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Functional and Clean Label Dry Fermented Meat Products: Phytochemicals, Bioactive Peptides, and Conjugated Linoleic Acid

Małgorzata Karwowska
Paulo E. S. Munekata
Jose M. Lorenzo
2,3,* and
Igor Tomasevic
Department of Animal Food Technology, Sub-Department of Meat Technology and Food Quality, University of Life Sciences in Lublin, Skromna 8, 20-704 Lublin, Poland
Centro Tecnológico de la Carne de Galicia, Adva. Galicia n° 4, Parque Tecnológico de Galicia, San Cibrao das Viñas, 32900 Ourense, Spain
Facultad de Ciencias de Ourense, Área de Tecnología de los Alimentos, Universidade de Vigo, 32004 Ourense, Spain
Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade, Serbia
Author to whom correspondence should be addressed.
Appl. Sci. 2022, 12(11), 5559;
Submission received: 18 April 2022 / Revised: 5 May 2022 / Accepted: 27 May 2022 / Published: 30 May 2022
(This article belongs to the Special Issue New Challenges in Improving the Quality and Safety of Meat Products)


Consumer demand for specific dietary and nutritional characteristics in their foods has risen in recent years. This trend in consumer preference has resulted in a strong emphasis in the meat industry and scientific research on activities aimed at improving the nutritional value of fermented meat products. These types of meat products are valued by modern consumers due to their nutritional value resulting, among others, from the method of production. One of the major focuses of the current innovations includes the incorporation of bioactive compounds from plant-based food, in relation to the replacement of additives that may raise concerns among consumers (mainly nitrate and nitrite) as well as the modification of processing conditions in order to increase the content of bioactive compounds. Many efforts have been focused on reducing or eliminating the presence of additives, such as curing agents (nitrite or nitrate) in accordance with the idea of “clean label”. The enrichment of fermented meat products in compounds from the plant kingdom can also be framed in the overall strategies of functional meat products design, so that the meat products may be used as the vehicle to deliver bioactive compounds that may exert benefits to the consumer.

1. Introduction

Fermentation and drying processes are likely the oldest forms of meat preservation [1]. Nowadays, there are many formulations of fermented meat products all over the world. The composition and ratio of raw materials, meat species, and salt as well as the addition of spices and seasonings and the processing procedures are key factors influencing the composition of the fermented meat product [2]. Sausages are one of the oldest foods processed by humanity, however, dry-fermented sausages are of particular importance to modern consumer in terms of their nutritional properties. These products are elaborated with meat and fat, mixed with salt, curing agents, sugar, and spices. Despite fermented sausages have their longest tradition in Southern and Central Europe, today many varieties of these products are produced all over the world. Many factors have been influencing this phenomenon: the availability of various meat raw materials, different climatic conditions at each location, cultural and religious factors, and the knowledge passed over generations about the production of these products [3,4,5]. In this context, the great variety of these products around the world makes fermented sausages a cultural heritage of many regions [6]. Generally, traditionally fermented meat products have become one of the most important components of the human diet and an important sector of the meat products market. In the recent years, the innovation on fermented meat products has been aiming to enhance their quality, especially health-related characteristics, while maintaining their sensory characteristics [6,7]. The sensory characteristics of fermented meat products are the result of a series of changes in the raw materials, ingredients, and additives due to the activity of meat enzymes and microorganisms (naturally occurring or added as starter cultures) [8].
The production of fermented sausages is characterized by key stages (fermentation, ripening, and drying) that modify carbohydrates, proteins, and lipids and generate the specific physicochemical parameters (pH, water activity, color) and sensory parameters (taste, aroma, flavor) [9]. Therefore, thanks to the selection of specific processing conditions, it is possible to obtain a wide variety of fermented meat products with specific, exclusive, and characteristic characteristics and sensory attributes [10]. The fermentation stage plays a central role in the production of fermented sausages by favoring the initial acidification of the meat mass. Naturally occurring or starter cultures produce organic acids (especially lactic acid) and reduce the pH of the sausage mass, which improves safety by inhibiting the growth of pathogenic and contaminant spoilage flora [5]. The production of H2O2 and bacteriocins from microbial activity are responsible for preventing the growth of spoilage microorganisms and food-borne pathogens in fermented sausages [10]. Additionally, specific starter cultures have been used to produce functional sausages due to their probiotic potential [11,12,13,14].
Natural peptides released during the fermentation of meat products are currently being studied due to their health-promoting characteristics, including immuno-modulatory activity and protection against oxidative stress [15,16,17]. Antioxidant activity is one of the potential health benefits associated with bioactive peptides from fermented meat products [18]. Their presence, as well as other bioactive ingredients including conjugated linoleic acid (CLA) can be changed by modifying the processing conditions or the strains composing the starter culture [16]. The type of starter culture included in the elaboration of fermented meat products affects the number of peptides produced and their properties [19].
Another strategy to obtain healthier and functional fermented meat products consist in the incorporation of bioactive compounds from plants [20]. The incorporation of bioactive ingredients from the plants has been effectively applied to fermented meat products [21,22,23]. The idea of producing foods with bioactive compounds has led to the development of “functional foods”. The use of these ingredients in food production affects not only the health of the consumer [24], but also makes it possible to produce products with better chemical properties and a longer shelf life [25]. Moreover, bioactive compounds from plants were evaluated as natural antioxidants in meat and meat products [25]. In this regard, extracts obtained from plants have been used to delay the lipid and/or protein oxidation of meat product [26,27,28].
The incorporation of plant materials rich in bioactive compounds in meat products is also related to the replacement of food additives that may raise concerns among consumers due to their perceived harmfulness, even though they are approved for use in accordance with food regulations [20]. This practice is in line with the strategies aiming to produce “clean label” foods. Food producers and consumers frequently use this term for food products in which the number of additives has been reduced. Food authorities have not yet provided a definition for the term “clean label”. Therefore, it can be interpreted differently among consumers and producers; however, any practices with the purpose of reducing the quantity of additives are especially appreciated by modern consumers and increase their confidence in the producer. The clean label trend is also of particular importance for the meat industry and consumers of meat products because the list of approved additives in the meat industry is long [29,30,31].
Fermented meat products are unique, thus often represented as elements of culinary heritage and identity. However, their nutritional importance has often been compromised throughout history due to safety and health issues. Therefore, novel strategies are emerging to improve their quality and healthiness. This review aims to discuss different strategies to improve the nutritious value of fermented meat products which are currently of interest to consumers, scientists, and food producers. Particular attention was paid to activities related to the incorporation of bioactive compounds from plant-based food, the modification of processing conditions in order to increase the content of bioactive compounds, and the elimination of nitrates in accordance with the idea of “clean label”.

2. Incorporation of Bioactive Compounds from Plant-Based Food

The inclusion of bioactive compounds from plant foods is an interesting strategy to obtain fermented meat products with bioactive compounds. Phenolic compounds and terpenoids have been indicated as relevant candidates due to their physiological effect and protective effect in different meat products [32,33,34,35,36,37]. These compounds can inhibit lipid peroxidation processes in meat products by acting according to various mechanisms: the primary antioxidants inhibit lipid peroxidation by preventing chain reactions, decreasing localized oxygen concentrations, reacting directly with lipid radicals, and converting them into relatively stable products; secondary antioxidants act by binding to catalysts such as metal ions, which in meat products include primarily heme iron [38,39].
The identification of main compounds varies among these plant-based food sources. For instance, some of the main compounds found in cranberries include protocatechuic acid, benzoic acid, p-coumaric acid, and peonidin-3-galactoside [40]. Likewise, rose extract is another source with phenolic acids (gallic acid and protocatechuic acid) and flavonoids (quercetin) [41]. These two classes of compounds can also be found in beer residue (catechin and p-coumaric acid) and peanut skin (catechin, epicatechin, and benzoic acid) [42]. In the case of chestnuts, some of the predominant polyphenols are cascalagin, vescalagin, and gallic acid [42,43]. Grape seeds are particularly rich in epicatechin gallate and its trimers [43]. Particularly from olive, the main compounds identified are hydroxytyrosol and tyrosol and their derived compounds [43].
In the case of essential oils, terpenoids are the principal class of bioactive compounds. In the case of rosemary, carnosic acid and carnosol have been indicated as key components [44]. Sage is another interesting source of bioactive compounds, wherein some of the main compounds found are α-thujone, camphor, and eucalyptol [45]. In the case of cinnamon essential oil, some of the major compounds identified were trans-cinnamaldehyde, α-calacorene, cis-cinnamaldehyde, copaene, α-guaiene, α-muurolene, and 2-propenal,3-(2-methoxyphenyl) [46]. The use of these compounds in the production of bioactive fermented meat products is presented in Table 1.
Regarding the effect of polyphenolic-rich sources in the properties of fermented meat products, one of the main outcomes is the reduction of oxidative reactions in key molecules and groups of compounds, particularly lipids, proteins, and myoglobin [44,45,47,48,49,50,51]. One relevant source is the cranberry extract (0.6 g/kg), which delayed the lipid oxidation of sausages, reduced the loss of heme-iron, and also increased the lightness of samples in fermented deer sausage [51]. A related study with cranberry pomace powder indicated that the reduction of Salmonella enterica and Staphylococcus spp. counts during processing (fermentation and ripening periods) was accelerated with the highest levels of extract (1.70 and 2.25 g/100 g) [49].
A similar protective effect was reported for the use of rose extract (1, 2, and 3 g/kg) in fermented pork sausages [47]. The pH, lipid oxidation, accumulation of biogenic amines, and microbial growth were reduced in relation to the control treatment (without natural extract), especially when using 3 g/kg of rose extract.
Table 1. Main bioactive compounds from plant origin used in fermented meat products.
Table 1. Main bioactive compounds from plant origin used in fermented meat products.
Meat ProductBioactive Compound, Source, and ConcentrationProcessing/Storage ConditionsEffect in Quality/Shelf LifeRef.
Fermented deer sausageCranberries extract (0.6 g/kg)Processing: 22 days at 16 °C and RH 80–90%↑ Heme iron and L* value
Fermented pork and beef sausageCranberry pomace powder (0.55, 1.70, and 2.25 g/100 g) and five strains of Salmonella entericaProcessing: 24 h at 25 °C and RH 88%, 24 h at 23 °C and RH 80%, 24 h at 21–20 °C and RH 78%, 24 h at 16 °C and RH 76–75%, and 28 days at 14 °C and RH 75%↑ a* value
Salmonella enterica and Staphylococcus spp. counts, L*, and hardness
Fermented pork sausageRose extract (1, 2, and 3 g/kg)Processing: 3 days at 20 °C and RH 90%, 5 days at 10 °C and RH 80%, and for 16 days at 10 °C and RH 70%↓ pH, TBARS, BAs, and TPC[47]
Harbin dry sausageAllium senescens seed extract (2, 4, 6, and 8 g/kg)Processing: 1 day at 25 °C and RH 30–50% and 1 day at 25 °C and RH 75–80%↑ aw, pH (2 and 4 g/kg), a* and b* values (2 and 4 g/kg)
↓ TBARS and carbonyl content
Cantonese sausagesMulberry extract (0.5 and 1.0 g/kg)Processing: 36 h at 50 °C↑ Moisture and pH
↓ L*, a*, and b* values, and TBARS (1.0 g/kg)
Fermented pork sausageBeer residue, chestnut leave and peanut skin extracts (2 g/kg)Processing: 2 days at 20 °C and RH 80–85% and for 49 days at 12 °C and RH 75–80%↑ Carbonyl content (chestnut leave and peanut skin) and FFA
↓ L* value and pH (chestnut leave)
Fermented pork sausageGrape seed or chestnut extracts combined with hydroxytyrosol and tocopherol (10 g/kg)Processing: 4 days at 28 °C and RH 85% and for 21 days at 13 °C and RH 70%↑ Cohesiveness, springiness, chewiness, and firmness (SA)
↓ a* (chestnut extract) and b* value (both); color uniformity and redness (SA)
Italian Cinta Senese sausageOlive pomace hydroxytyrosol with either grape seed extract or chestnut extract (10 g/kg)Processing: 4 days at 28 °C and RH 85% and 21 days at 13 °C and RH 70%Weissella, Enterococcus, Lactococcus, Bacillus, and Pluralibacter counts (grape seed extract)
↓ pH, chroma, and ash (grape seed extract)
Fermented beef sausagePistachio hull extract (500, 750, and 1000 ppm)Processing: 5 days at 23 °C and RH 85–95% followed by 23 days at 14 °C and RH 75–80%
Storage: 90 days at 4 °C
↑ pH, moisture, aw, and L* value
Fermented pork sausageRosemary oleoresin in oil-in-water emulsion (100 g/kg emulsion; concentration in meat mass 4.8 g emulsion/kg)Processing: 48 h at 22 °C and RH 93%, 60 h at 22 °C and RH 90%, 60 h at 21 °C and RH 90%, 120 h at 20 °C and RH 88%, 10 h at 18 °C and RH 85%, and 206 h at 15 °C and RH 85%
Storage: 49 days at 7 or 20 °C with MAP (60:25:15 for O2:CO2:N2)
↑ a* value (7 °C)
↓ POV, TBARS, hexanal accumulation; L* and a* values (20 °C), b* values, rancidity (SA)
Fermented pork sausageSage essential oil (0.01, 0.0.5, and 0.1 mL/kg)Processing: 21 days at 14–16 °C and RH 95–80%↑ pH and b* value
↓ L* value and TBARS
Concentration-dependent for sensory color, odor, and flavor
Fermented beef sausageEncapsulated (alginate with or without nanocrystal cellulose) cinnamon essential oil (2.25 g/100 g)Processing: 48 h at 25 °C and RH 90% and 5 days at 14 °C and RH 70%↑ L* and b* values
↓ Microbial load
Fermented pork sausageGround chia and black cumin (1 and 2 g/100 g)Processing: 3 days at 20–22 °C and RH 55–63%, 3 days at 14–16 °C and RH 68–75%, and 24 days at 13 °C and RH 76%↑ Fat, moisture (black cumin), volatile compounds (black cumin), and herbal odor and flavor (SA)
↓ Protein (black cumin) and overall quality (2 g/100 g black cumin)
a*: redness; aw: water activity; b*: yellowness; BA: biogenic amine; FFA: free fatty acids; L*: luminosity; MAP: modified atmosphere packaging; POV: peroxide value; RH: relative humidity; SA: sensory analysis; TBARS: thiobarbituric acid reactive substances; and TPC: total plate count. ↑: Increase and ↓: Decrease.
The study carried out by Qin et al. [52] indicated that Allium senescens seed can be explored as a natural ingredient to improve the lipid oxidative stability of Harbin dry sausage during processing. The authors also highlighted the concentration-dependent effect of this extract wherein the highest concentrations had the highest value for redness, yellowness, and pH. A related experiment with mulberry extract in Cantonese sausages indicated a different outcome in terms of color [50]. Both levels of inclusion reduced L*, a*, and b* values. Additionally, the moisture and pH were increased due to the mulberry extract incorporation into the sausages. A comparative study with beer residue, chestnut leave, and peanut skin extracts (2 g/kg) indicated differences in the protective effect of each extract rich in bioactive compounds in fermented pork sausages [42]. The use of chestnut extract increased the carbonyl content and free fatty acid content, and also caused significant reductions in the lightness and pH. No major effects were reported for the addition of beer residue and peanut skin extracts in relation to control sausage (with sodium nitrite).
The inclusion of grape seed or chestnut extracts combined with hydroxytyrosol and tocopherol (10 g/kg) caused significant reductions in the color of fermented pork sausages [43]. Another effect associated with the addition of these combined bioactive compounds was the increase of instrumental cohesiveness, springiness, and chewiness that was also perceived at sensorial level in terms of firmness. A related experiment combined olive pomace hydroxytyrosol with either grape seed extract or chestnut extract (10 g/kg) to produce Italian Cinta Senese sausages [53]. Different results were obtained from the incorporation of these combinations of natural compounds. The combination of hydroxytyrosol with grape seed extract caused significant differences in microbial community composition and ash content, whereas the combination of hydroxytyrosol with chestnut extract had similar results to control treatments for these indicators.
Pistachio hull extract is an interesting source of bioactive compounds with potential application in fermented beef sausage [48]. This extract (500, 750, and 1000 ppm) delayed lipid oxidation (during both processing and storage periods) and reduced the pH, lightness, moisture, and aw. Moreover, no significant differences in redness, microbial growth, and sensory analysis were reported between sausages produced with pistachio hull extract and control sausages (without natural extract). A similar protective effect during storage was observed in fermented pork sausages produced with rosemary oleoresin in oil-in-water emulsion (4.8 g emulsion/kg) [44]. In this case, the authors explored the effect of different emulsion droplet sizes (mean particle diameter of 0.2 or 4.6 µm) and observed a protective effect against lipid oxidation in all samples, regardless of storage temperature (7 vs. 20 °C). Interestingly, the preservation of redness was also affected by storage temperature wherein the protective effect against the loss of redness observed in samples stored at 7 °C was not observed in samples stored at 20 °C.
The use of essential oils (rich in terpenoids) obtained from plant-based foods are also interesting materials to produce fermented sausages. One relevant example about the incorporation of essential oils in fermented meat products is the study carried out by Šojić et al. [45] with sage essential oil. The inhibition of lipid oxidation was achieved (especially with the highest concentration of essential oil; 0.1 mL/kg), but a modification in luminosity and pH were also observed in the fermented sausages containing the essential oil. Another possible strategy to incorporate essential oils in fermented meat products consist in a previous encapsulation, which can improve the stability of active compounds during processing and shelf life [55]. In this sense, Ji et al. [54] tested two encapsulating/wall materials (alginate alone or combined with nanocrystal cellulose) in the encapsulation of cinnamon essential oil. Both wall materials had minor effect in the textural properties of sausages and inhibited the growth of microorganisms (especially total bacteria and lactic acid bacteria groups). However, the luminosity and yellowness of fermented sausages were increased.
It is also relevant to comment that the inclusion of plant-based food can influence the sensory properties of fermented meat products and aggregate specific odors and flavors. This aspect was reported in the study carried out by Borrajo et al. [21] who evaluated the incorporation of ground chia and black cumin (1 and 2 g/100 g). According to these authors, the herbal odor and flavor were more perceived in sausages containing ground black cumin (regardless of concentration). Moreover, the content of volatile compounds in these sausages was also superior to obtain for other treatment (control and ground chia). Another experiment with sage essential oil highlighted the relation between the concentration of plant-based ingredients with the sensory properties of fermented meat products [45]. The sausages produced with the highest concentration of sage essential oil caused the most intense modification in the perception of color, odor, and flavor of fermented pork sausages.

3. Bioactive Compounds from Plant-Based Food as “Clean Label” Ingredients in Fermented Sausages

The so-called “clean label” trend is one of fastest growing initiatives in term of food production aiming to enhance the health quality of foods. Although this concept has not yet been defined in food law, it is commonly used in the context of food with few and recognizable ingredients. The meat industry has an important challenge in the development of meat products produced according to the clean label trend, as meat products belong to the food group that is customarily produced with numerous functional additives [29,31,56]. Within the EU legislation, there is a list of permitted additives and their maximum level of use depending on the type of product [57]. According to this regulation, the list of permitted additives for meat products is long and include antimicrobials, antioxidants, and texturizers as the most used ones, but also some other additives (such as colorants, acidity regulators, and stabilizers). Although all the additives employed in meat products elaboration are considered safe within the established limits by the food safety authorities, the consumers perceive these additives as unhealthy and expect their elimination from the recipe of meat products [29].
The subject of increasing scientific controversy in the context of the impact on human health are compounds such as nitrites and nitrates that are commonly used as the main additives in the production of fermented meat products [58,59]. Nitrates and nitrites are authorized as additives in meat processing in the European Union under Commission Regulation (EU) No 1129/2011 at the maximum amount of 150 mg/kg; however, the addition of sodium nitrate is allowed only in uncooked meat products [57]. There are several reasons for using these compounds to enhance the meat products quality: participation in the formation and stabilization of red color named cured-meat color; inhibition of the growth of undesirable bacteria (especially Clostridium botulinum and its toxin); delay of the lipid oxidation; participation in the formation of the meat product flavor [6,60,61,62,63,64,65]. The mechanisms of these interactions described in the literature are presented in Table 2. From a different point of view, nitrites and nitrates are the subject of increasing scientific and consumers controversy in the context of the impact on human health as an increase in the content of reactive forms of nitrogen can result in a harmful process called nitrosation stress. This process can cause damage to cellular structures such as proteins, membranes, lipids, and DNA [59]. Some scientific evidence suggests a link between the consumption of foods containing nitrates and the risk of cancer including colorectal cancer, thyroid cancer, breast cancer, renal cell carcinoma, gastric cancer, and esophageal cancer [66,67,68,69,70,71].
The clean label trend in food production is very often involved with using plant-based ingredients/additives. In recent years, several studies in this area have been carried out with fermented meat products, mainly sausages [21,22,51,72]. The fundamental criterion for selecting an ingredient from plants as a component of a meat product without nitrates/nitrites is to show the same functions of nitrates. The best results are obtained for those that show both antimicrobial and antioxidant properties, additionally having a positive effect on the color, taste, and smell of products and their structure. According to Bernardo et al. [73] the inclusion or not of nitrite/nitrate depends on considering two crucial risks—the eventual formation of nitrosamines known as carcinogenic compounds to humans or the eventual out-growth of pathogens.
Table 2. The role of nitrates/nitrites in meat products.
Table 2. The role of nitrates/nitrites in meat products.
The formation and stabilization of red colorNitric oxide reaction with myoglobin (deoxymyoglobin and metmyoglobin) forms nitrosylmyoglobin complex, which generates the unique cured-meat color[59]
Preservative effectAntimicrobial effect due to the reduction in oxygen uptake, inactivation of some metabolic enzymes, and breakage of the electron transport chain[6]
Inhibition of growth of some anaerobic bacteria, including Clostridium botulinum and its toxins, Clostridium perfringens, Staphylococcus cereus, Bacillus cereus, and Listeria monocytogenes[61,62]
Improvement of the oxidative stabilityNitrite and nitrate delay the lipid oxidation through oxygen deletion
Nitric oxide can react with radicals (alkoxy radicals, hydroxyl radical, and peroxyl radicals) interrupting radical chain reactions and bind to transitional metals
Contribution to the flavor formationSeveral compounds are formed when nitrite is bound to proteins and lipids. For example, when nitrites are bound to sulfur-containing amino acids of meat proteins, SH-residues with a specific flavor and aroma are formed and contribute to the flavor of cured meat[64]
Potential formation of N-nitrosaminesThe addition of nitrates and nitrites to the processed meat can induce the development of N-nitroso compounds linked with metabolic and genotoxicity disturbances in the large intestine mucosa[65,74,75,76]
The specificity of the production of fermented meats poses a high risk of Listeria monocytogenes contamination, which is the main frequently detected pathogen in dry fermented sausages [77]. This pathogen is highly difficult to control in fermented sausage processing environments because of its high tolerance to a high salt content and low pH. The optimum growth conditions of L. monocytogenes include a temperature in the range of 30–37 °C, optimum pH between 6 and 8 (however, it can grow at pH ranges from 4.5 to 9.0), NaCl levels of 12%, and water activity values of 0.92 [78,79]. However, nitrites and nitrates have inhibitory effect on L. monocytogenes growth in dry-fermented sausage [80]. The research conducted by Hospital et al. [81] showed that a lower content of nitrite and nitrate would increase Listeria counts in case of contamination with this microorganism. Therefore, the elimination of nitrates requires alternative and efficient measures to inhibit the growth of this pathogen. In this sense, Dalzini et al. [82] investigated the inactivation of L. monocytogenes during the manufacturing process of Milano-type salami elaborated without nitrites/nitrates and with vegetable additives (carrot and Swiss chard juice concentrate powder) to assess the pathogens’ behavior when exposed to alternative additives or nitrite/nitrate replacements from vegetable origin. The results showed no growth of L. monocytogenes during the process, even in case when the salami was elaborated without nitrites/nitrates. The authors point to the key role of other barriers to inhibit the growth of pathogens in the elaboration of raw fermented meat products, such as pH and water activity. A related experiment indicated that the same protective effect of leaf celery and spinach extract encapsulated in tubular cellulose improved the preservation of redness and also reduced the load of microorganisms (enterobacteriaceae, lactic acid bacteria, yeasts, and molds) [83].
The scientific research for ingredients of plant origin with antimicrobial properties for incorporation into raw fermented meat products notably increased in recent years. Several natural extracts and ingredients (based on herbs and spices, vegetables, or fruits) were applied to fermented meat products as clean label alternatives to nitrites [55,84,85]. Tomovic et al. [86] indicated the efficiency of Juniperus communis L. essential oil application in dry fermented sausage and suggested that it could be considered as a potential partial replacement for sodium nitrite. Similarly, the research by García-Díez et al. [87] revealed that the L. monocytogenes count was reduced by the addition of essential oils of oregano, bay, rosemary, garlic, nutmeg, and thyme. Moreover, a higher antimicrobial effect was obtained by increasing the essential oil concentration.
In recent years, chia seeds (Salvia hispanica) have been frequently selected by researchers in the elaboration of fermented meat products as substitutes for nitrates/nitrites due to their properties [21,22]. Research by Duda-Chodak et al. [88] noticed that the main Salvia hispanica bioactive compounds (kaempferol and quercetin) could exert a strong negative impact on the development of pathogenic microorganisms. According to Munoz et al. [89] chia seeds can also positively promote consumers’ health due to their high amount of soluble dietary fiber.
However, some authors note that in some cases, the solution of using additives from the plant kingdom in nitrite-free fermented meat products only rely on pseudo-solutions, such as using plant extracts that naturally contain nitrate residues [73,90]. It is known that some vegetables (especially celery, spinach, beets, and chard) may contain a great content of nitrate [91], which can be converted to nitrite by the meat microflora or by added bacteria. According to Alahakoon et al. [60], using vegetables is a great solution to include the natural sources of nitrate into meat products that could satisfy both the consumer demand and technological requirements for natural products, even if natural nitrates do not avoid the formation of N-nitrosamines. A contrary opinion was provided by Delgado-Pando et al. [29], who suggested that nitrites of natural origin do not offer any healthier advantage towards synthetic nitrites and that they only provided a clean label option for the consumer.

4. Modification of Processing Conditions in Order to Increase the Content of Bioactive Compounds

Among the possible strategies to obtain fermented meat products with bioactive compounds, the use of starter cultures with proteolytic activity has been suggested to promote the accumulation of bioactive peptides [19]. Recent studies about the use of selected starter cultures to increase the content of bioactive peptides in fermented meat products are presented in Table 3. For instance, the use of Lactobacillus sakei JCM1157 and no. 23; and Lactobacillus curvatus NBRC15884 and no. 28 as individual starter cultures was associated with reduced angiotensin-converting enzyme activity (ACE; an enzyme involved in the constriction of blood vessels) in beef fermented sausages [92]. Additionally, an increase in the antioxidant activity (evaluated by the DPPH method) of the sausages was also observed for all samples except for those produced with Lactobacillus sakei JCM1157.
A recent study explored the effect of individual and combined use of a commercial starter culture (Pediococcus pentosaceus and Staphylococcus carnosus) and Lactobacillus plantarum KX881772 and the meat source (camel meat vs. beef) in the production of bioactive fermented sausages [93]. The authors indicated that the meat source affected the antioxidant activity of sausages (camel > beef), regardless of the starter culture composition. Conversely, the antidiabetic potential was affected by the starter culture strains. The sausages produced with Lactobacillus plantarum KX881772 (alone or combined with the commercial starter culture) displayed the highest inhibitory effects in α-amylase and α-glucosidase (enzymes involved in the digestion of carbohydrates). Particularly for the ACE activity test, the highest inhibitory activity was obtained from the sausage produced with camel meat and Lactobacillus plantarum KX881772 (alone or combined with the commercial starter culture). Moreover, starter cultures increased the degree of protein hydrolysis, which suggested the involvement of peptides in these potential biological effects. A related experiment reported the influence of different starter cultures and the average particle size in the biological activity of fermented camel sausage [94]. According to the authors, the fractions with less than 3 kDa displayed the highest antioxidant activity (especially from sausages produced with Staphylococci xylosus and Lactobacillus plantarum) and inhibitory activity against ACE (from all sausages produced with starter cultures). Additionally, the authors identified this effect was mainly observed with peptides in the 3 kDa fraction and indicated the presence of amino acids (such as glutamic acid, valine, and alanine) usually associated with antioxidant activity in their sequences.
Fermented pork sausages can also contain peptides with biological activity [95]. In this case, the combined use of Staphylococcus simulans NJ201 and Lactobacillus plantarum CD101 produced sausages with enhanced inhibitory activity against ACE and with high antioxidant activity. The authors also reported an increase in both the degree of protein hydrolysis and the content of free amino acids. Additionally, the identification of the main peptides in the <3 kDa fraction contained key amino acids such as proline, aspartic acid, alanine, and arginine. Similarly, the use of Lactobacillus plantarum CD101 and S. simulans NJ201 as a starter culture in the elaboration of fermented pork sausage increased the antioxidant activity of these meat products in comparison with control samples (without the starter culture) [96].
Another relevant bioactive compound for fermented meat products is CLA. The consumption of this fatty acid has been associated with anticarcinogenic effects and, to some extent, with cardiovascular protection and body weight control at the human level [97]. Increasing the content of CLA in fermented meat products is another interesting effect derived from changes in the processing of fermented meat products (Table 3).
Again, the use of starter cultures plays a central role in the elaboration of fermented meat products. A relevant starter culture for the elaboration of beef sausages with high CLA content was Lactobacillus plantarum in a recent study [98]. The strains AA1-2 and AB20-961 displayed capacity to produce CLA, especially during the fermentation stage of processing. This study also revealed that a small decrease in the content of CLA could be observed until the end of the processing period, but the levels remained stable until the end of the storage period. Interestingly, the concentration of CLA was strain-dependent, wherein the highest content was obtained from the use of the AB20-961 strain. In a posterior experiment, the use of Lactobacillus plantarum AB20–961 and DSM2601 strains in the production of fermented beef sausages was compared with hydrolyzed safflower oil [99]. Both strains produced CLA wherein the highest concentration was obtained from the DSM2601 strain.
It is also important to mention that the production of CLA in fermented meat is also dependent on other processing variables such as pH (4.3–9.3), time (0–120 h), temperature (4–37 °C), inoculum (6–8), and fatty acid/oil amount (1.44, 2.50, 3.56, and 5.00%) as reported in a recent experiment [100]. In this case, a central composite design was applied to optimize the production of CLA using Lactobacillus plantarum AB20–961 and DSM2601 strains. The optimum fermenting conditions for both microorganisms were 37 °C, 5% fatty acid, inoculum with 8 log CFU/g, safflower fatty acids. In the case of pH and fermenting time, the values were specific for each strain.
Table 3. Increasing the content of bioactive peptides and conjugated linoleic acid in fermented meat products.
Table 3. Increasing the content of bioactive peptides and conjugated linoleic acid in fermented meat products.
Meat ProductStater Culture Strains
(Inoculum Cell Count)
Processing/Storage ConditionsEffect in Biological Activity PotentialRef.
Increasing bioactive peptides
Fermented beef sausageLactobacillus sakei JCM1157 and no. 23 and Lactobacillus curvatus NBRC15884 and no. 28 as starter culture (inoculum: 108 CFU/mL)Processing: 2 days at 20 °C and RH 98–100% and for 19 days at 20 °C and RH from 90 to 65%↑ Antioxidant (all starters except Lactobacillus sakei JCM1157) and inhibition of ACE (all starters) activity[92]
Fermented camel or beef sausageCommercial starter (Pediococcus pentosaceus and Staphylococcus carnosus) and Lactobacillus plantarum KX881772 as starter culture (inoculum: 107–108 CFU/kg)Processing: 48 h at 30 °C and RH 90% and vacuum-packaged for 21 days at 15 °C↑ DH (combined starter culture in camel sausage), antioxidant (camel sausage), inhibition of α-amylase, α-glucosidase (L. plantarum combine or alone), and ACE activities (camel sausage and L. plantarum combined or alone)[93]
Fermented camel sausagesStaphylococci xylosus and Lactobacillus plantarum, Staphylococci xylosus with Lactobacillus pentosus, or Staphylococci carnosus and Lactobacillus sakei as starter cultures (inoculum: 107 CFU/g)Processing: 5 days at 24 °C and RH 85–90% and for 23 days at 14 °C and RH 75–80%↑ Antioxidant (fraction < 3 kDa with Staphylococci xylosus and Lactobacillus plantarum) and inhibition of ACE (fraction < 3 kDa from all starters) activities[94]
Fermented pork sausageLactobacillus plantarum CD101 and Staphylococcus simulans NJ201 as starter culture (inoculum: 107 CFU/g)Processing: 24 h at 30 °C and RH 80% and for 20 d at 15–12 °C and RH 72–75%↑ DH, FAA, inhibition of ACE activity, and antioxidant activity[95]
Fermented pork sausageLactobacillus plantarum CD101 and S. simulans NJ201 as starter culture (inoculum: 107 CFU/g)Processing: 1 day at 30 °C and RH 80%, 3 days at 15 °C and RH 75%, and for 19 days at 12 °C and RH 72%↑ Antioxidant activity[96]
Increasing CLA content
Fermented beef sausageLactobacillus plantarum AA1-2 and AB20-961 (inoculum: 105 CFU/kg)Processing: 7 days at 25–18 °C and RH 95–70%, 24 h at 24 °C and RH 95%, 24 h at 22 °C and RH 90%, 12 h at 20 °C and RH85%, 12 h at 20 °C and RH 80%, 48 h at 18 °C and RH 75%, and for 48 h at 18 °C and RH 70%
Storage: 30 days at 4 °C (vacuum-packaged)
Highest content was obtained with strain AB20-961 (4.2 mg CLA/g fat)[98]
Fermented beef sausageLactobacillus plantarum AB20–961 and DSM2601 (inoculum: 108 CFU/kg)Processing: 73 (DSM2601) or 79 (AB20–961) h at 24 °C and RH 90% and heat-treated until reaching internal temperature of 65 °C for 15 minHighest content was obtained with strain DSM2601 (7.5 mg CLA/g fat)[99]
Fermented beef meatLactobacillus plantarum AB20–961 and DSM2601Processing: pH (4.3–9.3), time (0–120 h), temperature (4–37 °C), inoculum (6–8), fatty acid/oil amount (1.44, 2.50, 3.56, and 5.00%) and added fatty acid source (linoleic acid, hydrolyzed sunflower oil, and hydrolyzed safflower oil)Optimum conditions: 37 °C, 5% fatty acid, inoculum 8 log CFU/g, safflower fatty acids, pH 7.94 and 78.78 h for Lactobacillus plantarum AB20–961) and pH 7.68 and 72.57 h for Lactobacillus plantarum DSM2601 (7.91 and 38.31 mg CLA/g for AB20–961 and DSM2601 strains, respectively)[100]
ACE: angiotensin-converting enzyme; CLA: conjugated linoleic acid; DH: degree of hydrolysis; FAA: free amino acids; and RH: relative humidity. ↑: Increase and ↓: Decrease.

5. Conclusions

One of the major focuses of the current innovation on fermented meat products involves the enhancement of health properties. The inclusion of plant material (rich in bioactive compounds) in the elaboration of fermented meat products as well as the modification of processing conditions (in order to increase the content of bioactive compounds) are key aspects to achieve progress in the development in these foods. The use of natural extracts from berries and agroindustry residues are among the sources that stand out, by delaying the oxidation of lipids, proteins, and myoglobin and the growth of pathogenic microorganisms. However, attention to sensory attributes is necessary to avoid a reduced acceptance of fermented meat products. Fermented sausages are complex food systems containing other components that can also be studied and considered to advance in terms of functional and clean label aspects. A more comprehensive study of these products is still necessary to integrate the development of fermented sausages containing bioactive compounds from plant origin and bioactive peptides with other key strategies, such as fat and sodium chloride reduction and replacement.
Exploring the use of starter cultures has proved to promote the necessary changes to obtain fermented sausages with enhanced bioactive peptides and CLA content. The Lactobacillus genus contains the most relevant strains, such as Lactobacillus plantarum KX881772, AB20–961, and DSM2601 strains. Advances are necessary to promote a more comprehensive characterization of the strains, especially in terms of safety and probiotic potential, in order to progress and expand the knowledge about functional fermented sausages and progressively meet consumers’ expectations.

Author Contributions

Conceptualization, M.K.; writing—original draft preparation, M.K., P.E.S.M., J.M.L. and I.T.; writing—review and editing, M.K. and J.M.L.; supervision, M.K. All authors have read and agreed to the published version of the manuscript.


This research received no external funding.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Not applicable.


Jose M. Lorenzo and Paulo E.S. Munekata are members of the HealthyMeat network, funded by CYTED (ref. 119RT0568). Thanks are extended to GAIN (Axencia Galega de Innovación) for supporting this review (grant number IN607A2019/01). Paulo E.S. Munekata acknowledges postdoctoral fellowship support from the Ministry of Science and Innovation (MCIN, Spain) “Juan de la Cierva” program (IJC2020-043358-I).

Conflicts of Interest

The authors declare no conflict of interest.


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MDPI and ACS Style

Karwowska, M.; Munekata, P.E.S.; Lorenzo, J.M.; Tomasevic, I. Functional and Clean Label Dry Fermented Meat Products: Phytochemicals, Bioactive Peptides, and Conjugated Linoleic Acid. Appl. Sci. 2022, 12, 5559.

AMA Style

Karwowska M, Munekata PES, Lorenzo JM, Tomasevic I. Functional and Clean Label Dry Fermented Meat Products: Phytochemicals, Bioactive Peptides, and Conjugated Linoleic Acid. Applied Sciences. 2022; 12(11):5559.

Chicago/Turabian Style

Karwowska, Małgorzata, Paulo E. S. Munekata, Jose M. Lorenzo, and Igor Tomasevic. 2022. "Functional and Clean Label Dry Fermented Meat Products: Phytochemicals, Bioactive Peptides, and Conjugated Linoleic Acid" Applied Sciences 12, no. 11: 5559.

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