## 3 Results and Discussion
The HPP of _Serra da Estrela_ cheese (0 months storage), independently of the treatment conditions, did not affect the total content of SFAs, MUFAs, and PUFAs in the EFAMEs fractions (values similar to those of Ch\({}_{\rm C}\) cheeses), except for HPP P2 cheeses, which revealed an increment of about 18% in relation to both Ch\({}_{\rm C}\) and other HPP (P1 and P3) cheeses (Figure 3 and Table S2). Upon 15 months of storage, the SFA content was numerically, although not significantly, slightly lower in HPP P2 and P3 cheeses (57.9 g FA/100 g and 53.5 g FA/100 g fat) than in Ch\({}_{\rm C}\) cheese (61.2 g FA/100 g fat), while HPP P1 chees revealed a higher content (64.5 g FA/100 g fat) (Figure 3). Analyzing the main SFAs at 15 months of storage, somewhat lower values were observed for HPP P3 cheeses in comparison with Ch\({}_{\rm C}\) cheeses (Figure 5A vs. Figure 5D) concerning palmitic (C16:0, 16.87 g FA/100 g fat vs. 19.26 g FA/100 g fat, respectively), myristic (C14:0, 8.84 g FA/100 g fat vs. 10.10 g FA/100 g fat), lauric (C12:0, 3.66 g FA/100 g fat vs. 4.18 g FA/100 g fat) and butyric acids (C4:0 2.68 g FA/100 g fat vs. 3.09 g FA/100 g fat). A further observation to be highlighted was the lower content of butyric acid in the HPP NEFAME P3 fraction (Figure 7 and Table S3) since butyric acid has been associated with a rancid-like cheese odor [33]. Similarly, lower amounts of butyric acid were reported in HPP (400 MPa and 600 MPa for 5 min and 20 min) processed Torta del Casar cheeses [13]. Nevertheless, sensory evaluation of those _Serra da Estrela_ cheeses did not reveal significant odor differences between Ch\({}_{\rm C}\) and HPP cheeses, and the characteristic lactic, acid, animal/stable, and short-chain fatty acids (vomit-like odor) attributes were detected [10]. Figure 6: Example of HPLC separation of CLA isomers of Ch\({}_{\rm C}\) (blue line) and HPP P1–600 MPa/6 min (black line) of _Serra da Estrela_ cheese at month 0 of storage. Nutritional indices are presented in Table 1. The PUFAs/SFAs ratio is measured to indicate the risk of incidence of CVD. It has also been discussed, although with some uncertainties, that single fatty acids might play negative roles in human health, in particular on the probability of increasing atheroma (C12:0, C14:0, and C16:0) and thrombus formation  
Figure 7: Evolution of the mean composition of the main SFAs, MUFAs, and PUFAs (mg FA/100 g of fat) found in the NEFAME fraction in control (**A**) and HPP cheeses (**B**) 600 MPa/6 min—P1, (**C**) 450 MPa/6 min—P2, and (**D**) 450 MPa/9 min—P3 _Serra da Estrela_ cheeses, during refrigerated storage. C9 with values outside the scale; data in Table S3. Data are expressed as mean (\(n=2\)). (C14:0, C16:0, and C18:0) [23]; hence, in many studies, the index of atherogenicity (IA) and the index of thrombogenicity (IT) are calculated (based on experimental values) to assess diet nutritional quality and the associated propensity to influence the incidence of CVD [23]. According to the results presented in Table 1, the EFAMEs fractions of all cheeses revealed similar atherogenic (between 2.32 and 2.40) and thrombogenic (between 2.55 and 2.64) indices, although the HPP P3 cheese revealed slightly lower (less 2%) values compared with the Ch\({}_{\text{C}}\) cheeses. Dairy products with lower atherogenic and thrombogenic indices are potentially healthier for humans [27]. The results reported herein are in fact lower than those reported in [PERSON] et al. 's study, where mean AI and TI values reported for _Serra da Estrela_ cheeses from different cheese producers were 2.92 \(\pm\) 0.73 and 2.73 \(\pm\) 0.52, respectively [8]. On the other hand, ripened buffalo cheeses revealed an IA index of 2.16 and an IT index of 1.20 [27], corroborating the importance of the product's origin and processing on lipid composition and consequent AI and TI. The balance of PUFAs omega n - 6/omega n - 3 is relevant, since PUFAs may suppress HDL, which has a protective action against CVD [23].