Bushnell haas broth

Samples collected from iron-rich stream sediment (Michigamme, MI [46.532, −88.141]), Lake Superior sediment (Bete Grise, MI [47.3723, −87.9529]), vermicompost (Calumet, MI [47.211, −88.553]), and a hydrocarbon seep in the Caspian Sea (39.7455, 50.4806) were used as starting material for enrichment cultures (17 (link)). Sediment and compost samples were collected using sterile containers and frozen for preservation. Detailed sample collection methods for Caspian Sea sediments can be found in Mahmoudi et al. (102 (link)) and Miller et al. (103 (link)).
Enrichment culture media consisted of 90 mL Bushnell-Haas broth (HiMedia Laboratories Pvt Ltd., Mumbai, India) amended with 0.25 g disodium terephthalate, 0.25 g terephthalamide, 2.5 mL BPA, and 0.25 mL of the alkene mixture. Duplicate biological replicates were set up in 250-mL Erlenmeyer flasks using 1 g of soil from the four different locations listed above. Cultures were placed in a shaking incubator at a speed of 200 rpm and at 30°C. Cultures were propagated every 14 days by transferring 10 mL of culture into 90 mL of fresh media. The culture medium composition and conditions remained consistent during the enrichment process. Biomass from cultures were collected after 10 (T5) and 22 (T11) weeks so the microbial community composition of enrichments could be assessed using 16S rRNA gene amplicon sequencing.

Samples collected from iron-rich stream sediment (Michigamme, MI [46.532, −88.141]), Lake Superior sediment (Bete Grise, MI [47.3723, −87.9529]), vermicompost (Calumet, MI [47.211, −88.553]), and a hydrocarbon seep in the Caspian Sea (39.7455, 50.4806) were used as starting material for enrichment cultures (17 (link)). Sediment and compost samples were collected using sterile containers and frozen for preservation. Detailed sample collection methods for Caspian Sea sediments can be found in Mahmoudi et al. (102 (link)) and Miller et al. (103 (link)).
Enrichment culture media consisted of 90 mL Bushnell-Haas broth (HiMedia Laboratories Pvt Ltd., Mumbai, India) amended with 0.25 g disodium terephthalate, 0.25 g terephthalamide, 2.5 mL BPA, and 0.25 mL of the alkene mixture. Duplicate biological replicates were set up in 250-mL Erlenmeyer flasks using 1 g of soil from the four different locations listed above. Cultures were placed in a shaking incubator at a speed of 200 rpm and at 30°C. Cultures were propagated every 14 days by transferring 10 mL of culture into 90 mL of fresh media. The culture medium composition and conditions remained consistent during the enrichment process. Biomass from cultures were collected after 10 (T5) and 22 (T11) weeks so the microbial community composition of enrichments could be assessed using 16S rRNA gene amplicon sequencing.

Bushnell-Haas broth (BHB), nutrient broth and nutrient agar were purchased from Himedia (Mumbai, India). All chemicals used in the study were of analytical and HPLC grade. Pyrene was purchased from Sigma-Aldrich^® (Bellefonte, Pennsylvania, USA) with 99% purity.