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Introduction
Pork plays a crucial role in the Mexican diet, with a per capita consumption of approximately 22 kg in 2023. In the same year, Mexico reported pork production of around 1.6 million metric tons (Mt), while imports and exports nearly reached 1.3 and 0.3 Mt, respectively (COMECARNE 2024, USDA 2024). Despite its popularity, pork quality remains a challenge due to the oxidation of lipids and proteins, as well as microbial spoilage, which negatively impacts shelf life, safety, and consumer acceptance (Liu et al. 2023, Papanagiotou et al. 2013).
To reduce these issues, synthetic antioxidants and antibacterial agents are commonly used in the meat industry. However, consumer concerns about the health risks and perceived unnaturalness of these additives have led to an increased demand for natural alternatives. Among natural sources, plant polyphenols have been extensively investigated for their antioxidant and antibacterial properties in meat products (Kane et al. 2024, Papuc et al. 2012).
Bee products such as propolis have gained attention due to their high content of bioactive compounds. The bioactivity of propolis depends on its botanical and geographical origin, which Influences Its polyphenol profile (Camacho-Bernal et al. 2021, Toreti et al. 2013). Previous studies have demonstrated that ethanolic propolis extracts can enhance the oxidative and microbial stability of raw beef and pork patties during refrigeration (Vargas-Sánchez et al. 2019). In particular, propolis samples collected in northwestern Mexico have been identified as bifloral, primarily composed of Mesquite and Catclaw (Vargas-Sánchez et al. 2020).
Although the antioxidant and antibacterial activity of propolis has been reported, there is limited information on specific effects of mesquite-derived propolis in thermally treated meat systems. Therefore, this study aimed to investigate the impact of Mesquite propolis extracts on the oxidative and microbial stability of a thermally treated pork meat homogenate.
Materials and methods
Materials and chemicals
Samples of propolis were acquired from two apiaries from Pueblo de Alamos (29.1476 N, -110.1239 O, 632 m; 29.1887 N, -110.1273 O, 632 m; respectively) and stored at -20 °C, in the dark. All the chemicals used were analytical grade and were purchased from Sigma Chemicals. At the same time, Brain Heart Infusion (BHI) and Plate Count Agar (PCA) were obtained from Merck.
Raw propolis characterization
The acetolysis method was used to determine the floral origin of propolis (Vargas-Sánchez et al. 2016), with slight modifications. Propolis was mixed with distilled water (1:10 w/v ratio) at 10 000 rpm (25 °C) for 1 min (Ultraturrax-T25, IKA, Germany) and centrifuged at 5 000 × g (4 °C) for 15 min (Sorvall ST18R, Thermo Fisher Scientific, USA). The precipitate was dehydrated with 1 mL of CH₃COOH, mixed with 1 mL of H2SO4 (9:1), centrifuged, and washed with distilled water (d-water). The sediment was mixed with 0.5 mL of glycerin-water solution (1:1), and 0.1 mL of the obtained suspension was placed on a microscope slide. Pollen grains were observed using an optical microscope (CX-31, Olympus®, Japan). At least 500 pollen grains were counted and assigned to four classes: minor (< 3%), important minor (3-15%), secondary (15-45%), and predominant (> 45%). Pollen slides, based on the plant species of the local region, were used to identify pollen grains.
The AOAC procedure was followed to measure the pH values (AOAC 2020), with slight modifications. Samples (1:10 ratio) were homogenized at 6 000 rpm (5 °C) for 1 min with d-water before pH measurements (pH211, Hanna Instruments Inc., USA).
Concerning the color values, L* (lightness), a*(redness), b* (yellowness), and RGB (red-green-blue) values were measured in the sample's surface (CM-508d, Konica Minolta Inc., Japan) (Hernández et al. 2016).
Regarding the sensory evaluation, a 15-person panel was used to measure sensory attributes of propolis (Habryka et al. 2020), with slight modifications. Color (brightness, intensity, and uniformity), aroma (floral, waxy, resinous, and sweet), flavor (acid, bitter, and sweet), and consistency (viscous, sticky, and solid) were the descriptors used, which were subjected to a hedonic scale.
Mesquite propolis extracts (MPE) obtention
Extracts were obtained from raw propolis samples with d-water (1:10 ratio) by maceration-assisted extraction at 150 rpm (25 °C) for 24 h in the dark (MaxQ-5000, Fisher Scientific, Canada). The resultant solution was filtered (Whatman no. 1 filter paper) under vacuum (FE-1500, Felisa, Mexico), and dried (DC401, Yamato, Japan). The obtained Mesquite propolis extracts (MPE) were stored at -20 °C in the dark (SAGARPA 2007).
Polyphenol’s content
The total phenolic content (TPC) was determined using the Folin-Ciocalteu method (Matić and Jakobek 2021). MPE (20 µL, 5 mg/mL) was mixed with 160 µL of d-water, 60 µL of sodium carbonate (7% w/v), and 40 µL of Folin-Ciocalteu reagent (2 M). The solution was incubated for 60 min (25 °C) in the dark, and the absorbance was read at 750 nm (Multiskan FC UV-Vis, Thermo Scientific, Japan), and the results were expressed as mg of gallic acid equivalents (GAE) g−1.
The flavone and flavanols content were measured by the aluminum chloride method (Matić and Jakobek 2021). MPE (10 µL, 5 mg/mL) was mixed with 130 µL of methanol and 10 µL of aluminum chloride (5%, w/v). The solution was incubated for 30 min (25 °C) in the dark, the absorbance was read (412 nm), and the results were expressed as mg of quercetin equivalent (QE) g−1.
The flavanone-dihydroflavonol content (FDC) was measured using the dinitrophenyl method (Isla et al. 2014). MPE (40 µL, 5 mg/mL) was mixed with 80 μL of dinitrophenyl solution, incubated for 50 min (50 °C) in the dark, and diluted with 280 μL of potassium hydroxide (10%, w/v). The obtained solution (30 μL) was mixed with 250 μL of ethanol, the absorbance was read (490 nm), and the results were expressed as mg of hesperidin equivalents (HE) g−1.
The chlorogenic acid content (CAC) was measured using the sodium nitrite method (Griffiths et al. 1992). MPE (100 µL, 5 mg/mL) was mixed with 200 µL of urea (0.7 M), 200 µL of acetic acid (0.1 M), and 500 µL of d-water. The obtained solution was mixed with 500 µL of sodium nitrite (0.14 M) and 500 µL of sodium hydroxide (0.5 M), and centrifuged at 2 250 × g (4 °C) for 10 min. The mixture was incubated for 10 min (25 °C) in the dark, the absorbance was read (510 nm), and the results were expressed as mg of chlorogenic acid equivalents (CGA) g−1.
Antioxidant activity
The free-radical scavenging activity was measured using the DPPH method (Ozgen et al. 2006). MPE (20 µL, 100 µg/mL) was mixed with 180 µL of DPPH solution (300 µM). The solution was incubated for 30 min (25 °C) in the dark, the absorbance was read (517 nm), and the results were expressed as inhibition percentage (%).
The radical cation scavenging activity was measured using the ABTS method (Ozgen et al. 2006). MPE (20 µL, 100 µg/mL) was mixed with 180 µL of ABTS solution (absorbance 0.8 nm in ethanol). The solution was incubated for 30 min (25 °C) in the dark, the absorbance was read (730 nm), and the results were expressed as inhibition percentage (%).
The reducing power ability (RPA) was measured using the Prussian-blue method (Işıl-Berker et al. 2010). MPE (100 µL, 100 µg/mL) was mixed with 300 µL of phosphate buffer (2 M) and 300 µL of potassium ferrocyanide (1%, w/v). The solution was incubated for 20 min (50 °C) in the dark (Aquabath, Thermo Scientific, USA). Subsequently, samples were mixed with 300 µL of TCA (10%, w/v) and centrifuged at 4 200 × g (4 °C) for 10 min. The supernatant (100 µL) was homogenized with 100 µL of d-water and 250 µL of FeCl3 (0.1%, w/v), the absorbance was read (700 nm), and the results were expressed as absorbance (abs).
The ferric reducing antioxidant power (FRAP) method was also measured (Işıl-Berker et al. 2010). MPE (20 µL, 100 µg/mL) was mixed with 150 µL of FRAP solution. The solution was incubated for 8 min (25 °C) in the dark, the absorbance was read (595 nm), and the results were expressed as mg of iron equivalents (Fe2+) g−1.
Antibacterial activity
The antibacterial activity was measured using the broth-microdilution method (Jorgensen et al. 1999). Staphylococcus aureus ATCC 29213B, Listeria monocytogenes ATCC 33090, Escherichia coli ATCC 25922, and Salmonella typhimurium ATCC 14028) were initially reactivated in BHI broth for 24 h (37 °C) in the dark (IC403C, Yamato, Japan). MPE (50 µL) was mixed with 50 µL of bacteria suspension (1.5 × 108 CFU mL−1) and incubated for 24 h (37 °C) in the dark. Gentamicin (25 µg mL−1) was used as a positive control, and BHI broth solution as the blank. The absorbance was read (630 nm), and the results were expressed as absorbance (abs).
Meat quality measurements
Fresh minced pork meat (Semimembranosus muscle) was purchased from a local processor (Norson®, Hermosillo, Mexico). The minced pork meat was mixed with salt (0.5%, w/v) and pork back fat (10%, w/v). A 1 g meat sample from the batch was homogenized with 10 mL of d-water at 6 000 rpm (5 °C) for 1 min, and 1 mL of the respective antioxidants: Control, without antioxidant; MPE1 and MPE2, extracts at mg kg−1; BHT, butylated hydroxytoluene at 500 mg kg−1. The obtained mixture was heated in a water bath for 0, 60, and 120 min (65 °C). After that, meat homogenates were subjected to meat quality assays.
The pH and color of meat homogenates were determined as previously described (AOAC 2020, Hernández et al. 2016). Additionally, the TBARS method was employed to measure lipid oxidation (Pfalzgraf et al. 1995). Meat homogenates (0.5 mL) were homogenized with 1 mL of TCA (10%, w/v) at 4 500 rpm (5 °C) for 1 min and centrifuged at 2 500 × g (5 °C) for 20 min. Then, 1 mL of the filtered supernatant was mixed with 1 mL of 2-TBA solution (20 mM) and incubated for 20 min (98 °C). After incubating, the absorbance was read (531 nm), and the results were expressed as mg of malondialdehyde (MDA) kg−1 of pork meat.
The pour-plate procedure measured the growth of psychrophilic and mesophilic bacteria (SS 1994). Meat product samples were aseptically homogenized with peptone water (0.1%, w/v) (Seward Stomacher® 400, UK); then, 1 mL of the appropriate dilutions was pour-plated using plate count agar as the standard, incubated during 48 h (37 °C) for mesophilic bacteria), as well during 10 days (5 °C) for psychrophilic bacteria and results expressed as log10 of colony-forming units (CFU) g−1.
Statistical analysis
The study employed a completely randomized design. Results were expressed as mean ± standard deviation (SD) of at least three independent experiments (n = 6). Data from physicochemical, sensory, and polyphenol content were subjected to a Student t-test to compare treatment groups. Data from bioactivity were subjected to a one-way analysis of variance (ANOVA). In contrast, data on oxidative and microbial stability were subjected to a two-way ANOVA, with the treatments and thermal process period as fixed effects. The interaction between these factors was also evaluated. Differences were considered significant at p ≤ 0.05 using the Tukey-Kramer post hoc test (NCSS ver21).
Results
Raw propolis characterization
As shown in Table 1, a total of 14 pollen types from eight botanical families were identified in raw propolis from both apiaries. The Fabaceae family showed the highest frequency of pollen grains (p ≤ 0.05). Prosopis velutina (Mesquite) was the predominant pollen type in both samples (p ≤ 0.05).
Table 1 Pollen types identified in propolis samples.
| Family | Pollen type | Apiary #1 | Apiary #2 | ||
|---|---|---|---|---|---|
| (%) | Classes | (%) | Classes | ||
| Agavaceae | Agave angustifolia | 3 | Important minor | 3 | Important minor |
| Asteraceae | Ambrosia | 3 | Important minor | 3 | Important minor |
| Burseraceae | Bursera laxiflora | 3 | Important minor | 3 | Important minor |
| Fabaceae | Acacia sp. | 10 | Secondary | 10 | Secondary |
| Havardia mexicana | 3 | Important minor | 3 | Important minor | |
| Mimosa distachya var. Laxiflora | 7 | Secondary | 7 | Secondary | |
| Olneya tesota | 10 | Secondary | 10 | Secondary | |
| Prosopis velutina | 49.8 | Predominant | 49.6 | Predominant | |
| Malvaceae | Ceiba acuminata | 3 | Important minor | 3 | Important minor |
| Herisantia crispa | 3 | Important minor | 3 | Important minor | |
| Myrtaceae | Eucalyptus sp. | 3 | Important minor | 3 | Important minor |
| Poaceae | Poaceae sp. | 0.2 | Minor | 0.2 | Minor |
| Sapindaceae | Cardiospermum halicacabum | 2 | Minor | 2 | Minor |
| Unidentified | 0 | 0.2 | Minor | ||
| Total | 100 | 100 | |||
Both apiaries were located in Pueblo de Álamos (t-test; p ≤ 0.05).
Table 2 presents the physicochemical and sensory properties of Mesquite propolis. The propolis sample from Apiary #1 showed significantly lower pH values compared to the sample from Apiary #2 (p < 0.05). Regarding color, Apiary #1 propolis also showed the lowest b* values, while no differences were observed in L* and a* values (p ≥ 0.05). Based on RGB values and HEX codes, the perceived colors were identified as Black Pepper (Apiary #1) and Dark Lava (Apiary #2). In terms of sensory attributes, Apiary #1 propolis received the highest scores (p ≤ 0.05) for color (brightness and uniformity), resinous aroma, bitter flavor, and sticky-solid consistency. In contrast, Apiary #2 propolis scored highest only in wax for aroma. Neither sample had a sweet flavor (p ≥ 0.05).
Table 2 Physicochemical and sensory properties of Mesquite propolis.
| Item | Apiary #1 | Apiary #2 | p-value |
|---|---|---|---|
| Physicochemical | |||
| pH | 4.51 ± 0.01 | 4.31 ± 0.02 | < 0.001 |
| L* | 28.43 ± 1.18 | 31.42 ± 1.49 | n.s. |
| a* | 1.82 ± 0.76 | 2.31 ± 0.47 | n.s. |
| b* | 3.16 ± 0.85 | 5.08 ± 0.66 | < 0.001 |
| RGB/HEX code | 72, 66, 62/#48423E | 81, 72, 66/#514842 | |
| Sensory | |||
| Color - brightness | 4.65 ± 0.47 | 2.75 ± 0.42 | < 0.001 |
| Color - uniformity | 4.95 ± 0.16 | 2.90 ± 0.32 | < 0.001 |
| Aroma - waxy | 2.10 ± 0.32 | 3.05 ± 0.16 | < 0.001 |
| Aroma - resinous | 4.85 ± 0.34 | 3.90 ± 0.32 | < 0.001 |
| Flavor - bitter | 4.95 ± 0.16 | 4.05 ± 0.16 | < 0.001 |
| Flavor - sweet | - | - | n.s. |
| Consistency - sticky | 4.85 ± 0.34 | 3.85 ± 0.34 | < 0.001 |
| Consistency - solid | 4.90 ± 0.21 | 3.95 ± 0.16 | < 0.001 |
Results expressed as mean ± SD of at least three independent experiments. Apiaries #1 and #2: Sample from Pueblo de Álamos. Lowercase letters indicate statistical differences between treatments (t-test, p ≤ 0.05).
Polyphenol content and bioactivity of propolis extracts
As shown in Table 3, MPE2 exhibited significantly higher values of TPC, FFC, FDC, and CAC than MPE1 (p ≤ 0.05). Regarding antioxidant activity, although the synthetic antioxidant BHT showed the highest efficacy, MPE2 showed higher RCSA, RPA, and FRAP values than MPE1 (p ≤ 0.05); no differences were observed in FRSA values (p ≥ 0.05). Both extracts demonstrated a higher antibacterial effect against Gram-positive (S. aureus and L. monocytogenes) than Gram-negative bacteria (E. coli and S. typhimurium) (p ≤ 0.05). However, gentamicin remained the most effective.
Table 3 Polyphenol content and bioactivity of Mesquite propolis extracts.
| Item | Assays | |||
|---|---|---|---|---|
| Polyphenols | TPC (mg GAE g-1) | FFC (mg QE g-1) | FDC (mg HE g-1) | CAC (mg CGA g-1) |
| MPE1 | 175.04 ± 1.53 | 29.38 ± 2.94 | 99.50 ± 2.59 | 6.83 ± 0.15 |
| MPE2 | 295.94 ± 8.65 | 69.44 ± 2.20 | 149.67 ± 1.86 | 13.28 ± 0.60 |
| p-value | < 0.001 | < 0.001 | < 0.001 | < 0.001 |
| Antioxidant | FRSA (%) | RCSA (%) | RPA (abs) | FRAP (mg Fe2+ g-1) |
| MPE1 | 89.77 ± 0.25 a | 91.07 ± 0.27 b | 0.30 ± 0.01 a | 1.02 ± 0.03 a |
| MPE2 | 89.47 ± 0.64 a | 90.78 ± 0.23 b | 0.34 ± 0.01 b | 1.38 ± 0.07 b |
| BHT | 91.20 ± 1.30 a | 64.60 ± 0.55 a | 1.08 ± 0.05 c | 1.40 ± 0.10 b |
| p-value | < 0.001 | < 0.001 | < 0.001 | < 0.001 |
| Antibacterial | S. aureus (%) | L. monocytogenes (%) | E. coli (%) | S. typhimurium (%) |
| MPE1 | 42.50 ± 2.89 a | 62.03 ± 2.84 a | 8.85 ± 2.67 a | 7.39 ± 3.22 a |
| MPE2 | 44.15 ± 3.54 a | 61.17 ± 1.85 a | 6.21 ± 1.19 a | 13.32 ± 3.75 a |
| Gentamicin | 67.31 ± 3.37 b | 71.43 ± 1.32 b | 67.28 ± 1.53 b | 68.32 ± 2.38 b |
| p-value | < 0.001 | < 0.001 | < 0.001 | < 0.001 |
Results expressed as mean ± SD of at least three independent experiments. MPE1 and MPE2: Mesquite propolis extract from Pueblo de Álamos (Apiaries #1 and #2, respectively). TPC, total phenolic content. FFC, flavone, and flavonol content. FDC, flavanone-dihydroflavonol content. CAC, chlorogenic acid content. FRSA, free-radical scavenging activity. RCSA, radical-cation scavenging activity. RPA, reducing power ability. FRAP, ferric-reducing antioxidant power. BHT, butylated hydroxytoluene. %: inhibition percentage. Lowercase letters indicate statistical differences between treatments (t-test; Tukey, p ≤ 0.05).
Oxidative and microbial stability of meat homogenates
The effect of treatment and thermal processing on pork meat homogenates is summarized in Table 4. A significant interaction was observed for pH, color, TBARS, and microbial count values (p ≤ 0.05). At 120 min, MPE1 maintained the highest pH values (p ≤ 0.05). In terms of color, at 120 min, MPE2 and BHT presented the lowest L* values (p ≤ 0.05), with no differences (p ≥ 0.05) in a* and b* values. Regarding TBARS, at 120 min, MPE1 showed the lowest TBARS values (p ≤ 0.05). For microbial stability, at 120 min, MP1 and MPE2 showed the lowest bacterial counts (p ≤ 0.05).
Table 4 Meat quality measurements of meat homogenates.
| Item | Treatment | Thermal process at 65 °C | ||
|---|---|---|---|---|
| 0 min | 60 min | 120 min | ||
| pH | Control | 5.62 ± 0.02 aA | 5.97 ± 0.01 aB | 6.09 ± 0.01 aB |
| MPE1 | 5.69 ± 0.01 bA | 6.15 ± 0.01 cB | 6.20 ± 0.02 cC | |
| MPE2 | 5.70 ± 0.01 bA | 6.03 ± 0.01 bB | 6.16 ± 0.01 bC | |
| BHT | 5.64 ± 0.01 aA | 5.99 ± 0.01 aB | 6.10 ± 0.01 aC | |
| L* | Control | 36.45 ± 1.69 aA | 55.49 ± 2.01 aB | 60.82 ± 1.44 bC |
| MPE1 | 36.69 ± 0.01 aA | 60.58 ± 0.84 bB | 61.35 ± 0.59 bB | |
| MPE2 | 36.44 ± 1.44 aA | 56.52 ± 0.92 aB | 56.18 ± 0.78 aB | |
| BHT | 37.28 ± 0.59 aA | 58.75 ± 3.18 abB | 57.80 ± 0.90 aAB | |
| a* | Control | 7.24 ± 0.45 aB | -2.17 ± 0.24 aA | -2.14 ± 0.24 aA |
| MPE1 | 6.88 ± 0.24 aB | -2.19 ± 0.14 aA | -2.50 ± 0.11 aA | |
| MPE2 | 6.51 ± 0.37 aB | -1.87 ± 0.24 aA | -2.24 ± 0.23 aA | |
| BHT | 6.79 ± 0.84 aB | -2.35 ± 0.12 aA | -2.23 ± 0.25 aA | |
| b* | Control | 9.42 ± 1.09 aB | 4.13 ± 0.74 aA | 5.67 ± 1.26 aA |
| MPE1 | 9.22 ± 0.75 aB | 5.27 ± 0.48 aA | 5.01 ± 0.37 aA | |
| MPE2 | 9.01 ± 0.58 aB | 4.95 ± 0.67 aA | 3.94 ± 0.89 aA | |
| BHT | 9.18 ± 0.61 aB | 5.04 ± 1.23 aA | 4.65 ± 0.55 aA | |
| TBARS | Control | 0.299 ± 0.013 cA | 0.537 ± 0.008 dB | 0.675 ± 0.015 dC |
| (mg MDA kg-1) | MPE1 | 0.004 ± 0.002 aA | 0.012 ± 0.004 aB | 0.031 ± 0.002 aC |
| MPE2 | 0.005 ± 0.001 aA | 0.030 ± 0.004 bB | 0.038 ± 0.004 bC | |
| BHT | 0.223 ± 0.006 bA | 0.439 ± 0.030 cB | 0.517 ± 0.019 cC | |
| Mesophilic | Control | 3.80 ± 0.09 aA | 3.83 ± 0.05 aA | 3.72 ± 0.08 bA |
| (Log10 CFU g-1) | MPE1 | 3.73 ± 0.08 aB | 3.78 ± 0.08 aB | 3.45 ± 0.08 aA |
| MPE2 | 3.75 ± 0.05 aB | 3.77 ± 0.08 aB | 3.47 ± 0.05 aA | |
| BHT | 3.85 ± 0.05 aA | 3.83 ± 0.08 aA | 3.73 ± 0.05 bA | |
| Psychrophilic | Control | 4.48 ± 0.08 aA | 4.45 ± 0.05 aA | 4.35 ± 0.05 bA |
| (Log10 CFU g-1) | MPE1 | 4.50 ± 0.06 aB | 4.48 ± 0.08 aB | 4.12 ± 0.08 aA |
| MPE2 | 4.48 ± 0.04 aB | 4.48 ± 0.04 aB | 4.10 ± 0.06 aA | |
| BHT | 4.52 ± 0.08 aA | 4.53 ± 0.05 aA | 4.33 ± 0.05 bA | |
Results expressed as mean ± SD of at least three independent experiments. MPE1 and MPE2: Mesquite propolis extract from Pueblo de Álamos (Apiaries #1 and #2, respectively). BHT, butylated hydroxytoluene. Capital letters indicate statistical differences in each treatment at different thermal process periods; lowercase letters indicate statistical differences between treatments (Tukey, p ≤ 0.05).
Discussion
Propolis is a resinous material processed by bees from plant resins, whose composition is closely linked to the vegetation surrounding the apiary (SAGARPA 2017). In this context, the Fabaceae family is frequently identified as a predominant pollen source (Temizer et al. 2017), with Mesquite pollen being a representative and particularly abundant component in raw propolis from the Sonoran Desert region (Vargas-Sánchez et al. 2016). The botanical origin of propolis is known to influence its physicochemical, sensory, and bioactive properties (Toreti et al. 2013, Vargas-Sánchez et al. 2020). Although the Mexican NOM-003-SAG/GAN-2017 regulation does not establish standard values for pH or color in propolis, previous research indicates that these parameters vary according to the botanical origin. For Instance, Quercus sp. propolis tends to present lower pH values compared to Populus sp., Pinus sp., and Castanea sativa (Dias et al. 2012). Botanical source also modifies sensory characteristics, which must align with the standards for color (red, reddish-yellow, dark yellow, brown-green, brown, or black), aroma (resinous), flavor (bitter or sweet), and consistency (solid) (SAGARPA 2017).
The presence and concentration of polyphenols, the primary contributors to propolis bioactivity, also depend on their biological origin (Kumazawa et al. 2012, Papuc et al. 2017, SAGARPA 2017). These compounds act as antioxidants by donating hydrogen atoms or electrons to stabilize free radicals and by chelating metal ions involved in oxidative reactions. They also exert antibacterial effects, possibly by altering membrane permeability or inhibiting nucleic acid synthesis (Papuc et al. 2017). In propolis from sources such as Cannabis sativa, Pine, Quercus spp., Helianthus annuus, the phenolic profile includes p-coumaric, ferulic, gallic, and chlorogenic acids, as well as flavonoids like quercetin, apigenin, and pinocembrin (Kekecoglu et al. 2021, Kolayli et al. 2023, Özkök et al. 2023).
Functionally, antioxidant activity is essential for determining the efficacy of propolis in preserving meat products. Regulatory standards require that propolis demonstrate free radical scavenging activity, although specific quantitative thresholds are not mandated (SAGARPA 2017). Previous studies have reported variable antioxidant activity, depending on the botanical source and extraction method. Notably, extracts from C. sativa and Eucalyptus sp., display high antiradical and reducing power activity, correlating with their phenolic content (Kumazawa et al. 2012, Castro-Falcón et al. 2016).
In terms of antibacterial properties, multiple studies have confirmed the Inhibitory effects of propolis against foodborne pathogens, particularly S. aureus and E. coli (SAGARPA 2017, Özkök et al. 2023). Interestingly, propolis also shows greater efficacy against Gram-positive bacteria, likely due to structural differences in bacterial cell walls that influence compound penetration (Kekecoglu et al. 2021).
The Incorporation of propolis extracts into meat systems has been increasingly studied as a natural alternative to synthetic antioxidants. Lipid oxidation and microbial growth are primary factors contributing to meat spoilage, and both are influenced by pH, thermal treatment, and packaging conditions (Papuc et al. 2017, Anton et al. 2019). Lipid oxidation is a radical-mediated chain reaction that generates primary products, such as hydroperoxides (ROOH), and secondary products, including alcohols and aldehydes. Phenolic compounds inhibit this process by donating hydrogen, which helps preserve both lipid Integrity (ROO• + ArOH → ROOH + ArO•) and meat color (MetMb3+ + ArOH → Mb2+ + ArO•) (Bai et al. 2025, Li et al. 2025, Pfalzgraf et al. 1995, Vargas-Sánchez et al. 2019).
In this study, propolis extract improves the oxidative stability of pork meat homogenates, consistent with findings from Kročko et al. (2014), who reported lower MDA values in cooked ham treated with ethanol extracts. Similarly, propolis extracts enhance oxidative stability, reducing pH, color, and lipid oxidation changes, as well as microbial stability, reducing microbial loads in sausages, ground beef, patties, and marinated chicken under various storage conditions (El-Demery et al. 2016, Vargas-Sánchez et al. 2019, López-Patiño et al. 2021, Fadhil 2023).
These results support the potential of propolis as a functional ingredient in meat preservation, aligning with consumer demand for naturally preserved products. However, variability in propolis composition due to geographic and botanical differences remains a limitation. Furthermore, while antioxidant and antibacterial effects were demonstrated in this study, further research is required to address sensory acceptability and scalability
Conclusions
The results demonstrated that raw Mesquite propolis meets the physicochemical and sensory standards required by Mexican regulations. Mesquite aqueous propolis extract exerts antioxidant and antibacterial activity, mainly associated with its polyphenol composition. Furthermore, the incorporation of this extract into pork meat homogenates reduced pH variation, lipid oxidation, color changes, and microbial growth after thermal processing. These results highlight that Mesquite propolis has great potential as a preservative for meat products. Its application may help extend shelf life by reducing synthetic additives. Future studies should evaluate its acceptability and scalability.
