Phage Enrichment from Aqueous Materials

Sewage treatment plants are ideal sources from which bacteriophages are isolated. Due to their different ecosystems including raw input sewage, and aerobic and anaerobic digestion tanks, they contain high numbers of diverse bacteria. Alternative sources include a pond, lake, river, or ocean waters. Creative Biolabs provides phage enrichment from water samples, such as pond water, pond sediment, raw sewage, etc. The goal of phage enrichment is to enrich your sample with phages that can infect the desired host. This is done by removing endogenous bacteria from your sample and adding them to the bacterial culture medium and the growing culture of the host, and then incubating them. Even if there is a single phage capable of infecting the inoculated bacterial strain in the sample, it will replicate to a level that should be detectable by normal plating techniques. The volume used for enrichment can vary, from 1 ml sample to many liters. Increasing the enrichment volume increases the sensitivity of the procedure, so if you want phage to be rare in the sample, you need a larger enrichment volume.

Our Services

At Creative Biolabs, we use membrane filter adsorption and elution to concentrate phages from water samples. The double agar overlay method was used to separate the phages from the water samples. The isolated phages were purified by diluting in SM buffer and filtering through a 0.22 μm filter. The purified lysate is further processed to analyze its host range using the dot method.

• Study design
• Bacteriophage isolation
• Purification of phages
• Determination of host range
• Results analysis

The Example of Phage Enrichment from Aqueous Materials

• Centrifuge at 10,000 * g for 10 minutes to clarify the water sample to remove large particles and bacteria.
• Add solid MgSO₄ to 50mM.
• Slowly filter the water sample through a 0.22 μm (47mm diameter) mixed cellulose ester GSWP filter.
• Cut the filter into several pieces and put them in a flask containing 5ml of the eluent.
• Put in the ultrasonic cleaning tank for 4 minutes to help clean the elution of virus.

The Advantages of Phage Enrichment

A generally more effective method of isolating lysed phages from the marine environment is to use enriched cultures. The two main advantages of the concentration method are

• It allows screening of phages in larger samples (usually 25-50 mL instead of 5-10 µL), thereby increasing the possibility of isolating rare phages.
• It allows different target hosts (e.g., different strains of specific target bacteria) to be combined in the same incubation, again increasing the possibility of phage isolation.

Creative Biolabs can meet the needs of customers by providing concentration of phages from water samples on time and on budget. We have in-depth knowledge and experience of the tools and processes involved in the phage projects. Our skilled and dedicated scientific researchers ensure that the most suitable methods and techniques are selected for your project. If necessary, please feel free to contact us.

Please kindly note that our services can only be used to support research purposes (Not for clinical use).