Effects of Bicarbonate on Growth of Neisseria gonorrhoeae: Replacement of Gaseous CO2 Atmosphere

The effect of NaHCO₃ on the growth of Neisseria gonorrhoeae cultures was studied in a liquid and a semisolid growth medium. With a broth culture, NaHCO₃ (0.009 M) greatly reduced the lag phase and also increased the total growth. The same concentration of bicarbonate supported rapid growth when added to the semisolid medium if the plates were individually incubated in sealed plastic bags. In a container with a large air space, a higher concentration of NaHCO₃ was necessary to support growth. The assimilation of ¹⁴C-labeled NaHo₃ by growing cultures was also investigated.     

Effect of Storage of Mueller-Hinton Agar Plates on Zone Sizes for Antimicrobial Testing

The length of time Mueller-Hinton agar plates can be stored at 4 C without affecting the size of zones of inhibition in susceptibility testing by the Bauer-Kirby method was studied. It was found that these plates can be stored for 3 weeks at 4 C without an appreciable affect on zone sizes. Storage of plates in sealed plastic bags did not alter the results significantly. The findings indicate that commercially prepared Mueller-Hinton agar plates, which may be several days old when received at the laboratory, are suitable for use in routine susceptibility tests by the Bauer-Kirby method.     

A SIMPLIFIED METHOD of COLONY HYBRIDIZATION USING RADIOLABELED PROBES IN SEALED PETRI DISHES

A simplified colony hybridization procedure was developed using sealed disposable plastic petri dishes in place of sealed plastic bags. Prior to hybridization, the dishes were sealed with successive layers of parafilm, plastic wrap and aluminum foil to prevent evaporation of the solution. This self-contained procedure eliminates some of the technical problems such as spilling of radioactive materials, leakage of solution, sealing of plastic bags and the formation of air bubbles. Therefore, this method allows for safer and easier handling of radioactive materials during hybridization procedures.     

Effect of conditions in sealed plastic bags on eclosion of mass‐reared Queensland fruit fly,Bactrocera tryoni

The sterile insect technique has been used for more than 50 years to control a range of insects around the world. Sterile insect technique is rapidly becoming a major component of many area‐wide fruit fly management programmes. Irradiation of immature life stages induces sterility in adults, which are then distributed over large areas to mate with wild flies, resulting in no viable offspring. However, irradiation in normal air results in declining adult quality. To optimize the quality of sterile adult flies, several techniques are available to lower the levels of oxygen in fruit fly tissues prior to irradiation. The simplest method is to seal pupae in plastic bags and allow the oxygen consumption of pupae to minimize oxygen in both the air and pupal tissue. Some fruit fly species have rapid decreases in eclosion as a result of low oxygen atmospheres. We tested the tolerance of Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), to low oxygen for the first time. In the first two experiments, unirradiated B. tryoni pupae were tested for different periods in sealed plastic bags at 17, 21, and 26 °C. Optimum eclosion occurred at 21 °C with the lowest eclosion at 26 °C. In general, mean full eclosion declined at ca. 0.1% eclosion per hour sealed in plastic bags during the first 96 h for all temperatures. In the third and fourth experiments at 17 °C, there was a decline in average eclosion for irradiated and unirradiated pupae of about 13.4% after they were sealed in plastic bags for 192 h. In general, B. tryoni eclosion declined at 0.1% per hour inside sealed plastic bags for periods up to 192 h at 17 °C. Queensland fruit flies can tolerate long periods of conditions found inside sealed plastic bags and current practices for sterile B. tryoni release programmes will result in minimum decrease in eclosion. The possible evolution of tolerance of these conditions is discussed.     

Dubos Oleic Acid Agar Medium in the Differentiation of Meningococci and Gonococci

Growth on Dubos oleic acid agar plates, incubated for 48 hr at 37 C in candle jars, provides a method for differentiating meningococci from gonococci. All of our 54 strains of meningococci, but none of the 49 strains of gonococci, grew on this medium.     

Flexible plastic bioreactors for photobiological hydrogen production by hydrogenase-deficient cyanobacteria

Uptake hydrogenase mutant cells of the cyanobacterium Nostoc sp. PCC 7422 photobiologically produced H(2) catalyzed by nitrogenase for several days in H(2)-barrier transparent plastic bags, and accumulated H(2) in the presence of O(2) evolved by photosynthesis. Their H(2) production activity was higher in the sealed flexible bags than in stoppered serum bottles of fixed gas volume.     

Survival and Outgrowth of Clostridium botulinum Type E Spores in Smoked Fish

Chub injected in the loin muscle with 10(6)Clostridium botulinum type E spores were smoked to an internal temperature of 180 F (82.2 C) for 30 min, sealed in plastic bags, and incubated at room temperature (20 to 25 C) for 7 days. Viable type E spores were found in practically all such fish. Toxin formation by the survivors in the smoked fish was dependent on the brine concentration of the smoked fish. A brine concentration of 3% or higher, as measured in the loin muscle, inhibited toxin formation. Six different type E strains gave similar results. Only a few hundred of the million spores in the inoculum survived the smoking. Moisture in the atmosphere during smoking did not reduce the incidence of fish with type E survivors.     

Stability of Antibiotics and Chemotherapeutics in Agar Plates

The stability of chemotherapeutic agents incorporated into agar plates was studied by comparison of minimum inhibitory concentrations on fresh and stored plates and by direct bioassay of the chemotherapeutic agar plates. Plates were stored in sealed bags at 4 C. No loss of bioactivity was demonstrated after 30 days of storage in plates containing methicillin, erythromycin, cephalothin, tetracycline, chloramphenicol, kanamycin, streptomycin, polymyxin B, or nalidixic acid. Penicillin G, ampicillin, and nitrofurantoin showed statistically significant losses of activity after 4 weeks. None of the chemotherapeutics tested showed significant loss in activity after 1 week.     

A device for cultivation of plant and animal cells

Summary A device is described for cultivation of suspension cultures of plant and animal cells in plastic bags placed on carrier plates in a thermostat box. Pendular motion of these plates ensures mixing of the fluid and aids in transfer of oxygen pumped above the fluid surface.     

Suspension cultures of mononuclear phagocytes in the teflon culture bag.

The Teflon culture bag (TCB) provides a cheap and simple method for culturing mononuclear phagocytes in suspension. The cells can easily be recovered intact and used in further experiments. The Teflon membrane is permeable to O₂, CO₂, and water vapor. Therefore, gas exchange is guaranteed when the bags are sealed after being filled with medium and cells. The risk of infection is minimized since the cultures are incubated in closed bags.     

A novel method for the transport and long-term storage of cultures and samples in an anaerobic atmosphere

Nylon-aluminium-polyethylene laminated, sealable bags provided a more economical and flexible method of transportation, incubation and storage than anaerobic jars. The bags were used with a special heat sealer and 'Anaerocult-A' sachets to remove oxygen. The environment inside the bags remained oxygen-free for at least 3 years.     

Effects of 1-methylcyclopropene alone and in combination with polyethylene bags on the postharvest life of mango fruit

Experiments were conducted to determine how 1‐methylcyclopropene (1‐MCP) treatments influence ethylene‐stimulated ripening of harvested mango cv. Zihua fruit at 20°C. The ripening response of fungicide (prochloraz) treated fruit was characterised following various 1‐MCP treatments in sealed jars followed by storage in polyethylene bags and/or subsequent ethephon (ethylene) exposure. Exposure of fruit to increasing concentrations of 1‐MCP for 12 h resulted in the reduced softening of produce when subsequently held in air for 7 days after ethephon treatment. Application levels of between 1 and 100 μl litre^?1 1‐MCP had increasing impact, while 200 μl litre^?1 1‐MCP apparently began to approach response saturation. Exposure of fruit to 50 or 100 μl litre^?1 concentrations of 1‐MCP for periods from 1 to 24 h subsequently resulted in reduced softening of produce when held in air for 7 days after ethephon treatment. Increasing periods of exposure from 1 to 12 h had increasing impact, while exposure times greater that 12 h appeared to reach saturation. In the absence of ethephon‐stimulation, the natural ripening of mangoes held in polyethylene bags was delayed by prior exposure to 100 μl litre^?1 1‐MCP for 12 h. Extended holding of 1‐MCP treated and non‐1‐MCP treated control fruit in polyethyene bags encouraged physiological and pathological deterioration. Following exposure to 100 μl litre^?1 1‐MCP for 12 h, mango fruit held for 10 days in polyethylene bags showed a delay in the onset of ripening relative to bagged but non‐1‐MCP treated control fruit. Treatment with 1‐MCP allowed storage of mango fruit in plastic bags at 20°C for 30 days. Observations suggest that 1‐MCP treatments do not adversely influence the quality of the post‐storage ethephon‐ripened fruit. Thus, application of 1‐MCP in combination with the use of polyethylene bags can extend the postharvest life of mango fruit at ambient temperature. Treatments that extend postharvest life are important in developing countries, such as China, where the cold chain infrastructure is often lacking.     

No difference in colony formation of Scenedesmus obliquus exposed to lakes with different nutrient levels

The algal genus Scenedesmus is famous for its highly phenotypic plasticity in response to various environmental factors. In laboratory, axenic cultures of Scenedesmus often fail to form colonies and remain in unicellular morph. To examine whether unicellular Scenedesmus can form colonies after exposure to natural lakes, dialysis bags and plastic bottles which contained the precultured Scenedesmus obliquus were exposed in two lakes with different nutrient levels for four weeks. Results showed S. obliquus grew well in dialysis bags but not in plastic bottles; S. obliquus can form colonies when incubated in dialysis bags which allow exchange with the aquatic environment of the substances, regardless of the nutrient levels of the two lakes. However, no colonies were observed in S. obliquus incubated in plastic bottles exposed in both lakes. This suggested that active growth and zooplankton infochemicals contributed together to the colony formation of S. obliquus exposed in situ environment.     

A streamlined technique for on-grid electron immunocytochemistry.

The unlabelled antibody enzyme technique for on-grid immunostaining demands exacting conditions of cleanliness. These can be met with minimal trouble by two technical modifications. Large supplies of all the solutions are millipore-filtered in advance and are then stored frozen in sealed plastic bags and allowed to thaw when required. A disposable, contamination-free surface with depressions to hold drops of staining solution is prepared from a strip of parafilm.     

A streamlined technique for on-grid electron immunocytochemistry

Summary The unlabelled antibody enzyme technique for on-grid immunostaining demands exacting conditions of cleanliness. These can be met with minimal trouble by two technical modifications.Large supplies of all the solutions are millipore-filtered in advance and are then stored frozen in sealed plastic bags and allowed to thaw when required.A disposable, contamination-free surface with depressions to hold drops of staining solution is prepared from a strip of parafilm.     

Rapid assessment of antimould efficacies of pressure-treated southern pine

A membrane-screening method was developed in conjunction with flow cytometric (FC) analysis for determining the efficacies of antimould pressure-treatment formulations for mould species of cosmetic significance on southern pine. Fusarium subglutinans, Aspergillus flavus, Penicillium chrysogenum, and Paecilomyces spp. were the predominant moulds colonizing surfaces of the variously treated pine stored in sealed plastic bags over 3- to 6-month periods. Nylon membranes placed directly on pressure-treated pine and membranes saturated with the various formulations were inoculated with the conidia of selected moulds. FC analysis of conidia stained with propidium iodide (PI) before and after exposure to the pressure-treatment formulations permitted a rapid assessment of the inocula and selection of those pressure-treatment formulations with probable inhibitory activity versus probable nonactive preparations. Recoveries of the fungi from the membranes over 9–14 days were in general agreement with the emergence of colonizing fungi on the similarly preserved uninoculated pine stored in sealed plastic bags for 6 months. This combination of procedures provided for a relatively rapid assessment of preservative formulations designed to provide enhanced efficacy against surface mould growth on lumber during storage and retail display. Received 21 December 2001/ Accepted in revised form 20 May 2002     

Effect of incubation atmosphere and temperature on isolation of Campylobacter jejuni from human stools

To determine the optimal conditions for isolation of Campylobacter jejuni from human fecal specimens, we compared incubation atmospheres that contained about 5, 10, and 15% oxygen with the 17% oxygen produced in candle jars and also compared incubation temperatures of 37 and 42 degrees C. At 42 degrees C, C. jejuni was isolated from all 16 specimens; however, colony sizes were larger when plates were incubated in 5 and 10% oxygen than in the other two atmospheres. At 37 degrees C some positive cultures were missed in 15% oxygen and in the candle jar. The largest colony sizes were obtained in 5% oxygen. For each atmospheric condition tested, the colonies were larger at 42 than at 37 degrees C. When incubation is done at 42 degrees C, use of a candle jar is adequate; however, at 37 degrees C candle jars should not be used for isolation of C. jejuni from human feces.     

Cultivation of mammalian cells in heat-sealable pouches that are permeable to carbon dioxide

Petri plates, 96-well plates, and other unsealed culture vessels are the chief cause of air-borne contamination of cell cultures. In this study, heat-sealable plastic pouches that are permeable to CO2 and other gases are used as a means to avoid contamination. Several applications of heat-sealable pouches are described. First, petri plates can simply be sealed in a wide variety of plastic films that are permeable to CO2. Second, a few CO2-permeable plastic films can be used directly as substrata for mammalian cell growth and can also be cut with a simple hole punch for the isolation of clones. Third, one can grow cells in chemically sterilized carbonic acid solutions and thereby avoid the use of a CO2 incubator entirely.     

Unicellular Microcystis aeruginosa cannot revert back to colonial form after short-term exposure to natural conditions

Colonial Microcystis aeruginosa isolates tend to lose their typical colonial morph after some generations under laboratory conditions, one interesting but yet important question is whether unicellular M. aeruginosa, originally isolated from the field, can revert back to colonial morphology when growing back in natural waters. Based on this idea, we employed dialysis bags and plastic bottles to conduct an in situ experiment. Each of the dialysis bags and plastic bottles was filled with unicellular M. aeruginosa with high concentrations and then submerged to two natural lakes (Lake Caiyue and Lake Taihu) for 40 and 28 days, respectively. Results showed M. aeruginosa grew well in dialysis bags but not in plastic bottles; no colonies were observed in M. aeruginosa incubated in either dialysis bags or plastic bottles exposed in both lakes. This suggests laboratory maintained unicellular M. aeruginosa cannot revert back to colonial form after short-term exposure in natural conditions.     

d- rice seedling development in periodically stirred water in sealed environments

Summary

Large numbers of germinating seedlings of two rice cultivars exhibited d^- type development consisting solely of completely white coleoptile growth after 15 days in the light in both static and periodically stirred water environments in completely sealed jars. Thus, whether germinated in static or agitated water in the sealed environments, coleoptile greening and subsequent normal seedling development were inhibited by hypoxic stress caused by the large numbers of germinating rice seedlings competing for the limited environmental oxygen supply. Consequently, the evidence presented points away from the formation of unfavourable oxygen diffusion gradients in static water environments in the sealed jars as having been responsible for d type seedling development observed in previous investigations. Continuous aeration of the 15-day-old static and periodically stirred aquatic environments, respectively, resulted in complete reversal of all d seedlings into normal green plants.