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Article

The E ffect of Irradiation on the Quality Properties of Tarhana

Nermin Ta¸so ˘gulları and Ömer ¸Sim¸sek *

Department of Food Engineering, University of Pamukkale, 20017 Denizli, Turkey;

ntasogullari09@posta.pau.edu.tr

* Correspondence: omers@pau.edu.tr; Tel.:+90-258-296-3015

Received: 27 May 2020; Accepted: 3 July 2020; Published: 10 July 2020 

Featured Application: Irradiation is a useful application to preserve tarhana by preventing microbiological risks and pest formation. An irradiation dose of 5 kGy and below can be applied for the preservation of the tarhana without quality loss.

Abstract:Tarhana is a traditional food produced by the fermentation, drying and grinding of dough prepared with wheat flour, yoghurt, various vegetables and spices. Microbiological risks and pest formation are the major problems encountered during the storage of tarhana. In this study, the effect of irradiation was determined in order to eliminate microbiological risks and pest formation while preserving the quality features during the storage of tarhana. Depending on the irradiation dose, microbial inhibition occurred in tarhana samples, and the maximum protection was achieved with 10 kGy. Nevertheless, doses of 2.5 and 5 kGy inhibited the growth of Bacillus cereus. Additionally, all irradiation doses prevented pest formation. The consistency coefficient of soups prepared with irradiated tarhana samples decreased depending on the irradiation doses. There was no difference in 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and total phenol content in the control with irradiated tarhana samples. However, the 10 kGy irradiated tarhana sample included higher thiobarbituric acid reactive substances. In conclusion, irradiation was applied for the first time to preserve tarhana by reducing the microbiological risk and preventing pest formation. Accordingly, a 5 kGy irradiation dose was recommended, with which the tarhana rheology was affected slightly.

Keywords: tarhana; food irradiation; microbiological safety; pest control; food safety

1. Introduction

Traditional fermented foods are a precious resource as they provide inexpensive, practical and convenient nutrients in the modern world under the threat of famine. Tarhana is a traditional fermented food that is prepared to consume as an instant soup by cooking a quantity of a dough powder in hot water [1,2]. It has nutritious and easy-to-digest properties, as well as having gained importance for infant nutrition in recent years [2,3]. Although there are slight differences in the production of tarhana, the tarhana dough is prepared by mixing wheat flour, yoghurt, sourdough and various vegetables and spices (tomato, red pepper, onion, mint, salt, etc.), and the dough is subsequently dried and ground after fermentation [1,4]. This fermented food is traditionally stored in a textile pouch, but plastic packages are preferred to avoid volatile loses [5]. However, it is difficult to keep the tarhana from moisture absorption, resulting in microbiological risks and pest formation problems during storage [6].

In fact, Turantas and Kemahlıoglu [7] showed the survival of the molds and some pathogenic bacteria such as E. coli O157:H7, S. aureus, S. typhimurium and B. cereus during the fermentation and storage period of tarhana. Additionally, aflatoxins ranging from 0.7–16.8 µg/kg were determined at 23.2%

of 138 collected tarhana samples, whereas 29 samples contained Aflatoxin B1 (AFB1) ranging from

Appl. Sci. 2020, 10, 4749; doi:10.3390/app10144749 www.mdpi.com/journal/applsci

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Appl. Sci. 2020, 10, 4749 2 of 10

0.2–13.2 µg/kg [8]. Irradiation may be one of the possible solutions to reduce the risk of microbiological and pest encroachment during the storage of tarhana [9].

The use of irradiation [9–11], which involves the exposure of food to gamma and X rays to ensure quality and safety, has gained importance in recent years due to the large increase in foodborne diseases.

Food irradiation is one of the prescribed methods for the control of the significant losses that can occur during the storage and distribution of products due to microorganisms, parasites and insects [9,11].

The high-energy rays applied to food systems stop the vitality of microorganisms or larvae in food by causing damage to their DNA. This sterilization of the food without the use of heat has led to the technique being referred to as “cold sterilization” [9]. The International Atomic Energy Agency (IAEA) concluded that energy beams emitted from food irradiated with doses below 60 kGy, with gamma rays from cobalt-60 and cesium 137, were less than 5 MeV in strength and could be considered insignificant [12].

The present study aims to determine the effect of irradiation on the quality and characteristics of tarhana to avoid microbiological risks and pest formation. Accordingly, the microbiological, chemical and physical effects of irradiation applied to packaged dried tarhana to prevent microbiological, physical and chemical losses during its storage is investigated.

2. Materials and Methods 2.1. Material

Five different flour type tarhana samples were purchased from local markets located in Denizli, Turkey in October 2015, which were blended and subsequently packaged in 5 kg plastic bags (LDPE, 30 × 52 cm, 60 micron thickness) for storage.

2.2. Irradiation and Storage Conditions of Tarhana Samples

The tarhana samples were irradiated by the GAMMAPAK Sterilization Ind. & Trd. Inc. (Tekirda ˘g, Turkey) with three different doses: 2.5, 5 and 10 kGy. Three packages of tarhana were irradiated for each dose, and three packages of tarhana were used as controls without irradiation. All the irradiated and control tarhana samples were stored in their packages at 25C for 5 months.

2.3. Microbiological and Pest Formation Analysis

The microbial analysis of tarhana samples (10 g) was carried out at the end of every month over 5 months of storage. The total aerobic mesophilic bacteria (TAMB) counts during the storage of tarhana samples were enumerated [13] on Plate Count Agar (PCA; Merck Co. Darmstadt, Germany) after 48 h incubation at 30C. The yeast grown in the tarhana samples was determined with a Dichloran Rose Bengal Chlortetracycline agar (DRBC; Merck Co., Darmstadt, Germany) at 25C after 5 days [14], and the Bacillus cereus was enumerated on Bacillus cereus agar (Merck Co., Darmstadt, Germany) supplemented with polymyxin B and trimethoprim at 30C after 72 h [15].

Pest analysis was performed by periodically observing the remaining material after 100 g of tarhana samples were sieved through a 2500 micron mesh during storage.

2.4. Acidity Degree, pH and Moisture Analysis

The acidity degree of the tarhana doughs was measured according to the TS2282 [5]. Accordingly, 10 g tarhana was homogenized with WiseStir mixer (HS-50A, DAIHAN, Korea) with 50 mL of 67%

neutralized ethanol (Sigma-Aldrich, St. Louis, MO, USA). Then, the mixture was filtered (cellulose filter paper, Fisher Scientific, Loughborough, UK) and finally titrated with 0.1 M NaOH (Sigma-Aldrich, USA). The spent NaOH amount was multiplied by 5 to reach the acidity value of the tarhana dough samples. The pH value of the dough samples was determined as follows: 25 mL of distilled water was added to a 5 g sample and homogenized using a WiseStir mixer (HS-50A, DAIHAN, Korea).

Distilled water was added to make a homogenous mixture of 50 mL, and the pH values were measured

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using a pH meter (Eutech Instruments, Singapore). The moisture contents of tarhana samples during storage were measured according to AOAC (Association of Official Analytical Chemists) [16].

2.5. Color and Viscosity Analysis

The color properties of the tarhana samples were determined with using the Hunter-Lab Mini Scan XE color measurement device (Reston, VA, USA, ABD). In order to determine the viscosity values of tarhana, 10 g of tarhana sample was weighed and a 10% (w/v) tarhana–water mixture was prepared by adding 90 mL of pure water. This mixture was heated in a mechanical shaker and boiled for 20 min.

The consistency coefficient (K) and flow behavior index (n) values of the tarhana–water mixtures were measured by a Brookfield programmable DV-II+ (Middleboro, MA, USA) viscometer. Samples were transferred to the sample container (Brookfield Accessories, SC4-13R) connected to a circulating water bath at 70C at 19 different speeds (5, 8, 10, 15, 17, 20, 25, 30, 35, 40, 50, 60, 70, 90, 105, 120, 140, 160 and 180 rpm) and K and n values were determined.

2.6. 2,2-Diphenyl-1-Picrylhydrazyl (DPPH) Radical Scavenging Analysis

The ability to scavenge the DPPH radical of tarhana samples was estimated by the methods of Wang et al. [17] and Fratianni et al. [18], respectively, with slight modifications. Aliquots of 0.1 mL tarhana extracts obtained from 1 g tarhana with 10 mL methanol (Sigma-Aldrich, USA) were mixed with 5 mL of 0.1 mM (prepared in methanol) DPPH radical in a test tube. The mixture was allowed to stand for 20 min at room temperature before the absorbance was measured at 517 nm spectrophotometrically (PG Instruments, Lutterworth, UK). The scavenging activity was calculated by the following equation:

Scavenging activity%= ((Absorbance of Blank − Absorbance of Sample))/(Absorbance of Blank) × 100 The “absorbance of blank” is the absorbance of the control reaction (containing all reagents except the test compound), and the “absorbance of sample” is the absorbance of the read test compound (517 nm).

2.7. Total Phenolic Content (TPC) Analysis

The tarhana samples were analyzed for total phenolic content by using the Folin–Ciocalteus assay after 1 g of tarhana sample was homogenized with 10 mL of methanol (Sigma-Aldrich, USA) and kept overnight for extraction at refrigeration temperature. The results were expressed in mg gallic acid (Sigma-Aldrich, USA) in 100 g of tarhana. [19].

2.8. Thiobarbituric Acid Reactive Substances (TBARS) Analysis

TBARS were determined using the extraction method described by Witte et al. [20]. TBARS numbers were calculated as mg of malonaldehyde per kg of tarhana (mg malondialdehyde/kg).

2.9. Statistical Analysis

All the experiments were carried out in triplicate. The microbiological, chemical and physical analyses of the tarhana samples, as well as the irradiation dose application, storage duration and intergroup differences, were determined a one-way analysis of variance using the MINITAB 17.1.0 program (State College, PA, USA).

3. Results and Discussion

3.1. Viable Organism Content of Tarhana Samples during Storage

In all irradiation doses applied, the formation of pests in tarhana samples could be prevented.

However, in the control group without irradiation, a significant number of pests were found (Figure1a).

These results showed that irradiation was effective at preventing pest formation. The major problem

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Appl. Sci. 2020, 10, 4749 4 of 10

for dried tarhana during storage is pest formation, which is a spontaneous biological problem due to the development of larval eggs over time if the tarhana is stored under unsuitable conditions with high moisture and temperature. This problem is recognized to cause economic losses for many dried foods [3]. In this study, to test the effectiveness of irradiation, irradiated and control tarhana samples were observed at a constant high temperature (25C). Normally, it is advised that tarhana should be stored under 20C with low humidity [2,6].

The irradiation widely and distinctly reduced the microbial load. However, microbiological activity increased with extended storage time dependent on the irradiation dose applied, meaning that higher doses retarded the microbial growth for longer. The TAMB amounts of tarhana samples were closely related with the applied irradiation dose. The TAMB content at approximately 7 log CFU/gr for control samples was reduced substantially after irradiation (Figure1b). However, the TAMB amounts significantly increased during storage (p< 0.05) for the 2.5 and 5 kGy irradiated tarhana samples.

In 10 kGy irradiated samples, the TAMB counts remained below 1 log CFU/gr until the fifth month of storage. Similarly, the yeast mold content of tarhana samples was considerably reduced at all irradiation doses. Only the 2.5 kGy irradiated sample remained above 1 log CFU/gr. Although the yeast mold content of the 2.5 kGy tarhana sample reached the same level as the control group at the end of storage (p> 0.05), the amount of yeast mold remained below 3 log CFU/gr for the 5 kGy irradiated tarhana sample. The lowest amount of yeast mold was found in the 10 kGy irradiated sample (Figure1c). After irradiation, the Bacillus cereus (BC) content was able to be reduced below 1 log CFU/gr at all doses applied. For the control tarhana sample, the BC content reached 5 log CFU/gr after 5 months.

However, the BC content increased above 1 log CFU/gr after the first month of storage for both 2.5 and 5 kGy irradiated samples as well as after the third month of storage for the 10 kGy irradiated sample;

however, the BC content was significantly below 2.5 log CFU/for at both irradiated tarhana samples (Figure1d). These results indicate that irradiation is important for ensuring microbiological safety in tarhana. In addition, while a similar level of microbiological protection was provided in tarhana at 2.5 and 5 kGy doses, more effective protection was achieved in the 10 kGy irradiated tarhana sample.

The effectiveness of irradiation is related with the microbial load of foods [9,11]. Indeed, tarhana is a fermented food which includes substantial amounts of lactic acid bacteria and yeast, although some of this flora are reduced during drying [1,2]. However, some spore-forming bacteria and molds could be contaminated according to the traditional drying conditions [6–8]. These microbiological results suggested that a minimum 5 kGy dose is required to eliminate the majority of fermenting and contaminating flora in dried tarhana.

Appl. Sci. 2020, 10, x FOR PEER REVIEW 4 of 10

problem due to the development of larval eggs over time if the tarhana is stored under unsuitable

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conditions with high moisture and temperature. This problem is recognized to cause economic losses

134

for many dried foods [3]. In this study, to test the effectiveness of irradiation, irradiated and control

135

tarhana samples were observed at a constant high temperature (25 °C). Normally, it is advised that

136

tarhana should be stored under 20 °C with low humidity [2,6].

137

The irradiation widely and distinctly reduced the microbial load. However, microbiological

138

activity increased with extended storage time dependent on the irradiation dose applied, meaning

139

that higher doses retarded the microbial growth for longer. The TAMB amounts of tarhana samples

140

were closely related with the applied irradiation dose. The TAMB content at approximately 7 log

141

CFU/gr for control samples was reduced substantially after irradiation (Figure 1b). However, the

142

TAMB amounts significantly increased during storage (p < 0.05) for the 2.5 and 5 kGy irradiated

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tarhana samples. In 10 kGy irradiated samples, the TAMB counts remained below 1 log CFU/gr until

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the fifth month of storage. Similarly, the yeast mold content of tarhana samples was considerably

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reduced at all irradiation doses. Only the 2.5 kGy irradiated sample remained above 1 log CFU/gr.

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Although the yeast mold content of the 2.5 kGy tarhana sample reached the same level as the control

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group at the end of storage (p > 0.05), the amount of yeast mold remained below 3 log CFU/gr for the

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5 kGy irradiated tarhana sample. The lowest amount of yeast mold was found in the 10 kGy

149

irradiated sample (Figure 1c). After irradiation, the Bacillus cereus (BC) content was able to be reduced

150

below 1 log CFU/gr at all doses applied. For the control tarhana sample, the BC content reached 5 log

151

CFU/gr after 5 months. However, the BC content increased above 1 log CFU/gr after the first month

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of storage for both 2.5 and 5 kGy irradiated samples as well as after the third month of storage for the

153

10 kGy irradiated sample; however, the BC content was significantly below 2.5 log CFU/for at both

154

irradiated tarhana samples (Figure 1d). These results indicate that irradiation is important for

155

ensuring microbiological safety in tarhana. In addition, while a similar level of microbiological

156

protection was provided in tarhana at 2.5 and 5 kGy doses, more effective protection was achieved

157

in the 10 kGy irradiated tarhana sample. The effectiveness of irradiation is related with the microbial

158

load of foods [9,11]. Indeed, tarhana is a fermented food which includes substantial amounts of lactic

159

acid bacteria and yeast, although some of this flora are reduced during drying [1,2]. However, some

160

spore-forming bacteria and molds could be contaminated according to the traditional drying

161

conditions [6–8]. These microbiological results suggested that a minimum 5 kGy dose is required to

162

eliminate the majority of fermenting and contaminating flora in dried tarhana.

163 164

(a) (b)

Ba

Aa

Aa

Cb Bb

Ab

Bb Ab Ac

0 2 4 6 8 10 12 14 16

0 1 3 5

Pest Formation (pcs/100 g)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Aa Aa Aa Aa

Bb

ABb

ABb

Ab

Cb

Bc

Ab

ABc

d

0 1 2 3 4 5 6 7 8

0 1 3 5

TAMB log CFU/g

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Figure 1. Cont.

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(c) (d)

Figure 1. The microbiological changes and pest formation during the storage of irradiated and control

165

tarhana samples. (a) Pest formation, (b) total aerobic mesophilic bacteria (TAMB), (c) yeast and (d)

166

Bacillus cereus counts. Capital letters represent the p < 0.05 level of statistical difference between the

167

storage times of the tarhana samples, while the lower-case letters show the p < 0.05 level of statistical

168

difference between the irradiation doses.

169

3.2. Physical Changes in Tarhana Samples During Storage

170

The irradiation doses applied had a slight effect on the pH, acidity degree and moisture content

171

of tarhana samples (Table 1). However, significant changes were observed in these parameters

172

independent of irradiation with storage time. The pH of the tarhana samples increased during the

173

storage (p < 0.05). There were no differences between tarhana samples at the beginning of storage,

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but the pH increase was slower (p < 0.05) in the 5 and 10 kGy irradiated tarhana samples for later

175

storage times. In contrast to the pH increase, the acidity degree of the tarhana samples decreased

176

during storage (p < 0.05); in particular, in the third month of storage, there was a significant decrease

177

in the acidity degree of all tarhana samples (p < 0.05). There was no difference during storage for the

178

acidity degree of tarhana samples with different irradiation doses. As expected, the moisture content

179

of tarhana increased depending on the storage time; in particular, in the third month, the amount of

180

moisture was increased significantly in the tarhana samples (p < 0.05). Moreover, the moisture

181

contents of 5 and 10 kGy irradiated tarhana samples were lower from the beginning to the end of

182

storage. Dried tarhana is hygroscopic and has a rapid moisture absorption ability [2,3]. Although

183

tarhana is traditionally stored in pouches or commercial polyethylene bags [1,4], the pH increase and

184

decreased acidity in addition to the increased moisture content might be the cause of microbial

185

development and pest formation, as shown above.

186

Table 1. The pH, acidity degree* and moisture content changes during the storage of

187

irradiated and control tarhana samples.

188 189

Samples Months pH Acidity Value* Moisture Content (%)

Control 0 4.28 ± 0.00Da 23.25 ± 4.11Aa 17.17 ± 0.05Ca 1 4.87 ± 0.28Ca 18.40 ± 2.52Aa 16.95 ± 0.24Bab 3 5.46 ± 0.57Db 11.71 ± 0.82Aa 18.44 ± 0.24Bc 5 6.16 ± 0.51Db 11.40 ± 1.67Aa 18.98 ± 0.15Bbc 2,5 kGy 0 4.20 ± 0.00Ca 20.92 ± 2.86Aa 17.14 ± 0.21Ca 1 5.08 ± 0.16Ca 17.20 ± 0.85Aa 16.97 ± 0.25Ba

Ba Ba

Aa Aa

Db

Cb

Bb

Ab

Bc

Ac

d 1

2 3 4 5 6 7

0 1 3 5

Yeast Log CFU/g

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Ca

Ba

Aa

Aa

Ab Ab

Bc

Ab Ac

0 1 2 3 4 5 6

0 1 3 5

Bacillus cereus Log CFU/g

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Figure 1.The microbiological changes and pest formation during the storage of irradiated and control tarhana samples. (a) Pest formation, (b) total aerobic mesophilic bacteria (TAMB), (c) yeast and (d) Bacillus cereus counts. Capital letters represent the p< 0.05 level of statistical difference between the storage times of the tarhana samples, while the lower-case letters show the p< 0.05 level of statistical difference between the irradiation doses.

3.2. Physical Changes in Tarhana Samples during Storage

The irradiation doses applied had a slight effect on the pH, acidity degree and moisture content of tarhana samples (Table 1). However, significant changes were observed in these parameters independent of irradiation with storage time. The pH of the tarhana samples increased during the storage (p< 0.05). There were no differences between tarhana samples at the beginning of storage, but the pH increase was slower (p< 0.05) in the 5 and 10 kGy irradiated tarhana samples for later storage times. In contrast to the pH increase, the acidity degree of the tarhana samples decreased during storage (p< 0.05); in particular, in the third month of storage, there was a significant decrease in the acidity degree of all tarhana samples (p< 0.05). There was no difference during storage for the acidity degree of tarhana samples with different irradiation doses. As expected, the moisture content of tarhana increased depending on the storage time; in particular, in the third month, the amount of moisture was increased significantly in the tarhana samples (p< 0.05). Moreover, the moisture contents of 5 and 10 kGy irradiated tarhana samples were lower from the beginning to the end of storage.

Dried tarhana is hygroscopic and has a rapid moisture absorption ability [2,3]. Although tarhana is traditionally stored in pouches or commercial polyethylene bags [1,4], the pH increase and decreased acidity in addition to the increased moisture content might be the cause of microbial development and pest formation, as shown above.

Table 1.The pH, acidity degree * and moisture content changes during the storage of irradiated and control tarhana samples.

Samples Months pH Acidity Value * Moisture Content (%)

Control 0 4.28 ± 0.00Da 23.25 ± 4.11Aa 17.17 ± 0.05Ca

1 4.87 ± 0.28Ca 18.40 ± 2.52Aa 16.95 ± 0.24Bab

3 5.46 ± 0.57Db 11.71 ± 0.82Aa 18.44 ± 0.24Bc

5 6.16 ± 0.51Db 11.40 ± 1.67Aa 18.98 ± 0.15Bbc

2.5 kGy 0 4.20 ± 0.00Ca 20.92 ± 2.86Aa 17.14 ± 0.21Ca

1 5.08 ± 0.16Ca 17.20 ± 0.85Aa 16.97 ± 0.25Ba

3 5.57 ± 0.03Cb 12.45 ± 1.32Aa 18.16 ± 0.42Bb

5 5.88 ± 0.56Cb 10.43 ± 0.36Aa 18.32 ± 0.36Bbc

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Appl. Sci. 2020, 10, 4749 6 of 10

Table 1. Cont.

Samples Months pH Acidity Value * Moisture Content (%)

5 kGy 0 4.18 ± 0.01Ba 22.57 ± 1.66Ba 16.87 ± 0.13Ba

1 4.83 ± 0.04Ba 19.35 ± 1.80Ba 16.76 ± 0.21Aab

3 5.11 ± 0,03Bb 12.33 ± 0.66Ba 17.65 ± 0.22Ac

5 5.45 ± 0.14Bb 10.70 ± 0.04Ba 17.70 ± 0.38Abc

10 kGy 0 4.15 ± 0.01Aa 21.36 ± 0.59Ba 16.98 ± 0.17Aa

1 4.54 ± 0.06Aa 21.71 ± 1.01Ba 16.64 ± 0.25Aab

3 5.21 ± 0.36Ab 13.43 ± 0.46Ba 18.09 ± 0.19Ac

5 5.68 ± 0.25Ab 10.55 ± 0.62Ba 18.36 ± 0.13Abc

Capital letters represent the p< 0.05 level of statistical difference between the storage times of the tarhana samples, while the lower-case letters show the p< 0.05 level of statistical difference between the irradiation doses. *: The acidity values of the tarhana samples were given according to the standard TS8222 [5] due to the amount of NaOH used to neutralize the acidity of 10 g tarhana extracted with 67% ethanol.

3.3. Color Changes in Tarhana Samples during Storage

The effect of irradiation on the color of tarhana is also of interest. In this sense, the effect of strong ionizing radiation should be evaluated in terms of the color components. The L value of the tarhana samples increased at the initial storage and stabilized after the third month. The rate of L value of tarhana increased depending on the irradiation dose applied, but there was no significant difference in the L value between the samples in terms of doses and storage time (Figure2a). The whitening at the end of the first month observed in this study is a common problem encountered in the storage of tarhana. The main factor here may be the loss of color due to the effect of light. The “a” value slightly decreased during the storage. This might be related to oxygen or light contact rather than the effect of irradiation. However, the decline rate of the “a” value of tarhana samples was dependent on radiation dose, where the 10 kGy dose did reduce faster than the other doses (Figure2b,c). In contrast, the “b” value of the tarhana samples increased in the first month and remained stable for a further four months. Tarhana has a characteristic yellowish color resulting from the paprika used in the ingredients, which is important for consumer perception [2]. These results clearly showed that irradiation has no detrimental effect on the color quality of tarhana samples, especially in terms of whitening, due to the loss of pigments arising from the paprika used in the production.

Appl. Sci. 2020, 10, x FOR PEER REVIEW 6 of 10

3 5.57 ± 0.03Cb 12.45 ± 1.32Aa 18.16 ± 0.42Bb 5 5.88 ± 0.56Cb 10.43 ± 0.36Aa 18.32 ± 0.36Bbc 5 kGy 0 4.18 ± 0.01Ba 22.57 ± 1.66Ba 16.87 ± 0.13Ba 1 4.83 ± 0.04Ba 19.35 ± 1.80Ba 16.76 ± 0.21Aab 3 5.11 ± 0,03Bb 12.33 ± 0.66Ba 17.65 ± 0.22Ac 5 5.45 ± 0.14Bb 10.70 ± 0.04Ba 17.70 ± 0.38Abc 10 kGy 0 4.15 ± 0.01Aa 21.36 ± 0.59Ba 16.98 ± 0.17Aa 1 4.54 ± 0.06Aa 21.71 ± 1.01Ba 16.64 ± 0.25Aab 3 5.21 ± 0.36Ab 13.43 ± 0.46Ba 18.09 ± 0.19Ac 5 5.68 ± 0.25Ab 10.55 ± 0.62Ba 18.36 ± 0.13Abc

190

Capital letters represent the p < 0.05 level of statistical difference between the storage times of the

191

tarhana samples, while the lower-case letters show the p < 0.05 level of statistical difference between

192

the irradiation doses.

193

*: The acidity values of the tarhana samples were given according to the standard TS8222 [5] due to

194

the amount of NaOH used to neutralize the acidity of 10 g tarhana extracted with 67% ethanol.

195

3.3. Color Changes in Tarhana Samples During Storage

196

The effect of irradiation on the color of tarhana is also of interest. In this sense, the effect of strong

197

ionizing radiation should be evaluated in terms of the color components. The L value of the tarhana

198

samples increased at the initial storage and stabilized after the third month. The rate of L value of

199

tarhana increased depending on the irradiation dose applied, but there was no significant difference

200

in the L value between the samples in terms of doses and storage time (Figure 2a). The whitening at

201

the end of the first month observed in this study is a common problem encountered in the storage of

202

tarhana. The main factor here may be the loss of color due to the effect of light. The “a” value slightly

203

decreased during the storage. This might be related to oxygen or light contact rather than the effect

204

of irradiation. However, the decline rate of the “a” value of tarhana samples was dependent on

205

radiation dose, where the 10 kGy dose did reduce faster than the other doses (Figure 2b and c). In

206

contrast, the “b” value of the tarhana samples increased in the first month and remained stable for a

207

further four months. Tarhana has a characteristic yellowish color resulting from the paprika used in

208

the ingredients, which is important for consumer perception [2]. These results clearly showed that

209

irradiation has no detrimental effect on the color quality of tarhana samples, especially in terms of

210

whitening, due to the loss of pigments arising from the paprika used in the production.

211 212

(a) (b)

Ba

Aa Aa Aa

Ba

Aa Aa Aa

Ba

Aa Aa Aa

Ba

Aa Aa Aa

0 10 20 30 40 50 60 70 80 90 100

0 1 3 5

Lightness (L)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Aa Ba BCa Ca

AaAb BaAa BCaBab CaCa

Aab Bb BCc

Cb

0 10 20 30 40 50

0 1 3 5

Redness (a)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Figure 2. Cont.

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(c)

213

Figure 2. The color changes during the storage of irradiated and control tarhana samples. (a) Lightness

214

L, (b) redness +a, (c) yellowness +b. Capital letters represent the p < 0.05 level of statistical difference

215

between the storage times of the tarhana samples, while the lower-case letters show the p < 0.05 level

216

of statistical difference between the irradiation doses.

217

3.4. Rheological Properties of Tarhana Samples During Storage

218

The consistency coefficients of soups prepared with tarhana powders decreased dramatically

219

depending on the irradiation dose applied because the consistency coefficient of the unirradiated

220

tarhana sample was higher than the irradiated samples during storage. However, 10 kGy irradiated

221

tarhana samples were less viscous (p < 0.05) than other 2.5 and 5 kGy irradiated tarhana samples

222

(Figure 3). However, there was no significantly consistent behavior between the 2.5 and 5 kGy

223

irradiated samples. The consistency coefficient of unirradiated samples increased in contrast with the

224

irradiated tarhana samples. Briefly, the irradiation destructively affected the viscosity of the tarhana

225

samples in a dose-dependent manner. This might be related with the degradation of the chain

226

structures of the amylose and amylopectin fractions of starch, which are mainly responsible with

227

hydration in tarhana. It was commented that gamma radiation may contribute to molecular changes

228

and the fragmentation of starch, leading to a progressive reduction in the molecular size of amylose

229

and amylopectin by random cleavage of the glycosidic chains and consequently changes to the

230

structural and physicochemical characteristics of starch [21]. However, the consistency coefficient of

231

tarhana samples was in the range reported by Ibanoglu and Ibanoğlu [22] except for the 10 kGy

232

irradiated tarhana sample.

233

234

AaAaAa AaAaAa AaAbAb AaAbAb

Aa Aa Ab Ab

0 10 20 30 40 50

0 1 3 5

Yellowness (b)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Da

Ca

Ba Aa

Bb Bb

Ab Ab ABb

Bb Bc Bb

Ac

Bc Bd

Bc

0 2 4 6 8 10 12 14 16

0 1 3 5

Consistency coefficient (Pa.Sn)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Figure 2.The color changes during the storage of irradiated and control tarhana samples. (a) Lightness L, (b) redness+a, (c) yellowness +b. Capital letters represent the p < 0.05 level of statistical difference between the storage times of the tarhana samples, while the lower-case letters show the p< 0.05 level of statistical difference between the irradiation doses.

3.4. Rheological Properties of Tarhana Samples during Storage

The consistency coefficients of soups prepared with tarhana powders decreased dramatically depending on the irradiation dose applied because the consistency coefficient of the unirradiated tarhana sample was higher than the irradiated samples during storage. However, 10 kGy irradiated tarhana samples were less viscous (p< 0.05) than other 2.5 and 5 kGy irradiated tarhana samples (Figure3). However, there was no significantly consistent behavior between the 2.5 and 5 kGy irradiated samples. The consistency coefficient of unirradiated samples increased in contrast with the irradiated tarhana samples. Briefly, the irradiation destructively affected the viscosity of the tarhana samples in a dose-dependent manner. This might be related with the degradation of the chain structures of the amylose and amylopectin fractions of starch, which are mainly responsible with hydration in tarhana.

It was commented that gamma radiation may contribute to molecular changes and the fragmentation of starch, leading to a progressive reduction in the molecular size of amylose and amylopectin by random cleavage of the glycosidic chains and consequently changes to the structural and physicochemical characteristics of starch [21]. However, the consistency coefficient of tarhana samples was in the range reported by Ibanoglu and Ibano ˘glu [22] except for the 10 kGy irradiated tarhana sample.

(c)

213

Figure 2. The color changes during the storage of irradiated and control tarhana samples. (a) Lightness

214

L, (b) redness +a, (c) yellowness +b. Capital letters represent the p < 0.05 level of statistical difference

215

between the storage times of the tarhana samples, while the lower-case letters show the p < 0.05 level

216

of statistical difference between the irradiation doses.

217

3.4. Rheological Properties of Tarhana Samples During Storage

218

The consistency coefficients of soups prepared with tarhana powders decreased dramatically

219

depending on the irradiation dose applied because the consistency coefficient of the unirradiated

220

tarhana sample was higher than the irradiated samples during storage. However, 10 kGy irradiated

221

tarhana samples were less viscous (p < 0.05) than other 2.5 and 5 kGy irradiated tarhana samples

222

(Figure 3). However, there was no significantly consistent behavior between the 2.5 and 5 kGy

223

irradiated samples. The consistency coefficient of unirradiated samples increased in contrast with the

224

irradiated tarhana samples. Briefly, the irradiation destructively affected the viscosity of the tarhana

225

samples in a dose-dependent manner. This might be related with the degradation of the chain

226

structures of the amylose and amylopectin fractions of starch, which are mainly responsible with

227

hydration in tarhana. It was commented that gamma radiation may contribute to molecular changes

228

and the fragmentation of starch, leading to a progressive reduction in the molecular size of amylose

229

and amylopectin by random cleavage of the glycosidic chains and consequently changes to the

230

structural and physicochemical characteristics of starch [21]. However, the consistency coefficient of

231

tarhana samples was in the range reported by Ibanoglu and Ibanoğlu [22] except for the 10 kGy

232

irradiated tarhana sample.

233

234

AaAaAaAa AaAaAaAa AaAbAbAb AaAbAbAb

0 10 20 30 40 50

0 1 3 5

Yellowness (b)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Da

Ca

Ba Aa

Bb Bb

Ab Ab ABb

Bb Bc Bb

Ac

Bc Bd

Bc

0 2 4 6 8 10 12 14 16

0 1 3 5

Consistency coefficient (Pa.Sn)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Figure 3. The consistency coefficient of soups prepared with using irradiated and control tarhana samples. Capital letters represent the p< 0.05 level of statistical difference between the storage times

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Appl. Sci. 2020, 10, 4749 8 of 10

of the tarhana samples, while the lower-case letters show the p< 0.05 level of statistical difference between the irradiation doses.

3.5. Antioxidant Properties of Tarhana Samples during Storage

The DPPH radical scavenging activity of tarhana samples, which were administered at 5 and 10 kGy irradiation doses at the beginning of storage, was found to be higher than that of the control and 2.5 kGy irradiated tarhana samples (p< 0.05). With continuing months of storage, no difference was observed between irradiated and control samples in terms of DPPH radical scavenging activity.

However, the scavenging activity of tarhana samples decreased with the storage time; in particular, a significant decrease in scavenging activity was observed in the third month (Figure4a). Similarly, there was no significant difference in the total phenol content between tarhana samples after irradiation and during storage. In the analyzes performed at the third and fifth months, only the 10 kGy irradiated tarhana sample had a lower total phenol content (p< 0.05, Figure 4b). As expected, the amount of TBARS in tarhana samples increased contrary to the decrease in antioxidant properties.

After irradiation, the 5 and 10 kGy irradiated tarhana samples contained higher amounts of TBARS (p< 0.05). During storage, the amount of TBARS in tarhana samples increased and more TBARS were detected in 10 kGy irradiated tarhana sample each time (Figure4c). The major concern regarding the adverse effect of irradiation in food systems is that it can trigger oxidation [9,11]. When evaluated in this respect, it was seen that irradiation did not cause a decrease in the antioxidant capacity of tarhana.

Even at the beginning of storage, higher levels of antioxidant properties were observed. This might be due to the effect of irradiation resulting in the release of the antioxidant substances. On the other hand, irradiation did not significantly affect oxidation in tarhana samples. Higher oxidative products were formed only when 10 kGy doses were applied.

Appl. Sci. 2020, 10, x FOR PEER REVIEW 8 of 10

Figure 3. The consistency coefficient of soups prepared with using irradiated and control tarhana

235

samples. Capital letters represent the p < 0.05 level of statistical difference between the storage times

236

of the tarhana samples, while the lower-case letters show the p < 0.05 level of statistical difference

237

between the irradiation doses.

238

3.5. Antioxidant Properties of Tarhana Samples During Storage

239

The DPPH radical scavenging activity of tarhana samples, which were administered at 5 and 10

240

kGy irradiation doses at the beginning of storage, was found to be higher than that of the control and

241

2.5 kGy irradiated tarhana samples (p < 0.05). With continuing months of storage, no difference was

242

observed between irradiated and control samples in terms of DPPH radical scavenging activity.

243

However, the scavenging activity of tarhana samples decreased with the storage time; in particular,

244

a significant decrease in scavenging activity was observed in the third month (Figure 4a). Similarly,

245

there was no significant difference in the total phenol content between tarhana samples after

246

irradiation and during storage. In the analyzes performed at the third and fifth months, only the 10

247

kGy irradiated tarhana sample had a lower total phenol content (p < 0.05, Figure 4b). As expected,

248

the amount of TBARS in tarhana samples increased contrary to the decrease in antioxidant properties.

249

After irradiation, the 5 and 10 kGy irradiated tarhana samples contained higher amounts of TBARS

250

(p < 0.05). During storage, the amount of TBARS in tarhana samples increased and more TBARS were

251

detected in 10 kGy irradiated tarhana sample each time (Figure 4c). The major concern regarding the

252

adverse effect of irradiation in food systems is that it can trigger oxidation [9,11]. When evaluated in

253

this respect, it was seen that irradiation did not cause a decrease in the antioxidant capacity of

254

tarhana. Even at the beginning of storage, higher levels of antioxidant properties were observed. This

255

might be due to the effect of irradiation resulting in the release of the antioxidant substances. On the

256

other hand, irradiation did not significantly affect oxidation in tarhana samples. Higher oxidative

257

products were formed only when 10 kGy doses were applied.

258 259

(a) (b)

b

a

a

a b

a

a

a a

ab

a

a a

ab

b

b

0 10 20 30 40 50 60

0 1 3 5

DPPH Scavenging Activity (%)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

a a

ab

a a

a

a

a

ab a

ab

b a

ab

b

b

0 20 40 60 80 100 120 140

0 1 3 5

TPC (mg Galc Acıd/100 g Tarhaha)

Storage (months) Control 2,5 kGy 5 kGy 10 kGy

Figure 4. Cont.

(9)

(c)

Figure 4. The antioxidant properties and changes of irradiated and control tarhana samples during

260

storage. (a) 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, (b) total phenolic

261

content (TPC), (c) thiobarbituric acid reactive substances (TBARS). Capital letters represent the p <

262

0.05 level of statistical difference between the storage times of the tarhana samples, while the lower-

263

case letters show the p < 0.05 level of statistical difference between the irradiation doses.

264

4. Conclusions

265

In this study, the use of irradiation for the preservation of tarhana was evaluated for the first

266

time. Irradiation was successfully applied to reduce microbiological risks as well as pest formation

267

in tarhana during storage. Although irradiation showed no adverse effect on the color and

268

antioxidation properties of tarhana, it reduced the consistency coefficient of the soups prepared from

269

tarhana. Accordingly, instead of the 10 kGy dose, it is recommended to use 5 kGy or lower doses in

270

order to preserve the quality features of tarhana.

271

Author Contributions: Conceptualization, Ö.Ş..; methodology, N.T.; validation, N.T., formal analysis, N.T.;

272

investigation, N.T; resources, Ö.Ş.; data curation, N.T..; writing—original draft preparation, N.T.; writing—

273

review and editing, Ö.Ş.; visualization, N.T.; supervision, Ö.Ş.; project administration, Ö.Ş..; funding acquisition,

274

Ö.Ş. All authors have read and agreed to the published version of the manuscript.

275

Funding: This research was funded by the University of Pamukkale, Scientific Research Projects Coordination

276

Unit, grant number 2016FEBE024.

277

Conflicts of Interest: The authors declare no conflict of interest.

278

References

279

1. Dağlıoğlu, O. Tarhana as a traditional Turkish fermented cereal food. Its recipe, production and

280

composition. Nahrung 2000, 44, 85–88.

281

2. Ö zdemir, S.; Gocmen, D.; Kumral, A.Y. A traditional Turkish fermented cereal food: Tarhana. Food Rev. Int.

282

2007, 23, 107–121.

283

3. Kabak, B.; Dobson, A.D.W. An introduction to the traditional fermented foods and beverages of Turkey.

284

Crit. Rev. Food Sci. Nutr. 2011, 51, 248–260.

285

4. Siyamoğlu, B. Türk Tarhanalarının Yapılışı ve Terkibi Üzerine Araştırma; Ege University, Faculty of

286

Agriculture Press: İzmir, Turkey, 1961; pp. 1–75.

287

5. Anonymous. TS 2282 Tarhana Standard; Turkish Standard Institute: Ankara, Turkey, 2004.

288

6. Dalgic, A.C.; Belibagli, K.B. Hazard analysis critical control points implementation in traditional foods: A

289

case study of tarhana processing. Int. J. Food Sci. Technol. 2008, 43, 1352–1360.

290

7. Turantaş, F.; Kemahlıoglu, K. Fate of some pathogenic bacteria and molds in Turkish tarhana during

291

fermentation and storage period. J. Food Sci. Technol. Mysore 2012, 49, 601–607.

292

b

b

b

b

b

b

ab

b

a

b

ab

ab

a

a

a

a

0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6

0 1 3 5

TBARS (mg malondialdehyde/kg)

Storage (months)

Control 2,5 kGy 5 kGy 10 kGy

Figure 4.The antioxidant properties and changes of irradiated and control tarhana samples during storage. (a) 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, (b) total phenolic content (TPC), (c) thiobarbituric acid reactive substances (TBARS). Capital letters represent the p< 0.05 level of statistical difference between the storage times of the tarhana samples, while the lower-case letters show the p< 0.05 level of statistical difference between the irradiation doses.

4. Conclusions

In this study, the use of irradiation for the preservation of tarhana was evaluated for the first time. Irradiation was successfully applied to reduce microbiological risks as well as pest formation in tarhana during storage. Although irradiation showed no adverse effect on the color and antioxidation properties of tarhana, it reduced the consistency coefficient of the soups prepared from tarhana.

Accordingly, instead of the 10 kGy dose, it is recommended to use 5 kGy or lower doses in order to preserve the quality features of tarhana.

Author Contributions: Conceptualization, Ö.¸S.; methodology, N.T.; validation, N.T., formal analysis, N.T.;

investigation, N.T; resources, Ö.¸S.; data curation, N.T.; writing—original draft preparation, N.T.; writing—review and editing, Ö.¸S.; visualization, N.T.; supervision, Ö.¸S.; project administration, Ö.¸S.; funding acquisition, Ö.¸S.

All authors have read and agreed to the published version of the manuscript.

Funding:This research was funded by the University of Pamukkale, Scientific Research Projects Coordination Unit, grant number 2016FEBE024.

Conflicts of Interest:The authors declare no conflict of interest.

References

1. Da ˘glıo ˘glu, O. Tarhana as a traditional Turkish fermented cereal food. Its recipe, production and composition.

Nahrung 2000, 44, 85–88. [CrossRef]

2. Özdemir, S.; Gocmen, D.; Kumral, A.Y. A traditional Turkish fermented cereal food: Tarhana. Food Rev. Int.

2007, 23, 107–121. [CrossRef]

3. Kabak, B.; Dobson, A.D.W. An introduction to the traditional fermented foods and beverages of Turkey.

Crit. Rev. Food Sci. Nutr. 2011, 51, 248–260. [CrossRef] [PubMed]

4. Siyamo ˘glu, B. Türk Tarhanalarının Yapılı¸sı ve Terkibi Üzerine Ara¸stırma; Ege University, Faculty of Agriculture Press: ˙Izmir, Turkey, 1961; pp. 1–75.

5. Anonymous. TS 2282 Tarhana Standard; Turkish Standard Institute: Ankara, Turkey, 2004.

6. Dalgic, A.C.; Belibagli, K.B. Hazard analysis critical control points implementation in traditional foods:

A case study of tarhana processing. Int. J. Food Sci. Technol. 2008, 43, 1352–1360. [CrossRef]

7. Turanta¸s, F.; Kemahlıoglu, K. Fate of some pathogenic bacteria and molds in Turkish tarhana during fermentation and storage period. J. Food Sci. Technol. Mysore 2012, 49, 601–607. [CrossRef] [PubMed]

(10)

Appl. Sci. 2020, 10, 4749 10 of 10

8. Colak, H.; Hampikyan, H.; Bingol, E.B.; Cetin, O.; Akhan, M.; Turgay, S.I. Determination of mould and aflatoxin contamination in Tarhana, a Turkish fermented food. Sci. World J. 2012, 2012, 218679. [CrossRef]

[PubMed]

9. Mahapatra, A.K.; Muthukumarappan, K.; Julson, J. Application of ozone, bacteriocins and irradiation in food processing; a review. Crit. Rev. Food Sci. Nutr. 2007, 45, 447–461. [CrossRef] [PubMed]

10. WHO. Food Irradiation-A Technique for Preserving and Improving the Safety of Food; WHO: Geneva, Switzerland, 1991.

11. Ravindran, R.; Jaiswal, A.K. Wholesomeness and safety aspects of irradiated foods. Food Chem. 2019, 285, 363–368. [CrossRef] [PubMed]

12. Agency, I.A.E. Natural and Induced Radioactivity in Food; International Atomic Energy Agency: Vienna, Austria, 2002.

13. ISO 15214:1998. Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Mesophilic Lactic Acid Bacteria—Colony-Count Technique at 30 Degrees C; International Organization for Standardization:

Geneva, Switzerlan, 1998.

14. ISO 21527-1:2008(en). Microbiology of Food and Animal Feeding Stuffs—Horizontal Method for the Enumeration of Yeasts and Moulds—Part 1: Colony Count Technique in Products with Water Activity Greater than 0.95;

International Organization for Standardization: Geneva, Switzerlan, 2008.

15. ISO 7932:1993. Microbiology—General Guidance for the Enumeration of Bacillus cereus—Colony-Count Technique at 30 Degrees C; International Organization for Standardization: Geneva, Switzerlan, 1993.

16. AOAC. Official Methods of Analysis, 15th ed.; Association of Official Analytical Chemists: Washington, DC, USA, 1990.

17. Wang, M.; Simon, J.E.; Aviles, I.F.; He, K.; Zheng, Q.Y.; Tadmor, Y. Analysis of antioxidative phenolic compounds in artichoke (Cynara scolymus L.). J. Agric. Food Chem. 2003, 51, 601–608. [CrossRef] [PubMed]

18. Fratianni, F.; De Martino, L.; Melone, A.; De Feo, V.; Coppola, R.; Nazzaro, F. Preservation of chicken breast meat treated with thyme and balm essential oils. J. Food Sci. 2010, 75, 528–535. [CrossRef] [PubMed]

19. Singleton, V.L.; Rossi, J.A. Colorimetry of total phenolics with phosphomolybdic-phosphotungistic acid reagents. Am. J. Enol. Vitic. 1965, 16, 144–158.

20. Witte, V.C.; Krause, G.F.; Bailey, M.E. A new extraction method for determining 2-thiobarbituric acid values of pork and beef during storage. J. Food Sci. 1970, 35, 582–585. [CrossRef]

21. Kong, X. Gamma Irradiation of Starch. In Physical Modifications of Starch; Sui, Z., Kong, X., Eds.; Springer:

Singapore, 2018; pp. 63–96.

22. Ibanoglu, S.; Ibanoglu, E. Rheological properties of cooked tarhana, a cereal-based soup. Food Res. Int. 1999, 32, 29–33. [CrossRef]

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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