Gaspak system

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

The GasPak System is a laboratory equipment designed for the generation and maintenance of anaerobic environments. It provides a controlled atmosphere for the cultivation and growth of anaerobic microorganisms. The system includes an airtight chamber and a gas generation mechanism to create and sustain the desired anaerobic conditions.

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Lab products found in correlation

Mrs broth, by Thermo Fisher Scientific (2 mentions) N hexadecane, by Merck Group (1 mentions) Bhi broth, by Thermo Fisher Scientific (1 mentions) Taurodeoxycholic acid, by Merck Group (1 mentions) Technical agar, by Thermo Fisher Scientific (1 mentions) Cacl2, by Merck Group (1 mentions) Peptone water, by Merck Group (1 mentions) Reinforced clostridial agar, by Thermo Fisher Scientific (1 mentions) Bhi broth, by Merck Group (1 mentions)

Spelling variants of this product

GasPak Anaerobic System (3 citations) AnaeroGen GasPak system (2 citations) Anaerobe Jar + GasPak System (2 citations)

9 protocols using gaspak system

Assessing Microbial Hydrophobicity: An Optimized Protocol

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The hydrophobicity, i.e., the ability to adhere to hydrocarbons, was assessed according to [20 (link)] with some modifications. Fresh cultures of the strains (24 h of incubation at 37 °C in MRS broth with 0.05% L-cysteine) in anaerobic conditions (GasPak System; Oxoid Ltd., Basingstoke, UK) were harvested in the stationary phase by centrifugation at 6000 rpm for 10 min. The pellet was resuspended in NaCl 0.9% isotonic solution and was subsequently diluted to the absorbance value of 1 at 560 nm using a spectrophotometer (model 6705, Jenway, Stone, UK). Then, 3 mL of the bacterial suspension was vortexed with 0.6 mL of n-hexadecane (Sigma-Aldrich, Milan, Italy) for 4 min. The two phases were allowed to separate for 1 h at 37 °C. The aqueous phase was removed, and the absorbance (A) at 560 nm was measured. Finally, the hydrophobicity percentage was calculated with the following formula: (A0 − At)/A0 × 100, where A0 represents the absorbance at time 0 and At represents the absorbance at 560 nm after 1 h of incubation at 37 °C.
Hydrophobicity was also evaluated for L. rhamnosus GG ATCC^® 53103™, a commercial probiotic strain used as reference strain.

D’Alessandro M., Parolin C., Bukvicki D., Siroli L., Vitali B., De Angelis M., Lanciotti R, & Patrignani F. (2021). Probiotic and Metabolic Characterization of Vaginal Lactobacilli for a Potential Use in Functional Foods. Microorganisms, 9(4), 833.

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Characterizing Probiotic Strains from Breast Milk

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For the present study, 10 Lactiplantibacillus, 4 Lactobacillus and 2 Bifidobacterium strains were used (Table 1). All these strains are part of the microbial culture collection of DISTAL (Department of Food science and Biotechnology, University of Bologna, Italy). Each strain was isolated from a breast milk sample different from the other samples. These strains were also compared with Lacticaseibacillus rhamnosus GG ATCC^® 53103™ and Bifidobacterium animalis subsp. lactis BB-12^® used as references. Lactobacilli and bifidobacteria were cultured in MRS broth (Oxoid Ltd., Basingstoke, UK) with 0.05% L-cysteine and incubated at 37 °C for 24 h in anaerobiosis (GasPak System; Oxoid Ltd., Basingstoke, UK). Moreover, in order to evaluate the potential antagonistic activity of the breast milk isolates, selected food spoilage strains, such as Listeria monocytogenes ATCC 13932 and Scott A, Listeria innocua ATCC 51742, Enterococcus faecium BC104, Escherichia coli 555, Staphylococcus aureus DSM 20231, Salmonella enteritidis E5 and MB1409, and the intestinal pathogens enterotoxigenic E. coli H10407, Salmonella choleraesuis serovar typhimurium, Yersinia enterocolitica [8 (link)], were chosen as target bacterial strains. The pathogenic and spoilage strains were cultured in Brain Heart Infusion (BHI) broth (Oxoid Ltd.) at 37 °C for 24 h.

D’Alessandro M., Parolin C., Patrignani S., Sottile G., Antonazzo P., Vitali B., Lanciotti R, & Patrignani F. (2022). Human Breast Milk: A Source of Potential Probiotic Candidates. Microorganisms, 10(7), 1279.

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Isolation of Anaerobic Dairy Microbes

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The pour plate technique was used to isolate the microorganisms [36 ]. For milk samples, 1 mL milk was inoculated into 9 mL of sterile saline solution (S.S.S) whereas, for the solid dairy products, 10 grams were aseptically homogenized in 90 mL of S.S.S. Samples were serially diluted (10 fold) then 1 mL aliquot of each dilution was plated into 9 mL of molten De Man, Rogosa, and Sharpe (MRS) medium agar (Sigma-Aldrich). Inoculated MRS plates were incubated anaerobically at 37°C for 48–72 hrs. in an anaerobic jar (GasPak System—Oxoid, Basingstoke Hampshire, England) using AnaeroPack™-Anaero anaerobic gas generator sachets (Mitsubishi, Japan). Typical colonies were selected according to the morphological characters described by [37 (link)], picked from the plates, and repeatedly sub-cultured to obtain pure isolates.

Ali S.M., Salem F.E., Aboulwafa M.M, & Shawky R.M. (2022). Hypolipidemic activity of lactic acid bacteria: Adjunct therapy for potential probiotics. PLoS ONE, 17(6), e0269953.

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Isolation and Characterization of Vaginal Lactobacilli

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For the present study, 15 Lactobacillus strains belonging to the collection of FABIT (Department of Pharmacy and Biotechnology, University of Bologna, Italy) were used (Table 1). The strains were isolated from the vagina of pre-menopausal Caucasian women (aged 18–45 years), with no symptoms of vaginal or urinary tract infections, in accordance with the Ethics Committee of the University of Bologna (52/2014/U/Tess). These vaginal strains were also compared with Lacticaseibacillus rhamnosus GG ATCC^® 53103™, used as a probiotic reference strain. Lactobacilli were cultured in MRS broth (Oxoid Ltd., Basingstoke, UK) with 0.05% L-cysteine and incubated at 37 °C for 24 h in anaerobiosis (GasPak System; Oxoid Ltd., Basingstoke, UK).

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Screening Lactobacilli for Bile Salt Hydrolase

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The ability of lactobacilli to deconjugate bile salts was investigated by modifying the procedure of [22 (link)]. Cultures were screened for bile salt hydrolase (BSH) activity by spotting 10 μL of a culture grown onto BSH agar plates. The plates were prepared with MRS (Oxoid Ltd., Basingstoke, UK) with 0.05% L-cysteine added with 16 g/L of Agar Technical (Oxoid Ltd., Basingstoke, UK), combined with 0.5% (w/v) sodium salt of taurodeoxycholic acid (Sigma, Milan, Italy) and 0.37 g/L of CaCl₂ (Sigma, Milan, Italy). Plates were incubated in anaerobic conditions (GasPak System; Oxoid Ltd., Basingstoke, UK) at 37 °C for 48 h; after this time, BSH activity of each strain was evaluated by measuring the diameter of precipitation on screening medium.

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Enumeration of L. plantarum B-59151

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The viable count of L. plantarum B-59151 in FO and HFO was enumerated following the standard plate count method according to Vinderola and Reinheimer [28 (link)]. Firstly, 10 mL of each sample was suspended in 90 mL of sterile peptone water (Merck, 0.1%), after which it was serially diluted. Aliquots of 1 mL of appropriate dilutions were inoculated on sterile plates of MRS agar. MRS agar-inoculated plates were incubated at 37 °C for 48–72 h in anaerobic jars (2.5 L) with a GasPak system (GasPak system-Oxoid, Basingstoke, Hampshire, UK) according to standard methods [28 (link)]. Data are expressed as the logarithm of colony forming units per mL (Log₁₀ CFU mL⁻¹).

Alharbi H.F., Algonaiman R, & Barakat H. (2022). Ameliorative and Antioxidative Potential of Lactobacillus plantarum-Fermented Oat (Avena sativa) and Fermented Oat Supplemented with Sidr Honey against Streptozotocin-Induced Type 2 Diabetes in Rats. Antioxidants, 11(6), 1122.

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Quantifying Microbial Populations in Wines

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The microbial population (yeast and bacteria) present in each wine was analysed. For this purpose, serial dilutions of the wines were plated onto different culture media. Total yeasts were measured by seeding onto a GYP culture medium (20 g/L glucose, 5 g/L yeast extract, 5 g/L peptone, 100 mg/L chloramphenicol and 0.1 g/plate biphenyl) and incubation at 28 ºC for 48 h. The presence of Dekkera/Brettanomyces yeasts was determined by seeding onto a DBDM culture medium (6.7 g/L YNB, 20 g/L agar, 60 mL/L ethanol, 10 mg/L cycloheximide, 100 mg/L p-coumaric, 22 mg/L bromocresol green and 0.1 g/plate biphenyl) and incubation at 25 ºC for 21 days in anaerobic conditions (Gas Pak System, Oxoid Ltd., Basingstoke, England). LAB were measured by seeding onto an MRS medium (52 g/L MRS broth, 20g/L agar, 50 mg/L pymaricine and 0.1 g/plate biphenyl) and incubation at 30 ºC under strict anaerobic conditions for at least ten days. Acetic acid bacteria were determined by seeding onto a Mann culture medium (25 g/L D-mannitol, 3 g/L peptone, 5 g/L yeast extract, 20 g/L agar, 3 U/mL penicillin, 50 mg/L pymaricine and 0.1 g/plate biphenyl) and incubation at 28 ºC for 48 h. When the incubation periods finished, the plates were examined and colony forming units per millilitre (CFU/mL) were counted in plates with counts between 30 and 300 colonies and expressed in logarithmic units (log).

González-Arenzana L., Santamaría R., Escribano-Viana R., Portu J., Garijo P., López-Alfaro I., López R., Santamaría P, & Gutiérrez A.R. (2020). Influence of the carbonic maceration winemaking method on the physicochemical, colour, aromatic and microbiological features of tempranillo red wines. Food chemistry, 319.

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Cultivation of P. avidum Isolates

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The three clinical P. avidum isolates T13, T14 and T15 obtained in this study were cultivated on Reinforced Clostridial Agar (Oxoid) plates for 3 days at 37°C under anaerobic conditions using the Gas-Pak™ system (Oxoid). For liquid cultures, brain heart infusion (BHI) broth (Sigma-Aldrich) was used and cultures were grown for 24 h at 37°C under anaerobic conditions.

Wildeman P., Brüggemann H., Scholz C.F., Leimbach A, & Söderquist B. (2016). Propionibacterium avidum as an Etiological Agent of Prosthetic Hip Joint Infection. PLoS ONE, 11(6), e0158164.

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Anaerobic Activation of Lactobacillus plantarum

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Lactobacillus plantarum LS5 was activated in MRS broth medium and incubated at 37°C/24 hr under anaerobic conditions using an anaerobic atmosphere generation system (GasPak system, anaerobic system, Oxoid).

Shariati Z., Jouki M, & Rafiei F. (2019). Flavored functional drinking yogurt (Doogh) formulated with Lactobacillus plantarum LS5, cress seed gum, and coriander leaves extract. Food Science & Nutrition, 8(2), 894-902.

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