Actual Application of Propolis

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This entry is adapted from the peer-reviewed paper 10.3390/plants12081654

Propolis is a natural hive product collected by honeybees from different plants and trees. The collected resins are then mixed with bee wax and secretions. Propolis has a long history of use in traditional and alternative medicine. Propolis possesses recognized antimicrobial and antioxidant properties. Both properties are characteristics of food preservatives. The antimicrobial and antioxidant properties of propolis were extensively studied. In addition, propolis is recognized as safe (GRAS) and can be considered as an alternative to chemical preservatives. Before its use in food products, propolis was in most cases extracted using different solvents such as ethanol, water, and glycerol. Each solvent offers advantages and disadvantages. Ethanol extracts (PEE) are reported to be the richest extracts in bioactive components. While water extracts (WPE) are the lowest extract on phenolic content. In contrast, the two extracts exhibited a strong flavor and aroma.

propolis antimicrobial antioxidant food preservative

1. Fruits and Vegetables

Several studies were focused on the effect of propolis extract on fruits and vegetables’ quality and storage. Propolis can be applied to the food surface or incorporated into the food formulation ^[1]. Propolis adjunction to food can provide health benefits. In addition, it was reported to prevent lipid oxidation and to improve shelf life ^[2]^[3]. The tested vegetables or fruits, propolis origin, and the used extracts are reported in Table 1.

In fresh whole-head and ready-to-eat lettuces (Lactuca sativa. L.), propolis was found to be efficient in reducing the microbial contamination ^[4]. Propolis used as an agricultural antimicrobial agent of cucumber crops and soybean reduced the disease severity of the two tested crops ^[5]. Similar results were reported for tomato fruits ^[6].

Propolis can be used alone or in combination with plant or essential oil. Propolis combined to gallic acid and tea essential oils was tested as a natural preservative for fresh celery, leek, and pumpkin. Both tea and propolis were found to control and reduce E. coli (endogenous and inoculated strains). In addition, propolis was reported to extend the shelf life and to improve the visual quality ^[7]. Similar results were reported for fresh-cut mixed vegetables for soup ^[8]. Another interesting combination is the use of propolis, chitosan, and thyme essential oil. The combination was effective on the “Vesuviono” tomato. However, chitosan was the most effective ^[9]. Propolis reduced the rate of weight loss and maintained fruit firmness (tomatoes) when applied as bio-coating ^[10]. In addition, propolis used as a coating agent can extend the shelf life of cherry tomatoes and delay the deterioration of their sensory traits ^[11]. In a recent investigation, pullulan film associated with ethanol extract of propolis was found to inhibit the growth of microorganisms in cherry tomatoes. The overall quality of tomatoes coated with propolis was reported to be very high, which could result in high consumer acceptance ^[12].

Propolis was found to decrease the population of spoilage microorganisms in potatoes ^[13] and to be effective in controlling potato soft rot. ^[14]. Propolis, cinnamon oil associated with gum Arabic were found to be an effective bio-fungicide for postharvest chili ^[15]. Different propolis extracts (hexane, dichloromethane, and ethanol) were reported to be efficient on molds and bacterial colonization inhibition during rice storage ^[16].

Postharvest storage of fruits is limited because of fruit spoilage, which can be caused by bacterial or fungal growth ^[1]. In addition, many other factors can be pointed out. For sweet cherries as well as many other fruits, fungal decay, weight and acidity losses, softening, stem browning, and color changes are of great importance ^[17].

The ability of propolis extracted in water and ethanol (70%) to prevent fungal decay in cherries stored at 0 °C for 4 weeks was tested alone and in combination. Cherries were dipped in different treatments. Ethanol treatment was the most active. In contrast, it affected stem cherries’ color and sensory quality ^[17]. In a recent study, propolis was stated to effectively reduce weight loss and respiration of sweet cherry and to delay soluble solids, titratable acid, and sweet cherry hardness ^[18]. Propolis was also active in the in vitro and in vivo inhibition of green and blue molds in Citrus (Pinicillium. Digitatum and Pinicillium. italicum). Moreover, no negative influence on the quality of Citrus fruit was reported ^[19]. Similar results were reported for propolis combined with S. vanrijiae ^[20]. Postharvest storage of several other Citrus species was also improved by propolis, such as mandarins ^[19] and oranges ^[21]^[22]^[23]^[24].

Propolis was effective in preservation of different fruits. Propolis protected Star Ruby grapefruit from fungal decay ^[25] and improved cold storage of table grape, cv. Muscatel. ^[26]. In addition, propolis decreased conidial germination and mycelial growth in dragon fruit ^[27] and control toxin production and penicillium decay ^[28]. Propolis was also reported to exhibit a significant antifungal activity on the growth of several strains—in particular, P. expansum, Fusarium sp, A. alternat, and A. niger isolated from apple fruit ^[29]. Propolis also reduced the development of anthracnose on mango fruit (variety Kent) ^[30] and caused a higher pulp firmness and a lower soluble solid in ‘Hindi-Besennara’ mango ^[31].

Banana is one of the most consumed fruits around the world. Different propolis types were tested in the post-harvested conservation of bananas as a coating and dipping solution. Propolis treatment caused a lower fresh weight loss in banana cv. Prata (Mica sapientum L.) ^[32]^[33]. In a more recent research, the crown of banana fruit was coated with 50% propolis and paraffin alone and mixed together to control crown diseases and compared to 250 ppm prochloraz and non-treated fruit. Simulation for transportation, storage, and the retail market was performed. The coating using propolis combined to paraffin was the most effective treatment showing the same results as using prochloraz ^[34].

In the last year, many investigations were performed on propolis and its application as a bio preservative of fruits. Pullulan, an exopolysaccharide produced by Aureobasidium pullulans (fungus) and one of the most used natural coatings substances, was mixed with 5% and 10% propolis ethanol extract (PEE) and tested on the shelf life of blueberry (Vaccinium corymbosum) fruit. The coating mixture reduced the number of molds and bacteria, delayed blueberry ripening, and decreased weight loss ^[35]. In the same context, edible coating with chitosan nanoparticles and 10, 20, and 30% propolis were stated to act positively on the antioxidant activity and the shelf life of strawberries. In addition, the coating did not affect the sensory characteristics of strawberries ^[36]. Chitosan and propolis were also effective in the preservation of quality and sensory in fig fruit. In addition, the coating substance inhibited the growth of Aspergillus flavus by 20–30% under laboratory and semi-commercial conditions. The coated fig fruit produced lower aflatoxin ^[37]. Propolis was reported to inhibit up to 90% of S. vesicarium mycelial growth in vitro. In artificially wounded and inoculated Rocha pear (Pyrus communis L. cv Rocha), propolis decreased the disease incidence and the diameter of lesions by 25% and up to 57%, respectively ^[38].

Table 1. Application of propolis extracts in fruits and vegetables.

Foods	Geographical Origin	Used Extracts	References
Vegetables
Lettuce	Portugal (Bragança, 41°48′ N; 06°45′ W)	2% Methanol extract	^[4]
Cucumber and soybean	Brazil (Pato Branco)	PEE	^[5]
Celery, leek and butternut squash	Argentine (Juricich, Mendoza)	PEE	^[7]^[8]
Tomato, lettuce seeds, onion roots	Argentine (Tucumán, Santiago del Estero, Chaco)	PEE	^[6]
Tomato	Italy	Glycolic extract	^[9]
Tomato	Bandung	PEE diluted with propylene glycol	^[10]
Cherry tomato	China	PEE	^[11]
Cherry tomato	Poland Toruń County (53.03′ N, 18.62′ E)	PEE + pullulan	^[12]
Mashed potatoes	Jordan (naour region)	Nd	^[13]
Potatoes	Argentine (semi-arid regions of Salta, Santiago del Estero and Tucumán)	PEE	^[14]
Chili	Brazil	PEE	^[15]
Rice	Brazil (Minas Gerais)	Ethanol, methylene chloride and hexane extract	^[16]
Fruits
Sweet cherry	Turkey (Hatay province (Eastern Mediterranean region of Turkey))	WPE and PEE	^[17]
Sweet cherry	China	Propolis fresh liquid	^[18]
Citrus	China (Baoding County, Hebei Province)	PEAE (propolis ethyl acetate extract)	^[19]
Mandarins	China (Baoding County, Hebei Province)	PEAE	^[19]
Orange	Egypt	PEE	^[21]
Orange	Iraq (Baghdad)	Hydro-alcohol extract	^[22]
Orange	Brazil (southern state of Paraná)	PEE	^[23]
Citrus reticulata Blanco	Indonesia	PEE	^[24]
Star Ruby grapefruit	Turkey (Hatay province)	PEE	^[25]
Table grape, cv. Muscatel	Spain (Bonamel Organic S.L. (Alquería de Aznar, Spain)).	PEE	^[26]
Dragon fruit	China	PEE	^[27]
Apple	Iraq (Baghdad)	PEE	^[28]
Apple	Egypt	PEE	^[29]
Mango	Brazil	PEE	^[30]
Mango	Saudi Arabia	PEE	^[31]
Banana	Brazil (Bambuí, Minas Gerais)	Hydro-alcohol extract and aqueous extract	^[32]
Banana	Saudi Arabia (Hada Al-Sham 21°48′3′′ N, 39°43′25′′ E)	PEE	^[33]
Banana	Thailand (Ratchaburi province)	PEE	^[34]
Blueberry	Poland (Torun County)	PEE	^[35]
Strawberry	Mexico	-	^[36]
Fig	Mexico	-	^[37]
Pear	Portugal (central coast and the northern region)	PEE	^[38]

2. Beverages

Pasteurization is one of the most commonly used techniques applied in food industries to improve the preservation and stability of fruit juices. As an advantage, pasteurization will prolong the duration of storage of fruit juices. However, pasteurization, in particular thermal, may accelerate the degradation of some functional compounds present in fruit juices, such as lycopene ^[39]^[40]. Chang et al. ^[40] used a multivariate statistical approach to analyze the previous experimental data and to demonstrate whether propolis could be used as a bio preservative in fruit juices. As a result, propolis could be used as a bio-pasteurization agent. Propolis, with its dual antioxidant and antibacterial effect, will be an interesting alternative to conventionally used techniques ^[40]. In this effect, propolis was effective and exhibited antifungal activity on three isolates of Penicillium spp. and one isolate of Zygomycetes spp. in four unpasteurized fruit juices (mandarin, grape, orange, and apple) ^[41]. Propolis was also reported to be as effective as sodium benzoate and potassium sorbate on yeast and mold inhibition ^[3]^[42]^[43], Propolis was also effective in apple juice ^[44]^[45], freshly squeezed juice ^[46], and red fruit juice ^[47]. Propolis was also reported to increase the phenolic content of strawberry juice. A positive effect on the color value was also observed. Sensory data determined that the effect of the addition of PE (propolis extract) on the taste, smell, color, sourness, and general acceptability of strawberry juice samples was statistically significant ^[48].

The effect of propolis on fruit juice is represented in Figure 1.

Figure 1. Effect of propolis on fruit juice.

The studies the researchers described in the entry indicated that propolis has been widely tested in the preservation of fruits, vegetables, and fruit juices. The results demonstrated the effectiveness of propolis and its positive effect on the preservation of the deterioration caused by oxidation process or food spoilage. Propolis also seems to increase the levels of flavonoids and polyphenols, resulting in an increase in the antioxidant effect which will in turn slow down the oxidation of the tested products. However, opinions on its effect on the physical and sensory aspect remain mixed. Propolis concentration or percentage, used method for preservation, and duration will be the main factors to consider.

3. Dairy

Dairy products are the most consumed food products due to their high nutritive values. Dairy food items are highly perishable foods with a very short shelf life. Therefore, quick and optimized processing and storage conditions are necessarily required after milking. Today, many studies are carried out to extend the shelf life of dairy products. The substances used in these studies should be natural, accessible, economical, and in a structure that does not change the physical, sensory, and chemical properties of the product used. Propolis is seen as a high-potential option that meets this requirement ^[49]^[50].

Shaban and Galal ^[51] investigated the fortification and preservative ability of PE in raw and pasteurized milk. Milk’s organoleptic properties were not affected. Moreover, Propolis increased flavonoids, phenolics, and antioxidant properties. Propolis addition to milk did not increase milk acidity compared to the control group. In addition, propolis reduced bacterial counts, and no mold and yeast were detected until the 14th day. The reported results are very promising and indicate that propolis had a positive effect on the shelf life of raw and pasteurized milk. Therefore, propolis is recommended and can be used as a natural preservative (Table 2). Propolis at 5% inhibited Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, and Pseudomonas fluorescence on pasteurized milk, skimmed milk, and cheese (prepared with goat’s and cow’s milk). The outcomes of the investigation revealed that PE could significantly be applied to ready-to-eat milk products to avoid the growth of L. monocytogenes ^[52]. The co-administration of PE with glycerol exhibited a potent inhibitory activity against L. monocytogenes in chocolate-flavored milk. Moreover, the chocolate-flavored milk administered with glycerol and deodorized PE gained a considerable consumer acceptance rate without any negative comments or complaints. As a conclusion, propolis may exhibit an auspicious role in preserving dairy products ^[53]. El-Deeb ^[54] also suggested that propolis can effectively preserve dairy products such as milk and yogurt with improved shelf life. Gunes-Bayir et al. ^[55] produced yogurts with starter cultures containing different concentrations of cinnamon and propolis. The chemical analysis showed that the fat ratio decreased in propolis and/or cinnamon-treated groups. While only propolis-applied yogurt had a lower pH value, yogurts with the highest percentage and only cinnamon had the highest pH value. In the titratable acidity data, there was an increase in the groups to which only propolis was applied. In contrast, the titratable acidity of yogurts decreased in the groups in which propolis and cinnamon were applied together. In addition, a significant decrease in the colony count of Lactobacillus acidophilus was measured in all groups. Propolis incorporation in yogurts increased the number of Streptococcus thermophilus colonies. Regarding sensory properties, Chon et Al. ^[56] reported that the taste values of market milk, kefir, and yogurt evaluated after various propolis addition in sensory properties were similar to the control group or lower. Bilici et al. ^[57] reported similar results. Propolis was used to produce yogurt with improved functional properties. No change was observed in taste, smell, or color (Table 3). Propolis tested on fruit yogurts affected the titratable acidity and increased total phenolic content and DPPH inhibition in a dose-dependent manner ^[58].

Propolis was found to completely inhibit the growth of S. thermophilus, B. bifidium, and L. bulgaricus in kareish cheese in the groups applied up to 600 and 1000 mg. Moreover, a negligible difference in the moisture content of the cheese after applying PE associated with a slight decrease in the amount of moisture and total protein was observed. According to the obtained results, propolis can be an ideal natural preservative ^[59].

4. Meat

Meat and numerous meat products are crucial sources of several nutrients such as proteins, amino acids, minerals, and different vitamins. Meat products are an essential part of the human diet due to their nutritional value. Several studies have reported propolis antimicrobial and antioxidant potential to improve the shelf life of meat and different meat products without causing adverse effects (Table 2). The effects of propolis on meat products are shown in Figure 2.

Figure 2. Effect of propolis on meat.

A chitosan (Ch)-based coating containing a mixture of Zataria multiflora essential oil (EO) and propolis (PE) was prepared and tested for its ability to prevent spoilage and enhance the shelf life of breast meat chicken ^[60]. The effect of packaging material was evaluated on the poultry meat’s microbial, chemical, and sensory properties. The coating mixture reduced the presence of psychrotrophic bacteria on all days of storage. Chitosan can cause a decrease in the pH of chicken meat. Propolis addition did not cause any change in color parameters ^[60]. Vargas-Sánchez et al. ^[61] aimed to evaluate the lipid oxidation and the antimicrobial effect of non-commercial and commercially available PE on beef patties stored in refrigerators. Raw beef patties were kept at 2 °C for a duration of 8 days with polyvinyl chloride (PVC) wrapping. The microbial count, lipid oxidation, color, and pH parameters were determined. The non-commercial PE showed the most effective results for all the tested parameters. In addition, lipid oxidation was inhibited, and microbial counts were reduced. No significant changes were observed in the pH of propolis extract treated groups. Moreover, the addition of PE to patties maintained the fresh red color for 8 days compared to the control. These results demonstrated that PE could be used as a natural antimicrobial and antioxidant agent for prolonging beef shelf life.

Similar results were reported by Gedikoğlu ^[62] for raw beef patties. Propolis was most active against S. epidermidis, followed by E. faecalis and L. monocytogenes. The PE decreased the total number of mesophiles. Color is an excellent marker of the freshness of meat products and is a critical factor for consumers’ choices. The microbiological and oxidative changes affect the meat color, leading to discoloration through metmyoglobin formation. In this analysis, all color values were reduced in the post-test time for metmyoglobin formation. In addition, the redness value was reduced with PE addition.

Propolis was added to Italian salami and kept for 90 days. Physical properties analysis showed that the inclusion of propolis did not affect the drying process of Italian-style salami, and no effect was observed on the lightness of the salami during its shelf life. The addition of propolis did not change the acceptability of the product. Regarding sensory properties, the judges rated the flavors as smooth and pleasant. Still, some judges detected the smell of propolis in the salami, which led to less acceptance of the salami. After the 90-day storage period, the majority of consumers did not comment on the presence of a sour flavor in the products. This confirmed that the salami remained stable in terms of lipid oxidation. Some consumers have noted that salami has an unpleasant, strong taste residue. However, they could not state that this aroma comes directly from propolis ^[63]. The organoleptic and physicochemical changes of propolis were investigated on sausages. The pH decreased until the 8th day, but after the 16th day, it began to increase due to enzymatic denaturation during storage. The volatile nitrogen bases (TVB-N) were also increased. A correlation was observed between the pH and TVB-N increase on the 16th day. In addition, TBA values showed that lipid oxidation increased up to the 24th day. Sensory investigations revealed that the taste and smell of the cooked product were good. Most of the scores were within the acceptable range. There was no significant difference in sensory evaluation between the control and propolis-treated groups. Consumers accepted the sensory properties of propolis sausages. However, they suggested improving the color. The physical properties analysis also suggested that propolis could be used as an alternative for the natural preservation of meat products ^[64].

5. Chicken

Chicken meat is the most economical and widely consumed meat source as compared to mutton or beef. Chicken meat containing proteins, vitamins (such as pantothenic acid, thiamin pyridoxine, and thiamine), fats, and minerals (Cu, Zn, Fe, etc.) is highly nutritive for the human body. The presence of high moisture levels and proteins in chicken meat facilitates microbial growth, reducing the quality and shelf life of chicken meat. Several preservatives are in use to avoid spoilage of chicken meat ^[60]. Propolis is emergingly being applied in the food industry to preserve food products with enhanced shelf life (Table 2).

Jonaidi Jafari et al. ^[65] reported that Ch and PE could be used together to extend the shelf life of chicken fillets. Propolis extract was sprayed on chicken fillets or combined with chitosan. If PE is sprayed, the concentration decreases over time due to its high volatility, resulting in a less antimicrobial effect. However, co-administration of PE and Ch helps the extract retain its antimicrobial properties for a long time. Color and texture changes in propolis-treated groups were not significantly different compared to the control group. However, the acceptance of smell and taste decreased. Higher amounts of propolis may reduce some sensory properties of fillets. Despite of some adverse effects on the sensory values of the groups, Ch combined with PE could be used to extend the shelf life of the chicken fillets.

Another investigation ^[66] evaluated the effects of propolis on immune response and meat quality in broilers infected with Escherichia coli. The number of Coliforms, Staphylococcus, and Psychrotrophic bacteria decreased in the chest muscle of the PE-treated group. Considering the sensory characteristics, the PE-treated group revealed the highest score in general acceptability. The PE-treated group revealed an increased body weight, decreased mortality, decreased rate of E. coli re-isolation from internal organs, and accelerated infection recovery. Pochop et al. ^[67] tested the effect of PE in chicken feeds against Salmonella spp. colonization on the gastrointestinal tract. The results showed that all experimental groups treated with PE could inhibit and eliminate Salmonella spp., suggesting that propolis positively impacts Salmonella spp. in the gastrointestinal tract. This study is not only beneficial to prevent humans from being infected with Salmonella but could also be interesting for food manufacturing companies and may be explored in the extension of the shelf life of food products and saving the cost of storing food products while awaiting pathogen test results.

Kročko et al. ^[68] studied antibiotic resistance changes in chickens after exposure to propolis and bee pollen. In general, Enterococci are found in the normal microbiota of the gastrointestinal tract of chickens but are not considered safe. Intermediate resistance to erythromycin disappeared in most E. faecalis isolates of broiler chickens fed propolis in their diet. Likewise, ampicillin and gentamicin resistance decreased after bee pollen application. However, it was noted that the sensitivity of E. faecalis to vancomycin and teicoplanin did not show any change in propolis and bee pollen compared to the control. The results showed that antibiotics could be used in association with propolis and bee pollen in broiler chickens’ diets. Mahdavi-Roshan et al. ^[69] reported that the use of PE (up to 8%) did not negatively affect the color and general acceptance of chicken breast. In addition, positive increases were observed in the texture values. The significant results of this investigation in the era of the poultry industry have opened the door to naturally effective preservatives. Solving poor quality and shelf life problems using PE-applied products will benefit public health and the economy.

6. Fish

The aquaculture is susceptible to develop different bacterial infections, particularly when nurtured in dense conditions. The occurrence of infections in aquaculture is responsible for causing a reduction in the production rate and an escalation in mortality, resulting in economic shortfalls ^[70]. Antibiotics were used to treat the bacterial infections in aquaculture. The escalated use of antibiotics in fish farms induced an antibiotic resistance in humans and animals. For example, the emergence of resistance was observed for Aeromonas salmonicida, A. hydrophila, Yersinia ruckeri, Vibrio salmonicida, V. anguillarum, Edwardsiella icttaluri, E. tarda, and Pasteuralla piscida ^[71]. Some fish microbes are highly pathogenic to cause diseases in humans via food-borne or zoonotic routes. Therefore, it is critically required to develop potential antimicrobials to treat microbial illness in fish. As natural products are a focus of researchers nowadays, propolis has been found to possess significant antimicrobial as well as antioxidant activity. Numerous studies have reported the significance of propolis in terms of providing preservative effects in food (Table 2).

The PE (2, 8, 16%) was used to prepare gelatin films ^[72] and evaluated for the preservation of trout fillets. At the end of storage, all PE-treated groups showed a significant reduction in the growth of mesophilic and psychrophilic bacteria. In addition, mold and yeast counts were also reduced. Based on the obtained results, it was concluded that the 16% concentration of PE showed the highest antimicrobial activity against all tested microorganisms. The sensory were affected by the increase in time. It can be suggested that PE-containing films can be used to maintain and enhance the quality of trout fish by controlling microbial deterioration and improving the sensory properties.

Table 2. Impact of propolis on different food products.

Food Product		Propolis Type	Treated Microbe	Origin	Chemical Composition	Reference
Dairy	Buffaloes’ milk	PE	Total bacterial count, coliforms, molds, and yeasts	Plant Protection Department at the Faculty of Agriculture, Mansoura University	Phenolic compounds (11.18 ± 0.511 mg/g) Flavonoids (7.716 ± 0.587 mg/g) Antioxidant activity (70.44 ± 0.327%)	^[54]
	Milk	PEE	L. monocytogenes, S. aureus, B. cereus	Val di Cecina (Tuscany, 50–450 m above sea level)	Flavonoids (2.3%)	^[52]
	Yogurt	PE	B. animalis ssp. lactis, L. delbrueckii subsp. bulgaricus, L. acidophilus, S. thermophilus	Purchased from Sepe Natural Organic Products (Turkey)	300 mg of Brazilian propolis (3.5 mg Gallic Acid and 2.1 mg Quercetin per mL product)	^[55]
	Gorgonzola type cheese	Brazilian green PEE	S. equorum, S. saprophyticus, C. parapsilosis, S. cerevisiae, C. flavescens	Campo das Vertentes (Latitude 21 140 S. Longitude 44 590) in the state of Minas Gerais, Brazil	ND	^[73]
	Kareish Cheese	PEE	S. thermophilus, B. bifidium, L. bulgaricus	Plant Protection Department at the Faculty of Agriculture, Mansoura University	Phenolic Compounds (13.64 ± 0.440 mg/g), Flavonids (10.57 ± 0.605 mg/g), Antioxidant activity (67.12 ± 0.288%)	^[59]
	Milk	PEE	L. monocytogenes	Collected in June from Megalopolis (MEG), in the region of Arcadia (central Peloponnese), Greece	Phenolic acids and derivatives (1.42 ± 0.04 mg/g), Flavones and flavonols (6.20 ± 0.32 mg/g), Flavanones and dihydroflavonols (3.56 ± 0.11 mg/g)	^[74]
	Kashkaval Cheese	PE	Lactic acid bacteria, yeast, and fungi	Purchased from producers from three different regions in the Western part of Bulgaria—town of Simitli, Blagoevgrad district (41°53′ N 23°7′ E), town of Bankya, Sofia district (42°42′ N 23°8′ E), and village of Vladimir, Pernik district (42°26′ N 23°5′ E).	ND	^[75]
	Pasteurized and raw milk	Crude organic propolis	Total microbial count	New Valley, Dakhla	ND	^[51]
Meat	Dry meat sausage	Raw dark brown propolis	S. aureus, P. aeruginosa	Honey Mahvin Company	Phenolic content (27.08 ± 2.4 mg/g)	^[76]
	Chicken breast meat	PE	Pseudomonas spp.	Natural propolis was gathered from four beehives.	Flavonoids (25.2%), Sesquiterpenes (9.5%), Aromatic acids (5.01%), Aliphatic hydrocarbons (4.87%), Aromatic hydrocarbons (5.80%), Fatty acids (2.24%), Alcohol (2.05%), Aldehydes (1.45%), Triterpenes (1.12%)	^[60]
	Beef patties	PE	Mesophilic and Psychrotrophic	Commercial propolis samples 1 and 2 Non-commercial propolis; Pueblo de Alamos, Sonora, Mexico (29°07′129′N)	Cinnamic acid, Rutin, Myricetin, Quercetin, Chrysin, Kaempferol, Apigenin, Pinocembrin, Luteolin, Acetin.	^[77]
	Minced beef	PE	L, monocytogenes, P. aeruginosa, F. oxysporum, S. cerevisiae	National Research Center in Cairo, Egypt	Pentacosane (0.36%), 5,5-D2-Trans-4,3-Dihydroxycylopentene (0.64%), Dodecane (1.04%), Trans-cyclohexanol,2-(methylaminomethyl) (7.30%), 4-aminocyclohepta[f]thieno[2,3-b]pyridine (66.02%), A-Neooleana-3(5), 12-diene (22.99%), 4-Methoxyamphetamine (1.66%)	^[78]
	Minced beef	PEE	S. aureus, B. subtilis, B. cereus, L. monocytogenes, S. typhimurium and E. coli O157:H7	Purchased from an apiary located in the suburb of Kermanshah, in the west of Iran	Lindane (11.96%), caffeic acid (11.14%), 3-cyano-5, 6-dihydro-4-(methylthio)- 2-phenylbenzo[h] quinoline (10.31%), buxozine-c (9.01%), 3,4-bis (3-byano-2-methylphenyl)-2,5-dimethylfuran (8.45%), 12-azabicyclo [9.2.2] pentadeca-1(14), 11(15)-dien-13-one (8.43%), and naringenin (7.31%)	^[79]
	Beef Meatballs	WPE	S. epidermidis, E. faecalis, L. monocytogenes	ND	Antioxidant Activity (IC₅₀ 38.025 ± 0.135 µg/mL) FRAP (23.27 ± 0.26 µM of Fe + 2/g)	^[62]
Chicken	Chicken breast fillet	PEE	Pseudomonas spp.	Natural propolis was gathered from four beehives	Flavonoids (25.2%), Sesquiterpenes (9.5%), Aromatic acids (5.01%), Aliphatic hydrocarbons (4.87%), Aromatic hydrocarbons (5.80%), Fatty acids (2.24%), Alcohol (2.05%), Aldehydes (1.45%)	^[60]
	Chicken fillet	PEE	S. aureus	Collected from different locations of Tehran Province in May 2016	ND	^[65]
	120-days old chicks	Egyptian propolis	E. coli	Dakahlia Governorate	ND	^[64]
	One day old chickens	PE	Salmonella spp.	ND	ND	^[67]
	Broiler chickens of hybrid combination Ross 308	Propolis and bee pollen	E. faecalis	ND	ND	^[68]
	Chicken breast	PE	Yeasts and molds, S. aureus, E. coli	Supplied from the mountainous area west of Guilan province	Phenolic compounds (5.83 ± 0.505 g/ 100 g), Flavonoids (4.92 ± 0.562 g/mL), Antioxidant activity (15.83 ± 2.341%)	^[69]
Fish	Nile tilapia (Oreochromis niloticus)	PEE and crude propolis	A. hydrophila	Collected in summer from Upper Egypt	ND	^[80]
	Fish	PEE	A. hydrophila, Y. ruckeri and S. iniae	Adana region	Propolis antioxidant activity value (569.68 µmol trolox/g), phenolic substance content (593.31 mg GAE/g)	^[72]
	Rainbow trout (Oncorhynchus mykiss)	PE	Mesophilic, psychrophilic, yeast and mold, coliform	Adana region	Propolis antioxidant activity value (569.68 µmol trolox/g), phenolic substance content (593.31 mg GAE/g)	^[72]
	Fish	PE	L. plantarum, E. cloacae, P. luteola, P. mirabilis, and P. damselea	Propolis was produced by Apis mellifera in January 2020, Adana, Turkey.	ND	^[81]
	Rainbow trout (Oncorhynchus mykiss)	PE	Mesophilic and psychrotrophic bacteria	Propolis was collected from a farm at village Kocaavsar in Balikesir, Turkey.	ND	^[82]
	Nemipterus japonicus fillets	PEE	Mesophilic and psychrotrophic bacteria	ND	ND	^[83]

References

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