test conditions. Results indicated that, as the crossflow velocities exceeded 100 fpm, the ability of the cabinet to protect either experiments or investigators decreased logarithmically with increasing crossdraft speed. Because 100 fpm is an airspeed easily achieved by some air conditioning and heating vents (open windows and doorways may create velocities far in excess of 200 fpm), the proper placement of a biological safety cabinet within the laboratory--away from such disruptive air currents--is essential to satisfactory cabinet performance.     

Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet.

BACKGROUND: The jet-in-air cell sorters currently available are not very suitable for sorting potentially biohazardous material under optimal conditions because they do not protect operators and samples as recommended in the guidelines for safe biotechnology. To solve this problem we have adapted a cell sorting system to a special biosafety cabinet that satisfies the requirements for class II cabinets. With aid of this unit, sorting can be performed in conformance with the recommendations for biosafety level 2. METHODS: After integrating a modified fluorescence-activated cell sorter (FACS) Vantage into a special biosafety cabinet, we investigated the influence of the laminar air flow (LAF) inside the cabinet on side stream stability and the analytical precision of the cell sorter. In addition to the routine electronic counting of microparticles, we carried out tests on the containment of aerosols, using T4 bacteriophage as indicators, to demonstrate the efficiency of the biosafety cabinet for sorting experiments performed under biosafety level 2 conditions. RESULTS: The experiments showed that LAF, which is necessary to build up sterile conditions in a biosafety cabinet, does not influence the conditions for side stream stability or the analytical precision of the FACS Vantage cell sorting system. In addition, tests performed to assess aerosol containment during operation of the special biosafety cabinet demonstrated that the cabinet-integrated FACS Vantage unit (CIF) satisfies the conditions for class II cabinets. In the context of gene transfer experiments, the CIF facility was used to sort hematopoietic progenitor cells under biosafety level 2 conditions. CONCLUSIONS: The newly designed biosafety cabinet offers a practical modality for improving biosafety for operators and samples during cell sorting procedures. It can thus also be used for sorting experiments with genetically modified organisms in conformance with current biosafety regulations and guidelines.     

The Influence of Light Quality on Levels of Ahscisic Acid in Tomato Plants, and Evidence for a Novel Abscisic Acid Metabolite

Tomato plants were grown in growth cabinets under two different light sources. One source consisted of white and red fluorescent tubes (Red cabinet), while the second consisted of both tungsten filament and fluorescent lamps (Far red cabinet). Energy fluence rates were adjusted to give equal photosynthetic rates in the two cabinets. Extension growth was approximately three times greater in the Far-red cabinet. No difference could be detected in abscisic acid (ABA) levels in leaves or petioles of plants grown under the two light regimes, but levels were 40–90% greater in stems of the slower growing plants illuminated with fluorescent light only. Exogenously applied ABA effectively reduced the growth rate of the plants in the Far-red cabinet to that of plants in the Red cabinet. However, it was shown that light-induced changes in growth rates occurred before any change in endogenous ABA could be demonstrated, thus precluding a role for ABA in the initial response to a change in light quality. The changed ABA levels appeared to be the result of a modified pathway of ABA degradation. Evidence is presented for a novel metabolite of ABA which yields free ABA on basic hydrolysis.     

Modified Laminar Flow Biological Safety Cabinet

Tests are reported on a modified laminar flow biological safety cabinet in which the return air plenum that conducts air from the work area to the high efficiency particulate air filters is under negative pressure. Freon gas released inside the cabinet could not be detected outside by a freon gas detection method capable of detecting 10(-6) cc/s. When T3 bacteriophage was aerosolized 5 cm outside the front opening in 11 tests, no phage could be detected inside the cabinet with the motor-filter unit in operation. An average of 2.8 x 10(5) plaque-forming units (PFU)/ft(3) (ca. 0.028 m(3)) were detected with the motor-filter unit not in operation, a penetration of 0.0%. Aerosolization 5 cm inside the cabinet yielded an average of 10 PFU/ft(3) outside the cabinet with the motor-filter unit in operation and an average of 4.1 x 10(5) PFU/ft(3) with the motor-filter unit not in operation, a penetration of 0.002%. These values are the same order of effectiveness as the positive-pressure laminar flow biological safety cabinets previously tested. The advantages of the negative-pressure return plenum design include: (i) assurance that if cracks or leaks develop in the plenum it will not lead to discharge of contaminated air into the laboratory; and (ii) the price is lower due to reduced manufacturing costs.     

A new controlled-rate cooling apparatus for freezing hematopoietic cells for storage at -196 degrees C

An apparatus has been constructed to cool biological material at a controlled rate. The material to be frozen is placed in glass ampuls which are immersed in an aluminum bath containing ethyl alcohol and the bath is placed inside a freezer cabinet. Liquid nitrogen is pumped intermittently into the cabinet by means of a single-speed electric pump. The rate of cooling is controlled by a device that varies the interval between successive pumping cycles. The temperature fall is monitored by thermocouples placed inside selected glass ampuls and recorded as a plot on moving graph paper.This simple instrument is capable of cooling at an accurately controlled rate over the range of 0 to 7 °C/min. We chose for our studies a cooling rate of 1 °C/min which we could maintain with an accuracy of ±0.1 °C. Temperature fluctuations were, however, observed at the freezing plateau and varied considerably in magnitude and temperature at onset even for the same material cooled under the same conditions. Mouse bone marrow cells frozen by our technique and stored for various periods of time may, on reconstitution, form colonies in vivo and in vitro identical in morphology and number to those from unfrozen control cells. Our results suggest that expensive and intricate devices may not be necessary to obtain optimal recovery of viable cells after storage in liquid nitrogen. The apparatus is now in regular use for the storage of human bone marrow cells intended for use in treatment of patients with leukemia refractory to conventional measures.     

Effect of amino acids,growth regulators and genotype on androgenesis in barley

The effects of an amino acid mixture and of plant growth regulators added to the FHG barley anther culture medium were examined using three barley cultivars (Cadette, Léger, and Igri) grown in two environments (growth cabinet and glasshouse). ‘Léger’ and ‘Igri’ were known as responsive, and ‘Cadette’ as recalcitrant to androgenesis. Our first experiment showed that the amino acid-supplemented medium was best for embryogenesis and regeneration of ‘Cadette’ and ‘Igri’ in both environments, and if ‘Léger’ in the growth cabinet. The addition of ABA and TDZ did not improve embryogenesis and plant regeneration, and PAA decreased them in the growth cabinet. The addition of the amino acid mixture in the FHG medium also reduced the percentage of albino plants in the growth cabinet, but growth regulators did not improve the percentage of albino plants, and in some cases increased it. In the growth cabinet, disregarding media, ‘Léger’ produced more embryos than ‘Cadette’ and ‘Igri’, and Léger' and ‘Igri’ produced more green plants than ‘Cadette‘. Percentages of albino plants were higher or ‘Cadette’ than for ‘Igri’ or ‘Léger’. In a second experiment, we compared seven hybrids with their parents for androgenic responsiveness. Hybrids had a higher ability to generate green plants than expected based upon the weighted average reflecting the contribution of each parent. This revised version was published online in June 2006 with corrections to the Cover Date.     

Trends in migrant and ethnic minority voting in Australia: findings from the Australian election study

A review of the research literature on migrant voting in Australia shows that the “ethnic vote” has almost disappeared now that migrants tend to vote in a similar way to the rest of the population according to traditional class cleavages. In addition, it is argued that migrants in Australia predominantly reside in safe Labor seats that are represented by cabinet or shadow cabinet ministers. As such their group-based interests are often neglected by the major parties. Using findings from the ABS census and the 1993–2013 AES, this article re-examines whether there is a migrant vote, and if so, the extent to which migrant voting patterns have changed since the 1990s when the migrant and ethnic vote reached its peak. This study reveals patterns of migrant voting among certain birthplace subgroups that are more volatile than in previous decades but at the same time distinctive.     

The total containment of a batch-type zonal centrifuge.

A batch-type zonal centrifuge has been modified and totally contained for use with biologically hazardous materials. A sealed cabinet encloses the centrifuge and the ancilliary equipment. It is operated with a flow of filtered air when the zonal system is on, decontaminated with ethylene oxide, and maintained at a negative pressure throughout. The centrifuge subsystems can be drained, flushed, and decontaminated with ethylene oxide before an engineer services the machine. The sample handling system within the cabinet is remotely controlled.