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Article

Nutritional Use of Greek Medicinal Plants as Diet Mixtures for Weaned Pigs and Their Effects on Production, Health and Meat Quality

by
Georgios Magklaras
1,
Athina Tzora
2,
Eleftherios Bonos
1,
Christos Zacharis
1,
Konstantina Fotou
2,
Jing Wang
3,
Katerina Grigoriadou
4,
Ilias Giannenas
5,
Lizhi Jin
6 and
Ioannis Skoufos
1,*
1
Laboratory of Animal Science, Nutrition and Biotechnology, Department of Agriculture, School of Agriculture, University of Ioannina, Kostakioi Artas, 47100 Arta, Greece
2
Laboratory of Animal Health, Food Hygiene and Quality, Department of Agriculture, School of Agriculture, University of Ioannina, Kostakioi Artas, 47100 Arta, Greece
3
Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, Nanjing Agricultural University, Nanjing 210095, China
4
Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization-DEMETER, 57001 Thessaloniki, Greece
5
Laboratory of Nutrition, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
6
Guangzhou Meritech Bioengineering Co., Ltd., Guangzhou 510300, China
*
Author to whom correspondence should be addressed.
Appl. Sci. 2025, 15(17), 9696; https://doi.org/10.3390/app15179696
Submission received: 1 July 2025 / Revised: 27 August 2025 / Accepted: 28 August 2025 / Published: 3 September 2025
Versions Notes

Abstract

Current consumer trends for meat production with reduced antibiotic use constitute huge challenges in animal farming. Using indigenous raw materials such as aromatic or medicinal plants or their extracts could positively affect or retain animals’ health. The present study aimed to evaluate the effects of medicinal plant extracts and essential oils on pig performance parameters, health indices and meat quality. A phytobiotic mixture (PM) consisting of oregano (Origanum vulgare subsp. hirtum) essential oil, rock samphire (Crithmum maritimum L.) essential oil, garlic flour (Allium sativum L.) and false flax flour (Camelina sativa L. Crantz) was used in pig diets, containing in the experimental trials two different proportions of the oregano essential oil (200 mL/t of feed vs. 400 mL/t of feed). Three groups of weaned pigs were fed either the control diet (CONT) or one of the enriched diets (PM-A or PM-B, 2 g/kg). After a 43-day feeding period, at 77 days of age, blood was taken from the jugular vein for biochemical and hematological tests, and eight pigs were humanely slaughtered. A microbiological analysis of intestinal digesta from the ileum and caecum was conducted. Additionally, meat tissue cuts (biceps femoris, external abdominal and triceps brachii) were collected for a chemical analysis, fatty acid lipid profile and oxidative stability testing. The statistical analysis revealed no differences (p > 0.05) in the body weights and growth rates among the groups. An increase (p < 0.05) in total aerobic bacteria was detected in the ileum of group PM-A, while Escherichia coli (E. coli) counts were reduced (p < 0.05) in group PM-B. In the caecum, reductions in Enterobacteriaceae and Lactobacillaceae counts were observed in groups PM-A and PM-B. Concentrations of malondialdehyde (MDA) as an indicator of lipid peroxidation were significantly reduced (p < 0.05) in triceps brachii and biceps femoris for both groups PM-A and PM-B (day 0). A reduction (p < 0.05) in MDA was noticed in triceps brachii and external abdominal meat samples (day 7) for groups PM-A and PM-B. In addition, the fatty acid profile of the meat lipids (ΣPUFA, h/H and PUFA/SFA ratios) was positively modified (p < 0.05) in the ham and belly cuts. The addition of the PM significantly (p < 0.05) affected the redness of the ham and shoulder meat (a* value increased), the yellowness of only the ham (b* value decreased) and the lightness of both belly (L* value increased) and ham samples (L* value decreased). The meat proximate analysis, as well as hematological and biochemical parameters, did not identify any differences (p > 0.05) between the groups. In conclusion, the two investigated mixtures could be used in weaned pigs’ diets, with positive results in intestinal microbial modulation, oxidative stability, fatty acid profile and color characteristics of the pork meat produced.

1. Introduction

There is a global need to identify new production systems in agriculture that will increase the efficiency of food produced from the animal production sector [1], and satisfy the increasing demand for meat and meat products. In addition to sustainability in production, limitations in global natural resources and nutritional quality have become decisive factors for consumer choice [2].
Furthermore, the growing movement to reduce antibiotic usage in livestock has underlined the need for efficient substitutes that improve growth and facilitate disease prevention in pigs while ensuring food security [3,4,5,6]. The reliance on antibiotics has raised worries about the emergence of antimicrobial resistance, prompting a trend towards the inclusion of natural additives, such as extracts from aromatic and medicinal plants (phytobiotics), in animal diets [7,8]. Phytobiotics are a type of nutraceuticals that consist of non-nutritive, plant-derived bioactive chemicals used as feed additives in swine diets [9,10,11]. These can be divided into four categories based on their origin and processing characteristics: herbs (blooming, nonwoody, and non-persistent plants), spices (plants with a strong odor or flavor), essential oils (volatile lipophilic compounds) and oleoresins (extracts obtained from non-aqueous solutions) [12]. This new generation of feed additives encompasses a diverse array of substances, including essential oils, herbal extracts and functional components derived from various plant species, which are acknowledged for their antimicrobial, antioxidant and anti-inflammatory properties [13,14,15,16,17], as well as their capacity to enhance growth [18,19,20] and positively influence the gut microbiome [21,22,23]. The favorable effects may be linked to improvements in intestinal architecture, enhanced antioxidant capacity and modification of microbial communities. Phytobiotics are rich in various bioactive chemicals responsible for their functional actions. Flavonoids, alkaloids (containing alcohols, aldehydes, ketones, esters, and lactones), phenols (tannins), glycosides, terpenoids and glucosinolates are the components that make up the bioactive chemicals. Although phytobiotics represent a wide spectrum of bioactive chemicals, their precise mechanisms of action, inclusion dosage and possible interactions with other feed components are yet unknown.
Several factors can affect the quantity and chemical composition of active compounds present in phytobiotics. These factors encompass the plant component utilized (e.g., seeds or leaves), the geographical region, the harvesting season and the time of year. An understanding of the specific processes through which phytobiotics exert their effects is quite complicated due to the factors mentioned [12,24,25,26]. The use of a blend of phytobiotic sources in pig diets is increasingly justified by the diverse bioactive properties these compounds offer. Individual phytobiotics—such as essential oils, plant extracts and polyphenols—possess antimicrobial, antioxidant, anti-inflammatory and gut-modulating effects. However, their efficacy can be limited when used alone due to variability in absorption, stability or target specificity [27,28]. Combining multiple phytobiotics can harness synergistic effects, where the combined action exceeds the sum of individual effects enhancing gut health, immune function and overall performance more effectively than single compounds [29,30]. Such blends may also reduce the risk of microbial resistance, offering a more robust and consistent alternative to antibiotics in sustainable pig production.
The present study evaluated, for the first time, a phytobiotic mixture of four different aromatic/medicinal plants as ingredients in weaned pig diets, in an effort to evaluate their potential benefits on production indices, animal health and meat quality attributes. The two phytobiotic mixtures (Table 1) were comprised of oregano (Origanum vulgare subsp. hirtum) essential oil, rock samphire (Crithmum maritimum L.) essential oil, camelina (Camelina sativa L. Crantz) and garlic (Allium sativum L.) flour, having been incorporated into pig feed at a concentration of 0.2%. Oregano essential oil, derived from the Greek aromatic plant Origanum vulgare subsp. hirtum, comprises a minimum of 24 chemical compounds, including carvacrol, thymol and β-caryophyllene [31,32,33]. Numerous studies indicate that the dietary inclusion of oregano essential oil enhances growth performance, immune function and antioxidative responses in weaned and finishing pigs [34,35,36,37]. It has also been utilized to improve meat quality characteristics in pigs and other animals [38,39,40,41,42]; however, the results were often inconsistent. Rock samphire (Crithmum maritimum L.), also known as sea fennel, is an edible halophyte and xerophyte that grows wildly on the Mediterranean and Atlantic coasts of Europe [43]. The leaves, flowers and schizocarps of C. maritimum L. consist predominantly of carbohydrates (>65%), wit h lesser amounts of ash, proteins and lipids. Sea fennel’s briny, tender leaves provide omega-6 and omega-3 polyunsaturated fatty acids, particularly linoleic acid. Extracts derived from flowers and fruits or schizocarps are abundant in antioxidants, polyphenols, vitamins and carotenoids, exhibiting antibacterial efficacy against Staphylococcus aureus, Staphylococcus epidermidis, Candida albicans and Candida parapsilosis [44,45]. Camelina (Camelina sativa L. Crantz), also known as false flax, is an oilseed crop belonging to the Brassicaceae family that can grow in different climatic and soil conditions. It has been evaluated in pig diets due to the fatty acid (FA) profile of its oil extract, which is exceptionally high in polyunsaturated FA and particularly alpha-linolenic acid (ALA), an essential omega-3 fatty acid that is beneficial for enhancing cardiovascular health [46,47], as well as its much higher protein and lysine contents than corn or other cereal grains. Blood plasma omega-3 fatty acid levels of pigs can rise when camelina is added to their food, and their serum triglyceride levels can drop [48]. Some types of camelina have compounds (glucosinolates, condensed tannins and sinapine) that make the feed taste bitter, which can potentially cause pigs to consume less [49,50]. Garlic (Allium sativum L.) has been used in pig diets due to its antifungal, antimicrobial and antioxidant properties [51,52]. Garlic extract contains compounds such as allicin, alliin, phenols and flavonoids, which have been demonstrated to exhibit antioxidant action through free radical scavenging mechanisms. Allium spp. derivatives, such as organosulfurate compounds, alter the gut microbiota by affecting both beneficial bacteria (Bifidobacterium spp. or Lactobacillus spp.) and pathogenic bacteria (Escherichia coli, Salmonella typhimurium or Clostridium spp.) [53,54]. To the best of our knowledge, it is the first time that such a combination of the previously mentioned four plant extracts and essential oils has been evaluated in weaned pig diets.

2. Materials and Methods

2.1. Experimental Design, Animals and Diets

All experimental protocols adhered to the National Guidelines for Animal Trials (PD, 2013) and received approval from the authorities of the School of Agriculture at the University of Ioannina, Greece (UOI University Research Committee research registration: 61291/135/10.06.2020. A veterinary surgeon and an animal scientist from the Department of Agriculture at the University of Ioannina oversaw the farm environment and the piglets throughout the entire experimental period. The collaborating commercial pig farm was situated in the region of Epirus, Greece, and fulfilled the national regulations and the European directive for the protection of animal welfare in research (Directive 2010/63/EU, European Commission, 2010). According to Eurostat data, the pig population in Greece was estimated at approximately 786,200 heads in 2024 [55]. Domestic pork production remains insufficient to meet national demand, covering only 30–35% of consumption, which necessitates substantial imports. Intensive production systems constitute the dominant farming model, particularly farrow-to-finish units, with the average farm maintaining around 200 sows. In these systems, pigs are generally provided with ad libitum dry feed composed primarily of cereal-based diets supplemented with essential amino acids, vitamins and minerals. Feed expenses represent a major cost component, accounting for approximately 55–70% of total production costs. Nevertheless, organic and pasture-based systems have been gaining momentum since the early 2000s, reflecting increasing interest in alternative and potentially more sustainable production practices.
Table 1 presents the composition of the phytobiotic mixture (PM) tested in the present trial, with the oregano oil added in a micro-encapsulated form, produced by the methodology described by Partheniadis et al. [56]. The liquid forms of the essential oils of oregano and rock samphire were mixed thoroughly with the powder form of camelina and garlic, which were previously ground to provide a fine flour (Table 2).
The Institute of Plant Breeding and Genetic Resources (IPB&GR) in Thessaloniki, Greece, provided plant samples of Origanum vulgare subsp. hirtum and Crithmum maritimum L. The biomass was dried at room temperature in the shade using a 50 L pilot-scale steam distillatory apparatus with a steam pressure of 1.2 atm. It was then distillated for 1.5 h for Origanum vulgare subsp. hirtum and 1 h for the other three species.
The dried plant components were hydro-distilled for two hours with a Clevenger-style apparatus coupled to a specially refrigerated EO container.
Essential oils (EOs) for the trial were extracted from Origanum vulgare subsp. hirtum (IPEN: GR-1-BBGK-03,2107) and Crithmum maritimum L. (IPEN: GR-1-BBGK-97,719), provided by ELGO-DIMITRA’s Institute of Plant Breeding and Genetic Resources. Additionally, Camelina sativa L. Crantz seeds and Allium sativum L. bulbs were supplied by a Union of Agricultural Cooperation (Vyssa, Greece). The methodology used to process the samples was based on that described by O’Fallon et al. [57]. The fatty acid composition was determined by gas chromatography; fatty acids methyl esters were obtained from the samples. Then, the separation and quantification of the methyl esters was carried out with a gas chromatographic system (TraceGC model K07332, ThermoFinnigan, ThermoQuest, Milan, Italy) equipped with a flame ionization detector and a fused silica capillary column (phase type SP-2380, Supelco, Bellefonte, PA, USA). Individually identified fatty acids were reported as percentages (%) of the total identified acids. The herbal mixture was formulated to provide in the feed: 100 or 200 mL/kg Origanum vulgare subsp. hirtum essential oil; 25 mL/kg Crithmum maritimum L. essential oil; 0.5 g/kg dried Camelina sativa L.; and 0.5 g/kg dried Allium sativum L. (Table 1).
A total of 45 crossbred weaned pigs (1/4 Large White, 1/4 Landrace, 1/2 Duroc) at 34 days old were randomly assigned to one of three treatment groups—CONT, PM-A or PM-B. The control treatment group (CONT) was fed a commercially formulated maize-based diet suitable for weaned pigs, adhering to the guidelines established by the National Research Council [58] and the Premier Nutrition database [59]. The remaining two treatments received diets containing 0.2% of the evaluated quadruple phytobiotic mixture (PM), PM-A with 200mL/t feed oregano oil or PM-B with 400mL/t feed oregano oil. All three diets were designed to be isonitrogenous and isocaloric. The ingredient composition and proximate analysis of the experimental diets are presented in Table 3.
The initial body weights of the pigs were comparable throughout the three groups, with an average mean body weight of 8.31 ± 0.94 kg. Each group had an numbers of females (7) and males (8). Each pig was distinctly identified with numbered plastic ear tags. The pigs in each treatment group (n = 15) were accommodated in pens with slated plastic flooring, and maintained under regulated environmental conditions (ambient temperature, average humidity, ventilation rate and animal density) appropriate for their production stage. All pigs were vaccinated against porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae and Aujeszky’s disease, under the farm’s regular management protocols. Access to feed and water was provided ad libitum.
The trial lasted for 43 days (from 34 to 77 days of age), during which pigs were weighed individually on the mornings of the 1st, 22nd and 43rd days using a Mini–L 3510 scale for animals (Zigisis S.A., Chalkidiki, Greece), while feed intake and mortality data were documented daily. To evaluate the impact of dietary interventions on the performance metrics of pigs, the average gain (AG, kg per period), average feed intake (AFI, kg feed intake per period) and feed conversion ratio (FCR, kg feed intake/kg live weight gain) were recorded. On the final day of the trial, blood samples were collected from eight pigs (four males and four females) in each group, after which these pigs were humanely slaughtered in a modern abattoir near the experimental farm.

2.2. Μicrobial Analysis

Digesta samples were taken aseptically from the ileum and caecum of eight animals (four males and four females) per treatment immediately after slaughter on day 43 of the trial. Regarding isolation, enumeration and identification of bacterial populations, 1 g of intestinal material was homogenized with 9 mL of a 0.1% sterile peptone water solution. Bacterial enumeration was based on the Miles and Misra plate method (surface drop). Each sample underwent repeated dilution through 12-fold dilutions (ranging from 10-1 to 10-12) utilizing typical 96-well plates for microdilution procedures. A total of 10 μL of each dilution was injected into the media and incubated properly. MacConkey agar and Kanamycin aesculin azide (KAA) agar (Merck, Darmstadt, Germany) were utilized for the isolation and enumeration of Enterobacteriaceae and Enterococci, respectively, while plates were incubated aerobically at 37 °C for 24 to 48 h. De Man, Rogosa and Sharpe (MRS) agar (Oxoid, Basingstoke, UK) and M17 agar (Lab M Limited, Lancashire, UK) were employed for the isolation and enumeration of Lactobacillaceae, while media were incubated at 37 °C for 48 h under anaerobic conditions. Total aerobic and anaerobic bacterial counts were assessed using plate count agar medium (Oxoid, Basingstoke, UK). Plates were incubated aerobically at 30 °C for 48 h and anaerobically at 37 °C for 48 to 72 h, respectively. Bacterial counts were determined by enumerating representative colonies from a suitable dilution using a microbial colony counter, with results represented as colony-forming units (CFU) x log per 1 g of wet weight material. Typical colonies grown on different media were then described and subcultured. All bacterial populations were classified using the automated Vitek 2 compact system (bioMérieux, Marcy l’Etoile, France), which yielded dependable and precise findings for a broad spectrum of Gram-positive and Gram-negative bacteria. The identification of Enterobacteriaceae, Enterococcaceae, Lactobacillaceae and Bifidobacteriaceae was conducted using the Vitek 2 compact system (bioMérieux, Marcy l’Etoile, France), as well as Gram-negative identification card (ID-GN), Gram-positive identification card (ID-GP) and CBC and ANC identification cards (bioMérieux, Marcy l’Etoile, France) [60].

2.3. Hematological and Biochemical Analysis of the Blood

Blood samples were obtained from eight pigs (four males and four females) per treatment before slaughter, for the assessment of hematological and biochemical parameters, on the final day of the trial (day 43). Feed was withdrawn from the feeders four hours prior to blood sampling. Initially, a sample of 4 mL blood was obtained from the pigs’ jugular vein and then transferred into vacutainer tubes containing ethylenediamine tetraacetic acid (EDTA). Hematological parameters (hemoglobin, erythrocytes, hematocrit, leucocytes, lymphocytes) were assessed using an MS4 automated analyzer (Melet Schloesing Lab, Osny, France), while biochemical parameters (albumin, alanine aminotransferase, aspartate aminotransferase, cholesterol, creatine kinase, glucose, total bilirubin, triglycerides) in serum were evaluated using the IDEXX VETTEST 8008 instrument (IDEXX LAB, Westbrook, ME, USA).

2.4. Chemical Analysis, pH Measurement and Color Analysis of the Meat

Meat samples from eight pigs per group were collected post-slaughter from the ham (biceps femoris), shoulder (triceps brachii) and belly (external abdominal muscles) and stored at –20 °C. At the same day, 200 g subsamples were homogenized using an industrial grinder (Bosch, Gerlingen, Germany). Moisture, crude protein, fat, collagen and ash contents were determined by near-infrared spectroscopy (FoodScan™ Lab, FOSS, Hillerød, Denmark) following AOAC 2007.04 [61]. The pH was measured in each muscle using a portable pH meter with a stainless-steel probe (HI981036, Hanna Instruments, Woonsocket, RI, USA). Meat color (L*, a*, b* values) was assessed with a CAM-System 500 Chromatometer (Lovibond, Amesbury, UK) according to the Hunter scale [62].

2.5. Oxidative Stability Analysis of the Meat

The lipid oxidation (malondialdehyde levels) of the meat samples was performed according to the method described by Florou-Paneri et al. [63] and Giannenas et al. [64]. In brief, the frozen specimens were thawed overnight at 4 °C, minced with a commercial food processor, wrapped in oxygen-permeable film and stored in a non-illuminated refrigerated cabinet at 4 °C for 7 days. On the 4th and 7th refrigeration days, subsamples were extracted from each sample and processed according to the methodology. Absorbance was measured at 532 nm relative to a blank sample utilizing a UV–Vis spectrophotometer (UV-1700 PharmaSpec, Shimadzu, Japan). Tetraethoxypropane at a concentration of 1.1 to 3.3 was utilized as a reference, and the results were quantified as nanograms of malondialdehyde (MDA) per gram of meat (ng/g).

2.6. Fatty Acid Analysis of the Meat

Samples for the fatty acid analysis of shoulder, ham and belly meat cuts were processed according to the guidelines established by O’Fallon et al. [57]. The separation and quantification of methyl esters were conducted using the methodology outlined by Skoufos et al. [25] utilizing a TraceGC (Model K07332, Thermofinigan, Thermoquest, Milan, Italy) fitted with a flame ionization detector. The fatty acid methylester retention time and elusion order were identified using as the reference standard the Supelco ‘37 Component FAME Mix’ (Sigma-Aldrich, Darmstadt, Germany). The percentages (%) of individually identified fatty acids were calculated as their peak areas divided by the total peak areas of all identified fatty acids. Additionally, the total polyunsaturated fatty acid-to-saturated fatty acid (PUFA/SFA) ratio, the PUFA n-6/n-3 ratio and the hypocholesterolemic/hypercholesterolemic fatty acid ratio (h/H) were calculated. The h/H index illustrates the connection between low-cholesterol fatty acids (C18:1n-9 + PUFA) and high-cholesterol fatty acids (C12:0, C14:0 and C16:0). This index can be utilized to assess the cholesterolemic effect of dietary lipids [65].

2.7. Statistical Analysis

The fundamental study design employed was a random complete block design (RCB), with each ear-tagged pig serving as the experimental unit. Microbiology data underwent log transformation (Log10) prior to the statistical analysis. Levene’s test was employed to assess data homogeneity. Experimental data were analyzed using a one-way analysis of variance (one-way ANOVA) or the Kruskal–Wallis test, contingent upon the data format, employing the SPSS v20 statistical program [66]. Tukey’s test was conducted for post hoc comparisons among the three treatments when significant effects were identified by the one-way ANOVA. The significance threshold for all tests was established at 5% (p ≤ 0.05).

3. Results

3.1. Performance Parameters

The performance parameters of pigs were influenced by the dietary use of the phytobiotic mixture, as illustrated in Table 4. The final body weight and average daily weight gain of the pigs exhibited no significant differences (p > 0.05) between the three treatments. The feed intake and feed conversion ratio were consistent with the anticipated values for the commercial pig farm conducting the experimental trial and exhibited no significant differences between treatments (p > 0.05), even though the piglets were in the critical post-weaning phase of their development. Concerning the carcass parameters, carcass weights and dressing percentages exhibited no significant differences (p > 0.05) among the three treatments.

3.2. Intestinal Microflora

The dietary inclusion of the phytobiotic mixture induced significant alterations in the intestinal microflora populations (Table 5). In the ileum digesta, total aerobic bacterial counts (TABCs) were significantly increased (p ≤ 0.05) in the PM-A group compared to the control. Although a similar trend was observed in the PM-B group, the increase did not reach statistical significance relative to either the control or PM-A groups. Furthermore, the Enterobacteriaceae population—composed of E. coli and E. fergusonii (Table 6)—was lower in the PM-B group, showing a significant reduction when compared to PM-A and control groups. Lactobacillaceae counts, predominantly consisting of Limosilactobacillus reuteri, Streptococcus alactolyticus, Streptococcus hyointestinalis and Lactobacillus johnsonii (Table 6), were not affected by the treatments in the ileum. In the caecum, both treatment groups exhibited reduced Enterobacteriaceae levels, with the most abundant species being E. coli and E. fergusonii (Table 6); however, statistical significance (p ≤ 0.05) was noted only for PM-B relative to the control. Lactobacillaceae counts were significantly reduced in both PM-A and PM-B groups (p ≤ 0.05), whilst the population of total anaerobe counts remained consistent (p > 0.05) across all treatments in both the ileum and caecum. The most abundant species Lactobacillaceae were Limosilactobacillus reuteri and Streptococcus alactolyticus, with the highest occurrence percentages in all experimental groups, followed by Lactobacillus johnsonii, Limosilactobacillus mucosae, Lactobacillus gasseri, Lactobacillus delbrueckii and Streptococcus hyointestinalis (Table 6). Finally, no statistically significant differences were observed in Enterococcaceae counts among the groups in either the ileum or caecum. The most abundant species was E. faecium, present in all samples, with only single isolates of E. durans and E. hirae detected in the PM-B and control groups of the caecum and ileum samples, respectively (Table 6).

3.3. Blood Parameters

The effects of the phytobiotic mixture on pig blood hematological and biochemical parameters are presented in Table 7. No significant differences (p > 0.05) were observed in hematological and biochemical parameters among the three groups.

3.4. Chemical Analysis of the Meat

Table 8 indicates that there were no significant differences (p > 0.05) in any of the assessed parameters (collagen, fat, moisture, protein, and ash) in shoulder meat (triceps brachii) cuts. In the case of ham meat (biceps femoris), the protein content was significantly lower (p ≤ 0.05) in group PM-A compared to the control group, while group PM-B exhibited protein levels comparable to both the control and PM-A groups. No significant differences (p > 0.05) were observed in any of the parameters examined for the belly meat cuts (external abdominal).
The pH measurements (Table 8) for shoulder (triceps brachii) and ham (biceps femoris) meat showed no significant differences between the treatments (p > 0.05). The PM-A group tended to have a lower pH level (0.05 < p ≤ 0.10) in the external abdominal belly meat, in comparison to the other two groups.
Regarding the color measurements (Table 9), the color of the shoulder meat (triceps brachii) samples was significantly (p ≤ 0.05) redder (increased a* value) in group PM-B, while L* and b* values did not differ between all groups. Ham meat (biceps femoris) color exhibited significantly (p ≤ 0.05) higher a* values and lower b* values for both treatment groups PM-A and PM-B, respectively. A similar pattern was observed for lightness (L* value), where the supplemented groups PM-A and PM-B were significantly darker (p ≤ 0.05), with group PM-B displaying the lowest (61.18) L* value. The belly meat (external abdominal) color analysis revealed significantly (p ≤ 0.05) lower a* values for both groups PM-A and PM-B and a higher L* value for group PM-A but only compared to the control group. Yellowness (b* value) did not differ (p > 0.05) between treatments.
Data on oxidative stability of meat samples are presented in Table 10. In the ham meat (biceps femoris), the malondialdehyde (MDA) level on the 4th day of refrigerating was the lowest (p ≤ 0.05) in the PM-B treatment (3.70 ng/g), intermediate in the PM-A treatment (4.35 ng/g) and highest in the control treatment (5.55 ng/g). Furthermore, the MDA level was significantly lower (p ≤ 0.05) in the shoulder meat (triceps brachii) of group PM-B (5.27 ng/g) compared to both group PM-A (7.36 ng/g) and control (9.94 ng/g). In the belly meat (external abdominal), the control group tended to have higher MDA levels (0.05 < p ≤ 0.10) compared to the phytobiotic-enriched groups (PM-A and PM-B). Malondialdehyde (MDA) levels on the 7th day of refrigerating shoulder meat (triceps brachii) and belly meat (external abdominal) samples were significantly (p ≤ 0.05) lower for treatment groups PM-B and PM-A (17.56 ng/g and 22.51 ng/g, respectively). In the ham meat (biceps femoris), the PM-B group tended to (0.05 < p ≤ 0.10) have the lowest MDA level (18.91 ng/g) among all groups.
The fatty acid analysis of shoulder meat cuts is presented in Table 11. The dietary supplementation of the examined phytobiotic mixture modified (p < 0.05) the fatty acid compositions of most examined fatty acids compared to the control treatment. Overall, the total saturated fatty acids (SFAs) were lowest (p ≤ 0.05) in the control treatment, intermediate in the PM-A treatment and highest in the PM-B treatment. The total monounsaturated fatty acids (MUFAs) were highest (p ≤ 0.001) in the PM-A treatment, intermediate in the control treatment and lowest in the PM-B treatment. The total polyunsaturated fatty acids (PUFAs) were highest (p ≤ 0.001) in the PM-B treatment, intermediate in the control treatment and lowest in the PM-A treatment. The total omega-6 fatty acids were highest (p ≤ 0.001) in the PM-B treatment, intermediate in the control treatment and lowest in the silage PM-A treatment. The polyunsaturated fatty acid (PUFA)-to-saturated fatty acid (SFA) ratio (PUFA/SFA) was highest (p ≤ 0.001) in the control treatment, intermediate in the PM-B treatment and lowest in the PM-A treatment. Finally, the hypocholesterolemic/hypercholesterolemic fatty acid ratio (h/H) tended to be higher (0.05 < p ≤ 0.10) in treatment PM-B compared to the other two treatments.
The fatty acid composition of the belly meat cuts is presented in Table 12. The phytobiotic mixture added in the pig diets significantly altered (p < 0.05) the fatty acid compositions of most of the analyzed fatty acids when compared to the control treatment. Concerning the saturated fatty acids (SFAs), the PM-A treatment exhibited the lowest levels (p ≤ 0.001), followed by the PM-B treatment, while the control treatment had the highest concentrations. In terms of total monounsaturated fatty acids (MUFAs), they were lower (p ≤ 0.05) in the control treatment compared to both PM-A and PM-B treatments. The total polyunsaturated fatty acids (PUFAs) were highest (p ≤ 0.001) in the PM-A treatment, intermediate in the control treatment and lowest in the PM-B treatment. Specifically, the levels of total omega-3 fatty acids were significantly greater (p ≤ 0.05) in the PM-A treatment compared to the control treatment. Conversely, total omega-6 fatty acids were lowest (p ≤ 0.001) in the PM-B treatment, intermediate in the control treatment and highest in the PM-A treatment. The omega-6-to-omega-3 fatty acid ratio was lowest (p ≤ 0.001) in the PM-B treatment, intermediate in the PM-A treatment and highest in the control treatment. The ratio of polyunsaturated fatty acid (PUFA)-to-saturated fatty acid (SFA) (PUFA/SFA) was highest (p ≤ 0.001) in the PM-A treatment, intermediate in the control treatment and lowest in the PM-B treatment. Lastly, the hypocholesterolemic-to-hypercholesterolemic fatty acid ratio (h/H) was highest (p ≤ 0.001) in the PM-A treatment, intermediate in the control treatment and lowest in the PM-B treatment.
The fatty acid composition of the ham meat cuts Is presented in Table 13. Dietary supplementation with the phytobiotic mixtures significantly altered (p < 0.05) the profiles of most fatty acids compared with the control group. In particular, total saturated fatty acids (SFAs) were lowest (p ≤ 0.001) in the PM-A treatment, intermediate in the control and highest in the PM-B treatment. Conversely, total monounsaturated fatty acids (MUFAs) were highest (p ≤ 0.001) in the PM-A group, intermediate in the control and lowest in the PM-B group. The total PUFAs were significantly higher (p < 0.05) in the control group but only compared to PM-A treatment. The total omega-6 fatty acids were significantly lower (p ≤ 0.05) in the PM-A treatment compared to the control group. The ratio of omega-6 to omega-3 fatty acids tended to increase (0.05 < p ≤ 0.10) in both treatments PM-A and PM-B. The polyunsaturated fatty acid (PUFA)-to-saturated fatty acid (SFA) ratio (PUFA/SFA) was significantly lower (p ≤ 0.05) in the PM-B treatment compared to both the PM-A treatment and the control group. Finally, the hypocholesterolemic/hypercholesterolemic fatty acid ratio (h/H) was highest (p ≤ 0.001) in the PM-A treatment, intermediate in the control group and lowest in the PM-B treatment.

4. Discussion

The growing demand for high-quality animal protein in human nutrition necessitates advancements in modern animal production. Sustainably produced food with advanced qualitative and health promoting characteristics is gaining global interest [67,68,69,70]. The economic potential and nutritional quality of pork have established it as a significant global commodity, evidenced by rising consumption [2]. The nutritional profile of pork supports a healthy, balanced and safe diet for the expanding global population [71,72]. Nevertheless, the concerns about using antibiotics as growth enhancers in the feeds of pigs and other productive animals have prompted the exploration of superior alternatives that can deliver comparable advantages and effectiveness without significant adverse consequences. As a result, numerous chemicals have been analyzed and recognized as advantageous for improving health and overall growth of livestock. These substances are essential for maintaining normal physiological functions and protecting the animals from infectious diseases. Assessing natural and phytogenic feed additives as potential substitutes for antibiotic growth promoters in swine production has led to encouraging results. Phytogenic products offer substantial benefits compared to antibiotic medications, being economically viable and demonstrating a lower propensity for resistance emergence; hence, research on phytogenic products is increasing.
Phytobiotics typically include vital nutrients such as carbohydrates, along with secondary elements such as essential oils and phenolic compounds. A range of studies have demonstrated the advantageous effects of their use in modern animal nutrition, highlighting numerous beneficial outcomes. The observed effects include growth enhancement, improved gut microbial balance and beneficial bacteria growth, in addition to antioxidative, antimicrobial and anti-inflammatory characteristics [12,26,73]. Various aromatic and medicinal plants and their essential oils have been reported in the literature to provide beneficial outcomes in pig producibility. Nonetheless, researchers are investigating the prospect of boosting essential oils’ effectiveness and general bioactivity through their combination, owing to identified synergistic reactions among different substances [74,75]. This synergistic effect has been demonstrated in many studies where combining multiple essential oils and herbs led to more effective results concerning improved performance, regulation of gut microbiota and nutrient absorption, immune status and antioxidative effects [16,76,77,78] than their individual components [26,27], while the mechanism of these results remain inadequately elucidated. A mixture of phytobiotics with the aforementioned quadruple synthesis has been tested as an aqueous or cyclodextrin extract in broiler chickens, with positive results on composition and oxidative status of the meat, growth performance and welfare [79].
In this trial, a novel quadraple mixture of phytobiotics consisting of oregano (Origanum vulgare subsp. hirtum) essential oil, rock samphire (Crithmum maritimum L.) essential oil, garlic flour (Allium sativum L.) and false flax flour (Camelina sativa L. Crantz) was used as a feed ingredient for the diet of weaned pigs, to study its effects on their performance, health and meat quality characteristics. The two tested mixtures differed in their dosages of oregano essential oil, provided in a micro-encapsulated form (Table 1). Essential oils are rapidly metabolized in the upper gastrointestinal tract, resulting in concentrations in the distal small intestine insufficient to enhance intestinal function [80]. Therefore, using essential oils processed with an innovative microencapsulation technology may protect against oxidation and sustained release of active ingredients in the pig gut [64,81,82] or even mask strong odors not acceptable by some animals [83]. Furthermore, we explored the potential synergistic effects of their bioactive compounds’ combination. Ingredient costs for phytobiotics and essential oils can vary considerably depending on factors such as raw material quality, geographical origin and procurement scale. Locally sourced oregano and garlic, particularly in Greece where oregano is of high quality, can significantly reduce expenses compared to imported products. In contrast, sea fennel (Crithmum maritimum L.) essential oil remains a niche and high-cost ingredient, necessitating low inclusion rates to maintain economic viability. Camelina sativa meal offers potential cost offsets when used as a partial replacement for protein sources such as soybean meal. Additionally, logistical considerations, the formulation form (e.g., pure oil versus encapsulated) and compliance with European Union feed additive regulations can substantially influence the final cost of incorporating these additives into pig diets. In this trial, for pigs aged 34–77 days consuming approximately 30 kg of feed per head for the experimental period, the inclusion of oregano essential oil (200 mL or 400 mL/t), sea fennel essential oil (50 mL/t), Camelina sativa flour (1 kg/t) and garlic flour (1 kg/t) resulted in additive costs of about €63.30/t (PM-A) and €73.30/t (PM-B), respectively. This translated to €1.90–€2.20 per pig for the whole feeding phase (≈€0.044–€0.051/day). Sea fennel EO accounted for the largest share of the cost, while the overall expenditure remained relatively low, making targeted application in the weaner or early grower stage a cost-effective strategy to support gut health and performance if proven by results.
Growth control and enhancement through improvements in digestion, absorption of nutrients and feed consumption are factors that decisively affect farm productivity and profitability indices. Numerous studies have revealed that phytobiotics may enhance pig growth by improving feed quality and consumption, increasing flavor and palatability and promoting anabolic activity similar to that of anabolic agents [15,84,85]. Suggested modes of action may include the improvement of nutritional digestion and absorption via stimulation of the synthesis of various digestive secretions such as saliva and bile, as well as digestive enzymes such as amylase, galactosidase, lactase and sucrase [12,23], leading to elevated digestibility of feed dry matter (DM), crude protein (CM), gross energy (GE) and ether extracts (EE) [86,87,88]. Further suggested mechanisms involve enhanced function of the intestinal barrier and an improved ratio of the gut villus height to crypt depth [23,89,90]. Sanchez et al. [76] tested whether an extract of Allium spp., which includes garlic and onion, could improve the growth of growing–finishing pigs by changing the microbiome and short-chain fatty acid metabolism in the digestive tract, with promising results for average daily gain (ADG). In another trial, weaned piglets fed an allium extract supplement were compared to control and antibiotic (colistin and zinc oxide)-treated piglets. The allium extract-fed piglets had higher body weight (BW), average daily weight gain (ADG) and feed conversion ratio (FCR) values that were only similar to the antibiotic-treated group [91]. Several other studies have attributed elevated growth performance to mixtures of plant and herbal extracts containing mainly carvacrol, thymol, cinnamaldehyde, thyme, garlic and other medicinal plants from various countries [19,92,93]. On the contrary, there is research indicating no significant influence of phytobiotic inclusion in pig diets regarding growth performance (weight gain and average daily feed intake) [37,94,95]. Our results, with the use of the phytobiotic mixture, are partly in agreement with the above findings, since it was noted that the final pig BW, ADWG, ADFI and FCR values exhibited no significant differences (p > 0.05) across the three treatments (Table 4). Similar findings were reported for carcass weight and carcass percentage at the end of the trial period. This reveals a degree of inconsistency in digestibility or palatability improvements, as reported previously. This may be due to endogenous loss caused by stimulated secretion of mucus induced by various plant extracts [96] or the fact that phytobiotics’ bioactive compound effects are a complex combination of the species variation, plant part or essential oil, chemical composition and dosage level.
The diet is a significant determinant influencing the makeup and functionality of the gut microbiota in pigs [97]. The composition of the gut microbiome and its associated bacterial metabolic products greatly affect intestinal health, nutritional absorption, overall pig health and meat quality [60,98,99]. Weaned pigs encounter stressors related to alterations in their diet and surroundings. Such changes may impede the formation of a stable gut environment and lead to increased diarrhea scores. An optimal diet directly influences digestibility and is crucial for intestinal barrier function, immune system development and feed utilization, thereby enhancing pig growth [100,101]. Numerous trials investigating phytobiotics and essential oils have reported enhancements, neutral outcomes or even detrimental effects on the regenerative capacity of pig intestinal epithelial cells regarding villus length, the ratio of the villus height to crypt depth, gastrointestinal health and absorptive efficiency [87,102]. Phytobiotics can aid in regulating and enhancing the digestive process, promote the establishment of a balanced intestinal microflora and ameliorate overall health and function of the gastrointestinal tract in pigs [103,104] through enzyme activation or intestinal microflora modification [22,105]. Several studies have suggested that gut microbes play a role in metabolizing ingested phytobiotics into simpler metabolites, thereby increasing the bioavailability of the phytobiotics in the gut of young pigs. Phytobiotic mixtures have also been shown to exhibit prebiotic and probiotic activities, promoting the growth and activity of beneficial gut bacteria [84,98]. Increased populations of Lactobacillus spp. and Bifidobacterium spp., increased total bacterial counts and decreased Enterobacteriaceae levels in the gastrointestinal tract [26,106,107] of pigs fed phytobiotics have been reported. Conversely, there is research that supports the incorporation of phytobiotics in pig diets, resulting in lowered total aerobe, E. coli, Campylobacter jejuni, Clostridium spp. and Salmonella spp. counts [23,76,108] or even exerting no influence on the microbial population and composition of the gastrointestinal tract [96]. In the present study, total aerobic bacterial counts (TABCs) in the ileum were significantly increased (p ≤ 0.05) in the PM-A group, and Enterobacteriaceae populations—predominantly composed of E. coli—were statistically decreased in the PM-B group (Table 5 and Table 6) This reduction in Enterobacteriaceae populations was also apparent in the caecum for the diets that contained the phytobiotic mixture, in agreement with the above-mentioned research. In contrast, although Lactobacillaceae counts were significantly reduced in the caecum for both groups PM-A and PM-B, their presence in all groups in the ileum and caecum promoted gastrointestinal health, as L. johnsonii and L. delbrueckii improve growth performance, intestinal barrier function and the ability of pigs to resist against pathogens, inhibiting the growth of harmful bacteria and enhancing the immunity response. Limosilactobacillus reuteri increases average daily gain and nitrogen digestibility levels, particularly in post-weaning piglets, and also stimulates the immune system and alters swine gut microbiota, leading to a more beneficial balance of bacteria in the gut. All three aforementioned Lactobacilli species have been proposed as future probiotics [109,110,111]. The combined results of lower counts of Lactobacillaceae and Enterobacteriaceae in our experiment indicate that the phytobiotic mixture used here had a direct impact against possible bacterial pathogens, as the production of lactic acid was degraded due to lower numbers of Lactobacilli present in the large intestine.
This research examined porcine hematological and biochemical markers as indicators of general health status [112]. These indices are critical markers of a pig’s physiological health, offering vital information regarding its capacity to adjust to diverse physiological obstacles or emerging nutrient deficiencies [113]. Numerous research studies have been undertaken to examine the impacts of phytobiotics on hematological and biochemical parameters of swine. Elevated serum levels of IgA, IgG, red blood cells (RBCs) and white blood cells (WBCs) were seen following the administration of herbal extracts to weaning and finishing pigs [114,115]. Correspondingly, enhancements in serum IgG, IgA, IgM, albumin (ALB) and alkaline phosphatase (ALP) and a decrease in aspartate transaminase (AST) were identified [93,116]. Our trial revealed no significant differences (p > 0.05) in hematological and biochemical parameters among the three experimental groups; all measured values remained within the defined physiological limits for pigs (Table 7) This strongly suggests that the animals were healthy and the examined diets well balanced to meet their physiological requirements and welfare.
Meat is an important component for a healthy and balanced diet. It is a rich source of bio-available iron and zinc, selenium and various vitamins and minerals. In modern societies, consumers seek meat and meat products with specific chemical compositions and organoleptic characteristics. These involve an optimum lean-to-fat percentage, increased shelf life, color and flavor. Dietary changes can significantly modify the meat composition in monogastric animals. Many studies have reported various effects of incorporating phytobiotics in pig diets on meat composition and characteristics. Increased crude protein (CP) and amino acid concentration, lowered drip loss, improved lean meat percentage, enhanced intramuscular fat and improved pH levels are proven [93,117,118]. In this research, adding the quadruple phytobiotic mixture to the diet did not change the chemical makeup of the meat in terms of fat, collagen, moisture or pH. The only apparent difference was a significant (p ≤ 0.05) reduction in protein in ham cuts (biceps femoris) in group PM-A compared to the control group (Table 8) Similarly, reports of lower levels of crude protein (CP) or even critical amino acids such as lysine following the addition of phytobiotics are reported [119,120]. Color is one of the most important quality characteristics of meat, used as an indirect basis for consumer preference. The depth of meat color is primarily caused by the myoglobin (Mb) content, and since meat components that affect its color are highly susceptible to oxidation, the various levels of lightness, redness and yellowness of storage meat cuts are mainly attributed to the above oxidative process [118,121]. In this study, the phytobiotic mixture affected color to some extent; we recorded elevated a* values (redness) in group PM-A of ham (biceps femoris) meat cuts and groups PM-B of both shoulder (triceps brachii) and ham (biceps femoris) meat cuts. This may impose an advantage to these meat cuts, since the redness of pork is perceived as a desirable trait by consumers [122]. Decreased lightness (L* value) was notable in both PM-A and PM-B groups of ham (biceps femoris) meat cuts with a simultaneous reduction in yellowness (b* value). The belly (external abdominal) meat cuts had reduced a* values and increased L* values in both experimental groups PM-A and PM-B. Inclusion of oregano essential oil in animal diets can alter the color of meat by reducing hemoglobin oxidation and accelerating pigment distribution [41,123]. The yellow color is primarily influenced by the meat’s pH, which affects redox processes [121]. A small drop in pH can increase oxidized oxymyoglobin, making the meat yellower and slightly lighter. Li et al. [124] reported that pig feed supplemented with oregano essential oil yielded advantages in color, brightness, yellowness and lean meat percentage. More studies revealed similar increases in redness and yellowness or reductions in L* and b* values after feeding young or finishing pigs with various herbal mixtures and essential oils [114,118,120]. Our findings are consistent with most of the available literature, keeping in mind the large variability and inconsistency of the results, an issue discussed earlier. The pig carcasses and meat cut quality parameters were within acceptable limits for commercial use.
Weaning is a critical event that can cause physiological, environmental and social stress in piglets, hence increasing their vulnerability to intestinal dysfunction and oxidative stress [125,126]. Lipid oxidation is closely associated with the control of pathogenic or spoilage bacteria in meat, as well as the quality and sensory qualities of meat products. This is regarded as a crucial indicator of quality degradation in food and meat products [127]. Essential oils can enhance the oxidative stability of tissues, leading to improved product quality [128]. Integrating natural antioxidants into swine diets is an efficacious approach to augment antioxidant stability, increase sensory qualities and prolong the shelf life of pork products [77,129]. In a trial that incorporated a mixture of eucalyptus, oregano, thyme, lemon, garlic and coconut essential oils in pig feed, a reduction in malondialdehyde (MDA) was observed in combination with elevated total antioxidant capacity (T-AOC) and total superoxide dismutase (T-SOD) levels and the well-desired ‘marbling fat’ trait attributed by consumers with enhanced flavor. Likewise, reduced TBARS and MDA contents; improvements in gene expression of oxidative stability; and increased T-SOD, T-AOC and GSH-Px activity levels were all reported in trials that evaluated the effects of including various phytobiotic mixtures and essential oils in pig diets on meat oxidative stability [118,130]. However, trials evaluating oregano essential oils in pig diets showed no positive effect on lipid oxidation [26,28]. In the present study, malondialdehyde (MDA) levels on the 4th and 7th day of refrigeration were significantly lower (p ≤ 0,05) or tended to be (0.05 < p ≤ 0.10) the lowest in group PM-B for all meat samples (ham, shoulder and belly meat cuts). This finding is significant, as lipid oxidation and rancidity directly influence meat quality and storage, particularly during refrigeration or freezing. The likely explanation is that the metabolized and assimilated phytogenics may act as an exogenous antioxidant, improving the T-SOD, T-AOC and GSH-PX activity, thereby significantly reducing lipid peroxidation, lowering MDA concentrations, maintaining cell membrane integrity and ultimately enhancing the color and water retention of pork.
Pigs are monogastric and an excellent model for the study of lipid metabolism, since their meat reflects a fatty acid (FA) deposition profile based on the fatty acid composition of their diet. Therefore, in pig production, specific dietary nutrients such as fats play a crucial role in determining the quality and nutritional profile of the meat [44,131,132]. The contemporary diet in many nations is marked by a significant consumption of fats, particularly saturated and n-6 polyunsaturated fatty acids (PUFAs) [133,134]. Saturated fatty acids are associated with an elevated risk of cardiovascular disease (CVD), whereas a substantial consumption of monounsaturated and n-3 polyunsaturated fatty acids has demonstrated a protective impact [135,136,137]. Conversely, a diet abundant in n-6 polyunsaturated fatty acids is deemed unbalanced. Robust scientific evidence supports a reduction in n-6 intake and the augmentation of n3 consumption to enhance health across the lifespan, highlighting the significance of the n-6/n-3 PUFA ratio over the absolute quantities of each fatty acid family in the diet [138,139]. The n-3/n-6 PUFA ratio is a critical determinant of cell function, impacting membrane dynamics and diverse cellular processes. Currently, the n-6/n-3 fatty acid ratios in Western diets range from about 15:1 to 16.7:1, in contrast to the recommended ideal ratio range of 1:1 to 4:1 [138,140]. The ratio of polyunsaturated fatty acids (PUFAs) to saturated fatty acids (SFAs) (PUFA/SFA) is a standard metric for assessing the impact of diet on cardiovascular disease (CVD), and is the primary index for evaluating the nutritional value of foods, such as meat (0.11–2.042), fish (0.50–1.62) and dairy products (0.02–0.175). Although a large proportion of PUFAs alone is not necessarily beneficial if the n-6/n-3 ratio is not balanced, a value for the PUFA/SFA ratio greater than 0.4 is recommended for healthy foods and diets [141]. In contrast to the PUFA/SFA ratio, the hypocholesterolemic/hypercholesterolemic fatty acid ratio (h/H) may more precisely indicate the influence of the fatty acid content on cardiovascular disease, as was utilized in this research. The recommended ranges for meat and dairy products are 1.27–2.786 and 0.32–1.29, respectively. Ratusz et al. [142] analyzed the FA contents in 29 cold-pressed camelina (Camelina sativa) oils using the h/H ratio as a nutritional quality index, reporting relatively high numbers ranging from 11.7 to 14.7, which is desirable.
In the present study, the meat fatty acid composition was modified to an extensive degree by the inclusion of the phytobiotic mixture in the piglet feed. In the shoulder meat (triceps brachii), myristic (C14:0) and palmitic (C16:0) acids, associated with coronary heart diseases [143], were significantly (p = 0.001) or tended to (0.05 < p ≤ 0.10) be lower, respectively, in the experimental groups PM-A and PM-B. Oleic acid (C18:1 cis n-9) levels, consisting of the largest amounts of MUFA fatty acids, were significantly (p ≤ 0.001) increased in group PM-A. The PUFA/SFA ratio was highest (p ≤ 0.001) in the control treatment. In the belly meat (external abdominal), stearic (C18:0) and oleic (C18:1 cis n-9) acids were significantly (p ≤ 0.005) lower and higher, respectively, both in group PM-B. The Σ PUFA, Σ n-3, PUFA/SFA ratio and h/H ratio values were significantly higher (p ≤ 0.05) in the PM-A group. The n-6/n-3 FA ratio was significantly lower (p = 0.001) in group PM-B. In the ham meat (biceps femoris), myristic (C14:0) and palmitic (C16:0) acids were significantly (p ≤ 0.05) decreased, while oleic acid (C18:1 cis n-9) was significantly increased, all in group PM-A. The PUFA/SFA ratio was the lowest (p = 0.006) in group PM-B, while the h/H ratio was significantly (p ≤ 0.001) the highest in group PM-A. Overall, the values of the PUFA/SFA and h/H ratios observed in our study fall within the range denoted for meat and meat products in previous studies [144], and indicate a positive effect on pig meat similar to other studies. It has been documented that the inclusion of various phytobiotic mixtures and essential oils in pig diets has positive effects on n-3 FA levels [120,145] and increases MUFA levels [118] while decreasing SFA levels [93], especially palmitic (C16:0) and lauric (C12:0) acids, in pig meat, and these effects are conducive to consumer health.

5. Conclusions

This study evaluated for the first time the impact of a phytobiotic mixture consisting of oregano (Origanum vulgare subsp. hirtum) essential oil in two different doses, rock samphire (Crithmum maritimum L.) essential oil, false flax flour (Camelina sativa L. Crantz) and garlic flour (Allium sativum L.) on the productive, health and meat quality parameters of weaned pigs. The results indicate that the tested quadruple herbal mixture, containing various bioactive compounds, had notable effects regarding meat oxidative stability and fatty acid profile and color, alongside a potentially beneficial change in intestinal microbial balance, especially reducing Enterobacteriaceae counts. Some results, such as Lactobacillaceae reductions in the caeca, are inconsistent with other similar trials; however, such discrepancies can be largely ascribed to the varied array of assessed herbal components and the differing integration rates. Given that the origin, fat content, color and welfare of pigs significantly influence consumers’ preferences for pork, further research studies akin to the present are required to evaluate complicated phytogenic mixtures in swine diets, utilizing various ratios and supplementation durations or research to effectively enhance desirable traits and deepen the understanding of the absorption, distribution, metabolism and excretion of phytobiotics and essential oils. Furthermore, the major concerns for the broad application of phytobiotic mixtures and essential oils in pig diets remain the cost of application, the identification of optimal inclusion levels and possible interactions with other feed ingredients.

Author Contributions

Conceptualization, I.S., I.G., J.W., L.J. and K.G.; methodology, I.G., G.M., C.Z. and E.B.; software, G.M. and E.B.; validation, I.S. and I.G.; formal analysis, G.M., I.S., E.B. and C.Z.; investigation, G.M. and K.F.; resources, I.S. and G.M.; data curation, G.M. and E.B.; writing—original draft preparation, G.M., E.B. and I.S.; writing—review and editing, C.Z., I.G., K.F. and A.T.; visualization, I.S., E.B. and I.G.; supervision, I.S. and I.G.; project administration, I.S., A.T. and I.G.; funding acquisition, I.S., A.T. and I.G. All authors have read and agreed to the published version of the manuscript.

Funding

This research has been co-financed by Greece and the European Union (European Regional Development Fund) in the context of the “Research–Create–Innovate” Operational Program “Competitiveness, Entrepreneurship and Innovation (EPAnEK)”, NSRF 2014-2020. Project Code: T7ΔKI-00313 (MIS 5050735). Acronym: GreenPro.

Institutional Review Board Statement

The experimental protocol for this trial was reviewed and approved by the Ethics and Research Ethics Committee of the University of Ioannina of Greece (protocol number 61291/135/10/06/2020).

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author.

Conflicts of Interest

Author Lizhi Jin was employed by the company Guangzhou Meritech Bioengineering Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Table 1. Composition of the evaluated phytobiotic mixture (PM).
Table 1. Composition of the evaluated phytobiotic mixture (PM).
Plant Material/Feed AdditivePM-APM-B
Oregano
(Origanum vulgare subsp. hirtum)
(Essential oil) *		100 mL
(25 mL methylcellulose/75 mL oil)		200 mL
(50 mL methylcellulose/150 mL oil)
Rock samphire
(Crithmum maritimum L.)
(Essential oil)		25 mL25mL
Camelina
(Camelina sativa L. Crantz)
(Dried and flour form)		0.5 kg0.5 kg
Garlic
(Allium sativum L.)
(Dried and flour form)		0.5 kg0.5 kg
PM-A: phytobiotic mixture in diet of pig group A; PM-B: phytobiotic mixture in diet of pig group B. * Microencapsulated form.
Table 2. Chemical composition of phytobiotic mixture (plant extracts and essential oils).
Table 2. Chemical composition of phytobiotic mixture (plant extracts and essential oils).
Garlic
(Allium sativum L.) 1		Oregano
(Origanum vulgare subsp. hirtum) 1		Rock Samphire
(Crithmum maritimum L.) 1		Camelina
(Camelina sativa L. Crantz) 2
Compound%Compound%Compound%FA%
Diallyl trisulfide58.46Carvacrol78.72β-phellandrene28.01C11:0 (Undecanoic) 0.01
Diallyl disulphide24.54p-cymene8.19Sabinene20.96C12:0 (Lauric) 0.01
Diallyl tetrasulphide4.73γ-terpinene2.11γ-terpinene18.69C14:0 (Myristic) 0.11
3-Vinyl-1,2-dithiocyclohex-5-ene0.64Myrcene1.641,8-cineol9.53C15:0 (Pentadecanoic) 0.03
N,N-dimethyl-Ethanethioamide0.63β-caryophyllene1.27Thymol methyl ether4.07C16:0 (Palmitic) 8.29
Allyl methyl trisulphide4.42α-terpinene1.01cis-β-ocimene3.68C16:1 (Palmitoleic) 0.14
Dimethyl trisulphide1.25α-pinene0.98p-cymene3.55C17:0 (Heptadecanoic) 0.05
Apiol0.26cis-sabinene hydrate0.62Terpinen-4-ol2.66C17:1 (cis-10-Heptadecenoic)0.05
(methylsulfinyl)(methylthio)-Methane0.24Terpinen-4-ol0.55α-pinene2.42C18:0 (Stearic)2.24
Carvacrol1.22α-thujene0.48α-terpinene1.64C18:1n9t (Elaidic) 0.03
Epiglobulol0.18Borneol0.42Myrcene1.44C18:1n9c (Oleic) 15.36
3-Vinyl-1,2-dithiocyclohex-4-ene0.171-octen-3-ol0.38α-terpinolene0.91C18:2n6t (Linolelaidic)0.01
Hinesol0.16α-humulene0.30α-thujene0.48C18:2n6c (Linoleic)22.31
Patchoulane0.15Thymol0.28α-phenalldrene0.44C18:3n6 (γ-Linolenic) 0.00
p-Cymene0.14Limonene0.27trans-β-ocimene0.24C20:0 (Arachidic)1.11
1-Docosanol0.12Camphene0.25Allo-ocimene0.23C18:3n3 (a-Linolenic) 34.54
3-(Methylthio)pent-4-yn-1-ol0.11Caryophyllene oxide0.24β-pinene0.20C20:1n9c (cis-11-Eicosenoic) 11.14
D-Limonene0.09β-phellandrene 0.23Bicyclogermacrene0.14C20:2 (cis-11,14-Eicossadienoic)1.59
Isobutyl isothiocyanate0.08α-phellandrene0.18cis-2-p-menthen-1-ol0.11C22:0 (Behenic)0.13
Linalool0.07β-pinene0.16α-terpineol0.08C20:4n6 (Arachidonic) 1.00
cis-2-Thiabicyclo [3.3.0]Octane0.06α-terpinolene0.15β-caryophyllene0.08C22:1n9 (Erucic)1.50
Eucalyptol0.05δ-cadinene0.13Camphene0.07C22:2 (cis-13,16-Docosadienoic)0.09
Camphor0.05δ-3-carene0.10cis-sabinene hydrate0.07C20:5n3 (cis-5,8,11,14,17-Eicosapentaenoic)0.02
p-Cymen-7-ol0.05trans-β-farnesene0.10Caryophyllene oxide0.02C24:0 (Lignoceric)0.04
Linalyl butyrate0.04β-bisabolene0.10 C24:1n9 (Nervonic)0.15
Butyl isothiocyanate0.02Germacrene D0.08 Σ SFA (Total Saturated FA)12.07
1,8-cineol0.07 Σ MUFA (Total Monounsaturated FA)28.38
Σ PUFA (Total Polyunsaturated FA)56.83
1 Bioactive compounds; 2 fatty acids (FAs).
Table 3. Compositions and the calculated proximate analysis of experimental diets.
Table 3. Compositions and the calculated proximate analysis of experimental diets.
Ingredients, %CONTPM-APM-B
Maize33.4833.2833.28
Barley34.8034.8034.80
Phytobiotic Mixture (PM)0.000.200.20
Soybean meal (47% CP)16.8116.8116.81
Fishmeal 62% CP3.003.003.00
Wheat middlings3.003.003.00
Soybean oil1.911.911.91
Vitamin and mineral premix 6% *6.006.006.00
Zinc oxide0.300.300.30
Benzoic acid0.300.300.30
Monocalcium phosphate0.200.200.20
Salt0.200.200.20
Total100.00100.00100.00
Calculated proximate analysis
Digestible energy, MJ/kg14.1814.1814.18
Crude protein, %18.7818.7818.78
Dry matter, %88.3188.3188.31
Ash, %5.265.265.26
Crude fat, %5.005.005.00
Crude fiber, %3.413.413.41
ADF, %3.943.943.94
NDF, %11.2511.2511.25
Ca, %0.200.200.20
Total P, %0.430.430.43
Lysine, %1.111.111.11
Methionine + Cystine, %0.490.490.49
CONT: control pig diet group; PM-A: phytobiotic mixture in diet of pig group A; PM-B: phytobiotic mixture in diet of pig group B. * Provided per kg of diet: 15,000 IU vitamin A, 50 mcg 25-hydroxycholecalciferol, 9.96 mg vitamin E, 10.02 mg vitamin K3, 3 mg vitamin B1, 10.02 mg vitamin B2, 6 mg pantothenic acid, 6 mg vitamin B6, 40.02 mcg vitamin B12, 100 mg vitamin C, 35 mg niacin, 300 mcg biotin, 1.5 mg folic acid, 375 mg choline chloride, 200 mg ferrous sulfate monohydrate, 90 mg copper sulfate pentahydrate, 60 mg manganese sulfate monohydrate, 100 mg zinc sulfate monohydrate, 2 mg calcium iodate, 300 mg sodium selenide, 150 mg L-selenomethionine–selenium, 1500 FYT 6-phytase, 80 U β-1,4-endoglucanase, 70 U β-1,3 (4)-endoglucanase, 270 U β-1,4-endoxylanase, 5000 mg benzoic acid, 40,8 mg butylated hydroxytoluene, 3.5 mg propyl gallate.
Table 4. Effects of the phytobiotic mixture supplementation on performance parameters of pigs.
Table 4. Effects of the phytobiotic mixture supplementation on performance parameters of pigs.
CONTPM-APM-BSEMp-Value
Body weight on day (kg)
18.388.408.160.9430.571
2014.5614.1914.280.1850.752
4326.8926.5527.150.3300.754
Weight gain for days (kg)
1–206.195.796.120.1760.632
20–4312.3312.3712.870.2760.712
1–4318.5118.1518.980.3310.573
Feed intake per group for days (kg)
1–20169.7161.6166.9NANA
20–43283.18290.36283.44NANA
1–43452.88451.96450.34NANA
FCR for days (g feed/g weight gain)
1–201.83 1.86 1.82 NANA
20–431.53 1.56 1.47 NANA
1–431.63 1.66 1.58 NANA
Carcass parameters
Carcass weight (kg)19.3018.9219.920.2830.368
Carcass dressing percentage (%)67.6766.6767.500.0040.505
N = 15 pigs per group. NA = not applicable.
Table 5. Effects of phytobiotic mixture supplementation on intestinal (ileum and caecum) microflora populations of pigs.
Table 5. Effects of phytobiotic mixture supplementation on intestinal (ileum and caecum) microflora populations of pigs.
CONTPM-APM-BSEMp-Value
Ileum microbes (Log10 CFU/g)
Total Aerobic Bacterial Count (TABC)6.99 a8.08 b7.21 ab0.2010.044
Total Anaerobes 7.1467.7957.2650.1900.374
Enterobacteriaceae5.24 b5.78 b4.69 a0.3520.020
Enterococcaceae4.7585.7754.5560.2950.218
Lactobacillaceae6.7956.6846.6600.1830.959
Caecum microbes (Log10 CFU/g)
Total Aerobic Bacterial Count (TABC)8.5457.8828.6440.1890.072
Total Anaerobes8.3948.0718.2090.1480.689
Enterobacteriaceae7.27 b5.937 ab4.235 a0.4460.004
Enterococcaceae5.0715.1464.8010.2570.872
Lactobacillaceae7.715 b6.88 a6.793 a0.1380.013
a,b Values with no common superscript differ significantly (p ≤ 0.05).
Table 6. Isolated bacteria and their distributions (log cfu/g) and mean counts in the ileum and caecum (8 in total for each group).
Table 6. Isolated bacteria and their distributions (log cfu/g) and mean counts in the ileum and caecum (8 in total for each group).
CONTPM-APM-B
Isolated Bacteria/Ileum Samples (%)CountLog10 Samples (%)CountLog10 Samples (%)CountLog10
Enterococcus faecium8 (100%)5.448 (100%)6.538 (100%)4.65
Enterococcus hirae2 (25%)6.00----
Escherichia coli8 (100%)6.068 (100%)7.108 (100%)5.52
Escherichia fergusonii4 (50%)4.716 (75%)7.022 (25%)4.38
Lactobacillus amylovorus--2 (25%)7.002 (25%)7.85
Lactobacillus crispatus--2 (25%)7.30--
Lactobacillus gasseri4 (50%)7.18----
Lactobacillus johnsonii2 (25%)5.482 (25%)7.08--
Lactobacillus kitasatonis--2 (25%)6.082 (25%)7.30
Lactobacillus ultunensis--2 (25%)6.90--
Ligilactobacillus murinus2 (25%)5.00----
Ligilactobacillus salivarius----2 (25%)6.95
Limosilactobacillus mucosae2 (25%)7.70--4 (50%)7.58
Limosilactobacillus reuteri8 (100%)7.148 (100%)7.556 (75%)6.80
Streptococcus alactolyticus4 (50%)8.224 (50%)7.702 (25%)5.70
Streptococcus hyointestinalis2 (25%)8.484 (50%)7.702 (25%)5.30
Streptococcus infantarius--2 (25%)7.95--
Streptococcus oralis----2 (25%)7.85
Streptococcus pneumoniae----2 (25%)3.70
CONTPM-APM-B
Isolated bacteria/CaecumSamples (%)CountLog10Samples (%)CountLog10Samples (%)CountLog10
Enterococcus durans--- 2 (25%)5.00
Enterococcus faecium8 (100%)5.298 (100%)6.258 (100%)5.25
Escherichia coli8 (100%)7.588 (100%)6.148 (100%)4.79
Escherichia fergusonii--4 (50%)5.012 (25%)4.30
Lactobacillus delbrueckii--2 (25%)6.48-
Lactobacillus gasseri--- 2 (25%)5.95
Lactobacillus johnsonii2 (25%)7.004 (50%)6.932 (25%)6.00
Ligilactobacillus salivarius--- 2 (25%)6.30
Limosilactobacillus mucosae2 (25%)7.902 (25%)7.004 (50%)8.19
Limosilactobacillus reuteri8 (100%)7.658 (100%)6.748 (100%)7.01
Streptococcus alactolyticus8 (100%)8.628 (100%)7.998 (100%)7.72
Streptococcus hyointestinalis--2 (25%)5.30-
Table 7. Effects of phytobiotic mixture supplementation on blood hematological and biochemical parameters of pigs.
Table 7. Effects of phytobiotic mixture supplementation on blood hematological and biochemical parameters of pigs.
Hematological ParametersCONTPM-APM-BSEMp-Value
WBC (103/μL)23.4722.0321.701.3380.87
Lymphocytes (%)33.6537.2234.471.2460.50
Monocytes (%)6.276.537.550.5000.58
Granulocytes (%)61.1255.6256.631.4310.27
RBC (106/μL)6.066.036.930.4330.70
HCT (%)33.3032.8333.301.9101.00
HB (g/dL)11.7710.9511.520.6930.89
THR (m/mm3)271.17277.67287.1710.0070.811
Blood biochemical parametersCONTPM-APM-BSEMp-Value
ALB (g/dL)3.333.083.280.1500.796
ALP (UL)285.17278.33310.3313.6210.631
ALT (U/L)165.00168.00175.507.5660.854
AST (U/L)129.00125.50133.335.7750.86
CHOL (mg/dL)117.17113.50116.673.0290.887
GLU (mg/dL)130.00128.00131.004.2740.961
TBIL (mg/dL)2.121.902.351.3320.378
TRIG (mg/dL)59.6757.8358.172.1550.943
N = 8 pigs per group. WBCs: white blood cells; RBCs: red blood cells; HCT: hematocrit; HB: hemoglobin; THR: transient hyperemic response; ALB: albumin; ALP: alkaline phosphatase; ALT: alanin aminotransferase; AST: aspartate aminotransferase; CHOL: cholesterol; GLU: glucose; TBIL: total bilirubin; TRIG: triglycerides.
Table 8. Effects of phytobiotic mixture supplementation on shoulder, ham and belly meat chemical compositions and pH.
Table 8. Effects of phytobiotic mixture supplementation on shoulder, ham and belly meat chemical compositions and pH.
CONTPM-APM-BSEMp-Value
Shoulder meat (triceps brachii)
chemical composition (%)
Collagen1.631.601.650.5170.912
Fat6.096.276.230.2290.953
Moisture74.1675.6674.620.4430.379
Protein17.5618.0417.540.1130.163
Ash1.101.061.110.3040.776
pH5.825.855.850.2310.841
Ham meat (biceps femoris)
chemical composition (%)
Collagen1.341.431.240.37110.108
Fat6.046.146.140.16590.970
Moisture74.3475.4375.420.46420.628
Protein18.88 b18.15 a18.56 ab0.11890.028
Ash1.081.021.110.02470.361
pH5.685.785.610.03380.109
Belly meat (external abdominal) chemical composition (%)
Collagen1.861.851.810.4450.920
Fat8.978.198.290.3850.708
Moisture71.4971.4971.620.5700.955
Protein17.1117.0617.160.1410.954
Ash1.051.091.100.1600.409
pH5.675.785.770.2300.085
a,b Values with no common superscript differ significantly (p ≤ 0.05).
Table 9. Effects of phytobiotic mixture supplementation on shoulder, ham and belly meat colors.
Table 9. Effects of phytobiotic mixture supplementation on shoulder, ham and belly meat colors.
CONTPM-APM-BSEMp-Value
Shoulder meat
(triceps brachii) color
L*62.7462.0661.100.3220.105
a*10.30 a11.10 ab11.77 b0.2280.020
b*9.349.879.600.1690.464
Ham meat
(biceps femoris) color
L*65.85 b62.66 a61.18 a0.6890.008
a*9.01 a11.67 b12.09 b0.4350.002
b*11.07 b9.88 a9.68 a0.2200.011
Belly meat
(external abdominal) color
L*64.41 a68.63 b66.38 ab0.6740.026
a*11.42 b8.4 a9.3 a0.4430.007
b*9.8810.4510.520.2970.654
L*: lightness; a*: redness; b*: yellowness. a,b Values with no common superscript differ significantly (p ≤ 0.05).
Table 10. Effects of phytobiotic mixture supplementation on ham, shoulder and belly meat oxidative stability levels.
Table 10. Effects of phytobiotic mixture supplementation on ham, shoulder and belly meat oxidative stability levels.
Day 4, MDA (ng/g)CONTPM-APM-BSEMp-Value
Ham meat (biceps femoris)5.55 b4.35 a3.70 a0.250.003
Shoulder meat (triceps brachii)9.94 b7.365 ab5.27 c0.720.020
Belly meat (external abdominal)6.31 x4.71 y4.33 y0.390.085
Day 7, MDA (ng/g)CONTPM-APM-BSEMp-Value
Ham meat (biceps femoris)22.69 y20.45 xy18.91 x0.690.073
Shoulder meat (triceps brachii)24.28 b19.50 a17.56 a1.140.036
Belly meat (external abdominal)28.88 b23.25 a22.51 a0.970.006
MDA: malondialdehyde. a–c Values with no common superscript differ significantly (p ≤ 0.05). x,y Means (n = 8 per treatment) with no common superscript tend to (0.05 < p ≤ 0.10).
Table 11. Effect of phytobiotic mixture supplementation on shoulder meat fatty acid composition.
Table 11. Effect of phytobiotic mixture supplementation on shoulder meat fatty acid composition.
Shoulder Meat (Triceps Brachii)CONTPM-APM-BSEMp-Value
FA (%)
C8:0 (Caprylic 0.010.010.010.0020.808
C10:0 (Capric)0.04 a0.10 b0.07 ab0.0080.005
C12:0 (Lauric)0.07 a0.10 b0.07 ab0.004<0.001
C14:0 (Myristic)1.72 b1.76 b1.53 a0.0250.001
C14:1 (Myristoleic) 0.020.020.010.0020.321
C15:0 (Pentadecanoic) 0.05 b0.03 a0.05 b0.0030.003
C16:0 (Palmitic)27.46 xy27.69 y26.95 x0.1440.092
C16:1 cis (Palmitoleic)2.70 c2.46 b1.97 a0.0830.002
C17:0 (Heptadecanoic)0.37 ab0.20 a0.39 b0.0570.014
C17:1 (cis-10 Heptadecenoic cis)0.22 a0.25 b0.31 c0.010<0.001
C18:0 (Stearic) 11.05 a11.88 b12.96 c0.204<0.001
C18:1n-9t (Elaidic) 0.12 b0.06 a0.07 a0.007<0.001
C18:1 cis n-9 (Oleic) 39.18 b42.12 c38.03 a0.432<0.001
C18:2n-6t (Linolelaidic) 0.020.020.030.0020.233
C18:2 n-6c (Linoleic)14.85 b10.86 a15.17 b0.484<0.001
C18:3n-6 (γ-Linolenic) 0.09 b0.08 b0.04 a0.005<0.001
C20:0 (Arachidic)0.04 a0.07 b0.11 c0.008<0.001
C18:3n-3 (a-Linolenic) 0.78 b0.65 a0.93 c0.029<0.001
C20:1 cis n-9 (cis-11 Eicosenoic) 0.22 a0.55 c0.39 b0.033<0.001
C21:0 (Henicosanoic)0.020.020.020.0020.918
C20:2 cis n-6 (cis-11,14-Eicosadienoic)0.30 a0.37 b0.45 c0.016<0.001
C20:3 cis n-3 (cis-11-14-17-Eicosatrienoate) 0.190.430.250.0910.154
C20:4 cis n-6 (Arachidonic)0.73 c0.54 b0.36 a0.037<0.001
C23:0 (Tricosanoic)0.020.020.010.0020.905
C20:5 cis n-3 (Cis-5,8,11,14,17-Eicosapentaenoic)0.020.020.020.0020.926
C24:1n-9 (Nervonic)0.07 xy0.25 y0.06 x0.0570.080
C22:6 cis n-3 (cis-4,7,10,13,16,19-Docosahexaenoic)0.030.040.030.0030.419
Σ SFA (Total Saturated FA)40.84 a41.86 ab42.17 b0.2270.033
Σ MUFA (Total Monounsaturated FA)42.53 b45.71 c40.84 a0.508<0.001
Σ PUFA (Total Polyunsaturated FA)17.00 b13.00 a17.29 b0.487<0.001
Σ n-3 (Total omega-3 FA)1.021.131.230.0890.208
Σ n-6 (Total omega-6 FA)15.98 b11.87 a16.06 b0.483<0.001
Ratio n-6/n-3 FA15.8212.2513.910.8220.236
PUFA/SFA0.42 b0.31 a0.41 b0.012<0.001
h/H c1.92 xy1.87 x1.94 y0.0140.066
FA: fatty acids; ΣSFA = (C10:0) + (C12:0) + (C14:0) + (C16:0) + (C17:0) + (C18:0); ΣMUFA = (C16:1 cis) + (C17:1 cis-10) + (C18:1 cis n-9) + (C18:1 n-7) + (C20:1 cis n-9); ΣPUFA = (C18:2 n-6c) + (C18:4n-3) + (C20:2 cis n-6) + C20:3 cis n-6) + (C22:5 cis n-3) + (C20:3 cis n-3) + (C20:4 cis n-6) + (C20:5 cis n-3) + (C21:5 n-3) + (C22:6 cis n-3). a,b,c Values with no common superscript differ significantly (p ≤ 0.05). x,y Values with no common superscript tend to (0.05 < p ≤ 0.10). c Hypocholesterolemic/hypercholesterolemic ratio = (cis-C18:1 + ΣPUFA)/(C12:0 + C14:0 + C16:0).
Table 12. Effect of phytobiotic mixture supplementation on belly meat fatty acid composition.
Table 12. Effect of phytobiotic mixture supplementation on belly meat fatty acid composition.
Belly Meat (External Abdominal)
FA (%)		CONTPM-APM-BSEMp-Value
C8:0 (Caprylic)0.020.020.020.0030.649
C10:0 (Capric)0.14 b0.11 a0.12 a0.0050.001
C12:0 (Lauric)0.120.100.100.0030.220
C14:0 (Myristic)1.87 a1.91 a2.06 b0.025<0.001
C14:1 (Myristoleic)0.020.020.020.0030.901
C15:0 (Pentadecanoic)0.050.040.040.0030.191
C16:0 (Palmitic)30.88 a30.35 a32.51 b0.272<0.001
C16:1 cis (Palmitoleic)2.06 a2.24 b2.67 c0.067<0.001
C17:0 (Heptadecanoic)0.23 a0.29 b0.22 a0.009<0.001
C17:1 cis-10 (Heptadecenoic cis) 0.15 a0.26 ab0.37 b0.0550.001
C18:0 (Stearic) 14.91 b11.85 a12.10 a0.3570.002
C18:1n-9t (Elaidic)0.09 a0.21 b0.22 b0.015<0.001
C18:1 cis n-9 (Oleic) 35.07 a35.98 b35.91 b0.1610.024
C18:2n-6t (Linolelaidic)0.020.020.020.0020.869
C18:2 n-6c (Linoleic)12.01 b13.40 c10.55 a0.2900.001
C18:3n-6 (γ-Linolenic)0.04 b0.05 b0.03 a0.0030.008
C20:0 (Arachidic)0.090.110.100.0040.140
C18:3n-3 (a-Linolenic)0.700.770.750.0140.175
C20:1 cis n-9 (cis-11-Eicosenoic) 0.310.310.320.0050.384
C21:0 (Henicosanoic)0.020.020.020.0020.580
C20:2 cis n-6 (cis-11,14-Eicosadienoic)0.32 b0.32 b0.30 a0.0050.015
C20:3 cis n-3 (cis-11-14-17-Eicosatrienoate) 0.05 a0.07 b0.07 b0.0040.023
C20:4 cis n-6 (Arachidonic)0.31 b0.50 c0.25 a0.026<0.001
C20:5 cis n-3 (Cis-5,8,11,14,17-Eicosapentaenoic)0.030.040.020.0050.443
C24:1n-9 (Nervonic)0.05 b0.06 b0.04 a0.0030.007
C22:6 cis n-3 (cis-4,7,10,13,16,19-Docosahexaenoic)0.020.040.030.0040.428
Σ SFA (Total Saturated FA)48.47 c44.79 a47.27 b0.422<0.001
Σ MUFA (Total Monounsaturated FA)37.76 a39.08 b39.55b0.2420.003
Σ PUFA (Total Polyunsaturated FA)13.50 b15.19 c12.00 a0.3240.001
Σ n-3 (Total omega-3 FA)0.80 a0.91 b0.87 ab0.0200.043
Σ n-6 (Total omega-6 FA)12.70 b14.28 c11.13 a0.319<0.001
Ratio n-6/n-3 FA16.13 b15.76 b12.87 a0.4620.001
PUFA/SFA0.28 b0.34 c0.25 a0.009<0.001
h/H c1.48 b1.58 c1.38 a0.021<0.001
FA: fatty acids; ΣSFA = (C10:0) + (C12:0) + (C14:0) + (C16:0) + (C17:0) + (C18:0); ΣMUFA = (C16:1 cis) + (C17:1 cis-10) + (C18:1 cis n-9) + (C18:1 n-7) + (C20:1 cis n-9); ΣPUFA = (C18:2 n-6c) + (C18:4n-3) + (C20:2 cis n-6) + C20:3 cis n-6) + (C22:5 cis n-3) + (C20:3 cis n-3) + (C20:4 cis n-6) + (C20:5 cis n-3) + (C21:5 n-3) + (C22:6 cis n-3). a,b,c Values with no common superscript differ significantly (p ≤ 0.05). c Hypocholesterolemic/hypercholesterolemic ratio = (cis-C18:1 + ΣPUFA)/(C12:0 + C14:0 + C16:0.
Table 13. Effect of phytobiotic mixture supplementation on ham meat fatty acid composition.
Table 13. Effect of phytobiotic mixture supplementation on ham meat fatty acid composition.
Ham Meat (Biceps Femoris)
FA (%)		CONTPM-APM-BSEMp-Value
C10:0 (Capric)0.09 a0.07 a0.11 b0.005<0.001
C12:0 (Lauric)0.120.110.110.0030.338
C14:0 (Myristic)2.01 b1.69 a1.94 b0.036<0.001
C14:1 (Myristoleic)0.03 b0.01 a0.05 c0.004<0.001
C15:0 (Pentadecanoic)0.06 a 0.04 a0.10 b0.007<0.001
C16:0 (Palmitic)28.83 b26.78 a28.99 b0.2570.002
C16:1 cis (Palmitoleic)3.40 b3.22 ab3.13 a0.0430.022
C17:0 (Heptadecanoic)0.27 a0.30 b0.30 b0.0050.018
C17:1 cis-10 (Heptadecenoic cis) 0.31 c0.25 a0.28 b0.007<0.001
C18:0 (Stearic) 8.63 a8.69 a9.85 b0.160<0.001
C18:1n-9t (Elaidic)0.10 b0.11 b0.08 a0.0060.019
C18:1 cis n-9 (Oleic) 29.91 b33.67 c29.13 a0.492<0.001
C18:2n-6t (Linolelaidic)0.020.030.030.0030.363
C18:2 n-6c (Linoleic)22.87 c20.87 a22.28 b0.2110.001
C18:3n-6 (γ-Linolenic)0.05 a0.15 b0.20 c0.016<0.001
C20:0 (Arachidic)0.20 b0.06 a0.05 a0.017<0.001
C18:3n-3 (a-Linolenic)1.41 b1.22 a1.24 a0.022<0.001
C20:1 cis n-9 (cis-11-Eicosenoic) 0.20 a0.28 b0.18 a0.012<0.001
C21:0 (Henicosanoic)0.030.020.030.0030.133
C20:2 cis n-6 (cis-11,14-Eicosadienoic)0.41 b0.41 b0.36 a0.0070.003
C20:3 cis n-3 (cis-11-14-17-Eicosatrienoate) 0.19 b0.22 c0.17 a0.006<0.001
C20:4 cis n-6 (Arachidonic)1.01 a1.41 c1.12 b0.044<0.001
C20:5 cis n-3 (cis-5,8,11,14,17-Eicosapentaenoic)0.070.070.250.0570.388
C22:6 cis n-3 (cis-4,7,10,13,16,19-Docosahexaenoic)0.170.170.170.0030.962
Σ SFA (Total Saturated FA)40.25 b37.77 a41.48 c0.396<0.001
Σ MUFA (Total Monounsaturated FA)33.96 b37.55 c32.85 a0.499<0.001
Σ PUFA (Total Polyunsaturated FA)26.22 b24.55 a25.82 b0.1980.019
Σ n-3 (Total omega-3 FA)1.851.681.830.0560.418
Σ n-6 (Total omega-6 FA)24.37 b22.87 a24.00 b0.1680.002
Ratio n-6/n-3 FA13.20 x13.66 y13.54 y0.2880.052
PUFA/SFA0.65 b0.65 b0.62 a0.0050.006
h/H c1.81 b2.04 c1.77 a0.029<0.001
FA: fatty acids; ΣSFA = (C10:0) + (C12:0) + (C14:0) + (C16:0) + (C17:0) + (C18:0); ΣMUFA = (C16:1 cis) + (C17:1 cis-10) + (C18:1 cis n-9) + (C18:1 n-7) + (C20:1 cis n-9); ΣPUFA = (C18:2 n-6c) + (C18:4n-3) + (C20:2 cis n-6) + C20:3 cis n-6) + (C22:5 cis n-3) + (C20:3 cis n-3) + (C20:4 cis n-6) + (C20:5 cis n-3) + (C21:5 n-3) + (C22:6 cis n-3). a,b,c Values with no common superscript differ significantly (p ≤ 0.05). x,y Values with no common superscript tend to (0.05 < p ≤ 0.10). c hypocholesterolemic/Hypercholesterolemic ratio = (cis-C18:1 + ΣPUFA)/(C12:0 + C14:0 + C16:0).
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Magklaras, G.; Tzora, A.; Bonos, E.; Zacharis, C.; Fotou, K.; Wang, J.; Grigoriadou, K.; Giannenas, I.; Jin, L.; Skoufos, I. Nutritional Use of Greek Medicinal Plants as Diet Mixtures for Weaned Pigs and Their Effects on Production, Health and Meat Quality. Appl. Sci. 2025, 15, 9696. https://doi.org/10.3390/app15179696

AMA Style

Magklaras G, Tzora A, Bonos E, Zacharis C, Fotou K, Wang J, Grigoriadou K, Giannenas I, Jin L, Skoufos I. Nutritional Use of Greek Medicinal Plants as Diet Mixtures for Weaned Pigs and Their Effects on Production, Health and Meat Quality. Applied Sciences. 2025; 15(17):9696. https://doi.org/10.3390/app15179696

Chicago/Turabian Style

Magklaras, Georgios, Athina Tzora, Eleftherios Bonos, Christos Zacharis, Konstantina Fotou, Jing Wang, Katerina Grigoriadou, Ilias Giannenas, Lizhi Jin, and Ioannis Skoufos. 2025. "Nutritional Use of Greek Medicinal Plants as Diet Mixtures for Weaned Pigs and Their Effects on Production, Health and Meat Quality" Applied Sciences 15, no. 17: 9696. https://doi.org/10.3390/app15179696

APA Style

Magklaras, G., Tzora, A., Bonos, E., Zacharis, C., Fotou, K., Wang, J., Grigoriadou, K., Giannenas, I., Jin, L., & Skoufos, I. (2025). Nutritional Use of Greek Medicinal Plants as Diet Mixtures for Weaned Pigs and Their Effects on Production, Health and Meat Quality. Applied Sciences, 15(17), 9696. https://doi.org/10.3390/app15179696

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