Gaspak ez container system

Manufactured by BD

Sourced in United States

The BD GasPak™ EZ container systems are airtight, resealable containers designed for microbial cultivation. They provide a controlled atmosphere for the growth of anaerobic and microaerophilic bacteria.

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Lab products found in correlation

Bbl co2 gas generators, by BD (2 mentions) Gaspak ez campy sachets, by BD (2 mentions) Purelink genomic dna mini kit, by Thermo Fisher Scientific (1 mentions) Quant it picogreen dsdna assay kit, by Thermo Fisher Scientific (1 mentions) Lactose, by BD (1 mentions) M17 agar, by BD (1 mentions) Blank disc, by Thermo Fisher Scientific (1 mentions) Etest, by bioMérieux (1 mentions) Nacl 0.9, by Fresenius (1 mentions) Gaspak ez anaerobe container system, by BD (1 mentions) Dry anaerobic indicator strips, by BD (1 mentions) Hemin, by Hardy Diagnostics (1 mentions) Vitamin k, by Hardy Diagnostics (1 mentions)

19 protocols using gaspak ez container system

Growth and Culturing of Haemophilus and Porphyromonas

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Haemophilus influenzae Rd [KW20] was obtained from ATCC (51907). Standard growth and culturing techniques were followed as described previously^{92 (link)}. Cultures were grown in Brain Heart Infusion broth (BHI) supplemented with 7.5 μM of hemin and 2 μg/ml NAD with or without the addition of peptide for 24 hours. The number of viable cells for every reaction mixture was then determined by serially diluting and spotting 10-μl aliquots in triplicates on BHI agar plates supplemented with 15 μM hemin and 3 μM NAD.
Porphyromonas gingivalis 2561 was obtained from ATCC (33277). Pre–reduced, anaerobically sterilized Brucella Broth and BRU - Brucella Blood Agar – were purchased from Anaerobe systems (CA, USA). They were opened just before use. Static cultures and plates were incubated at 37°C in an incubation chamber from BD GasPak™ EZ Container Systems. Anaerobic conditions were maintained by using BD BBL CO₂ gas generators and BD BB GasPak CO₂ indicators. Cultures were grown anaerobically in Brucella Broth with or without addition of peptides for 48 hours and viable cells for every reaction mixture were then determined by serially diluting and spotting 10-μl aliquots in triplicates on Brucella Blood Agar.

Sankari S., Babu V.M., Bian K., Alhhazmi A., Andorfer M.C., Avalos D.M., Smith T.A., Yoon K., Drennan C.L., Yaffe M.B., Lourido S, & Walker G.C. (2022). A haem-sequestering plant peptide promotes iron uptake in symbiotic bacteria. Nature microbiology, 7(9), 1453-1465.

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Bifidobacterium animalis Effect on C. elegans

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Bifidobacterium animalis was grown for 72 h on anoxic BSM broth at 37°C. Afterward, 150 uL of bacterial culture was spread onto medium-sized NGM plates (4 plates) and incubated for 24 h at 37°C in an anaerobic container (BD GasPak™ EZ container systems) prior to worm addition. E. coli OP50 was grown on NGM at normoxic conditions. C. elegans N2 population was synchronized as described above and approximately 150 worms were seeded on either B. animalis or OP50 plates. Plates were incubated for 24 h at 20°C in the anaerobic container. Developmental assay was carried out as previously described.^{122 (link)} In brief, 30 worms per bacterium were put individually onto small UV-killed OP50 plates and incubated for 48 h at 20°C before developmental stage of each worm was visually assessed. Reproductive aging assay was carried out as described previously.^{123 (link)} In brief, after incubation with B. animalis or OP50 the worms were washed with M9 and let to develop until L4 stage on UV-killed OP50 plates at 20°C, normoxia. At this moment 25 randomly picked worms (per condition) were transferred individually onto small-sized UV-killed OP50 plates. Every day the brood size of each worm was determined (sum of eggs and L1s) and parent worm was transferred to new plate until egg laying ceased. These experiments were performed 3 times.

Marfil-Sánchez A., Zhang L., Alonso-Pernas P., Mirhakkak M., Mueller M., Seelbinder B., Ni Y., Santhanam R., Busch A., Beemelmanns C., Ermolaeva M., Bauer M, & Panagiotou G. (2021). An integrative understanding of the large metabolic shifts induced by antibiotics in critical illness. Gut Microbes, 13(1), 1993598.

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Enrichment and Detection of Thermophilic Campylobacter

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The enrichment was processed according to Jeffrey et al. with modifications [23 (link)]. Briefly, SAEM tubes containing faecal samples were vortexed and incubated at 37 °C for 24 h. Swabs were washed in 10 mL PBS and then 5 mL of that rinsate was transferred into SAEM tubes. The enriched SAEM tubes were centrifuged at 5000 RPM for 20 min (Thermo Scientific Sorvall RC-6 plus, Ashville, NC) and 50 µL of supernatant was plated onto Campy-Cefex Agar (Acumedia, Lansing, MI) followed by incubation in anaerobic jars at 42 °C for 48 h under microaerophilic conditions (10% CO₂, 5% O₂ and 85% N₂) using BD GasPak EZ container systems (BD, Sparks, MD) [28 (link)]. Presumptive thermophilic Campylobacter spp. positive colonies were confirmed using a real-time PCR as described previously [29 (link)]. Campylobacter spp positive isolates were subjected to an additional multiplex real-time PCR to identify C. coli and C. jejuni [30 (link)].

Pires A.F., Patterson L., Kukielka E.A., Aminabadi P., Navarro-Gonzalez N, & Jay-Russell M.T. (2019). Prevalence and risk factors associated with Campylobacter spp. and Salmonella enterica in livestock raised on diversified small-scale farms in California. Epidemiology and Infection, 147, e321.

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Isolation and Quantification of Bovine Respiratory Pathogens

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P. multocida, M. haemolytica, and H. somni were isolated and final DNA concentrations were quantified according to Mohan et al. [16 (link)]. Specifically, all pathogens were streaked on tryptic soy agar plates supplemented with defibrinated sheep blood (blood agar). P. multocida and M. haemolytica were incubated aerobically at 37 °C for 16–18 h while H. somni was incubated in a 5% CO₂ atmosphere at 37 °C for 2–3 days by using BD GasPak™ EZ container systems (BD 260672) with BD BBL™ CO₂ gas generators (BD 260679). Individual colonies of each bacterial species were picked from the blood agar plates. P. multocida and M. haemolytica were inoculated into brain–heart infusion (BHI) broth and H. somni was inoculated into tryptic soy broth (TSB). All were incubated in the same conditions as the plates.
DNA extraction of each pathogen was carried out by taking 2 mL of saturated liquid culture and following the PureLink™ Genomic DNA Mini Kit (Catalog #K182002, Invitrogen, Waltham, MA, USA) procedure. Final DNA concentrations (ng/µL) of eluted extracts were measured using the Quant-iT PicoGreen dsDNA Assay Kit (Invitrogen P11496).

Pascual-Garrigos A., Maruthamuthu M.K., Ault A., Davidson J.L., Rudakov G., Pillai D., Koziol J., Schoonmaker J.P., Johnson T, & Verma M.S. (2021). On-farm colorimetric detection of Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in crude bovine nasal samples. Veterinary Research, 52, 126.

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Antimicrobial Activity of B. lactis HN019

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The in vitro antimicrobial activity of B. lactis HN019 was assessed against the following periodontopathogens using the agar diffusion method, as described by Zhu et al. [32 (link)]: P. gingivalis (W83), Prevotella intermedia (ATCC 25611), Fusobacterium nucleatum (ATCC 25586), and Aggregatibacter actinomycetemcomitans (ATCC 33393). For that purpose, 200-μL aliquots (10⁹ CFU/mL) of B. lactis HN019, previously grown on MRSA (Difco Laboratories, Detroit, MI, USA), were inoculated into 15-mm wells in Tryptic soy agar—TSA (Difco), supplemented with 5 μg/mL of hemin, 1 μg/mL of menadione, and 5% of defibrinated sheep blood, previously seeded (1.5 x 10⁸ CFU/mL) with indicator microorganisms. After pre-incubation for 30 min at room temperature, TSA plates were incubated at 37° C for 72 h under anaerobic (BD GasPak^™ EZ container systems, Becton, Dickinson and Company, Franklin Lakes, NJ, USA) (P. gingivalis, P. intermedia, and F. nucleatum) or microaerophilic (A. actinomycetemcomitans) conditions. Thereafter, the diameter (mm) of inhibition halos was measured using a digital caliper. Each indicator strain was tested three times in duplicate.
The means and standard deviations of the zones of inhibition observed in the sensitivity of different periodontopathogens to B. lactis HN019 were calculated.

Invernici M.M., Furlaneto F.A., Salvador S.L., Ouwehand A.C., Salminen S., Mantziari A., Vinderola G., Ervolino E., Santana S.I., Silva P.H, & Messora M.R. (2020). Bifidobacterium animalis subsp lactis HN019 presents antimicrobial potential against periodontopathogens and modulates the immunological response of oral mucosa in periodontitis patients. PLoS ONE, 15(9), e0238425.

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Enumeration and Detection of Campylobacter in Poultry

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After rinses were homogenized, 30 mL of the rinse was combined with 30 mL of pre-warmed (42 °C) CampyQuant solution (Hygiena, Camarillo, CA, USA) for poultry rinses and immediately incubated at 42 °C for 20 h for recovery under microaerophilic conditions (6 to 16% O₂ and 2 to 10% CO₂) using BD GasPak EZ Campy Sachets (Becton Dickinson and Company, Franklin Lakes, NJ, USA) in BD GasPak EZ Container Systems (Becton Dickinson and Company, Franklin Lakes, NJ, USA). After recovery, the BAX^®-System-CampyQuant™ (Hygiena, Camarillo, CA, USA) methodology was followed for the enumeration of Campylobacter. Samples were then incubated for 28 h at 42 °C for enrichment. Samples that were not positive for enumeration using BAX^®-System-CampyQuant™ (Hygiena, Camarillo, CA, USA) were tested for prevalence analysis using the BAX^® System RT-Campylobacter Assay (Hygiena, Camarillo, CA, USA).

Vargas D.A., Betancourt-Barszcz G.K., Chávez-Velado D.R., Sánchez A., Bueno López R, & Sanchez-Plata M.X. (2023). Bio-Mapping of Microbial Indicators and Pathogen Quantitative Loads in Commercial Broiler Processing Facilities in South America. Foods, 12(19), 3600.

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Enumeration of Lactic Acid Bacteria by Pour Plating

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After serial dilutions, one mL of sample was inoculated on a petri dish and pour plated with 20 mL of Mann–Rogosa–Sharpe Agar (MRS) in duplicates. Plates were placed in BD GasPak EZ Container Systems (Becton Dickinson and Company, Franklin Lakes, NJ, USA) and incubated at 48 ± 3 h at 35 ± 1 °C under microaerophilic conditions (6 to 16% O₂ and 2 to 10% CO₂) using BD GasPak EZ Campy Sachets (Becton Dickinson and Company, Franklin Lakes, NJ, USA), [14 (link),15 (link)]. Enumeration was conducted using a Q-Counter (Spiral Biotech Inc, Norwood, MA, USA)

Vargas D.A., Miller M.F., Woerner D.R, & Echeverry A. (2021). Microbial Growth Study on Pork Loins as Influenced by the Application of Different Antimicrobials. Foods, 10(5), 968.

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Enumeration of Probiotic Bacteria in Yogurt

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On each sampling day, 1 ml of the yogurt sample was serial diluted (1:10) with 0.1% sterile peptone water (Becton Dickinson and Company (BD), Sparks, MD). The aliquots of 100 µL of the diluted samples were plated out onto Difco De Man, Rogosa, and Sharpe (MRS) agar (BD, Sparks, MD) with pH adjusted to 5.2 and M17 agar (BD, Sparks, MD) with 10% w/v lactose (BD, Sparks, MD) to enumerate Lactobacillus delbrueckii subsp. Bulgaricus and Streptococcus thermophiles, respectively. The MRS plates were anaerobically incubated for 72 h at 37 °C using GasPak™ EZ Container Systems (BD, Sparks, MD), and the M17 plates were incubated at 42 °C for 72 h. The counts were presented as logarithms of colony-forming units per mL of yogurt (log CFU/mL). The analyses were carried out in duplicate.

Sarker A, & Ali Siddiqui R. (2023). Effects of ultrasonic processing on the quality properties of fortified yogurt. Ultrasonics Sonochemistry, 98, 106533.

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Bacterial Community Structure Changes Induced by Diatom

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To examine bacterial community structure changes induced by the presence of the diatom, ~1 × 10⁶ P. tricornutum cells ml⁻¹ were placed in the center well and the results were compared to control incubations where sterile media was placed in the center (Fig. 1d). Phycosphere enrichment samples [11 , 16 (link)] (with the algal cells removed with a 0.8 μm filter) were used as a bacterial community inoculant where 100 µl samples were placed in the surrounding wells in the array. Layer 1 and Layer 2 refer to bacterial wells with a distance from the centre well of 8 mm and 16 mm, respectively. On the outmost well, 100 µl sterile f/2-Si medium was inoculated as a blank control for 16 S rRNA community analysis.
The devices were incubated in a container (GasPak™ EZ container systems, BD) which was filled with f/2-Si medium. All preparation steps were performed in a laminar flow hood to prevent any bacterial contamination. The container was incubated under the same condition as described above. After 7 days, each well sample was collected and filtered into 96-well filter plate (0.2 μm pore size, Pall AcroPrep) to remove liquid, and was stored at −20 °C until further analysis.

Kim H., Kimbrel J.A., Vaiana C.A., Wollard J.R., Mayali X, & Buie C.R. (2021). Bacterial response to spatial gradients of algal-derived nutrients in a porous microplate. The ISME Journal, 16(4), 1036-1045.

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Peptide Effects on Iron Oxide Crystals

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All of the peptides were dissolved in Milli-Q-purified water. A solution of Fe₃Cl and Fe₂Cl, at a ratio of 2:1, was continuously sparged with N₂, stirred (300 rpm) and slowly titrated with 0.1 M NaOH (100 mL/h) at room temperature. At pH 5, the peptide was added into the iron solution to a final concentration of 100 μM to detect the effect of the peptide on crystal size and shape. In control samples, an equal amount of Mili-Q-purified water (400 μl) was added at pH 5 to eliminate the effect of the liquid that the peptides were dissolved in. In all samples, titration was stopped between pH 9 and 10. All samples were stored in a BD GasPak EZ container system (Becton Dickinson and Company, Franklin Lakes, NJ, United States).

Nudelman H., Lee Y.Z., Hung Y.L., Kolusheva S., Upcher A., Chen Y.C., Chen J.Y., Sue S.C, & Zarivach R. (2018). Understanding the Biomineralization Role of Magnetite-Interacting Components (MICs) From Magnetotactic Bacteria. Frontiers in Microbiology, 9, 2480.

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