All surgical guides were investigated at different time intervals; after one day, one week, and two weeks from the production stage. Each first half of the three study groups was immersed in one of the three disinfectants; 100%VCO, 2% GA, and 70% EA for 20 min, left to dry for an additional 10 min then soaked in sterile glass containers containing 100 ml. of sterile distilled water for 10 min, and each second half of the three control groups was immersed in sterile glass containers containing 100 ml. of sterile distilled water for 20 min. Three samples were pipetted and cultured on three microbiological media, in which 50 µl (µl) were spread over the surface of Blood (Oxoid, CM0271) and MacConkey agar plates (Oxoid, CM0115) to be incubated at 37 °C for 24 h, and over the surface of Sabouraud dextrose agar plates (HiMEDIA, M063) to be incubated at 37 °C for 48–72 h. After the incubation period, all plates were examined, and the microbial count was done and expressed as colony-forming units per plate (CFU/plate). The percentage (%) of reduction was calculated by the following equation [18 (link)]:
Cm0271
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The CM0271 is a laboratory equipment product from Thermo Fisher Scientific. It is a centrifuge designed for general laboratory applications. The product description and specifications are limited to the core function of the equipment without any interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Cm0007, by Thermo Fisher Scientific (2 mentions)
Cm1135, by Thermo Fisher Scientific (2 mentions)
Cm0099, by Thermo Fisher Scientific (2 mentions)
Cm0115, by Thermo Fisher Scientific (1 mentions)
Bactalert blood culture bottles, by bioMérieux (1 mentions)
Defibrinated horse blood, by Thermo Fisher Scientific (1 mentions)
Vitek densichek, by bioMérieux (1 mentions)
Cm0739, by Thermo Fisher Scientific (1 mentions)
Cm085, by Thermo Fisher Scientific (1 mentions)
11 protocols using cm0271
1
Disinfection Efficacy on Surgical Guides
2
COVID-19 Bacterial Co-Infection Identification
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Microbiological examination was only done for patients with suspected co-infection. Under proper infection control measures, sputum or endotracheal aspirates were collected from COVID-19 patients throughout the course of illness. Blood samples were inoculated directly into BacT/ALERT blood culture bottles (BioMérieux, Marcy l’Etoile, France) and monitored for bacterial growth. Samples were streaked out on Blood agar (Oxoid, CM0271) with 5% mammalian blood, initial identification of the bacterial isolates was done by Gram staining and growth characteristic on Mannitolsalt agar (Oxoid, PO0151) and MacConkey agar (Oxoid, CM0007). Microorganisms and their antimicrobial sensitivity patterns were identified using the VITEK2 Microbiology automated system (BioMerieux’s). To distinguish between colonization and infection, 10 μL of the respiratory specimen were inoculated on appropriate bacteriological media and incubated at 37◦C for 24–48 h. Then, the number of CFU per mL of the sample was determined. Presence of ≥105 CFU/mL in sputum or endotracheal aspirates indicated bacterial co-infection based on the American Thoracic Society definitions.21 (link),22 (link)
3
Campylobacter Phage Isolation and Propagation
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Campylobacter jejuni PT14 (Brathwaite et al., 2013 (link)) was used as a reference strain and also for phage isolation and propagation. Campylobacter jejuni HPC5 (Loc Carrillo et al., 2005 (link)) and C. jejuni 81–176 (Korlath et al., 1985 (link)) were used as controls in the chicken colonisation experiments and the phage treatments of contaminated chicken livers. All Campylobacter isolates were cultured on blood agar base no. 2 CM0271 (Oxoid, Basingstoke, United Kingdom) supplemented with 5% defibrinated horse blood (TCS, Buckingham, United Kingdom) under microaerobic conditions (5% O2, 5% H2, 10% CO2, 80% N2) at 42 °C for 18–24 h. Campylobacter phages CP30A (GenBank accession number JX569801 ) and CPX (GenBank accession number JN132397 ) were propagated on C. jejuni PT14 or a contemporary Campylobacter isolate using the soft agar overlay method (Atterbury et al., 2003 (link)). Phages from the UK typing scheme (ɸ1 to ɸ16) were propagated as described by Frost et al. (1999) (link). In order to obtain high titre stocks of bacteriophage, 30 ml volumes of plate lysates were centrifuged at 40,000g for 2 h at 4 °C. The pellets obtained were re-suspended in 1 ml of SM buffer (50 mM Tris·HCl (pH 7.5), 100 mM NaCl, 8 mM MgSO4, 0.01% Gelatin) to give a phage suspension containing approximately 10 log10 PFU/ml.
4
Listeria monocytogenes Inoculation of Cucumber
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A suspension of L. monocytogenes (serotype1/2a lineage II, CCUG 15527, Culture Collection University of Göteborg, Sweden) was prepared by swabbing fresh colonies, grown on blood agar at 37 ± 1°C for 24 h, into sterile saline water (0·9% NaCl). A theoretical bacterial concentration of 1·5 × 108 cells ml−1 (0·5 McFarland) was obtained using VITEK DENSICHEK (bioMerieux, Marcy l'Etoile, France). To ensure a more accurate quantification of the suspension, 0·1 ml of ten-fold dilutions were plated onto blood agar (CM 0271; Oxoid, Basingstoke, UK) and incubated at 37 ± 1°C for 24 h prior to enumeration. Bacterial suspensions of L. monocytogenes were used in the experiment the same day they were made.
Pieces of cucumber (Cucumis sativus) were sliced and kept at room temperature overnight resulting in a drier texture capable of some absorption. Small squares were then cut from the outer cucumber layer, with the bottom of each piece consisting of peel to prevent penetration of fluid. A small well was made with a scalpel in each cucumber segment prior to application of the L. monocytogenes suspension. Each cucumber segment weighed 0·2–0·3 g and was inoculated with 0·02 ml of suspension corresponding to ≅4·0 × 105 CFU L. monocytogenes (blood agar enumeration).
Pieces of cucumber (Cucumis sativus) were sliced and kept at room temperature overnight resulting in a drier texture capable of some absorption. Small squares were then cut from the outer cucumber layer, with the bottom of each piece consisting of peel to prevent penetration of fluid. A small well was made with a scalpel in each cucumber segment prior to application of the L. monocytogenes suspension. Each cucumber segment weighed 0·2–0·3 g and was inoculated with 0·02 ml of suspension corresponding to ≅4·0 × 105 CFU L. monocytogenes (blood agar enumeration).
5
Blood Culture and Bacterial Isolation
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Two milliliters of venous blood sample taken from patients was inoculated into 20 ml brain-heart infusion broth (BHI) (CM1135; Oxoid Ltd., England, UK). The culture bottles were incubated at 37 °C for 7 days and examined daily for evidence of bacterial growth, including turbidity and hemolysis. If bacterial growth was observed, a subculture was made after 24 h on blood agar (CM0271, Oxoid Ltd., England UK), Salmonella Shigella agar (SS) (CM0099, Oxoid Ltd., England, UK), and MacConkey agar (CM0007, Oxoid Ltd., England, UK).
6
Isolating Diverse Campylobacter Strains
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An isolate collection was assembled, from the Swansea University archive, including 102 C. jejuni and C. coli isolates from various host species and clinical samples, including all isolates sequenced in Sheppard et al. (2013a , 2013b ) (Table S1 ). This collection contained strains representing much of the genetic diversity among agricultural C. jejuni and the three major C. coli clades (Parkhill et al., 2000 ; Fouts et al., 2005 ; Pearson et al., 2007 ; Sheppard et al., 2013a , 2013b ). These were supplemented with Campylobacter whole genome sequences from the NCBI database for genomic analyses (Genbank accession numbers: NC_009839; NC_008787; NC_003912; NC_002163) (Table S1 ). The isolate genomes are archived in the Dryad data repository (doi.org/10.5061/dryad.215jd/1 and doi:10.5061/dryad.28n35 ). Isolates were stored in a 20% (v/v) glycerol medium mix at minus 80°C and subcultured onto Campylobacter selective blood‐free agar (mCCDA, CM0739, Oxoid). Plates were incubated at 42°C for 48 h under microaerobic conditions (5% CO2, 5% O2) generated using a CamyGen (CN0025, Oxoid) sachet in a sealed container. Subsequent phenotype assays were performed on Brucella agar (CM0271, Oxoid).
7
Antimicrobial Susceptibility Testing Protocol
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MICs were determined by agar dilution (CLSI VET01-A4).14 Prior to MIC determination, strains were subcultured twice on blood agar base (CM0271, Oxoid) containing 5% sheep blood (TCS Biosciences, Buckingham, UK) at 35°C for 24 h. Stock solutions were prepared at 10× final concentration in distilled water (fusidic acid sodium salt F0881, chlorhexidine digluconate C9394; Sigma-Aldrich Inc.) or 1% DMSO (miconazole nitrate PHR1163; Sigma), adjusted for drug potency.14 Final concentrations of the active fraction ranged from 0.015 to 2048 mg/L for fusidic acid and from 0.03 to 256 mg/L for chlorhexidine, miconazole and a 1:1 combination of chlorhexidine and miconazole.14 Discrepancy between duplicate MICs was accepted provided they varied by only one dilution; in such cases, the higher value was identified as the final MIC. S. aureus ATCC 25923, S. aureus ATCC 29663 and S. pseudintermedius LMG 22219 were included for quality control purposes. Three MRSA strains previously reported with low, medium and high MICs of fusidic acid6 (link) were also included for comparative purposes.
8
Campylobacter Strains as Phage Hosts
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Campylobacter strains used as bacteriophage hosts were as follows: C. jejuni NCTC 12662, C. jejuni NC3142 (Farm C isolate sourced from re-use litter in 2011), and C. coli NC2934 (Farm C isolate sourced from non-reused litter in 2011). These were grown on Blood agar No 2 (Oxoid CM0271) with 5% (v/v) lyzed horse blood added (LHB; Oxoid Australia), for 24 h at 42°C, under microaerobic conditions, produced by using Campygen gas packs (Oxoid, CN0025A; Basingstoke, United Kingdom). Isolates used for resistance testing were sourced from chicken ceca and carcasses pre- and post-initiation of the study.
9
Quantifying Staphylococcus aureus in Rinse Fluids
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Using spread plate method, 0.5 ml of each 4 consecutive 10 fold serial dilutions (10 -1 to 10 -4 ) of the rinse fluid were inoculated in triplicates into mannitol salt agar (Oxoid CM 085, UK). The plates were incubated at 37°C for 24 h and the number of yellow colonies recorded per dilution. CFU/ml, CFU/cm 2 and the respective logs were then calculated. For confirmation of coagulase positive S. aureus, four suspect colonies per dilution were sub-cultured onto blood agar (OXOID CM0271, UK) and incubated at 37°C for 24 h. Golden yellow and cream white colonies were tested for catalase and coagulase activity.
10
Anaerobic Isolation of C. perfringens
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Isolation of C. perfringens was carried out according to Roberts and Greenwood (2003) . Briefly, each sample was inoculated into freshly prepared cooked meat medium (Oxoid, CM0081), and anaerobically incubated at 37 °C for 24 hrs in an anaerobic jar. A loopful from the cooked meat medium is streaked onto neomycin sheep blood agar (Oxoid, CM0271) plate, followed by incubation at 37 °C for 18-24 hrs. under complete anaerobic condition. The characteristic colonies failed to be detected (double zone of hemolysis around colonies on blood agar).
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