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Changes in free amino acids and hardness in round of Okinawan delivered cow beef during dry- and wet-aging processes

Changes in free amino acids and hardness in round of Okinawan delivered cow beef during dry- and... Background: Aging trials are conducted to determine characteristics associated with dry- and wet-aging processes of beef from delivered cows grown in Okinawa, i.e., dams that have finished giving birth (Okinawan delivered cow beef). Changes in free amino acids, hardness, and other factors were analyzed in round of Okinawan delivered cow beef during dry- and wet-aging processes along with a comparison with characteristics of beef imported from Australia. Results: Functional amino acids did not increase during both dry- and wet-aging processes. However, proteinogenic amino acids increased significantly (P < 0.05) and hardness tended to decrease during both dry- and wet-aging processes. On comparison between dry- and wet-aging processes by analysis of variance, drip and cooking losses were significantly lower during the dry-aging process than during the wet-aging process. However, there was no significant difference in free amino acids or hardness in this comparison. Conclusion: There was no significant difference between dry- and wet-aging methods for all studied variables related to free amino acids or hardness in this study. Keywords: Dry aging beef, Okinawan delivered cow, Free amino acids and hardness Background Western countries but also in Asian countries, such as The livestock industry is a major part of the Okinawan South Korea, Japan, Singapore, Taiwan, and Hong Kong [3]. economy, with the Prefecture selling the fourth highest Over 2.5 million foreign tourists visited Okinawa in number of calves in Japan. In Okinawa, there are numer- 2017, which is the highest number recording in any year, ous dams, some of which have finished giving birth and according to the official figures of the Okinawa prefectural thus, are termed as Okinawan delivered cows. The turn- government. Tourists from Asian countries, particularly over for Okinawan delivered cows is approximately 5000 those from Taiwan, Hong Kong, South Korea, and China, heads per year; these slaughtered cows are generally sold accounted for approximately 90% of this number. In this at a lower price than fattened cattle and heifers. The rea- context, there is an urgent need to boost the production son for them being less valuable is their less tender meat. and awareness of Okinawa’sunique food brands.Thus,we However, there is significant potential for adapting these hope to create an Okinawan brand of dry-aged beef by cows’ meat and increasing their value. adding value to Okinawan beef because it is produced in Dry aging is one aging method that is said to improve Okinawa and has high potential to become very popular meat with respect to some characteristics. Recently, in- in view of the region’s tourist industry. creased interest in dry-aged beef has emerged not only in During the dry-aging process, juices are absorbed into the meat and chemical breakdown of protein occurs, giving amoreintense nuttyand beefyflavor[3]. Moreover, during aging, the endogenous enzymes break down myofibrillar * Correspondence: hangskit@pref.okinawa.lg.jp proteins in the muscle, which leads to more tender beef [1, Okinawa prefectural Industrial Technology Center, 12-2 Suzaki, Uruma-shi, Okinawa 904-2234, Japan 2]. Because of this, it is considered that lean meat with low Present address: Okinawa prefectural Agricultural Research center, 820 fat content is more suitable for dry aging and has higher Makabe, Itoman city, Okinawa 901-0336, Japan potential to undergo a change in its quality. Specifically, Full list of author information is available at the end of the article © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 2 of 9 round of beef (i.e., from the rear leg of the cow) is most each aging experiment. With regard to Australian likely to be appropriate for dry aging. Indeed, dry-aging beef, six chunks of meat (approximately 6 kg/each) products using round of beef have already been commer- from the round of each individual were grouped into cialized in Okinawa. dry-aging and wet-aging experiments, namely, three In general, the dry-aging process is performed under in each group. Each chunk of meat was divided into aerated conditions, whereas the wet-aging process es- five pieces by cutting for experiments involving 0, 1, sentially involves vacuuming and packaging, which 2, 3, or 4 weeks of aging. means that the conditions do not involve aeration. Savell [7] reported a survey on consumer preferences Methods between dry-aged and wet-aged beef in an executive Aging environment summary of the National Cattlemen’s Beef Associa- The dry-aging environment was created in a refriger- tion’s Center in the United States of America (herein- ator (Showa Denko K.K., Tokyo, Japan) in Okinawa after referred to as “USMEF”). There is only a little Industrial Technology Center at a temperature of 2 °C. thing known about specific differences by scientific ap- Dry boxes were placed in the refrigerator for the proach between beef aged by these two methods. War- dry-aging experiment. Three pieces of meat were put ren and Kastner [11] obtained the result that dry-aged in the dry box without a fan for each week under steaks had significantly higher beefy and brown/ maintained conditions of approximately 80% relative roasted flavor intensities than the unaged or humidity and no air flow. The pieces of meat for the vacuum-aged steaks, whereas vacuum-aged steaks had wet-aging experiment, which were vacuumed and significantly higher bloody/serumy and sour flavor in- packaged, were placed in a refrigerator set at 2 °C tensities than the unaged or dry-aged steaks. King et during the aging process, the same as that used for al. [4] stated wet-aged beef had significantly greater the dry-aging experiment. percentages of acids than dry-aged beef. In terms of hardness, Parrish Jr. et al. [6] found that rib and loin Measurement of moisture and trim losses steaks from their wet aging treatment were signifi- Moisture loss is the weight of water lost from meat and cantly more tender than the rib and loin steaks from is determined by measuring the difference in meat their dry aging treatment. However, Warren and weight between before and after it has been subjected to Kastner [11] found that both vacuum aging and dry dry aging. Trim loss is weight of the trimming part of aging for 11 days resulted in tenderness scores that meat that is discolored and dehydrated. Productive loss were significantly higher than the unaged controls. To is the sum of moisture and trim losses. shed light on this issue, in this study, scientific data, Samples for these measurements were cut to a size such as free amino acids and hardness, were analyzed of approximately 2.5 cm (length) × 2.5 cm (width) × in the round of Okinawan delivered cow beef during 1 cm (height) at a depth of over 1 cm from the sur- dry-aging and wet-aging processes along with a com- face of the meat and were frozen until subsequent parison with beef imported from Australia (hereinafter analyses. Prior to analyses, these frozen samples were referred to as Australian beef). kept at a normal temperature for 2.5 h and weighed before their drip was removed. The weight of these Methods samples was measured again after their drip had been Animals and muscle samples removed. The weight difference was calculated and Two types of beef were used for the aging experi- was denoted as a ratio relative to the initial weight. ments: one from Okinawan delivered cows that The percentage of drip loss was calculated in this were > 10 years old and thus, had already finished way. In terms of cooking loss, samples that had giving birth and the other one Australian beef. Type already undergone drip loss measurement were put of Okinawan delivered cow beef was a kind of Japa- into a plastic bag and incubated at 70°C in a water nese cow called black-haired Japanese cow. Type of bath for 1 h. After they had been cooled and their Australian beef was crossbreed thought to be in the drip had been removed, the weight of these samples family lineage of black cattle mainly, which was was measured. The weight difference between before raised in pastures. With regard to Okinawan deliv- and after incubation was determined as a ratio rela- ered cow beef, two chunks of meat (approximately tive to the weight before incubation [5]. 6 kg/each) from the round in the same position of both right and left sides of the carcass from one in- Quantitative analysis of amino acids dividual were used for both dry-aging and wet-aging First, sampling for the dry-aging experiment for 2, 3, experiments. The same approach was also applied to and 4 weeks was performed to a depth of over 1 cm two other individuals, so there were three pieces for from the surface of the edible part, after it had been Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 3 of 9 Fig. 1 Changes in moisture and trim losses during the dry-aging process. Different letters in the same group indicate significant differences trimmed. The sampling for the dry-aging experiment formic acid (9/75/16/0.3, v/v/v/v). Solvent B was aceto- for 0 and 1 week and the wet-aging experiment for 0, nitrile/100 mM ammonium formate (20/80, v/v). The gra- 1, 2, 3, and 4 weeks was performed to a depth of dient program was as follows: 0, A 100%; 2.75, A 100%; over 1 cm from the surface of the meat without trim- 7.75, A 83%; and 7.76 min, A 0%. Analyses were moni- ming. These were cut and homogenized. Extract solu- tored in the positive-ion mode using an ESI source at tions were obtained from these homogenized samples 350 °C and MRM. after protein was removed with acetonitrile and perchlo- Amino acids were sorted according to their features rate and fat was removed with hexane. Sample solutions into four groups.Glycine,alanine,threonine,serine, for LC/MS were prepared after extract solutions had been and proline were classified as sweet-tasting amino filtered. Sample solutions were injected onto an Intrada acids. Aspartic acid, glutamic acid, glutamine, and as- Amino Acid column (3 × 100 mm, Imtakt Corp., Kyoto, paragine were classified as umami-tasting amino Japan) at a flow rate of 0.6 ml/min. The separation was acids. Methionine, lysine, isoleucine, leucine, phenyl- performed with a two-pump gradient. Solvent A was alanine, tyrosine, valin, histidine, arginine, and cystine acetonitrile/tetrahydrofuran/25 mM ammonium formate/ were classified as savory-tasting amino acids. Finally, Fig. 2 Changes in drip and cooking losses during both dry- and wet-aging processes Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 4 of 9 Fig. 3 Change in levels of each amino acid group in both types of beef during both dry- and wet-aging processes. Amino acid groups are classified as sweet-, umami-, and savory-tasting and functional. The sum of sweet-, umami-, and savory-tasting amino acids constitute the proteinogenic category. Changes in their levels during the dry-aging process are shown by solid lines. Changes in their levels during the wet- aging process are shown by dotted lines carnosine, anserine, taurine, ornithine, and GABA Institute Inc., Cary, NC, USA). Tukey’s test was used for were classified as functional amino acids. identifying differences (P < 0.05) between in the same beef sample during the same aging process. Rheological properties Breaking stress, shearing stress, and other rheological Results properties of each meat sample were measured using a It was thought that moisture loss occurred immediately rheometer RE-3305S (Yamaden Co. Ltd., Tokyo, Japan) after the initiation of the dry-aging process, whereas trim and a breaking strength analyzer BAS-33005-16 (Yama- loss occurred approximately 10 days after the dry-aging den Co. Ltd., Tokyo, Japan). Samples for these measure- process had started. Levels of both losses increased in ments were those that had already undergone both drip both beef types as the number of days of dry aging in- and cooking loss measurements as described above. creased (Fig. 1). In Australian beef, drip loss significantly With regard to the measurement of breaking stress, this decreased and cooking loss (P = 0.086) showed a decreas- was performed on a sample of approximately 1 cm in ing trend during the dry-aging process. In addition, drip height using a rheometer with a plunger No. 5 stick-type at loss (P = 0.07) showed a decreasing trend and cooking loss a speed of 1 mm/s. Approximately seven runs were per- significantly increased during the wet-aging process. In formed for each measurement and the average was calcu- Okinawan delivered cow beef, drip loss significantly de- lated. The measurement of shearing stress was performed creased and cooking loss (P = 0.42) appeared to remain in an almost similar manner. Specifically, it was performed unchanged during the dry-aging process (Fig. 2). on samples of a size of approximately 1 cm (width) × 1 cm The results also showed that sweet-, savory-, and (height) using a rheometer with a plunger No. 21 knife-type umami-tasting amino acids increased in both beef types dur- at a speed of 1 mm/s. These samples were cut vertically on ing dry- and wet-aging processes (Fig. 3 and Table 1). Their muscle fiber. Approximately five runs were performed for sum reflects the level of proteinogenic amino acids, which each measurement and the average was calculated. also increased. However, levels of functional amino acids did not increase in both beef types during dry- and wet-aging Statistical analysis processes, but rather decreased in Okinawan delivered cow Two-way analysis of variance (ANOVA) was used for beef. Total amino acids, including functional amino acids, in statistical analysis with the program JMP® 13 (SAS both beef types significantly increased during dry- and Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 5 of 9 Table 1 Results of the average value ± standard deviation and Tukey’s test for each amino acid group listed in Fig. 3 Australian beef week 0 1 2 3 4 proteinogenic dry 206±22 a 214±9 a 228±32 a 243±31 a 399±29 b wet 246±24 a 301±26 a 319±36 a 330±34 a 467±44 b function dry 239±15 a 243±22 a 233±15 a 225±16 a 210±21 a wet 238±16 a 243±11 a 239±22 a 224±11 a 209±14 a umami dry 107±9 a 105±6 a 105±15 a 106±9 a 151±12 b wet 121±3 a 118±9 a 116±1 a 119±5 a 175±6 b sweet dry 53±4 a 56±1 a 59±7 a 63±7 a 104±11 b wet 61±4 a 76±5 ab 81±9 b 84±6 b 121±10 c savory dry 46±10 a 53±16 a 64±11 a 75±20 a 144±30 b wet 64±19 a 108±14 ab 122±28 ab 128±28 ab 170±37 b Okinawan delivered cow week 0 1 2 3 4 proteinogenic dry 205±19 a 223±22 a 334±18 b 367±18 bc 423±42 c wet 186±12 a 202±28 a 230±43 a 321±41 b 404±24 b function dry 215±7 a 206±16 ab 193±12 ab 186±6 bc 164±2 c wet 200±8 a 176±3 b 170±4 b 178±10 b 167±0 b umami dry 129±13 b 120±14 a 131±5 a 135±2 a 139±18 a wet 115±8 ab 108±5 ab 95±6 a 108±4 ab 129±16 b sweet dry 47±3 a 50±4 a 75±2 b 72±6 b 78±15 b wet 42±3 a 44±6 a 48±11 a 53±9 ab 69±3 b savory dry 29±2 a 54±9 a 128±12 b 160±14 b 206±21 c wet 29±2 a 50±25 a 88±26 a 160±32 b 207±22 b Different letters in the same group represent significant differences (P < 0.05) wet-aging processes (data not shown); this increase was al- breaking point and shearing point were also determined most the same as that observed for proteinogenic amino (breaking point in Australian beef during dry- and acids. GABA and cystine were not detected at all in all sam- wet-aging processes: P =0.52 and P = 0.7, respectively; ples. Representative sweet-, savory-, and umami-tasting shearing point in Australian beef during dry- and wet-aging amino acids, namely, alanine, leucine, and glutamic acid, re- processes: P =0.8 and P = 0.1, respectively; breaking point in spectively, are shown (Fig. 4 and Table 2). Levels of leucine Okinawan delivered cow beef during dry- and wet-aging and glutamic acid increased stably in both beef types during processes: P =0.054 and P = 0.059, respectively; and shear- both aging processes. In terms of alanine, it showed no in- ing point in Okinawan delivered cow beef during dry- and crease in Okinawan delivered cow beef after dry- and wet-aging processes: P = 0.055 and P = 0.658, respectively). wet-aging processes. In terms of ornithine, it was initially at Australian beef has a decreasing trend about only shearing a low level and remained constant in both beef types during point during wet-aging process, contrary to the case for both aging processes. Okinawan delivered cow beef (Figs. 5 and 6). Therewereapproximatelydecreasingtrendsinbreaking Upon comparison of drip loss, cooking loss, and the stress [dry aging of Australian beef: P < 0.05, wet aging of sum of these between beef types and between aging Australian beef: P = 0.09, dry aging of Okinawan delivered methods (Table 3), there were significant differences (P < cow beef: (P < 0.05), and wet aging of Okinawan delivered 0.01) between beef types for all these variables, which was cow beef: P <0.05] (Fig. 5). In addition, there were approxi- also the case (P < 0.01) between the aging methods. In the mately decreasing trends in sharing stress [dry aging of comparison of each amino acid group between beef types Australian beef: P = 0.1 and wet aging of Australian beef: P and between aging methods (Table 4), there were signifi- < 0.05 and dry aging of Okinawan delivered cow beef: P = cant differences between beef types regarding function (P 0.086 and wet aging of Okinawan delivered cow beef: P = <0.05), sweetness (P < 0.01), and total (P < 0.05). However, 0.136] (Fig. 6). Probability values for the strain regarding there was no significant difference between the aging Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 6 of 9 Fig. 4 Changes in levels of alanine, glutamic acid, leucine and ornithine in both types of beef during both dry- and wet-aging processes. Changes in their levels during the dry-aging process are shown by solid lines. Those during the wet-aging process are shown by dotted lines methods for all studied variables even for all amino acids Discussion (data of amino acids not shown). Comparison of rheo- The present study demonstrated the effects of both dry- logical properties between beef types and between aging and wet-aging processes on Okinawan delivered cow methods showed there was a significant difference be- beef and Australian beef. Productive loss increased as tween beef types regarding breaking stress alone (Table 5). the number of days of dry aging increased. However, Table 2 Results of the average value ± standard deviation and Tukey’s test for each amino acid listed in Fig. 4 Australian beef week 0 1 2 3 4 Alanine dry 37±3 a 37±2 a 38±6 a 40±4 a 58±4 b wet 40±0 a 43±1 ab 44±2 b 45±2 b 62±2 c Glutamic acid dry 13±2 a 15±1 ab 16±2 ab 18±3 b 29±2 c wet 16±2 a 22±3 ab 24±3 ab 25±3 b 34±3 b Leucine dry 12±2 a 14±5 a 17±2 a 20±5 a 34±7 b wet 16±5 a 25±2 ab 29±6 ab 30±6 b 39±6 b Ornithine dry 7±0 a 7±1 a 7±0 a 7±0 a 8±0 a wet 7±1 ab 7±0 a 7±1 ab 7±1 a 10±2 b Okinawan delivered cow week 0 1 2 3 4 Alanine dry 33±2 ab 32±3 a 38±1 ab 36±2 ab 40±4 b wet 30±2 ab 29±1 ab 28±4 a 29±2 ab 35±1 b Glutamic acid dry 16±1 a 17±2 a 24±1 ab 26±3 b 38±5 c wet 13±1 a 16±1 a 17±4 a 24±2 b 31±3 c Leucine dry 4±1 a 11±1 a 25±2 b 32±4 b 42±5 c wet 5±1 a 10±5 ab 18±5 b 33±7 c 44±5 c Ornithine dry 7±0 a 7±0 a 7±0 a 7±0 a 7±0 a wet 6±0 ab 6±0 a 6±0 a 6±0 a 7±1 b Different letters in the same group represent significant differences (P < 0.05) Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 7 of 9 Fig. 5 Changes in breaking stress and strain of breaking point in both types of beef during both dry- and wet-aging processes there was no productive loss, such as moisture and trim the flavor [3, 7, 11]. Therefore, it is considered that losses, during the wet-aging process. This shows that the amino acids in beef are also concentrated during the price of dry-aged beef must be higher than that of dry-aging process. On the contrary, as described in wet-aged beef to compensate for this loss. In our study, Introduction, wet-aged steaks had significantly higher productive loss from both beef types increased to > 30% sour flavor than dry-aged steaks [11] and wet-aged beef under conditions of approximately 80% relative humidity had significantly greater percentages of acids than for 4 weeks. Many of the compounds responsible for fla- dry-aged beef [4]. Free amino acid is one kind of acids vor are concentrated by the dry-aging process, according and possibly contributes to sour flavor of wet-aged beef. to USMEF [7]. In other words, the distinguishing effect In our study, the change in levels of amino acids during of the dry-aging process on beef is that it concentrates the dry-aging process was compared with that during Fig. 6 Changes in shearing stress and strain of shearing point in both types of beef during both dry- and wet-aging processes Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 8 of 9 Table 3 Comparison of drip loss, cooking loss, and their sum between beef types and between aging methods Beef type Aging method SEM† P values in ANOVA‡ Australian Okinawan wet aging dry aging Beef type Aging method Drip loss 4.7 2.2 4.1 2.8 0.1 ** ** Cooking loss 32.3 27.2 30.7 28.8 0.3 ** ** Sum 37.0 29.4 34.8 31.6 0.4 ** ** Values are least-square means (n = 3). †Pooled standard error of the mean. ‡Asterisks indicate **P < 0.01 in analysis of variance Table 4 Comparison of levels of each amino acid group between beef types and between aging methods Beef type Aging method SEM† P values in ANOVA‡ Australian Okinawan wet aging dry aging Beef type Aging method Function 230.3 185.5 204.4 211.4 3.3 * ns Sweet 61 42.9 52.7 51.2 2.5 ** ns Savory 97.4 111.2 112.7 95.9 10.6 ns ns Umami 122.3 120.7 120.3 122.7 3.7 ns ns Proteinogen ic amino acids 295.4 289.6 300.8 284.2 16.1 ns ns Total amino acids 525.7 475 505.2 495.6 14.9 * ns Values are least-square means (n = 3). †Pooled standard error of the mean. ‡Asterisks indicate **P < 0.01 and *P < 0.05 in analysis of variance Table 5 Comparison of rheological properties between beef types and between aging methods Beef type Aging method SEM† P values in ANOVA‡ Australian Okinawan wet aging dry aging Beef type Aging method Breaking stress (Pa) 1.3E+06 1.1E+06 1.2E+06 1.3E+06 6.9E+04 * ns Strain of breaking point (%) 68.4 64.7 66.8 66.3 1.6 ns ns Sharing stress (N) 20.6 19.5 19.7 20.5 9.6E+05 ns ns Strain of sharing point (%) 74.5 69.4 72.8 71.1 2 ns ns Values are least-square means (n = 3). †Pooled standard error of the mean. ‡Asterisks indicate *P < 0.05 in analysis of variance the wet-aging process. The meat from Okinawan delivered during both types of aging. Sugioka et al. [9] reported cow beef seemed to have a higher increase in proteino- that meat from Japanese brown cattle have high levels of genic amino acids during the dry-aging process than dur- glutamic acid and leucine, which increase stably during ing the wet-aging process, particularly in the middle of the the aging process. However, alanine did not increase but aging process, contrary to the case for Australian beef. fluctuated sharply and non-proteinogenic amino acids However, finally, 4 weeks after aging, these increases dur- also did not increase during the aging process. The ing both aging processes were about the same in each type results in our study were very similar to these results. In of beef. On ANNOVA, there was no significant difference particular, the results in Okinawan delivered cow beef between aging methods for all studied variables related to were exactly the same as these. free amino acids. The trend of a decrease in hardness during the It is known that there are many steps in the degradation dry-aging process is almost the same as that during the of proteins to produce free amino acids. The way to pro- wet-aging process. It is difficult to analyze a non-uniform duce free proteinogenic amino acids assumes to be from tissue, such as a piece of meat, using a hardness test short peptide not directly from protein. Therefore, it is diffi- because of its scattered value of measurement. At this cult to elucidate the difference in the underlying mechan- point, the only assertion that can be safely made is ism of amino acid production between dry- and wet-aging that there were similar decreasing trends in hardness processes by molecular biological techniques. As such, in during both aging processes. However, the tender I the current study, we cannot draw definitive conclusions bit them is the clear difference between both dry- that the dry-aging process can concentrate amino acids to a and wet-aged beef. Upon chewing wet-aged beef, its greater extent than the wet-aging process and vice versa. tenderness seemed to remain for longer than that of The results showed that glutamic acid increased stably, dry-aged beef, whereas dry-aged beef was much easier which is essential for the umami taste of meat, for beef to cut with the teeth. Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 9 of 9 The hardness of dry-aged beef gradually decreases as Consent for publication Not applicable. the number of days of aging increases [10]. In addition, Sitz et al. [8] reported that for both the Prime and Competing interests Choice comparisons, Warner-Bratzler shear force values The authors declare that they have no competing interests. did not differ between the dry- and wet-aged steaks [8]. These are similar to the results in our study. From the Publisher’sNote Springer Nature remains neutral with regard to jurisdictional claims in results of the strain regarding breaking point and shear- published maps and institutional affiliations. ing point, deformation process of meats in both type beefs while being pressed seem to be totally different. Author details Okinawa prefectural Industrial Technology Center, 12-2 Suzaki, Uruma-shi, The positive effects, such as an increase in free amino Okinawa 904-2234, Japan. Okinawa prefectural Livestock Research Center, acids and a decrease in hardness, in dry-aged beef were 3 2009-5 Shoshi, Nakizin-son, Okinawa 905-0426, Japan. Present address: almost the same as those found in wet-aged beef in our Okinawa prefectural Agricultural Research center, 820 Makabe, Itoman city, Okinawa 901-0336, Japan. Present address: Livestock Division of Okinawa study. In fact, there were no significant differences be- prefectural government, 1–2-2 Izumizaki, Naha city, Okinawa 900–8570, tween the two aging methods for all studied variables, Japan. except drip, cooking, and productive losses. On the Received: 6 March 2018 Accepted: 30 August 2018 other hand, some significant differences between the beef types were identified on ANOVA, particularly re- garding free amino acids. However, it is difficult to study References 1. Baird B. Dry aging enhances palatability of beef, Beef safety and quality. why these differences occurred because these types of 2008. https://www.beefresearch.org/CMDocs/BeefResearch/PE_Issues_ beef have different genetic factors and have undergone Update/Dry_aging_enhances_palatability_of_beef.pdf. Accessed March April different fattening methods and for different periods 2008. 2. Campbell RE, Hunt MC, Levis P, Chambers Iv E. Dry-aging effects on since slaughter. palatability of beef longissimus muscle. J Food Sci. 2001;66:196–9. Thepresent studyshows theresults ofthe 3. Dashmaa D, Tripathi VK, Cho S, Kim Y, Hwang I. Dry aging of beef; review. J dry-aging process under conditions without the inocu- Anim Sci Technol. 2016;58:20. 4. King MF, Matthews MA, Rule DC, Field RA. Effect of beef packing method lation of microorganisms. However, in actual condi- on volatile compounds developed by oven roasting or microwave cooking. tions, certain molds can grow on the surface of the J Agric Food Chem. 1995;43:773–8. meat during the dry-aging process, which is one fac- 5. Muramoto T, Maeno K, Okada Y, Tezuka S, Kamata T. Relationship between shear force and tenderness of Japanese shorthorn beef. Tohoku J Anim Sci tor that confers the taste of dry-aged beef [3]. There- Technol. 2014;64(1):7–12. fore, further studies are needed in demonstrate the 6. Parrish FC Jr, Boles JA, Rust RE, Olson DG. Dry and wet aging effects on effects of mold on dry-aged beef. palatability attributes of beef loin and rib steaks from three quality grades. J Food Sci. 1991;56:601–3. 7. Savell JW. Dry-aging of beef, exective summery: National Cattlemen’s Beef Conclusion Association; 2008. https://www.beefresearch.org/CMDocs/BeefResearch/ Dry%20Aging%20of%20Beef.pdf There was no significant difference between dry- and 8. Sitz BM, Calkins CR, Feuz DM, Umberger WJ, Eskridge KM. Consumer sensory wet-aging methods for all studied variables related to acceptance and value of wet-aged and dry-aged beef steaks. J Anim Sci. free amino acids or hardness in this study. 2006;84:1221–6. 9. Sugioka K, Tasaki T, Sakai H, Ieiri S, Araki T. Study of free amino acid in aging Acknowledgements of beef. Proceedings of the annual meeting 2015 Okayama of Japan Society We appreciate the technical supports offered by Ms. Sakima. Without her for Bioscience, biotechnology, and Agrochemistry. 2015: 2F26P18. persistent help this paper would not have been possible. 10. Tsuchiya T, Ukai Y, Saitou Y. Change in quality and productive loss of beef We are very grateful to Mr. Higa, Managing Director, and Mr. Gushiken, meat during dry-aging process. Bulletin of shizuoka prefecture livestock factory manager, Uehara Meat Co., Ltd., for providing meat and helping in research institute. 2013;6:12–4. preparations for the aging experiments. We express our sincere thanks to Dr. 11. Warren KE, Kastner CL. A comparison of dry aged and vacuum-aged beef Kobayashi, Livestock Improvement Association of Japan, Inc., Maebashi striploins. J Muscle Foods. 1992;3:151–7. Institute of Animal Science, for advice on the mechanism of the dry-aging process of beef, particularly about the process of producing free amino acids. We also gratefully acknowledge the help by Mr. Tamaki, President, and Mr. Uezu, Executive Managing Director, Okinawa Aging Company, for helpful discussions. Funding Funded by Science and Technology Promotion Division of Okinawa prefectural government. Authors’ contributions TH was concerned all things about this research and drafted and wrote this manuscript, NA supported everything about analysis and statistical work. Both authors read and approved the final manuscript. Ethics approval and consent to participate Not applicable. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Animal Science and Technology Springer Journals

Changes in free amino acids and hardness in round of Okinawan delivered cow beef during dry- and wet-aging processes

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Copyright © 2018 by The Author(s).
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Life Sciences; Animal Genetics and Genomics; Agriculture
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Abstract

Background: Aging trials are conducted to determine characteristics associated with dry- and wet-aging processes of beef from delivered cows grown in Okinawa, i.e., dams that have finished giving birth (Okinawan delivered cow beef). Changes in free amino acids, hardness, and other factors were analyzed in round of Okinawan delivered cow beef during dry- and wet-aging processes along with a comparison with characteristics of beef imported from Australia. Results: Functional amino acids did not increase during both dry- and wet-aging processes. However, proteinogenic amino acids increased significantly (P < 0.05) and hardness tended to decrease during both dry- and wet-aging processes. On comparison between dry- and wet-aging processes by analysis of variance, drip and cooking losses were significantly lower during the dry-aging process than during the wet-aging process. However, there was no significant difference in free amino acids or hardness in this comparison. Conclusion: There was no significant difference between dry- and wet-aging methods for all studied variables related to free amino acids or hardness in this study. Keywords: Dry aging beef, Okinawan delivered cow, Free amino acids and hardness Background Western countries but also in Asian countries, such as The livestock industry is a major part of the Okinawan South Korea, Japan, Singapore, Taiwan, and Hong Kong [3]. economy, with the Prefecture selling the fourth highest Over 2.5 million foreign tourists visited Okinawa in number of calves in Japan. In Okinawa, there are numer- 2017, which is the highest number recording in any year, ous dams, some of which have finished giving birth and according to the official figures of the Okinawa prefectural thus, are termed as Okinawan delivered cows. The turn- government. Tourists from Asian countries, particularly over for Okinawan delivered cows is approximately 5000 those from Taiwan, Hong Kong, South Korea, and China, heads per year; these slaughtered cows are generally sold accounted for approximately 90% of this number. In this at a lower price than fattened cattle and heifers. The rea- context, there is an urgent need to boost the production son for them being less valuable is their less tender meat. and awareness of Okinawa’sunique food brands.Thus,we However, there is significant potential for adapting these hope to create an Okinawan brand of dry-aged beef by cows’ meat and increasing their value. adding value to Okinawan beef because it is produced in Dry aging is one aging method that is said to improve Okinawa and has high potential to become very popular meat with respect to some characteristics. Recently, in- in view of the region’s tourist industry. creased interest in dry-aged beef has emerged not only in During the dry-aging process, juices are absorbed into the meat and chemical breakdown of protein occurs, giving amoreintense nuttyand beefyflavor[3]. Moreover, during aging, the endogenous enzymes break down myofibrillar * Correspondence: hangskit@pref.okinawa.lg.jp proteins in the muscle, which leads to more tender beef [1, Okinawa prefectural Industrial Technology Center, 12-2 Suzaki, Uruma-shi, Okinawa 904-2234, Japan 2]. Because of this, it is considered that lean meat with low Present address: Okinawa prefectural Agricultural Research center, 820 fat content is more suitable for dry aging and has higher Makabe, Itoman city, Okinawa 901-0336, Japan potential to undergo a change in its quality. Specifically, Full list of author information is available at the end of the article © The Author(s). 2018 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 2 of 9 round of beef (i.e., from the rear leg of the cow) is most each aging experiment. With regard to Australian likely to be appropriate for dry aging. Indeed, dry-aging beef, six chunks of meat (approximately 6 kg/each) products using round of beef have already been commer- from the round of each individual were grouped into cialized in Okinawa. dry-aging and wet-aging experiments, namely, three In general, the dry-aging process is performed under in each group. Each chunk of meat was divided into aerated conditions, whereas the wet-aging process es- five pieces by cutting for experiments involving 0, 1, sentially involves vacuuming and packaging, which 2, 3, or 4 weeks of aging. means that the conditions do not involve aeration. Savell [7] reported a survey on consumer preferences Methods between dry-aged and wet-aged beef in an executive Aging environment summary of the National Cattlemen’s Beef Associa- The dry-aging environment was created in a refriger- tion’s Center in the United States of America (herein- ator (Showa Denko K.K., Tokyo, Japan) in Okinawa after referred to as “USMEF”). There is only a little Industrial Technology Center at a temperature of 2 °C. thing known about specific differences by scientific ap- Dry boxes were placed in the refrigerator for the proach between beef aged by these two methods. War- dry-aging experiment. Three pieces of meat were put ren and Kastner [11] obtained the result that dry-aged in the dry box without a fan for each week under steaks had significantly higher beefy and brown/ maintained conditions of approximately 80% relative roasted flavor intensities than the unaged or humidity and no air flow. The pieces of meat for the vacuum-aged steaks, whereas vacuum-aged steaks had wet-aging experiment, which were vacuumed and significantly higher bloody/serumy and sour flavor in- packaged, were placed in a refrigerator set at 2 °C tensities than the unaged or dry-aged steaks. King et during the aging process, the same as that used for al. [4] stated wet-aged beef had significantly greater the dry-aging experiment. percentages of acids than dry-aged beef. In terms of hardness, Parrish Jr. et al. [6] found that rib and loin Measurement of moisture and trim losses steaks from their wet aging treatment were signifi- Moisture loss is the weight of water lost from meat and cantly more tender than the rib and loin steaks from is determined by measuring the difference in meat their dry aging treatment. However, Warren and weight between before and after it has been subjected to Kastner [11] found that both vacuum aging and dry dry aging. Trim loss is weight of the trimming part of aging for 11 days resulted in tenderness scores that meat that is discolored and dehydrated. Productive loss were significantly higher than the unaged controls. To is the sum of moisture and trim losses. shed light on this issue, in this study, scientific data, Samples for these measurements were cut to a size such as free amino acids and hardness, were analyzed of approximately 2.5 cm (length) × 2.5 cm (width) × in the round of Okinawan delivered cow beef during 1 cm (height) at a depth of over 1 cm from the sur- dry-aging and wet-aging processes along with a com- face of the meat and were frozen until subsequent parison with beef imported from Australia (hereinafter analyses. Prior to analyses, these frozen samples were referred to as Australian beef). kept at a normal temperature for 2.5 h and weighed before their drip was removed. The weight of these Methods samples was measured again after their drip had been Animals and muscle samples removed. The weight difference was calculated and Two types of beef were used for the aging experi- was denoted as a ratio relative to the initial weight. ments: one from Okinawan delivered cows that The percentage of drip loss was calculated in this were > 10 years old and thus, had already finished way. In terms of cooking loss, samples that had giving birth and the other one Australian beef. Type already undergone drip loss measurement were put of Okinawan delivered cow beef was a kind of Japa- into a plastic bag and incubated at 70°C in a water nese cow called black-haired Japanese cow. Type of bath for 1 h. After they had been cooled and their Australian beef was crossbreed thought to be in the drip had been removed, the weight of these samples family lineage of black cattle mainly, which was was measured. The weight difference between before raised in pastures. With regard to Okinawan deliv- and after incubation was determined as a ratio rela- ered cow beef, two chunks of meat (approximately tive to the weight before incubation [5]. 6 kg/each) from the round in the same position of both right and left sides of the carcass from one in- Quantitative analysis of amino acids dividual were used for both dry-aging and wet-aging First, sampling for the dry-aging experiment for 2, 3, experiments. The same approach was also applied to and 4 weeks was performed to a depth of over 1 cm two other individuals, so there were three pieces for from the surface of the edible part, after it had been Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 3 of 9 Fig. 1 Changes in moisture and trim losses during the dry-aging process. Different letters in the same group indicate significant differences trimmed. The sampling for the dry-aging experiment formic acid (9/75/16/0.3, v/v/v/v). Solvent B was aceto- for 0 and 1 week and the wet-aging experiment for 0, nitrile/100 mM ammonium formate (20/80, v/v). The gra- 1, 2, 3, and 4 weeks was performed to a depth of dient program was as follows: 0, A 100%; 2.75, A 100%; over 1 cm from the surface of the meat without trim- 7.75, A 83%; and 7.76 min, A 0%. Analyses were moni- ming. These were cut and homogenized. Extract solu- tored in the positive-ion mode using an ESI source at tions were obtained from these homogenized samples 350 °C and MRM. after protein was removed with acetonitrile and perchlo- Amino acids were sorted according to their features rate and fat was removed with hexane. Sample solutions into four groups.Glycine,alanine,threonine,serine, for LC/MS were prepared after extract solutions had been and proline were classified as sweet-tasting amino filtered. Sample solutions were injected onto an Intrada acids. Aspartic acid, glutamic acid, glutamine, and as- Amino Acid column (3 × 100 mm, Imtakt Corp., Kyoto, paragine were classified as umami-tasting amino Japan) at a flow rate of 0.6 ml/min. The separation was acids. Methionine, lysine, isoleucine, leucine, phenyl- performed with a two-pump gradient. Solvent A was alanine, tyrosine, valin, histidine, arginine, and cystine acetonitrile/tetrahydrofuran/25 mM ammonium formate/ were classified as savory-tasting amino acids. Finally, Fig. 2 Changes in drip and cooking losses during both dry- and wet-aging processes Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 4 of 9 Fig. 3 Change in levels of each amino acid group in both types of beef during both dry- and wet-aging processes. Amino acid groups are classified as sweet-, umami-, and savory-tasting and functional. The sum of sweet-, umami-, and savory-tasting amino acids constitute the proteinogenic category. Changes in their levels during the dry-aging process are shown by solid lines. Changes in their levels during the wet- aging process are shown by dotted lines carnosine, anserine, taurine, ornithine, and GABA Institute Inc., Cary, NC, USA). Tukey’s test was used for were classified as functional amino acids. identifying differences (P < 0.05) between in the same beef sample during the same aging process. Rheological properties Breaking stress, shearing stress, and other rheological Results properties of each meat sample were measured using a It was thought that moisture loss occurred immediately rheometer RE-3305S (Yamaden Co. Ltd., Tokyo, Japan) after the initiation of the dry-aging process, whereas trim and a breaking strength analyzer BAS-33005-16 (Yama- loss occurred approximately 10 days after the dry-aging den Co. Ltd., Tokyo, Japan). Samples for these measure- process had started. Levels of both losses increased in ments were those that had already undergone both drip both beef types as the number of days of dry aging in- and cooking loss measurements as described above. creased (Fig. 1). In Australian beef, drip loss significantly With regard to the measurement of breaking stress, this decreased and cooking loss (P = 0.086) showed a decreas- was performed on a sample of approximately 1 cm in ing trend during the dry-aging process. In addition, drip height using a rheometer with a plunger No. 5 stick-type at loss (P = 0.07) showed a decreasing trend and cooking loss a speed of 1 mm/s. Approximately seven runs were per- significantly increased during the wet-aging process. In formed for each measurement and the average was calcu- Okinawan delivered cow beef, drip loss significantly de- lated. The measurement of shearing stress was performed creased and cooking loss (P = 0.42) appeared to remain in an almost similar manner. Specifically, it was performed unchanged during the dry-aging process (Fig. 2). on samples of a size of approximately 1 cm (width) × 1 cm The results also showed that sweet-, savory-, and (height) using a rheometer with a plunger No. 21 knife-type umami-tasting amino acids increased in both beef types dur- at a speed of 1 mm/s. These samples were cut vertically on ing dry- and wet-aging processes (Fig. 3 and Table 1). Their muscle fiber. Approximately five runs were performed for sum reflects the level of proteinogenic amino acids, which each measurement and the average was calculated. also increased. However, levels of functional amino acids did not increase in both beef types during dry- and wet-aging Statistical analysis processes, but rather decreased in Okinawan delivered cow Two-way analysis of variance (ANOVA) was used for beef. Total amino acids, including functional amino acids, in statistical analysis with the program JMP® 13 (SAS both beef types significantly increased during dry- and Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 5 of 9 Table 1 Results of the average value ± standard deviation and Tukey’s test for each amino acid group listed in Fig. 3 Australian beef week 0 1 2 3 4 proteinogenic dry 206±22 a 214±9 a 228±32 a 243±31 a 399±29 b wet 246±24 a 301±26 a 319±36 a 330±34 a 467±44 b function dry 239±15 a 243±22 a 233±15 a 225±16 a 210±21 a wet 238±16 a 243±11 a 239±22 a 224±11 a 209±14 a umami dry 107±9 a 105±6 a 105±15 a 106±9 a 151±12 b wet 121±3 a 118±9 a 116±1 a 119±5 a 175±6 b sweet dry 53±4 a 56±1 a 59±7 a 63±7 a 104±11 b wet 61±4 a 76±5 ab 81±9 b 84±6 b 121±10 c savory dry 46±10 a 53±16 a 64±11 a 75±20 a 144±30 b wet 64±19 a 108±14 ab 122±28 ab 128±28 ab 170±37 b Okinawan delivered cow week 0 1 2 3 4 proteinogenic dry 205±19 a 223±22 a 334±18 b 367±18 bc 423±42 c wet 186±12 a 202±28 a 230±43 a 321±41 b 404±24 b function dry 215±7 a 206±16 ab 193±12 ab 186±6 bc 164±2 c wet 200±8 a 176±3 b 170±4 b 178±10 b 167±0 b umami dry 129±13 b 120±14 a 131±5 a 135±2 a 139±18 a wet 115±8 ab 108±5 ab 95±6 a 108±4 ab 129±16 b sweet dry 47±3 a 50±4 a 75±2 b 72±6 b 78±15 b wet 42±3 a 44±6 a 48±11 a 53±9 ab 69±3 b savory dry 29±2 a 54±9 a 128±12 b 160±14 b 206±21 c wet 29±2 a 50±25 a 88±26 a 160±32 b 207±22 b Different letters in the same group represent significant differences (P < 0.05) wet-aging processes (data not shown); this increase was al- breaking point and shearing point were also determined most the same as that observed for proteinogenic amino (breaking point in Australian beef during dry- and acids. GABA and cystine were not detected at all in all sam- wet-aging processes: P =0.52 and P = 0.7, respectively; ples. Representative sweet-, savory-, and umami-tasting shearing point in Australian beef during dry- and wet-aging amino acids, namely, alanine, leucine, and glutamic acid, re- processes: P =0.8 and P = 0.1, respectively; breaking point in spectively, are shown (Fig. 4 and Table 2). Levels of leucine Okinawan delivered cow beef during dry- and wet-aging and glutamic acid increased stably in both beef types during processes: P =0.054 and P = 0.059, respectively; and shear- both aging processes. In terms of alanine, it showed no in- ing point in Okinawan delivered cow beef during dry- and crease in Okinawan delivered cow beef after dry- and wet-aging processes: P = 0.055 and P = 0.658, respectively). wet-aging processes. In terms of ornithine, it was initially at Australian beef has a decreasing trend about only shearing a low level and remained constant in both beef types during point during wet-aging process, contrary to the case for both aging processes. Okinawan delivered cow beef (Figs. 5 and 6). Therewereapproximatelydecreasingtrendsinbreaking Upon comparison of drip loss, cooking loss, and the stress [dry aging of Australian beef: P < 0.05, wet aging of sum of these between beef types and between aging Australian beef: P = 0.09, dry aging of Okinawan delivered methods (Table 3), there were significant differences (P < cow beef: (P < 0.05), and wet aging of Okinawan delivered 0.01) between beef types for all these variables, which was cow beef: P <0.05] (Fig. 5). In addition, there were approxi- also the case (P < 0.01) between the aging methods. In the mately decreasing trends in sharing stress [dry aging of comparison of each amino acid group between beef types Australian beef: P = 0.1 and wet aging of Australian beef: P and between aging methods (Table 4), there were signifi- < 0.05 and dry aging of Okinawan delivered cow beef: P = cant differences between beef types regarding function (P 0.086 and wet aging of Okinawan delivered cow beef: P = <0.05), sweetness (P < 0.01), and total (P < 0.05). However, 0.136] (Fig. 6). Probability values for the strain regarding there was no significant difference between the aging Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 6 of 9 Fig. 4 Changes in levels of alanine, glutamic acid, leucine and ornithine in both types of beef during both dry- and wet-aging processes. Changes in their levels during the dry-aging process are shown by solid lines. Those during the wet-aging process are shown by dotted lines methods for all studied variables even for all amino acids Discussion (data of amino acids not shown). Comparison of rheo- The present study demonstrated the effects of both dry- logical properties between beef types and between aging and wet-aging processes on Okinawan delivered cow methods showed there was a significant difference be- beef and Australian beef. Productive loss increased as tween beef types regarding breaking stress alone (Table 5). the number of days of dry aging increased. However, Table 2 Results of the average value ± standard deviation and Tukey’s test for each amino acid listed in Fig. 4 Australian beef week 0 1 2 3 4 Alanine dry 37±3 a 37±2 a 38±6 a 40±4 a 58±4 b wet 40±0 a 43±1 ab 44±2 b 45±2 b 62±2 c Glutamic acid dry 13±2 a 15±1 ab 16±2 ab 18±3 b 29±2 c wet 16±2 a 22±3 ab 24±3 ab 25±3 b 34±3 b Leucine dry 12±2 a 14±5 a 17±2 a 20±5 a 34±7 b wet 16±5 a 25±2 ab 29±6 ab 30±6 b 39±6 b Ornithine dry 7±0 a 7±1 a 7±0 a 7±0 a 8±0 a wet 7±1 ab 7±0 a 7±1 ab 7±1 a 10±2 b Okinawan delivered cow week 0 1 2 3 4 Alanine dry 33±2 ab 32±3 a 38±1 ab 36±2 ab 40±4 b wet 30±2 ab 29±1 ab 28±4 a 29±2 ab 35±1 b Glutamic acid dry 16±1 a 17±2 a 24±1 ab 26±3 b 38±5 c wet 13±1 a 16±1 a 17±4 a 24±2 b 31±3 c Leucine dry 4±1 a 11±1 a 25±2 b 32±4 b 42±5 c wet 5±1 a 10±5 ab 18±5 b 33±7 c 44±5 c Ornithine dry 7±0 a 7±0 a 7±0 a 7±0 a 7±0 a wet 6±0 ab 6±0 a 6±0 a 6±0 a 7±1 b Different letters in the same group represent significant differences (P < 0.05) Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 7 of 9 Fig. 5 Changes in breaking stress and strain of breaking point in both types of beef during both dry- and wet-aging processes there was no productive loss, such as moisture and trim the flavor [3, 7, 11]. Therefore, it is considered that losses, during the wet-aging process. This shows that the amino acids in beef are also concentrated during the price of dry-aged beef must be higher than that of dry-aging process. On the contrary, as described in wet-aged beef to compensate for this loss. In our study, Introduction, wet-aged steaks had significantly higher productive loss from both beef types increased to > 30% sour flavor than dry-aged steaks [11] and wet-aged beef under conditions of approximately 80% relative humidity had significantly greater percentages of acids than for 4 weeks. Many of the compounds responsible for fla- dry-aged beef [4]. Free amino acid is one kind of acids vor are concentrated by the dry-aging process, according and possibly contributes to sour flavor of wet-aged beef. to USMEF [7]. In other words, the distinguishing effect In our study, the change in levels of amino acids during of the dry-aging process on beef is that it concentrates the dry-aging process was compared with that during Fig. 6 Changes in shearing stress and strain of shearing point in both types of beef during both dry- and wet-aging processes Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 8 of 9 Table 3 Comparison of drip loss, cooking loss, and their sum between beef types and between aging methods Beef type Aging method SEM† P values in ANOVA‡ Australian Okinawan wet aging dry aging Beef type Aging method Drip loss 4.7 2.2 4.1 2.8 0.1 ** ** Cooking loss 32.3 27.2 30.7 28.8 0.3 ** ** Sum 37.0 29.4 34.8 31.6 0.4 ** ** Values are least-square means (n = 3). †Pooled standard error of the mean. ‡Asterisks indicate **P < 0.01 in analysis of variance Table 4 Comparison of levels of each amino acid group between beef types and between aging methods Beef type Aging method SEM† P values in ANOVA‡ Australian Okinawan wet aging dry aging Beef type Aging method Function 230.3 185.5 204.4 211.4 3.3 * ns Sweet 61 42.9 52.7 51.2 2.5 ** ns Savory 97.4 111.2 112.7 95.9 10.6 ns ns Umami 122.3 120.7 120.3 122.7 3.7 ns ns Proteinogen ic amino acids 295.4 289.6 300.8 284.2 16.1 ns ns Total amino acids 525.7 475 505.2 495.6 14.9 * ns Values are least-square means (n = 3). †Pooled standard error of the mean. ‡Asterisks indicate **P < 0.01 and *P < 0.05 in analysis of variance Table 5 Comparison of rheological properties between beef types and between aging methods Beef type Aging method SEM† P values in ANOVA‡ Australian Okinawan wet aging dry aging Beef type Aging method Breaking stress (Pa) 1.3E+06 1.1E+06 1.2E+06 1.3E+06 6.9E+04 * ns Strain of breaking point (%) 68.4 64.7 66.8 66.3 1.6 ns ns Sharing stress (N) 20.6 19.5 19.7 20.5 9.6E+05 ns ns Strain of sharing point (%) 74.5 69.4 72.8 71.1 2 ns ns Values are least-square means (n = 3). †Pooled standard error of the mean. ‡Asterisks indicate *P < 0.05 in analysis of variance the wet-aging process. The meat from Okinawan delivered during both types of aging. Sugioka et al. [9] reported cow beef seemed to have a higher increase in proteino- that meat from Japanese brown cattle have high levels of genic amino acids during the dry-aging process than dur- glutamic acid and leucine, which increase stably during ing the wet-aging process, particularly in the middle of the the aging process. However, alanine did not increase but aging process, contrary to the case for Australian beef. fluctuated sharply and non-proteinogenic amino acids However, finally, 4 weeks after aging, these increases dur- also did not increase during the aging process. The ing both aging processes were about the same in each type results in our study were very similar to these results. In of beef. On ANNOVA, there was no significant difference particular, the results in Okinawan delivered cow beef between aging methods for all studied variables related to were exactly the same as these. free amino acids. The trend of a decrease in hardness during the It is known that there are many steps in the degradation dry-aging process is almost the same as that during the of proteins to produce free amino acids. The way to pro- wet-aging process. It is difficult to analyze a non-uniform duce free proteinogenic amino acids assumes to be from tissue, such as a piece of meat, using a hardness test short peptide not directly from protein. Therefore, it is diffi- because of its scattered value of measurement. At this cult to elucidate the difference in the underlying mechan- point, the only assertion that can be safely made is ism of amino acid production between dry- and wet-aging that there were similar decreasing trends in hardness processes by molecular biological techniques. As such, in during both aging processes. However, the tender I the current study, we cannot draw definitive conclusions bit them is the clear difference between both dry- that the dry-aging process can concentrate amino acids to a and wet-aged beef. Upon chewing wet-aged beef, its greater extent than the wet-aging process and vice versa. tenderness seemed to remain for longer than that of The results showed that glutamic acid increased stably, dry-aged beef, whereas dry-aged beef was much easier which is essential for the umami taste of meat, for beef to cut with the teeth. Hanagasaki and Asato Journal of Animal Science and Technology (2018) 60:23 Page 9 of 9 The hardness of dry-aged beef gradually decreases as Consent for publication Not applicable. the number of days of aging increases [10]. In addition, Sitz et al. [8] reported that for both the Prime and Competing interests Choice comparisons, Warner-Bratzler shear force values The authors declare that they have no competing interests. did not differ between the dry- and wet-aged steaks [8]. These are similar to the results in our study. From the Publisher’sNote Springer Nature remains neutral with regard to jurisdictional claims in results of the strain regarding breaking point and shear- published maps and institutional affiliations. ing point, deformation process of meats in both type beefs while being pressed seem to be totally different. Author details Okinawa prefectural Industrial Technology Center, 12-2 Suzaki, Uruma-shi, The positive effects, such as an increase in free amino Okinawa 904-2234, Japan. Okinawa prefectural Livestock Research Center, acids and a decrease in hardness, in dry-aged beef were 3 2009-5 Shoshi, Nakizin-son, Okinawa 905-0426, Japan. Present address: almost the same as those found in wet-aged beef in our Okinawa prefectural Agricultural Research center, 820 Makabe, Itoman city, Okinawa 901-0336, Japan. Present address: Livestock Division of Okinawa study. In fact, there were no significant differences be- prefectural government, 1–2-2 Izumizaki, Naha city, Okinawa 900–8570, tween the two aging methods for all studied variables, Japan. except drip, cooking, and productive losses. On the Received: 6 March 2018 Accepted: 30 August 2018 other hand, some significant differences between the beef types were identified on ANOVA, particularly re- garding free amino acids. However, it is difficult to study References 1. Baird B. Dry aging enhances palatability of beef, Beef safety and quality. why these differences occurred because these types of 2008. https://www.beefresearch.org/CMDocs/BeefResearch/PE_Issues_ beef have different genetic factors and have undergone Update/Dry_aging_enhances_palatability_of_beef.pdf. Accessed March April different fattening methods and for different periods 2008. 2. Campbell RE, Hunt MC, Levis P, Chambers Iv E. Dry-aging effects on since slaughter. palatability of beef longissimus muscle. J Food Sci. 2001;66:196–9. Thepresent studyshows theresults ofthe 3. Dashmaa D, Tripathi VK, Cho S, Kim Y, Hwang I. Dry aging of beef; review. J dry-aging process under conditions without the inocu- Anim Sci Technol. 2016;58:20. 4. King MF, Matthews MA, Rule DC, Field RA. Effect of beef packing method lation of microorganisms. However, in actual condi- on volatile compounds developed by oven roasting or microwave cooking. tions, certain molds can grow on the surface of the J Agric Food Chem. 1995;43:773–8. meat during the dry-aging process, which is one fac- 5. Muramoto T, Maeno K, Okada Y, Tezuka S, Kamata T. Relationship between shear force and tenderness of Japanese shorthorn beef. Tohoku J Anim Sci tor that confers the taste of dry-aged beef [3]. There- Technol. 2014;64(1):7–12. fore, further studies are needed in demonstrate the 6. Parrish FC Jr, Boles JA, Rust RE, Olson DG. Dry and wet aging effects on effects of mold on dry-aged beef. palatability attributes of beef loin and rib steaks from three quality grades. J Food Sci. 1991;56:601–3. 7. Savell JW. Dry-aging of beef, exective summery: National Cattlemen’s Beef Conclusion Association; 2008. https://www.beefresearch.org/CMDocs/BeefResearch/ Dry%20Aging%20of%20Beef.pdf There was no significant difference between dry- and 8. Sitz BM, Calkins CR, Feuz DM, Umberger WJ, Eskridge KM. Consumer sensory wet-aging methods for all studied variables related to acceptance and value of wet-aged and dry-aged beef steaks. J Anim Sci. free amino acids or hardness in this study. 2006;84:1221–6. 9. Sugioka K, Tasaki T, Sakai H, Ieiri S, Araki T. Study of free amino acid in aging Acknowledgements of beef. Proceedings of the annual meeting 2015 Okayama of Japan Society We appreciate the technical supports offered by Ms. Sakima. Without her for Bioscience, biotechnology, and Agrochemistry. 2015: 2F26P18. persistent help this paper would not have been possible. 10. Tsuchiya T, Ukai Y, Saitou Y. Change in quality and productive loss of beef We are very grateful to Mr. Higa, Managing Director, and Mr. Gushiken, meat during dry-aging process. Bulletin of shizuoka prefecture livestock factory manager, Uehara Meat Co., Ltd., for providing meat and helping in research institute. 2013;6:12–4. preparations for the aging experiments. We express our sincere thanks to Dr. 11. Warren KE, Kastner CL. A comparison of dry aged and vacuum-aged beef Kobayashi, Livestock Improvement Association of Japan, Inc., Maebashi striploins. J Muscle Foods. 1992;3:151–7. Institute of Animal Science, for advice on the mechanism of the dry-aging process of beef, particularly about the process of producing free amino acids. We also gratefully acknowledge the help by Mr. Tamaki, President, and Mr. Uezu, Executive Managing Director, Okinawa Aging Company, for helpful discussions. Funding Funded by Science and Technology Promotion Division of Okinawa prefectural government. Authors’ contributions TH was concerned all things about this research and drafted and wrote this manuscript, NA supported everything about analysis and statistical work. Both authors read and approved the final manuscript. Ethics approval and consent to participate Not applicable.

Journal

Journal of Animal Science and TechnologySpringer Journals

Published: Sep 25, 2018

References