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1.
A disposable anaerobic system which is characterized by its light weight and its compactness is described. The system consists of a multilayer plastic bag with a unique sealing device. A collapsible impregnated cardboard container is fitted with a catalyst and holders for a disposable hydrogen generator and an anaerobic indicator. The catalyst is active at room temperature and requires no heat activation. This system, which lends itself readily to compact storage, quick assembly, and ease of operation, is disposable after use. 相似文献
2.
S ummary : Comparable results were obtained with a commercially available disposable system for the production of H2 and CO2 in an anaerobic jar (the Gaspak system) and with a standard anaerobic jar procedure. Both systems were used with the Baird & Tatlock anaerobic jar and a room temperature catalyst. The test organisms included strict anaerobes such as Clostridium oedematiens type D and Cl. tetani. Comparative tests were also made with Bacteroides species from laboratory stock cultures and with freshly isolated strains of Bacteroides. On occasion, higher recoveries were obtained with the standard system when relatively demanding models were set up but it has limitations, having been developed initially for research purposes. The Gaspak system probably gives more reliable and more reproducible results in a laboratory in which experience with strict anaerobes is limited or sporadic. 相似文献
3.
Practical Method for Isolation of Anaerobic Bacteria in the Clinical Laboratory 总被引:1,自引:0,他引:1
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William J. Martin 《Applied microbiology》1971,22(6):1168-1171
An inexpensive but practical and simple method is described for providing an environment suitable for the maintenance and survival of clinically significant anaerobic bacteria, consistent with the needs of a large-volume laboratory. This modification of the GasPak procedure is based on two concepts. One involved the storage of "freshly prepared" plating medium under a constant stream of carbon dioxide. The other concept was to use an anaerobe jar, continuously flushed with oxygen-free carbon dioxide, as a "holding receptacle" for the inoculated media Experience with this modification has shown that the number of isolations of anaerobic bacteria from clinical material can be increased significantly. 相似文献
4.
Isolation of Anaerobic Bacteria from Human Gingiva and Mouse Cecum by Means of a Simplified Glove Box Procedure 总被引:52,自引:49,他引:52
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Alexander Aranki Salam A. Syed Ernest B. Kenney Rolf Freter 《Applied microbiology》1969,17(4):568-576
An anaerobic glove box constructed of clear flexible vinyl plastic is described. It is sufficiently inexpensive and simple in operation to be used not only in research but also in a clinical laboratory by technicians without special training. Conventional bacteriological techniques may be used inside the glove box for culturing and transferring anaerobic bacteria. The box may be heated to 37 C and thus serve as an anaerobic incubator as well, permitting inspection of cultures at any time. Media may be prepared and agar plates may be poured on the laboratory bench in the conventional manner. An overlay of trace amounts of palladium black catalyst over plated agar media reduces the medium to an oxidation-reduction (O-R) potential of - 300 mv within 2 days after introduction into the glove box. In spite of its greater simplicity, the system matched or excelled the roll tube method with respect to all parameters tested, including O-R potential obtainable in the media, O(2) concentration in the gas phase, and efficiency in isolating anaerobic bacteria from the mouse cecum. Comparative studies indicate that the conventional anaerobic jar method was inadequate for the isolation of strict anaerobes from human gingival specimens and from the mouse cecum. This was due to the exposure of specimens and media to air during plating on the open laboratory bench. Anaerobic jars were adequate for maintaining the proper conditions for growth of anaerobic bacteria once these had been established in the glove box. 相似文献
5.
An Evaluation of the 'GasKit'Disposable Hydrogen and Carbon Dioxide Generator for the Culture of Anaerobic Bacteria 总被引:3,自引:0,他引:3
The new GasKit disposable hydrogen and carbon dioxide generator is a reliable and convenient method for producing anaerobiosis in standard anaerobic jars when compared with a conventional procedure. The generator produces an atmosphere containing 10–12% carbon dioxide by volume which is satisfactory for the culture of exacting anaerobes. 相似文献
6.
A method for the concentration of cells from cerebrospinal fluid is described. An adaptation of a commercial cytochamber, consisting of a holder that fixes a disposable chamber directly on a microscope slide, was used. The cells were spun down in a conventional swing-out centrifuge, which was provided with a bucket for the cytochamber system. After removing most of the supernatant with a pipette, the remaining fluid was absorbed by means of a suction device consisting of a disposable pipette tip covered with a piece of Leukopor and filled with Sephadex G10 beads. The method gives a high recovery of cells (90%), together with a good preservation of cell morphology, and leaves about 80% of the fluid available for analysis of the soluble components. 相似文献
7.
Yuji Kikuchi Hiromi Sasaki Yukie Furuhata Yoko Tazawa Hajime Horiuchi Jun Okada 《Rinshō Biseibutsu Jinsoku Shindan Kenkyūkai shi》2007,18(1):11-17
Anoxomat Mart II (Mart Microbiology BV, Lichtenvooorde, Netherlands, Central Scientific Commerce Inc., Tokyo, Japan) is an anaerobic jar apparatus which uses a vacuum pump in combination with catalyst as gas replacement procedure to remove all traces of oxygen. As we had a chance to use Anoxomat Mart II, we compared it with other two anaerobic culture methods; namely AnaeroPack anaero (Mitsubishi Gas Chemical Co., Tokyo, Japan) which employs anaerobic jar method, and Concept400 (RUSKINN TECHNOLOGY LTD, England; Central Scientific Commerce INc., Tokyo, Japan) which uses anaerobic chamber method. We used 10 different species of anaerobic bacteria obtained from ATCC. One strain each of 10 species was cultured and examined for measurement of the sensitibity of an anaerobic indicator, th number of bacteria after 48 hour culture, the diameter of colonies, and MIC value. As a result, the time to reach the anaerobic condition was around 30 minutes by the Mart II against around 60 minutes by the AnaeroPack anaero. There was no difference concerning the number of bacteria after 48 hour culture among three methods. But anaerobic bacteria cultured by Mart II tended to make bigger colonies compared to other two methods in the 5 strains out of 9, except for one strain in which the diameter of colonies could not be measured. On the other hand, the comparison of MIC value showed good correlation in 11 antibiotics out of 12 among three methods. The MIC value of 11 antibiotics fitted within 1-fold difference, and 2-fold difference was observed in only one antibiotic. Mart II is so small that it does cheep consumables. From these reasons, we concluded that Mart II can be one of the useful anerobic culture methods. 相似文献
8.
Effect of Culture Medium and Carbon Dioxide Concentration on Growth of Anaerobic Bacteria Commonly Encountered in Clinical Specimens 总被引:14,自引:1,他引:13
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Representative strains of anaerobic bacteria from human infections were used to evaluate broth media, gas mixtures, and inocula for use in developing a procedure for performing minimal inhibitory concentration antimicrobic susceptibility tests. Nine commercially available media, including two that were chemically defined, were tested. Tests were performed in atmospheres with carbon dioxide concentrations between 2.5 and 10% and also in the GasPak system (BBL) that had a disposable hydrogen-carbon dioxide generator. Growth curves on each organism grown in schaedler broth and a 5% carbon dioxide atmosphere were used to determine growth characteristics, equate time of the particular growth phases to turbidity readings, and determine the numbers of viable organisms present in the culture. Schaedler broth proved to be most advantageous in combination with an atmosphere of 5% carbon dioxide, 10% hydrogen, and 85% nitrogen. The growth curve studies yielded valuable data on the rapidity and quantity of growth under these conditions. We believe these data have provided information which can be used as the basis for developing a standardized procedure for antimicrobic susceptibility testing for anaerobic bacteria. 相似文献
9.
Tomoo Kamiya A-Hon Kown Toshiki Kanemaki Yoichi Matsui Shouji Uetsuji Tadayoshi Okumura Yasuo Kamiyama 《In vitro cellular & developmental biology. Animal》1998,34(2):131-137
Summary The Anaeropack system for cell culture, which was originally designed for the growth of anaerobic bacteria, was used to produce
a hypoxic atmosphere for cultured hepatocytes. We measured changes in the oxygen and carbon dioxide concentrations and the
atmospheric temperature in an airtight jar. We also measured changes in the pH of the medium during hypoxia to assess the
accuracy of this system. Moreover, we used three durations (2, 3, and 4 h) of hypoxia and 8 h of reoxygenation in cultured
rat hepatocytes, and then measured the lactate dehydrogenase (LDH), ketone body concentration (acetoacetate + β-hydroxybutyrate),
and the ketone body ratio (KBR: acetoacetate/β-hydroxybutyrate) in the medium in order to assess the suitability of this system
as a model for reperfusion following liver ischemia. The oxygen concentration dropped to 1% or less within 1 h. The concentration
of carbon dioxide rose to about 5% at 30 min after the induction of the hypoxic conditions, and was maintained at this level
for 5 h. No effect of the reaction heat produced by the oxygen absorbent in the jar was recognized. The extent of cell injury
produced by changing the hypoxic parameters was satisfactorily reflected by the KBR, the ketone body concentration, and the
LDH activity released into the medium. Because this model can duplicate the conditions of the hepatocytes during revascularization
following ischemic liver, and the Anaeropack system for cell culture is easy to manipulate, it seems suitable for the experimental
study of hypoxic injury and revascularization in vitro. 相似文献
10.
Comparison of Atmospheric Conditions for Culture of Clinical Specimens of Neisseria gonorrhoeae 总被引:8,自引:1,他引:7
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Ann N. James-Holmquest Reuben D. Wende Robie L. Mudd Robert P. Williams 《Applied microbiology》1973,26(4):466-469
We cultured 55 clinical specimens of Neisseria gonorrhoeae in the following atmospheric conditions: (i) 10% carbon dioxide in a CO(2) incubator; (ii) a candle extinction jar; (iii) an air convection incubator; and (iv) an anaerobic jar without added CO(2). The number and size of colonies growing on modified Thayer-Martin medium were evaluated after incubation of cultures for 24 and 48 h at 36 C. After 24 h, the specimens from the candle extinction jar had the greatest number and size of colonies, but after 48 h growth was approximately equal for specimens from the candle jar and the CO(2) incubator. Only 19 of 55 specimens grew in the air convention incubator. None of 55 clinical specimens or of 10 laboratory strains grew anaerobically. Development of colonial morphology for colony types 1, 2, 3, and 4 was studied at 24 h on a base medium that contained no hemoglobin. The relative numbers of the four colony types in specimens were comparable after 24 h of incubation in any of the three atmospheric conditions under which growth occurred, but the different types were distinguished most readily when grown in the candle extinction jar. 相似文献
11.
Hydrogen sulfide is produced by many anaerobic bacteria and can irreversibly damage the palladium catalyst used for oxygen removal in anaerobic jars and cabinets. The present study used direct measurements of oxygen concentration to quantify catalyst activity following exposure to H2S and volatile fatty acids in the presence of eight potential H2S adsorbents. Most adsorbents were unsatisfactory, but two compounds not tested previously afforded full protection against the effects of H2S alone and provided protection in the presence of volatile fatty acids. The investigation demonstrated the importance both of selecting an adsorbent suitable for use in anaerobic conditions and of heating anaerobic jar catalysts in order to maintain activity. 相似文献
12.
Four types of hydrosol filters, two reusable (diatomaceous cylinder and fritted-glass funnel) and two disposable (asbestos pad and membrane filter) were challenged with a heavy bacterial suspension to assess their ability to produce sterile filtrates. Two of the four diatomaceous earth filters, the four fritted-glass funnels, and all of the asbestos pads tested generally gave sterile filtrates. However, only one type of filter, one of the membranes in its manufacturer's own holder, consistently gave sterile filtrates. The two other types of membranes usually gave sterile filtrates if tested in one manufacturer's holder, but all types invariably gave contaminated filtrates when tested in another manufacturer's holder. Contaminated filtrates were generally attributed to a poor reusable filter or to a faulty holder used with a disposable filter. If a high degree of certainty is required for sterile heat-labile filtrate, it is suggested that the liquid be passed through two or more filters in a previously tested and proven system. 相似文献
13.
We describe the application and evaluation of a widely available commercial jar as an anaerobic container suitable for the growth of a wide variety of anaerobes. A system for generating stable anaerobiosis was developed by combining standard anaerobic environment generators with Click-Clack jars produced by Click-Clack Ltd. This system was simple, reliable, and reduced capital outlay on anaerobic jars by at least an order of magnitude. 相似文献
14.
15.
G Clevstr?m H Ljunggren S Tegelstr?m K Tideman 《Applied and environmental microbiology》1983,46(2):400-405
In a previous experiment on the preservation of hay of high moisture content with formic acid, among other agents, aflatoxin was formed in the hay, and aflatoxin-forming strains of Aspergillus flavus were isolated from this hay after incubation in air as well as in an anaerobic jar. One isolate from the anaerobic jar was cultivated in a chemostat (Bioflo model C 30; New Brunswick Scientific Co.) in a defined medium with added B vitamins, yeast extract, or formic acid, with or without gas flow (air or nitrogen). In all cases where spore germination occurred, aflatoxin was formed in the cultures with gas flow, and small quantities of aflatoxins B1 and B2 occurred even in an atmosphere of nitrogen. Addition of B vitamins and supply of traces of air gave an approximately 15-fold increase in the amount of aflatoxin in 2 days. Carbon dioxide enrichment hindered aflatoxin formation on the defined medium even in the presence of B vitamins, but when formic acid was added, small quantities (5 to 15 micrograms/liter) were formed, and this low level remained constant until the gas flow was started. 相似文献
16.
Production of aflatoxin by an Aspergillus flavus isolate cultured under a limited oxygen supply. 总被引:1,自引:0,他引:1
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In a previous experiment on the preservation of hay of high moisture content with formic acid, among other agents, aflatoxin was formed in the hay, and aflatoxin-forming strains of Aspergillus flavus were isolated from this hay after incubation in air as well as in an anaerobic jar. One isolate from the anaerobic jar was cultivated in a chemostat (Bioflo model C 30; New Brunswick Scientific Co.) in a defined medium with added B vitamins, yeast extract, or formic acid, with or without gas flow (air or nitrogen). In all cases where spore germination occurred, aflatoxin was formed in the cultures with gas flow, and small quantities of aflatoxins B1 and B2 occurred even in an atmosphere of nitrogen. Addition of B vitamins and supply of traces of air gave an approximately 15-fold increase in the amount of aflatoxin in 2 days. Carbon dioxide enrichment hindered aflatoxin formation on the defined medium even in the presence of B vitamins, but when formic acid was added, small quantities (5 to 15 micrograms/liter) were formed, and this low level remained constant until the gas flow was started. 相似文献
17.
We designed, fabricated and tested a novel compact fluorescence analysis system for quantification of uric acid (UA) in clinical samples at the point-of-care. To perform an analysis, diluted saliva, urine or blood samples are simply placed in a disposable thin-film sample holder using a dropper. A new enzyme immobilization technique was developed to retain within the sample holder two enzymes and a molecule, which transforms into a fluorescer in amounts depending on the UA concentration. The small instrument (7.5 cm × 5 cm × 5 cm) into which the sample holder is placed for analysis contains an LED, a narrow-band filter and an amplified photodiode. The analysis time is 30s, and the dynamic range of the system is 4-400 μM of UA. The calibration curve for transparent saliva and urine was made using solutions of UA. The calibration curve for opaque blood was obtained with spiked samples of blood. The three different types of clinical samples were collected from three subjects and simply diluted before their measurements. Analysis with our instrument yielded UA concentrations within the expected concentration ranges. Development of instruments based on the current laboratory prototype is expected to result in products for clinical trials and point-of-care. 相似文献
18.
The Anoxomat system provides an automated evacuation-replacement technique to create an anaerobic or microaerophilic environment in a jar. We evaluated the Anoxomat system for the growth of obligate anaerobes and for the recovery of anaerobic organisms from clinical specimens, and compared its performance to that of an anaerobic chamber and the GasPak System. Of the 54 stock strains tested, the Anoxomat, the chamber, and the GasPak recovered 95%, 95% and 93% at 24 h, respectively. On 29 occasions (51%), the colonies on the Anoxomat plates were slightly larger than those in the chamber and on 17 (30%) occasions larger than the colonies on the GasPak jar plates. At 48 h, the Anoxomat, the chamber, and the GasPak recovered 93.5%, 94.4% and 88.9%, respectively; of 108 anaerobes isolated from 31 clinical specimens. Methylene blue indicators became decolorized (average of 10 tests) within 2 h inside the Anoxomat jars, 2 h 10 min inside the anaerobic chamber, and 2 h 30 min inside the GasPak jars. 相似文献
19.
A prerequisite for successful identification of anaerobic pathogenic bacteria from samples of clinical material is the method of cultivation. Currently, several methods of cultivation in anaerobic environment are used: cultivation in anaerobic box, anaerobic jar, and in nonrecurring cultivation system. Here, we determined the suitability of the above methods of cultivation using the estimation of the growth (diameters of colony size) of commonly isolated anaerobic pathogens (Bacteroides fragilis, Clostridium difficile, and Clostridium perfringens). The tested bacterial strains were exposed to atmospheric oxygen for various time periods and then they were cultivated using different anaerobic cultivation systems. Maximum growth differed, depending on the type of cultivation and the strain used. Thus, largest zone diameters, in the majority of measurements, were achieved in the anaerobic box. However, nonrecurring cultivation system seemed better in several cases; this applied to the cultivation of C. perfringens after 15, 30, and 60 min exposure to atmospheric oxygen as well as the cultivation of B. fragilis after 30 and 60 min of oxygen exposure. The cultivation in anaerobic box was the most convenient method for growth of C. difficile. In almost all cases, higher growth was observed in nonrecurring cultivation system than in the system of anaerobic jar. On the other hand, no significant differences were observed among these anaerobic cultivation systems which confirmed their applicability (taking into account some individual features concerning the optimization of cultivations) for identification of pathogenic anaerobes. 相似文献
20.
The GasPak disposable hydrogen and carbon dioxide generator is a convenient and reliable method for producing anaerobiosis in the GasPak system. As a source of carbon dioxide, however, the generator is inadequate, uncontrolled and unreliable. 相似文献