3.2.2. Microbiological Content

Antimicrobial capacity of different extract was also proved in fish products elaborated from thawed hake. Consequently, microbiological results of fish patties at days 0, 4, 7 and 11 from elaboration, are shown in Table 5.


**Table 5.** Microbiological results (cfu/g) of fish patties analysis at days 0, 4, 7 and 11 under refrigerated storage.

TVC: Total Viable Count; TCC: Total Coliform Count P: Pomegranate extract, RA: Rosemary extract rich in Rosmarinic Acid; NOS: Rosemary extract rich in diterpenes; NOVS: Rosemary extract rich in diterpenes and with lecitin as emulsifier; HYT-L: Hydroxytyrosol extract obtained from olive leaf; HYT-F: Hydroxytyrosol extract obtained from olive fruit.

As it can be appreciated, TVC results after 11 days of refrigerated storage were lower in samples that incorporated HYT-F and HYT-L in their formula, followed by P, control sample, NOVS, NOS and RA. While, TCC results showed as both pomegranate and rosemary extracs, diterpens rich extracts (NOS and NOVS) and rosmarinic acid rich extract (RA) obtained the lowest results in comparison with the control sample, or samples that incorporated HYT to their formulas. These last results can be related with antimicrobial activity of all the extracts against *E. Coli* O157:H7ATCC 25922 CECT 434. On the other hand, results obtained from analysis of *E. Coli* and *L. monocytogenes* did not present significant differences (*p* < 0.05) among incorporation of natural extracts. In addition, it must be taken into account that only samples enriched with HYT, both from fruit or leaf, did not exceed the limit stablished by European legislation regarding to TVC in fish products (5,000,000 cfu/g). In parallell, only P and rosemary extracts (RA, NOS, and NOVS) kept fish product samples below legal microbiological safety limits of TCC (50,000 cfu/g). However, all the natural extracts, including the control sample prevented against microbiological growth of *E. Coli* and *L. monocytogenes.*
