Potential metal-resistant LAB isolates were biochemically characterized by API 50 CHL test kits (BioMérieux, Marcy I´Etoile, France) according to the manufacturer’s instructions and interpretation of results was performed using the computer-aided database API-WEB™V.5.0 software as mentioned by Khalil et al.43 . Furthermore, selected isolates were identified by 16S rRNA gene sequencing, following Hamdan et al.44 (link), analyzed in the Genbank DNA database using the online tool (BLAST) at NCBI (http://www.ncbi.nlm.nih.gov/BLAST/ ), and deposited in the DNA Data Base of Japan (DDBJ) for serial accession numbers.
Api 50 chl test kit
Manufactured by bioMérieux
Sourced in France
The API 50 CHL test kits are a standardized identification system for the biochemical characterization of microorganisms. The kits provide a standardized and reproducible method for the identification of Gram-positive and Gram-negative bacteria, based on the observation of bacterial growth and carbohydrate fermentation in a series of miniaturized biochemical tests.
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Lab products found in correlation
4 protocols using api 50 chl test kit
1
Identification of Metal-Resistant Lactic Acid Bacteria
2
Isolation and Identification of Lactobacillus Strains from Algerian Poultry
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The lactobacilli strains were isolated from gizzard contents of Algerian local poultry. Decimal dilution of these samples were mixed with MRS medium and incubated at 37°C for 48 h under anaerobiosis (7 ). Selected colonies were picked from the higher dilutions and sub-cultured in MRS broth. The identification of the isolates was performed according to the criteria of Bergey’s Manual of Determinative Bacteriology and using the methods and criteria of Sharpe (8 ). The isolates were initially subjected to Gram staining and catalase test (3% H2O2). Only the Gram positive, catalase negative isolates were further identified. Growth at different temperatures was determined in MRS broths (10°C, 15°C, 40°C and 45°C). Hydrolysis of arginine was also recorded. The fermentative type was determined on agar.
The ability of the isolated strains to produce acid from different carbohydrates was determined by API 50 CHL test kits (BioMerieux, S.A., France). The results were loaded onto the API system software, which used the phenotypic data to predict a species identity (%) for each isolate.
The ability of the isolated strains to produce acid from different carbohydrates was determined by API 50 CHL test kits (BioMerieux, S.A., France). The results were loaded onto the API system software, which used the phenotypic data to predict a species identity (%) for each isolate.
3
Pediococcus Strain Identification Using 16S rDNA and Genomic Sequencing
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The species identification of the Pediococcus strain from the 16S rDNA was conducted using an API 50 CHL test kit (bioMérieux, Marcy l´Etoile, France) according to the
manufacturer’s instructions. Next, we used whole-genomic sequence analysis, which was carried out by Takara Bio Inc. (Shiga, Japan). Briefly, the sequence was analyzed by using a PacBio
RS II (Pacific Biosciences, Menlo Park, CA, USA), and the obtained reads were assembled by a genome assemble algorithm, SMRT Analysis v2.3.0 (Pacific Biosciences, Menlo
Park, CA, USA). The obtained contig sequences including the 16S rRNA gene sequence were further analyzed by NCBI BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi ).
manufacturer’s instructions. Next, we used whole-genomic sequence analysis, which was carried out by Takara Bio Inc. (Shiga, Japan). Briefly, the sequence was analyzed by using a PacBio
RS II (Pacific Biosciences, Menlo Park, CA, USA), and the obtained reads were assembled by a genome assemble algorithm, SMRT Analysis v2.3.0 (Pacific Biosciences, Menlo
Park, CA, USA). The obtained contig sequences including the 16S rRNA gene sequence were further analyzed by NCBI BLAST (
4
Bacterial Catalase and Carbohydrate Utilization
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The presence of catalase from a bacterial isolate is obvious when a small amount of bacterial culture from their agar selective medium developed bubbles (effervescence) at addition of 3% hydrogen peroxide (3%, v/v) (MacFaddin, 2000) .
The API 50 CHL All strains were identified (genus and species) and characterized based on biochemical evaluation tests using a bioMerieux API 50 CHL test kit according to the manufacturer's instructions (BioMerieux, Marcy l'Etoile, France). The test API 50 CHL put in evidence the enzymatic equipment bacteria, which differs from one species to another. The API 50 CHL consists of 50 microtubes used to study carbohydrates fermentation. The fermentation tests are inoculated with bacterial strain suspension in API 50 CHL medium which rehydrates the substrates. During incubation, the ability of isolate to ferment carbohydrates is indicated by the colour change of the basal medium used, caused by the anaerobic production of acid and detected by the pH indicator present in the selective medium. The strips are visually examined after 24-72 h. A positive or negative result was determined from the colour change from purple to yellow. The results were then introduced to the API databases using APIwebTM 50 CHL V5.1 and ABIS online software (Stoica and Sorescu, 2017) .
The API 50 CHL All strains were identified (genus and species) and characterized based on biochemical evaluation tests using a bioMerieux API 50 CHL test kit according to the manufacturer's instructions (BioMerieux, Marcy l'Etoile, France). The test API 50 CHL put in evidence the enzymatic equipment bacteria, which differs from one species to another. The API 50 CHL consists of 50 microtubes used to study carbohydrates fermentation. The fermentation tests are inoculated with bacterial strain suspension in API 50 CHL medium which rehydrates the substrates. During incubation, the ability of isolate to ferment carbohydrates is indicated by the colour change of the basal medium used, caused by the anaerobic production of acid and detected by the pH indicator present in the selective medium. The strips are visually examined after 24-72 h. A positive or negative result was determined from the colour change from purple to yellow. The results were then introduced to the API databases using APIwebTM 50 CHL V5.1 and ABIS online software (Stoica and Sorescu, 2017) .
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