Comprehensive details are provided in the supplementary file . Briefly, samples were immediately placed into an anaerobic pouch (AnaeroGen™ COMPACT, Oxoid Limited, Hampshire, UK) and processed within an anaerobic cabinet. Quantitative culture was performed and the total viable count (TVC; colony forming units per gram of sputum [CFU/g]) of all distinct colony morphologies were enumerated and identified to the genus-level using near full-length 16S rRNA sequencing.
Anaerogen compact
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
AnaeroGen Compact is a self-contained, single-use sachet that generates an anaerobic atmosphere when placed in an airtight container. It is designed to create an oxygen-depleted environment suitable for the cultivation of anaerobic microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Campygen compact, by Thermo Fisher Scientific (1 mentions)
Dm2500, by Leica (1 mentions)
Dm1000 photonic microscope, by Leica (1 mentions)
Tecnai g20 electron microscope, by Thermo Fisher Scientific (1 mentions)
Columbia agar, by bioMérieux (1 mentions)
Milli q water, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Columbia 5 sheep blood agar, by BD (1 mentions)
Columbia blood agar plates bbl, by BD (1 mentions)
Tryptic soy broth (tsb), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Azelaic acid, by Merck Group (1 mentions)
Potassium dihydrogen phosphate, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Dichloromethane, by Merck Group (1 mentions)
Ethanol 96, by Merck Group (1 mentions)
Propylene glycol, by Dow (1 mentions)
Rnaprotect bacterial reagent, by Qiagen (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Ribo zero rrna removal kit bacteria, by Illumina (1 mentions)
19 protocols using anaerogen compact
1
Quantitative Anaerobic Sputum Microbiome
2
Anaerobic Duodenal Fluid Sampling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The large bowel was cleansed using polyethylene glycol (Laxabon®) the day before endoscopy of the small and large intestines. Duodenal fluid was aspirated via a sterile tube before the biopsies were sampled using the working channel of the gastroscope; no wash step was used in the fluid sampling procedure. All samples were immediately placed in a sterile gas-tight sachet (AnaeroGen Compact, Oxoid Ltd, Basingstoke, UK) in which an anaerobic milieu was created using an anaerobic generator (Benex Ltd., Shannon, County Clare, Ireland) [21 (link)]. Anaerobiosis was confirmed upon arrival to the laboratory by checking the anaerobic indicator included in the sachet. The fluids were stored frozen at -80°C within 24 hours after sampling.
3
Microbial Profiling of Animal Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rectal swabs and pulmonary, central nervous system (CNS), and hepatic samples were aseptically collected for standard bacteriological examinations (Table 1 ). The samples were placed on MacConkey, mannitol salt agar, and sheep blood agar plates for aerobic incubation at 37 °C for 3 days. Suspicious colonies were isolated and then identified by biochemical reactions (API® ID, bioMérieux, Craponne, France). For anaerobic incubation, samples were also spread on sheep blood agar plates placed in an anaerobic jar with AnaeroGen Compact (Oxoid Limited®, Basingstoke, England) and incubated at 37 °C for 18–24 h. Incubation was prolonged for one more day if no colonies appeared within 24 h.
Viral isolation was carried out from oral swabs on Vero cell cultures (ATCC CRL-1586 VERO C1008) at the Biosafety Level 3 (BSL-3) laboratory at the Istituto Zooprofilattico Sperimentale Abruzzo e Molise ‘Giuseppe Caporale’ (IZSAM). Vero cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with antibiotics and 10% foetal calf serum. Monolayer cultures were inoculated with swab material and examined daily for the evidence of cytopathic effect (CPE) for 5–6 days. Three subsequent blind passages were carried out for each sample to allow CPE to appear.
Viral isolation was carried out from oral swabs on Vero cell cultures (ATCC CRL-1586 VERO C1008) at the Biosafety Level 3 (BSL-3) laboratory at the Istituto Zooprofilattico Sperimentale Abruzzo e Molise ‘Giuseppe Caporale’ (IZSAM). Vero cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with antibiotics and 10% foetal calf serum. Monolayer cultures were inoculated with swab material and examined daily for the evidence of cytopathic effect (CPE) for 5–6 days. Three subsequent blind passages were carried out for each sample to allow CPE to appear.
4
Hypoxia-Reoxygenation Cycling in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AnaeroGen Compact (Oxoid; Hampshire, UK) was used for hypoxic episode application. The system encompasses a tightly sealed bag with a gas-generating sachet. Each sachet contains activated carbon and ascorbic acid, which combine with the air when opened, consuming oxygen and reducing its concentration within the bag to less than 1%. Culture flasks and opened sachets were placed inside plastic pouches, which were then sealed. PANC-1 and MCF7 cells were exposed to a 72-hour hypoxic cycle using the abovementioned system followed by 24-48 hours of reoxygenation. Cells were subjected to 20 cycles of hypoxia. Apart from these signified cycles, control cells were split beside their hypoxic counterparts and were subjected to similar conditions except hypoxic cycles (10 (link), 11 (link)).
5
Comprehensive Characterization of Strain Marseille-P2645
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Different growth conditions were tested on a 5% sheep's blood–enriched Columbia agar (bioMérieux) for strain Marseille-P2645T. Five temperatures (room temperature, 28, 37, 45 and 55°C) and three atmospheres—anaerobic (anaeroGen Compact; Oxoid), microaerophilic (campyGen Compact; Oxoid) and aerobic (in a plastic pouch to maintain a humid atmosphere)—were evaluated. Tolerance of this strain to salt was tested using 5%, 7.5%, 10%, 15% and 20% of NaCl, and the pH tolerance (5, 5.5, 6, 6.5, 7, 7.5 and 8) was also tested. Individual cells of strain Marseille-P2645T were visualized using a Tecnai G20 electron microscope (FEI Company, Limeil-Brevannes, France). Gram staining was performed and observed using a photonic microscope Leica DM2500 (Leica, Wetzlar, Germany) with a 100× oil-immersion objective. Motility testing was performed by observation of a fresh colony between the blades and slats using a DM1000 photonic microscope (Leica) at 40×. To check the ability to sporulate, strain Marseille-P2645T was grown on 5% sheep's blood–enriched Columbia agar (bioMérieux) for 2 days, and then a heat-shock test (20 minutes at 80°C) was performed.
6
Hypoxia Induction and Redox Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Leaf discs were placed on Milli-Q water with 0.05% Tween-20 (Sigma-Aldrich) +/− MV. Unless specified otherwise, 1 µM MV was used. Final concentration of antimycin A (AA) was 2.5 µM [3 (link)]. Dark pre-treatment with MV and AA was overnight. To generate hypoxic atmosphere, nitrogen gas was flushed inside a custom-built chamber containing plant material. Imaging was performed through the glass cover. Alternatively, plant material was placed into the AnaeroGen Compact anaerobic gas generator bag (Oxoid). AnaeroGen decreases oxygen concentration below 0.5% producing 9–13% CO2 [32 (link)]. CO2 accumulation was prevented by LoFloSorb non-caustic containing carbon dioxide absorbent (Intersurgical). O2 was controlled with resazurin anaerobic indicator (Oxoid).
7
Identification of Clinical Isolates via MALDI-TOF MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All clinical samples were inoculated onto aerobic and anaerobic blood agar (BD Columbia Agar 5% Sheep Blood, Becton Dickinson, Franklin Lakes, NY), chocolate agar (BD Choco Agar, Becton Dickinson), mannitol agar (BD Mannitol Salt, Becton Dickinson), Mac Conkey agar (BD Mac Conkey II, Becton Dickinson), and thioglycolate broth (BD Fluid Thioglycolate Medium, Becton Dickinson), incubating all media at 35–37 °C for 5 days. Anaerobic plates were incubated in an anaerobic atmosphere generated with the AnaeroGen Compact anaerobic system (Oxoid Ltd., Wide Road, Basingstoke, England) at 35–37 °C. Identification of all isolates was carried out using MALDI-TOF MS (Bruker Biotyper, Bellerica, MA, USA), following the manufacturer’s recommendations. Only strains with a log (score) ≥2.0 were included and interpreted as high confidence [24 (link)] and identified only with this technique. Two isolates with the lowest log (score) were further identified by 16S rRNA gene sequencing [25 (link)].
8
Evaluating Bacillus cereus Interactions with Gut Microbiome
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 100 µL of fecal suspension, prepared as reported in Section 2.1 , was inoculated upon sterile EGM structures in each well of the 24-well microplates. A total of 1.9 mL of RPMI 1640 was added to a final volume of 2 mL. Plates were incubated for 24 h at 37 °C in an anaerobic atmosphere and generated using AnaeroGenTM Compact (Thermo Fisher Scientific). After incubation, 100 µL of supernatants were removed from all wells. Half of the wells were inoculated with 100 µL of the suspension of B. cereus prepared as described above (Section 2.3 ). In the remaining wells, 100 µL of B. cereus supernatant obtained in Section 2.3 were added. Plates were incubated for 24 h at 37 °C in an anaerobic atmosphere. After incubation, culture supernatants (2 mL) were removed from all wells and replaced with 2 mL of fresh RPMI 1640. This grossly eliminates planktonic B. cereus vegetative cells and their secreted virulence factors. Plates were incubated for a further 24 h at 37 °C in an anaerobic atmosphere. In parallel, sterility control wells were included, constituted by sterile EGM structures and RPMI 1640 medium without fecal samples and B. cereus vegetative cells and supernatant. After incubation, supernatants were discarded, and EGM scaffolds were collected for DNA extraction.
9
Cloning and Characterization of Giardia lamblia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Giardia lamblia assemblage A1 isolate WB (ATCC® 50803) clones and clone GS/M-83 (ATCC® 50581) (assemblage B) were cultured in TYI-S-33 medium, as previously described55 (link). Clones expressing different VSPs were obtained by limiting dilution in 96-well culture plates placed in anaerobic chambers (AnaerogenTM Compact, Thermo Scientific® Oxoid®, Cat. # AN0010C) at 37 °C for 5 days and positive clones were then selected using specific anti-VSP mAb by immunofluorescence assay (IFA) as described55 (link). Reactive clones were expanded in a culture medium overnight and tested for homogeneity before use56 (link). For in-cell studies, these clones were re-cloned as described above, but in the presence of 50 nM of their specific anti-VSP mAb.
10
Anoxia Tolerance in Head Lice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate the effect of anoxia on head lice survival, ten groups of 30 lice each were used. Each group (n = 30) was placed in a plastic Petri dish (5 cm in diameter) and was placed in an AnaeroGenTM Compact plastic pouch (Oxoid Ltd, Basingstoke, England, UK). Then, the AnaeroGenTM Compact sachet (Oxoid Ltd, Basingstoke, England, UK) was quickly placed in the pouch. The system was tied shut with a clip. Each group endured anoxia for one time point (H1, H3, H6, H8, H10, H12, H14, H16, H18, and H24). Each test group had a control group (n = 30) that was placed in a plastic Petri dish under ambient conditions (25 ± 2 °C, 20-50% relative humidity). The pouches were then opened and viable and dead lice were counted separately (≤ 1h) for the test and control groups, respectively. Death was defined as the absence of vital signs such as movement of antennae, movements of legs, and gut peristalsis. Each experiment was performed in triplicate.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!