P U B L I C A T I O N S CODON Italian Journal of Food Science, 2021; 33 (SP1): 163–169 ISSN 1120-1770 online, DOI 10.15586/ijfs.v33iSP1.2093 163 Replacement of meat by mycoproteins in cooked sausages: Effects on oxidative stability, texture, and color Narges Shahbazpour1, Kianoush Khosravi-Darani2*, Anousheh Sharifan1, Hedayat Hosseini2 1Department of Food Science and Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran; 2Department of Food Science and Technology, Faculty of Nutrition Science and Food Technology, National Nutrition and Food Technology Research Institute, Shahid Beheshti University of Medical Sciences, Tehran, Iran *Corresponding Author: Kianoush Khosravi-Darani, Department of Food Science and Technology, Faculty of Nutrition Science and Food Technology, National Nutrition and Food Technology Research Institute, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Email: k.khosravi@sbmu.ac.ir, kiankh@yahoo.com Received: 23 June 2021; Accepted: 16 November 2021; Published: 12 December 2021 © 2021 Codon Publications OPEN ACCESS PAPER Abstract Processed meat is one of the most consumed products worldwide. Naturally, production of proteins with ani- mal origins includes limitations such as costs, energy, time, and environmental problems. Thus, replacement of meats by alternative biomaterials such as mycoproteins can be promising. Mycoproteins with hyphal mor- phologies, including branches and lengths, have close structures to meat and can be a potential alternative for meat products. Therefore, the major objectives of this study included complete replacement of sausage meats by mycoproteins and comparing characteristics of the novel formula with those of meat. In general, physicochemi- cal, microbial, nutritional, and mechanical characteristics of the formulas were assessed. Results showed that the mycoprotein substitution improved the nutritional and health effects due to the higher valuable protein and lower lipid contents. Besides, it had a high content of essential amino acid and unsaturated fatty acid, compared to meat sausage. Absence of yeasts, molds, Salmonella spp., Eshrichia (E.) coli, and Staphiloccocus (S.) aureus verified the effectiveness of the heat treatment and also the effectiveness of the hygienic procedures in both samples. With regard to phycicochemical properties, more contents of moisture and lipids in sausages containing mycoprotein were linked to further water binding capacity (WBC) (P < 0.05) and oil binding capacity (OBC) in them, compared to beef samples. Besides, the mycoprotein sample had lower (P < 0.05) values of carbohydrates, ash, and pH, com- pared to the beef sample. In contrast, beef sausages had better textural characteristics, such as hardness, cohesive- ness, gumminess, and springiness indexes, compared to mycoprotein sausages. Higher water and OBC values of the mycoproteins led to the filling of the protein interstitial spaces as well as decreasing of the textural attributes. Thus, it resulted in the use of less oil and water in mycoprotein formulations. In conclusion, mycoproteins can be addressed as appropriate replacements for meats in sausages. Keywords: meat alternatives; mycoproteins; nutritional values; sausages; textural properties Introduction In recent decades, the world population has significantly increased from 2.6 to 8 billion individuals (Gabriel et al., 2014). If the global population grows at the recent rate, it may reach 9 billion individuals by 2042, which could pose serious problems in providing food to all (Upadhyaya et  al., 2016). Approximately, 1 billion people globally will not be able to properly access food sources with suf- ficient energy and proteins (Godfray et al., 2010). This can result in serious medical problems such as defective immune system and stunted growth. In contrast, high 164 Italian Journal of Food Science, 2021; 33 (SP1) Shahbazpour N et al. blood cholesterol. Toxin analysis and allergy assays have shown no general concerns (Hashempour‐Baltork et al., 2020a). However, a little information is available on var- ious formulations for the replacement of food meats by mycoproteins. Hence, the major objective of the current study was to compare physicochemical, microbial, nutri- tional, and mechanical characteristics of sausages con- taining mycoproteins with those containing meat. Materials and Methods Preparation of sausages Sausage samples (40% red meat) were prepared in three replications in a famous meat production factory, Tehran, Iran. Frozen beef samples were defreezed at 4°C for 16 h before use. Then, beef samples were minced twice using laboratory mincer (Model MK-G1800, Panasonic, Japan) equipped with 6–10 mm steel plates. Mycoprotein masses were provided by Ghazabon Paya, Iran. Two sau- sages of mycoproteins and meats were prepared for 4 kg batters (Table 1) based on the guidelines from meat pro- ducers. Formulations of both samples were mostly simi- lar, with differences in meat and mycoprotein contents. To prepare sausages, minced beef/mycoprotein was transferred into a bowl chopper (Robot Coupe Model R-10, France) and mixed slowly with the dry ingredients, except spices. Ice was continuously added to the mix- ture in the chopping process to control the temperature. Then, oil and spices were added to the mixture, respec- tively. The total time of mixing was 10 min while the final temperature of the batters was set below 12°C. Then, the batters were stuffed into impermeable cellulose casings using a hydraulic piston-type stuffer. Then, the sausages were cooked at 76°C for 60 min using a steam chamber and then cooled down to a final temperature of 10°C using ice-water bath and stored at 4°C overnight (Kamani et al., 2019). In general, each sausage type was prepared in two batches. Totally, two sausages from each batch were chosen for further analysis. consumption of meat products can pose serious health problems as well (Qian et al., 2020). Research and devel- opment of meat replacements majorly focuses on the production of products that imitate the physical char- acteristics of meat such as appearance, taste, and tex- ture, as well as providing its nutritional values. Muscle products, such as chicken and steaks, minced products, such as burgers and nuggets, and emulsion products, such as Frankfurter and Mortadella sausages, are the major meat replacements (Kyriakopoulou et al., 2021). However, excessive meat consumption can significantly affect the global climate change (Hashempour-Baltork et al., 2020b). Thus, the quest for novel substitutes for ani- mal proteins requiring less financial resources, energy, and time consumption can be promising. One of the best meat substitutes includes mycoproteins with relatively similar textures to meat (Upadhyaya et al., 2016). The common source of mycoproteins is usually Fusarium (F.) venenatum, a filamentous fungus generally recognized as safe (GRAS) (Hashempour-Baltork et al., 2020b). Generally, mycoproteins contain 10 g of carbohydrates, 13 g of fats, 25 g of fibers, and 45 g of proteins, as well as various vitamins, carotenes, minerals, and essential amino acids (EAA) in 100 g of dry matter (Finnigan et al., 2019). Recent studies on human volunteers have demon- strated that biological values of proteins from mycopro- teins are similar to biological values of proteins from milks. Commonly, sensory characteristics, such as texture, taste, and overall appearance of the final products, are critical for their overall acceptance. Fusarium biomass is virtually odorless and tasteless and is appropriate to imitate the consistency and taste of regular meats. Comparing myco- proteins with textured soya and poultry muscle tissues, recent studies using microscopy techniques have shown that large cable-like fibers (especially in vegetable protein textures) result in rubbery textures, which are unfavor- able when chewing. Technically, further fibrous and rub- bery eating qualities occur in poultry and mycoprotein products due to their tight packaging laminations, com- pared to those that occur in products of vegetable pro- tein origins (Hashempour-Baltork et al., 2020b). Ideally, mycoproteins can be good replacements for meat since their dry weights include nearly 50% of proteins, sim- ilar to grilled steaks. However, the fungi include lower fat quantities (∼13%) than those steaks do. Furthermore, this is a vegetable fat with cholesterol alternative (ergos- terin) and good fiber content (∼25%), which are increas- ingly accepted by the health-conscious people (Finnigan et al., 2017). Based on the literature, mycoproteins can mitigate the problem of food unavailability worldwide. Moreover, even routine production of mycoproteins would need only lower water resources and occupy less  land (Hashempour-Baltork et al., 2020b). The use of mycoproteins can limit foodborne diseases and lower Table 1. Sausage ingredients. Ingredient Content (% w/w) Meat/mycoprotein 40 Sunflower oil 10 Ice 20 Mixed spices 3.5 Soy protein isolate 5 Gluten 10 Flour 10 Salts 1.5 Italian Journal of Food Science, 2021; 33 (SP1) 165 Physicochemical assessment of sausages containing mycoproteins One-hundred microliters of this hydrolysate were care- fully injected into the HPLC instrument (Waters, USA) at 40°C. The HPLC instrument was equipped with Zorbax Eclipse-AAA Column (4.6 mm × 150 mm, 5 μm) (Agilent Technologies, USA) as well as a fluorescence detector. Furthermore, sodium dihydrogen phosphate (NaH2PO4) solution (40 mmol l−1) was used in the instrument as Mobile Phase A and acetonitrile:methanol:water solution (45:45:10 v/v/v) as Mobile Phase B. Fatty acids Generally, the Folch method (chloroform:methanol 2:1 v/v) was used for the sample lipid extraction (Folch et  al., 1957). Fatty acid methyl esters (FAME) were methylated based on the European Official Methods of Analysis (Godfray et al., 2010; Hashempour-Baltork et al., 2018). The FAME were analyzed using gas chro- matography (GC) (Agilent 6890, Agilent Technologies, USA) equipped with capillary column (30 m per 0.25 mm ID, 0.25-μm film thickness) and flame ionization detector (FLD) (Thermo TR-5, ThermoFisher Scientific, USA). The instrument used helium (He) as the carrier gas with 0.2 mL min−1 flow rate based on a method described by Hashempour-Baltork et al. (2017). Fatty acid (FA) was identified by comparing the sample and the reference methyl ester chromatograms (Sigma- Aldrich, USA). Mechanical characteristics Texture profile analysis (TPA) was performed using Stable Micro Systems Texture Analyzer Model TA.XT Plus (Stable Micro Systems, UK). The analyzer was equipped with a 50-kg load cell. The sample was cut into pieces of 25 mm and axially fixed on the platform. Two- cycle compression assay was carried out with up to 50% of the strain compression of the original height using steel probes. Return speed, distance, and contact force were 2 mm s−1, 50 mm, and 20 g, respectively. The vari- ous attributes of the food, including cohesiveness, hard- ness (N), adhesiveness (N.S), springiness (%), gumminess, chewiness, and springiness, were assessed (Kamani et al., 2019). Statistical analysis Descriptive data of the study were recorded as mean ± SD (standard deviation) using SPSS Software v.17 (IBM Analytics, USA). Duncan’s test was used for the compar- ison of means and different letters represent significant statistical differences (P < 0.05). For each sample of each test, three replicates were used. Physicochemical characteristics Proximate pH, moisture, protein, lipids, carbohydrate, ash, peroxide value, water holding capacity (WHC), oil binding capacity (OBC), and water binding capacity (WBC) of the two sample types were assessed using offi- cial methods (AOCS, 2017). Microbial characteristics The count of microorganisms, Escherichia (E.) coli, Salmonella spp., Staphylococcus (S.) aureus, Bacillus (B.) cereus, Clostridium (C.) perfringens, yeasts, and molds were enumerated based on the ISO protocols (ISO, 2013). Nutritional characteristics Vitamins Briefly, 10 g of the ground samples were weighed using a 50-mL glass beaker. Then, 20 mL of fresh 5% (w/v) met- aphosphoric acid solution were added to the vessel and mixed well. The mixture was then homogenized by stir- ring at room temperature (RT) for 2 min. The homog- enate was centrifuged at 3000 rpm for 5 min; then, the upper solution was filtered using Albet no. 1305 filter papers and re-filtered using 0.45-µm Millipore filters for liquid chromatography (LC) analysis (Valls et al., 2001). Then, stock solutions of 100 µg mL−1 vitamin B5 (panto- thenic acid), vitamin B9 (folic acid), vitamin B2 (ribofla- vin), and vitamin B7 (biotin) (Sigma-Aldrich, St. Louis, MO, USA) were prepared and stored in 4  °C until use. The vitamin content of the sample was assessed using the standard curve, and high-performance liquid chro- matography (HPLC) (Waters, USA) was used for the assessment of vitamin B according to Sasaki et al. (2020). The method included the use of Capcell Pak C18 SG120 HPLC Column (250 nm × 4.6 mm with 5-μm particle sizes) (Osaka Soda, Japan), gradient elution of phosphate buffer-acetonitrile (pH 3), ion-pairing reagent (mobile phase) with 1.0 mL min−1 flow rate and UV detection (210 nm). The vitamin B compound was separated within 60 min. The value of 0.01 µg g−1 was set as the detection limit. Amino acids Briefly, 10 g of dried sausage sample were hydrolyzed using 1 N hydrochloric acid (50 mL) based on an origi- nal protocol by Czauderna et al. (2003). Amino acid (AA) standards were purchased as a cell-free AA mixture from Sigma-Aldrich, St. Louis, Missouri, USA. The sample was then centrifuged (10,000 g) to collect the hydrolysate. 166 Italian Journal of Food Science, 2021; 33 (SP1) Shahbazpour N et al. of yeasts, molds, Salmonella spp., and E. coli verified the effectiveness of the heat treatment in both samples. Due to the absence of S. aureus, the hygienic proce- dures seemed to be effectively preventive. The number of B. cereus and C. perfringens were similarly reported to be less than 10 cfu g−1 in both samples. Researchers demon- strated that the presence of NaCl and phosphate might inhibit the bacterial growth (Kim et al., 2021). Moreover, a similar report has been published on increased load of Pseudomonas spp. in fish sausages enriched with mycoproteins during refrigerated storage (Bahmani and Movanes, 2021). Nutritional assessments Vitamins Levels of vitamins B2, B5, B7, and B9 were assessed in both samples (Table 3). Contents of vitamin B9 showed no significant difference (P < 0.05) between the sam- ples. For other vitamins, mycoprotein sausages signifi- cantly achieved higher scores (P < 0.05). According to Hashempour-Baltork et al. (2020), who comprehen- sively compared the vitamin B content in meats and mycoproteins, contents of riboflavin, niacin, pyridox- ine, and pantothenic acid was 9, 14, 5, and 10 µg g−1 in mycproteins and 0.018, 0.5, 0.052, and 0.35 µg g−1 in meats. These differences were seen in the formulated products as well. Results and Discussion Physicochemical assessments Proximate analyses of sausages formulated with myco- proteins and beef are shown in Table 2. Results of mois- ture contents demonstrated that moisture in sausages formulated with mycoproteins significantly was higher (P < 0.05) than moisture in sausages formulated with beef. Protein and lipid contents seemed to increase with the substitution of mycoproteins in the formulation of sausages. Higher contents of moisture and lipids were linked to further WBC (P < 0.05) and OBC in mycopro- tein samples, compared to beef samples. Based on the results, mycoprotein formulation included lower values of carbohydrates, ash, and pH, verified by the previous studies (Hashempour-Baltork et al., 2020). The low pH in mycoprotein samples was associated with low pH of mycoproteins (4.7) in comparison to pH of meat (5.6). In another report, use of mycoproteins significantly increased nutritional values of fish sausages (e.g., ash, carbohydrate, fat, and protein) (P < 0.05) (Bahmani and Movanes, 2021). In this study, the pH of fish sausages enriched with mycoproteins increased during storage (Bahmani and Movanes, 2021). No significant difference (P < 0.05) was generally reported between the two for- mulations of sausages in WHC and peroxide values (P > 0.05). Characterization of these two sausage samples indicated that lesser oil and water should be used in the formulation due to the higher WBC and OBC values in mycoproteins than beef. In addition to higher OBCs, higher proteins and lower carbohydrates could be used in mycoprotein samples as appropriate meals for obese people. These findings verified previous findings, which addressed mycoproteins as healthy nutritious proteins (Finnigan et al., 2019). Microbial assessments Microbiological analysis was carried out on Day 1 after cooking to understand heat behaviors of mycoproteins and compare microbial patterns of the samples. Absence Table 2. Proximate analysis and pH of beef sausage. Treatment Moisture (%) Protein (%) Lipid (%) Carbohydrate (%) Ash (%) pH Peroxide (mEq/kg) WHC** (%) WBC** (mL/g) OBC** (mL/g) Mycoprotein- sausage 59 ± 1.9a* 12.5 ± 1.0a 13.9 ± 1.2a 9.8 ± 1.1b 2 ± 0.5b 5.7 ± 0.01b 1.2 ± 0.2a 54 ± 2.5a 0.98 ± 0.09a 0.37 ± 0.08a Beef sausage 47.5 ± 1.5b 11.6 ± 1.1b 8.9 ± 1.3b 10.9 ± 1.2a 3 ± 0.4a 6.5 ± 0.01a 1.3 ± 0.2a 55 ± 3.3a 0.79 ± 0.1b 0.3 ± 0.08b *Mean ± SD, Different letters represent significant differences (P < 0.05). **WHC: water holding capacity, WBC: water binding capacity, OBC: oil binding capacity. Table 3. The level of vitamin B group in mycoprotein/beef sausages (µg/g). Treatment Vit B2 Vit B5 Vit B7 Vit B9 Mycoprotein sausage 3.31 ± 0.5a* 0.02 ± 0.05a 8.81 ± 1.45a 0.48 ± 0.05a Beef sausage 1.51 ± 0.5b 0.01 ± 0.04b 2.57 ± 1.5b 0.51 ± 0.03a *Mean ± SD, Different letters represent significant differences (P < 0.05). Italian Journal of Food Science, 2021; 33 (SP1) 167 Physicochemical assessment of sausages containing mycoproteins Fatty acids The FA composition of the sausage samples are pre- sented in Table 5. The total SFAs in mycoprotein and beef sausages were 29.7 and 60.7% (w/w), respectively. In fact, unsaturated fatty acid (UFA) levels in mycopro- tein sausages (65.44) were significantly (P < 0.05) higher than UFA levels in beef sausages (35.54% w/w). These contents as well as previous contents highly verified the results, especially for the ratio of UFA to saturated fatty acids (SFA) of 2:1 (Reihani and Khosravi-Darani, 2018). In 2009, Hosseini et al. (2009) reported 3.2–3.5:1 ratio of UFA to SFA. Higher consumption of USFAs provides health benefits to patients with cardiovascular diseases (CVD). Naturally, the ratio of polyunsaturated fatty acid (PUFA) to SFA in beef is typically 0.1. However, the ratio decreases with an increase in meat fats (Vahmani et  al., 2015). Naturally, the ratio reaches 1.44 in mycopro- teins. Chicken fat naturally includes 30% of SFAs, 45% of monounsaturated fatty acids (MUFA), and 21% of PUFAs (Hashempour‐Baltork et al., 2020; USDA, 2008). These values are close to those of mycoproteins (Table 5). Mechanical assessments Table 6 represents mechanical properties of the cooked samples. Hardness of mycoprotein sausages was signifi- cantly lower than that of meat sausages (P < 0.05). Similar decreases were reported for cohesiveness when meat was totally replaced by mycoproteins. It was reported that beef sausages needed a greater force of chewing, com- pared to that of nonmeat samples. This was possibly due to the occurrence of stronger networks in myofibril pro- teins, increasing the product resistance to compression. Mycoprotein sausages showed lower values for gummi- ness and springiness (P < 0.05). Lower springiness val- ues were reported by Youssef and Barbut (2011), when soy protein extracts were used as meat alternatives in emulsified meat batters. Kamani et al. (2019) recorded lower levels of food hardness, cohesiveness, gumminess, and springiness by replacement of meats by proteins of Amino acids Based on the nutritional and physiological roles, AAs can be differentiated as EAAs, including valine, tryp- tophan, threonine, phenylalanine, methionine, lysine, isoleucine, leucine, histidine (essential for infants), arginine (semi-essential), and nonessential amino acids (NEAA), including tyrosine, serine, proline, glycine, glutamine, glutamic acid, cysteine, asparagine, aspar- tic acid, and alanine (Damodaran and Parkin, 2017). The AA profiles showed that mycoprotein sausages included almost all EAAs, compared to beef sausages (Table 4). These findings were similar to those of other studies, reporting the presence of EAAs in single-cell proteins (SCP) of F.  venenatum (Hashempour‐Baltork et al., 2020). In fact, three food categories totally pro- vide 80.9% of daily protein needs of humans (Górska- Warsewicz et al., 2018). The significance of protein nutrition includes EAA content, biological value, digestibility, net protein use, and protein efficiency ratio. Hashempour-Baltork et al. (2020) compared the quality of the mycoproteins with that of meat proteins and demonstrated that nutritional indices of these two sources were almost similar. Monteyne et al. (2020) reported that mycoproteins were good food sources enriched with EAAs. Table 4. The amino acid profile in mycoprotein/beef sausages (µg/g). Amino acid Content in mycoprotein- sausage (%g per 100 g protein) Content in beef- sausage (mg per 100 g protein) L-Alanine 4.84 ± 0.99b* 14.90 ± 0.59a* L-Arginine 6.74 ± 0.07a 6.0 ± 0.18a Aspartic acid 5.25 ± 0.03b 15.20 ± 0.10a L-Cystine 11.21 ± 0.15b 96 ± 0.05a L-Glutamic 12.92 ± 0.52b 29.02 ± 0.12a Glycine 4.15 ± 0.23b 9.05 ± 0.35a L-Histidine 3.20 ± 0.75a 1.10 ± 0.85b L-Isolucine 5.90 ± 0.63a 3.00 ± 0.53b L-Leucine 7.80 ± 0.35a 5.01 ± 0.33b L-Lysine 7.50 ± 0.61a 3.0 ± 0.66b L-Methionine 1.90 ± 0.80a 1.2 ± 0.11b L-serine 5.35 ± 0.23a 1.04 ± 0.17b L-Theronine 13.75 ± 0.77a 11.95 ± 0.95b L-Tyrosine 4.35 ± 0.97a 4.950 ± 0.907a L-Valine 5.90 ± 0.70a 4.10 ± 0.07b Phenyl Alanin 9.78 ± 0.87a 3.15 ± 0.50b Proline 5.30 ± 0.77b 17.1 ± 0.60a Tryptophan 6.7 ± 0.60a 1.5 ± 0.70b *Mean ± SD, Different letters represent significant differences (P < 0.05). Table 5. Fatty acid profile of mycoprotein/beef sausage. Fatty acid Content in mycoprotein- sausage (% w/w) Content in beef- sausage (% w/w) Palmitic (C16:0) 18.8 ± 0.20b 32.4 ± 0.29a* Stearic (C18:0) 10.90 ± 0.16a 28.3 ± 0.20b Oleic (C18:1) 24.95 ± 0.11a 9.13 ± 0.365b Linoleic (C18:2) 25.35 ± 0.15a 21.31 ± 0.353b α-Linolenic (C18:3) 15.14 ± 0.18a 5.10 ± 0.748b *Mean ± SD, Different letters represent significant differences (P < 0.05). 168 Italian Journal of Food Science, 2021; 33 (SP1) Shahbazpour N et al. The production of meat alternatives seems necessary due to the preference of consumers for vegetarian diets, and the increasing nutritional awareness of the popu- lace. Like other functional foods that were unknown in the past, after numerous studies and production of var- ious products, today there is a unique response to these products. In the past 2 years, the COVID-19 pandemic has drawn the public attention to food security and meat supply worldwide with further global demands for meat alternatives with plant origins. Acknowledgment This study was supported financially by the National Nutrition and Food Technology Research Institute, School of Nutrition Sciences and Food Technology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Conflict of interest The authors report no conflict of interest. References AOCS, 2017. Official methods and recommended practices of the aocs, reapproved 2017. Available at: https://www.aocs.org/ attain-lab-services/methods/methods/search-results?subject=B. Bahmani, Z. and Movanes, F., 2021. Enrichment of carp sausage with Fusarium venatum mycoprotein. Utilization and Cultivation of Aquatics 10(11–25). Czauderna, M., Kowalczyk, J., Niedzwiedzka, K. and Wasowska, I., 2003. A highly efficient method for determination of some amino acids and glutathione by liquid chromatography. Journal of Animal and Feed Sciences 12(1): 199–214. https://doi. org/10.22358/jafs/67697/2003 Damodaran, S. and Parkin, K.L., 2017. Amino acids, peptides, and proteins. In: Fennema’s food chemistry. CRC Press, pp. 235–356. Edited By Srinivasan Damodaran, Kirk L. Parkin. Boca Raton. Finnigan, T., Needham, L. and Abbott, C., 2017. Mycoprotein: a healthy new protein with a low environmental impact. In: Sustainable protein sources. Elsevier, pp. 305–325. Sudarshan R. Nadathur Givaudan Flavors, Cincinnati, OH, United States; Janitha P.D. Wanasundara Agriculture and Agri-Food Canada, Saskatoon SK, Canada; Laurie Scanlin Colorado State University, Fort Collins, CO, United States plant origin in chicken sausages. Researchers concluded that proteins of nonmeat origin could include further fat and water contents, which might fill the protein interstitial spaces and decrease the product springiness (Kamani et al., 2019; Youssef and Barbut, 2011). This is also addressed for mycoprotein replacement regarding WBC and OBC (Table 2). Textural analysis demonstrated association of sample meats with lower values of adhe- siveness. It could be interpreted that a decrease in meat quantity might lead to a significant decrease in the con- sistency of the cooked emulsions. Conclusion This study was carried out to investigate the appropri- ateness of mycoproteins as complete substitutes for meat in beef sausages. The results showed that mycoprotein substitution improved nutritional and health effects due to the high-value proteins with EAAs and less lipid content (mostly UFAs). Also, it has high contents of EAA and UFA, compared to meat sausages. Absence of yeasts, molds, Salmonella spp., Eshrichia (E.) coli, and Staphiloccocus (S.) aureus verified the effectiveness of heat treatment and also hygienic procedures in sam- ples. Phycicochemical evaluations show higher contents of moisture and lipids in sausages containing mycopro- tein due to WBC and OBC, compared to beef samples. Besides, mycoprotein samples had lower values (P < 0.05) of carbohydrates, ash, and pH, compared to beef samples. However, mycoprotein sausages achieved lower scores of hardness, cohesiveness, gumminess, and springiness in mechanical assessments, compared to beef sausages. Mycoproteins could hold excessive water and fat caused by higher WBC and OBC, filling the protein interstitial spaces and decreasing the springiness. This suggested less use of oil and water in mycoprotein formulations. This study has provided valuable information for increasing public awareness on the characteristics of mycoprotein products, including nutritional, textural, and formulation characteristics. This is the first study that substituted meat with myco- proteins in sausages. However, further studies are nec- essary to optimize mycoprotein sausages using texture improvement ingredients to enhance their gel-forming and textural characteristics. These are currently the major problems in the manufacturing of meat-free sausages. Table 6. The hardness, adhesiveness, springiness, cohesiveness, chewiness and gumminess of mycoprotein/beef sausages. Treatments Hardness (N) Adhesiveness (N.S) Springiness (%) Cohesiveness Gumminess Chewiness Mycoprotein-sausage 23 ± 1.1b* 1.5 ± 0.19a 0.49 ± 0.21b 0.19 ± 0.02b 20.1 ± 1.2b 1.0 ± 0.01b Beef-sausage 37.12 ± 1.5a 0.5 ± 0.10b 0.75 ± 0.2a 0.24 ± 0.01a 25.4 ± 1.5a 2.1 ± 0.2a *Mean ± SD, Different letters represent significant differences (P < 0.05). Italian Journal of Food Science, 2021; 33 (SP1) 169 Physicochemical assessment of sausages containing mycoproteins and Technology 56(5): 2660–2669. https://doi.org/10.1007/ s13197-019-03754-1 Kim, T.-K., Yong, H.-I., Jung, S., Kim, H.-W. and Choi, Y.-S., 2021. Technologies for the production of meat products with a low sodium chloride content and improved quality characteristics—a review. Foods 10(5): 957. https://doi.org/10.3390/foods10050957 Kyriakopoulou, K., Keppler, J.K. and van der Goot, A.J., 2021. 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