antioxidant and antimicrobial activities of pomegranate (punica ...

Доклади на Българската академия на науките Comptes rendus de l’Acad´ emie bulgare des Sciences Tome 67, No 1, 2014

SCIENCE AGRAIRE Culture des plantes

ANTIOXIDANT AND ANTIMICROBIAL ACTIVITIES OF POMEGRANATE (PUNICA GRANATUM L.) SOUR SAUCE EXTRACTS Hilal Yildiz, Memnune Sengul∗, Bulent Cetin∗, Neva Karatas∗, Sezai Ercisli∗∗, Zuhal Okcu, Ahmed Dzubur∗∗∗, Semina Hadziabulic∗∗∗ (Submitted by Academician A. Atanassov on September 10, 2013)

Abstract In this study, nine-pomegranate sour sauce (PSS) obtained from market in Turkey was tested for their total phenolic content (TPC), antioxidant and antimicrobial activity and selected physico-chemical properties. Methanolwater extracts of PSS were found more effective than water extracts against microorganisms. In particular, Pseudomonas pseudoalkaligenes BC 3445 and Staphylococcus aureus ATCC 29213 showed the greatest sensitivity to the extracts. Total soluble solids, total sugar, reducing sugar, sucrose, ash, pH, titratable acidity, colour (L, a, b) of PSS were determined between 69.5–73.3◦ Brix, 21.89–53.44 g/100 g, 20.97–50.62 g/100 g, 0.75–2.81 g/100 g, 0.05–0.58 g/100 g, 2.64–2.91, 4.30–7.97, 16.58–55.38, (+3.94)–(+38.54), (−6.98)–(−38.36). βcarotene bleaching assay indicated that PSS possessed considerable antioxidant activities. The higher antioxidant capacities of PSS implied that they might be potential resources for the development of functional food. Key words: pomegranate, Punica granatum L., total phenolic content, antioxidant activity, antimicrobial activity

Introduction. The pomegranate (Punica granatum L.) is an ancient fruit that has been widely consumed in many different cultures for thousands of years, largely without incidents, and thus is considered generally safe by the general public [1 ]. Turkey is one of the native lands of the pomegranate and it is widely grown in the Mediterranean, Aegean, southeast and some of the microclimates of Turkey [2 ]. The edible part of the pomegranate fruit contains considerable 10

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amount of acids, sugars, vitamins, polysaccharides, polyphenols and important minerals. Pomegranate also contains coloured arils that give a delicious juice [1 ]. Pomegranate has been used in folk medicine for many centuries. In traditional medicine pomegranate fruits have been used to treat acidosis, dysentery, microbial infections, diarrhoea, helminthiasis, haemorrhage and respiratory pathologies. Pomegranate phytochemicals also show potential in chemoprevention of various types of cancers, by exerting antiproliferative effects on tumour cells [3 ]. Pomegranate juice has been proposed as a chemopreventive, chemotherapeutic, antiatherosclerotic and anti-inflammatory agent. Pomegranate juice has demonstrated high antioxidant activity and is effective in the prevention of atherosclerosis, low-density lipoprotein oxidation, prostate cancer, platelet aggregation and various cardiovascular diseases [4 ]. The health benefits of pomegranate have been attributed to its wide range of phytochemicals. The phytochemicals found in pomegranate are predominantly polyphenols, including flavonoids (anthocyanins, catechins and other complex flavonoids) and hydrolyzable tannins (punicalin, pedunculagin, punicalagin, gallagic and ellagic acid esters of glucose), which account for 92% of its antioxidant activity [5 ]. Ellagitannins found in the outer part of the fruit are largely responsible for the antioxidant activity of the pomegranate juice. It has been demonstrated that one of the ellagitannins, punicalagins (punicalagin anomers A and B), are responsible for over 50% of the antioxidant activity of the pomegranate juice [4 ]. The pomegranate is popularly consumed as fresh fruit, beverage (juice and wine), as a food product (jams and jellies) [5 ], grenadine or wine. The pomegranate sour sauce is commonly used for salads and many dishes in Turkey. In this study we have investigated the total phenolic content, antioxidant activity, antimicrobial activity and selected physico-chemical properties in some commonly consumed pomegranate sour sauces in Turkey. Materials and methods. Materials. Pomegranate sour sauce, (PSS) belonging to nine different commercial trademarks, was purchased from local markets in Turkey. PSS stored at 4 ◦ C until analysis. Preparation of the extracts. 5 g sample was mixed with 150 ml methanolwater or water. Then stirred for eighteen hours on a magnetic stirrer. The suspension was filtered through Whatman filter paper (No 1) and then concentrated in vacuo at 40 ◦ C using a Rotary evaporator. The residues obtained were stored in a freezer at −80 ◦ C until further tests. Determination of total phenolics and antioxidant activity in PSS. The concentration of total phenolics in the methanol-water (1:1, v/v) and water extracts of PSS was determined colourimetrically using Folin–Ciocalteau reagent as described by Gulcin et al. [6 ]. Gallic acid was used as the standard. The antioxidant activity in the methanol-water (1:1, v/v) and water extracts of PSS was determined according to the β-carotene bleaching method described by 146

H. Yildiz, M. Sengul, B. Cetin et al.

Kaur and Kapoor [7 ]. In the β-carotene bleaching assay, antioxidant capacity is determined by measuring the inhibition of the volatile organic compounds and the conjugated hydroperoxides arising from linoleic acid oxidation [7 ]. Chemical analyses. Total soluble solid (TSS), ash, pH and titratable acidity were determined according to standard AOAC [8 ]; pH was determined with an ATI ORION 420A model pH meter; titratable acidity, expressed as percentage of citric acid, was determined with 0.1 N NaOH up to pH 8.1; solubles dry matter was determined with an Abbe-Zeis refractometer. Total sugar, reducing sugar and sucrose contents were analysed by the Lane-Eynon method [9 ]. Reducing sugar concentration was measured before inversion, whereas total sugar was determined after inversion. Sucrose was calculated by subtracting the reducing sugar concentration from the total invert sugar and multiplying the result by 0.95. Colour of PSS was determined by measuring Hunter L (brightness; 100:white, 0:black), a (+: red; –: green) and b (+: yellow; –: blue) parameters with a colourimeter (Model CR 200, chromometer, minolta, Japan). Bacterial strains. Twenty five bacterial strains (target-strains) (Acinetobacter lwoffi BC 2819, Bacillus cereus BC 6830, Bacillus subtilis ATTC 6633, Enterobacter cloacea BC 3213, Enterobacter feacalis ATTC 29122, Escherichia coli BC 1402, Escherichia coli BC 1818, Escherichia coli BC 2326, Flavobacterium indologenes BC 1520, Klebsiella pneumoniae BC 1749, Klebsiella pneumoniae BC 3126, Listeria monocytogenes BC 8353, Proteus mirabilis BC 2644, Proteus vulgaris KUKEM 1329, Providencia alkalifaciens BC 0236, Pseudomonas aeruginosa ATCC 27859, Pseudomonas fluorescens BC 7324, Pseudomonas pseudoalkaligenes BC 3445, Pseudomonas putida BC 1617, Salmonella Typhimurium RSSK 95091, Staphylococcus aureus ATCC 29213, Staphlococcus hominis BC 2288, Streptococcus pyogenes ATCC 176, Streptococcus thermophilus BC 6453, Yersinia enterocolitica BC 0184) – pathogenic and nonpathogenic – were used in this study. The microorganisms maintained on Nutrient Agar (Merck, Darmstadt, Germany) were supplied by the Department of Food Engineering, Food Microbiology Laboratory, Faculty of Agriculture at Atat¨ urk University, Erzurum, Turkey. Identity of the bacteria used in this study was confirmed by Microbial Identification System in Biotechnology Application and Research Center at Atat¨ urk University, API and Biolog identification kits and conventional (morphological and biochemical) methods. The food-associated bacteria were selected because they are frequently reported in foods [10 ]. Disc-diffusion assay. The sour pomegranate extracts were dissolved in dimethylsulfoxide (DMSO) (Sigma) to a final concentration of 30 mg/mL and sterilized by filtration through 0.45 µm Millipore filters (Schleicher & Schuell, Microscience, Dassel, Germany). The antimicrobial activity of the extracts was carried out by disc diffusion test [11 ] using 100 µl of suspension containing 108 CFU/ml of bacteria spread on nutrient agar (NA) medium. Sterile 6 mm diameter filter paper discs were impregnated with 300 µg of the sterile test material and Compt. rend. Acad. bulg. Sci., 67, No 1, 2014

147

placed onto nutrient agar. SAM20 (10 µg sulbactam + 10 µg ampicillin/disc), OFX10 (10 µg ofloxacin/disc), AMC30 (20 µg amoxicillin + 10 µg clavulanic asit/disc), KF30 (30 µg cephalothin/disc), TE30 (30 µg tetracycline/disc), AZM15 (15 µg azithromycin/disc) were used as positive reference standards to determine the sensitivity of one strain/isolate in each microbial species tested. The inoculated plates were incubated for 24 h at 30 ◦ C for mesopfilic bacterial strains, 20 ◦ C for 48 h for psychrotrophic bacterial strains. Antimicrobial activity was evaluated by measuring the DD (Disk Diffusion) zone of inhibition against the test organisms. Statistical analysis. Analysis of variance was performed by ANOVA procedures (SPSS 9.0 for Windows). Significant differences between means were determined by Duncan’s Multiple Range tests. P values < 0.05 were regarded as significant. Results and discussion. Table 1 summarizes the physicochemical properties of PSS. There were statistical differences among PSS in terms of total soluble solids, total sugar, sucrose, reducing sugar, ash, titratable acidity, pH and colour (L, a, b) of concentrates (p < 0.01). Total soluble solids ranging from 69.5 for sample 4 to 73.3 for sample 8 (Table 1). As seen from Table 1, pH in PSS ranged from 2.64 to 2.91 and titratable acidity of PSS varied from 4.30 to 7.97 (% citric acid). Citric acid levels of seven different commercial pomegranate juices (PJs) were reported by Tezcan et al. [12 ] as between 0.393–1.306 g/100 mL. It was found that pomegranate juice concentrate (PC) possessed low levels of ash (0.05– 0.58 g/100 g). In this study total sugar, reducing sugar, sucrose of PSS was determined between 21.89–53.44 g/100 g, 20.98–50.62 g/100 g, 0.75–2.81 g/100 g, respectively. In our study, total ash content in PSS was found to be 0.05– 0.58 g/100 g. A previous study using pomegranate juice from Turkey reported titratable acidity between 8.3 and 17.4 g/l, sucrose 0–1.5 g/l, and soluble solids 14 ◦ Brix [13 ]. There are wide variations in colour of the PSS. The highest L value was in sample 2 (55.38), followed by sample 8 (39.25) and sample 3 (35.63). The lowest a value was recorded in sample 4 (3.95) and sample 9 (3.96). b value was determined between (–6.98)–(–38.36). The Folin–Ciocalteu assay was used for determination of total phenolic contents (TPC) in this study using gallic acid as a standard phenolic compound. The results were expressed as µg of gallic acid equivalents per milligram of sample (µg GAE/ mg of sample). TPC in PSS are given in Table 2. The PSS were found to have widely varying levels of phenolics, ranging from 37.65 to 383.95 and from 52.86–461.99 µg GAE/mg of sample for water and methanol-water extracts, respectively. Sample 6 is the highest in TPC of both methanol-water and water extracts at about 461.99 and 383.95 µg GAE /mg of sample, respectively (Table 2). Earlier, TPC in pomegranate juice (PJs) was reported 112.3 ± 6.4 mg/100 g [14 ]. TPC of seven commercial PJs are reported between 144 ± 80 and 10086 ± 85 mg GAE/L [12 ]. Gokoglu et al. [1 ] determined TPC contained 148


Compt. rend. Acad. bulg. Sci., 67, No 1, 2014

149

5.95c 55.38c 11.96b 36.51e

7.97e 17.28a 4.98a 7.15a

Titratable acidity, g/100 g

L

a

b

3

16.04b

27.17d

35.63b

5.98c

2.79c

34.38b

1.55d

32.83c

0.16bc

72.50f

7.49a

3.95a

16.58a

4.30a

2.82d

35.02c

1.04c

33.98d

0.09ab

69.50a

4

24.03c

21.36c

36.84b

6.94d

2.69b

34.35b

2.75g

31.60b

0.27d

72.30e

5

6.88a

4.44a

17.03a

5.11b

2.91e

21.89a

0.91b

20.98a

0.58f

70.00b

6

Pomegranate sour sauce samples 7

7.44a

4.16a

16.92a

5.26b

2.64a

34.93c

0.75a

34.18e

0.09ab

72.50f

Means within the same line followed by same letter are not statistically significant (P < 0.05)

2.75c

53.44g

2.81g

50.62i

0.22cd

2.77c

39.14f

2.59f

36.55h

2 71.70c

pH

Total reducing sugar, g/100 g

Sucrose, g/100 g

Reducing sugar, g/100 g

0.43e

Ash, g/100 g

1 72.00d

TSS, g/100 g

Parameters

1

Phycochemical properties of pomegranate sour sauce

Table

8

24.53c

36.58e

39.25b

5.35b

2.70b

37.47e

2.11e

35.36f

0.05a

73.30g

9

7.48a

3.96a

16.85a

7.69e

2.70b

36.97d

1.01bc

35.97g

0.24cd

72.00d

150

H. Yildiz, M. Sengul, B. Cetin et al. 82.28ef

87.74g

220.47f

187.21f

1

70.70b

81.48f

318.95g

221.57g

2

81.18def

78.51e

52.86a

46.56b

3

73.16bc

73.70c

76.34cd

55.69c

4

83.01f

81.10f

53.73ab

62.86cd

5

62.53a

34.43a

461.99h

383.95h

6

7

74.63bcd

78.51e

68.08bc

37.65a

Pomegranate sour sauce samples

Means within the same line followed by same letter are not statistically significant (P < 0.05)

(methanol+water), %

Antioxidant activity

(water), %

Antioxidant activity

(µg GAE/mg of sample)

(in metanol+water)

Total phenolic content

(µg GAE/mg of sample)

Total phenolic content (in water)

Parameters

2

Phenolic contents and antioxidant activity of pomegranate sour sauce

Table

76.10bcde

75.55d

86.56d

66.34d

8

77.94cdef

71.85b

125.03e

96.99e

9

phenolic substances 56mg GAE/100 g. Phenolics are aromatic secondary plant metabolites, and widely exist in the plant kingdom. They have been associated with colour and sensory qualities, nutritional and antioxidant properties of food. The phenolic compounds may contribute directly to antioxidative action. They may contribute to reducing human diseases such as cancer, arteriosclerosis, brain disorders and hearth diseases [15 ]. The total antioxidant activity of the PSS was determined in this work by measuring the ability of the extract to inhibit the bleaching of β-carotene; the result was then compared with that of the control that contained no antioxidant compound. β-carotene shows strong biological activity and is a physiologically important compound [7 ]. In the β-carotene bleaching assay, linoleic acid produces hydroperoxides as free radicals during incubation at 50 ◦ C. The presence of antioxidants in the extract will minimize the oxidation of β-carotene by hydroperoxides. Hydroperoxides formed in this system will be neutralized by the antioxidants from the extracts. Thus, the degradation rate of β-carotene depends on the antioxidant activity of the extracts. As shown on Table 1, there is large variation in the antioxidant activity of PSS investigated ranging from 34.40 to 87.74%, and from 62,53 to 83,01%, for those of water and methanol+water extractions, respectively. The lowest levels of antioxidant activity of water extracts were obtained from Sample 6, 34.43a%. The highest levels of antioxidant activity were obtained from the water extracts of Sample 1 (87.74%) and the methanol+water extracts of sample 5 (83.01%). The β-carotene assay indicated that PSS possessed considerable antioxidant activities. All samples showed lower antioxidant activity than BHA (91.7%). The active compounds present in them mainly contribute to the antioxidant activity of plants. These results suggest that the higher levels of antioxidant activity were due to the presence of phenolic components. Pomegranate fruit has valuable functional and medicinal effects such as confirmed antioxidant, anticancer and anti-atherosclerotic effects. Pomegranate juice is nutritionally an important beverage and is consumed for its phenolic compounds (such as anthocyanins, ellagic acid, phytoestrogenic flavonoids and tannins) [16 ]. Lu et al. [17 ] reported that commercial PJ had one of the highest antioxidant activities compared to other fruit juices, red wine and green tea. This can be attributed to its high content of polyphenols including ellagic acid in its free and bound forms, gallotannins and anthocyanins, and other flavonoids [17 ]. De Nigris et al. [18 ] reported that the antioxidant level in PJ was higher than that in other natural juices such as blueberry, cranberry, and orange, as well as in red wine. Twenty five microorganism strains were tested for their sensitivity to water and methanol-water extracts. Table 3 presents diameters of inhibition zones (clear zones around wells) exerted by the various extracts towards challenged microorganisms. Negative control was inactive against tested microorganisms. As shown in Table 3, methanol-water extracts of PSS were found more effective than water extracts against microorCompt. rend. Acad. bulg. Sci., 67, No 1, 2014

151

152


31(OFX10) 32(OFX10) 16(TE30) 12(TE30) 34(TE30) 24(KF30) 25(OFX10) 34(KF30) 26(AZM15)

22(OFX10) 12(AZM15) 26(AZM15) 27(AZM15) 30(OFX10) 25(OFX10) 22(SAM20) 28(OFX10) 18(AMC30) 33(OFX10)

Positive Controla 18(OFX10) 14(SAM20) 36(AMC30) 24(KF 30)

extracts

a

SAM20 (10 µg sulbactam + 10 µg ampicillin/disc), OFX10 (10 µg Ofloxacin/disc), AMC30 (20 µg amoxicillin + 10 µg clavulanic asit/disc), KF30 (30 µg cephalothin/disc), TE30 (30 µg tetracycline/disc), AZM15 (15 µg azithromycin/disc) were used as positive reference standards antibiotic discs, –, no inhibition zone

Inhibition zone in diameter (mm/sensitive strains) Sour pomegranate Sample 1 Sample 2 Sample 3 Aqueous MeOH Aqueous MeOH Aqueous MeOH Acinetobacter lwoffi BC 2819 – – – – – – Bacillus cereus BC 6830 – 8.30±0.20b – 8.03±0.25b – – Bacillus subtilis ATTC 6633 – – – – – – Enterobacter cloacea BC 3213 – – – – – – Enterococcus feacalis ATTC 29122 – – – – – – Escherichia coli BC 1402 – – – – – – Escherichia coli BC 1818 – – – – – – Escherichia coli BC 2326 – – – – – – – – – – – – Flavobacterium indologenes BC 1520 Klebsiella pneumoniae BC 1749 – – – – – – Klebsiella pneumoniae BC 3126 – – – – – – Listeria monocytogenes BC 8353 – – – – – – Proteus mirabilis BC 2644 – – – – – – ¨ Proteus vulgaris KUKEM 1329 7.27±0.51b 8.40±0.36b 7.27±0.25b 7.33±0.42c – 7.30±0.20b – – – – – – Providencia alkalifaciens BC 0236 Pseudomonas aeruginosa ATCC 27859 – – – – – – Pseudomonas fluorescens BC 7324 – – – – – – – – – Pseudomonas pseudoalkaligenes BC 3445 8.13±0.21a 18.33±0.51a – Pseudomonas putida BC 1617 – – – – – – – – – – – – Salmonella Typhimurium RSSK 95091 Staphylococcus aureus ATCC 29213 – 8.30±0.40b 10.13±0.21a 12.27±0.25a – 7.30±0.20b – – – – – – Staphlococcus hominis BC 2288 Streptococcus pyogenes ATCC 176 – 7.17±0.31c – 8.13±0.20b 7.27±0.15a 8.30±0.20a 7.23±0.23b 7.17±0.15c – 7.13±0.15d – 7.37±0.47b Streptococcus thermophilus BC 6453 Yersinia enterocolitica BC 0184 – – – – – –

Microorganisms

3

Antibacterial activity of pomegranate sour sauce

Table

Compt. rend. Acad. bulg. Sci., 67, No 1, 2014

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in diameter (mm/sensitive Sample 5 Aqueous MeOH – – – 8.32±0.56a – – – – – – – – – – – – – – – – – – – – – – 7.20±0.30a 7.10±0.70b – – – – – – – – – – – – – – – – 7.03±0.32b 7.17±0.31b 7.07±0.11ab 7.23±0.21b – –

3

strains) Sour pomegranate Sample 6 Aqueous MeOH – – – – – – – – – – – – – – – – – – – – – – – – – – 7.03±0.25a 8.13±0.55a – – – – – – – – – – – – – 7.07±0.51b – – – 7.17±0.15b – 7.30±0.26b – – 31(OFX10) 32(OFX10) 16(TE30) 12(TE30) 34(TE30) 24(KF30) 25(OFX10) 34(KF30) 26(AZM15)



extracts


a

Inhibition zone Sample 4 Aqueous MeOH Acinetobacter lwoffi BC 2819 – – Bacillus cereus BC 6830 – 7.07±0.25b Bacillus subtilis ATTC 6633 – – – – Enterobacter cloacea BC 3213 Enterobacter feacalis ATTC 29122 – – Escherichia coli BC 1402 – – Escherichia coli BC 1818 – – Escherichia coli BC 2326 – – – – Flavobacterium indologenes BC 1520 Klebsiella pneumoniae BC 1749 – – – – Klebsiella pneumoniae BC 3126 Listeria monocytogenes BC 8353 – – Proteus mirabilis BC 2644 – – ¨ Proteus vulgaris KUKEM 1329 – – Providencia alkalifaciens BC 0236 – – Pseudomonas aeruginosa ATCC 27859 – – Pseudomonas fluorescens BC 7324 – – Pseudomonas pseudoalkaligenes BC3445 – – Pseudomonas putida BC 1617 – – Salmonella Typhimurium RSSK 95091 – – – – Staphylococcus aureus ATCC 29213 Staphlococcus hominis BC 2288 – – Streptococcus pyogenes ATCC 176 – – 8.40±0.44a 8.20±0.17a Streptococcus thermophilus BC 6453 Yersinia enterocolitica BC 0184 – –

Microorganisms

(continue)

Table

154


in diameter (mm/sensitive Sample 5 Aqueous MeOH – – – – – – – – – – – – – – – – – – – – – – – – – – 8.17±0.35b 8.37±0.50c – – – – – – 14.27±0.3a 16.40±0.5a – – – – – – – – 7.10±0.87c 7.23±0.21d 7.10±0.17c 9.33±0.15b – –

3

strains) Sour pomegranate Sample 6 Aqueous MeOH – – – – – – – – – – – – – – – – – – – – – – – – – – 8.07±0.57a 8.27±0.31a – – – – – – 7.23±0.31b 8.20±0.10a – – – – – – – – 7.00±0.36c 7.37±0.25b 7.23±0.53b 7.17±0.38c – – 31(OFX10) 32(OFX10) 16(TE30) 12(TE30) 34(TE30) 24(KF30) 25(OFX10) 34(KF30) 26(AZM15)



extracts


a

Inhibition zone Sample 4 Aqueous MeOH Acinetobacter lwoffi BC 2819 – – Bacillus cereus BC 6830 – – Bacillus subtilis ATTC 6633 – – – – Enterobacter cloacea BC 3213 Enterobacter feacalis ATTC 29122 – – Escherichia coli BC 1402 – – Escherichia coli BC 1818 – – Escherichia coli BC 2326 – – – – Flavobacterium indologenes BC 1520 Klebsiella pneumoniae BC 1749 – – – – Klebsiella pneumoniae BC 3126 Listeria monocytogenes BC 8353 – – Proteus mirabilis BC 2644 – – ¨ Proteus vulgaris KUKEM 1329 7.50±1.00a 7.20±0.3bc Providencia alkalifaciens BC 0236 – – Pseudomonas aeruginosa ATCC 27859 – – Pseudomonas fluorescens BC 7324 – – Pseudomonas pseudoalkaligenes BC3445 – – Pseudomonas putida BC 1617 – – Salmonella Typhimurium RSSK 95091 – – – 7.33±0.15b Staphylococcus aureus ATCC 29213 Staphlococcus hominis BC 2288 – – Streptococcus pyogenes ATCC 176 – 7.07±0.25c – 8.20±0.26a Streptococcus thermophilus BC 6453 Yersinia enterocolitica BC 0184 – –

Microorganisms

(continue)

Table

ganisms. The samples showed antimicrobial activity against 6 within 25 tested bacteria (18 gram negative and 7 gram positive). Gram positive bacteria were more sensitive than negatives but Pseudomonas pseudoalkaligenes BC3445 had the highest DD zone. Staphylococcus aureus is one of the most common of the gram-positive bacteria causing food poisoning [19 ]. The symptoms of Staphylococcal food poisoning usually develop within 4 h of the ingestion of contaminated food, although a range of 1–6 h has been reported. The symptoms are nausea, vomiting, abdominal cramps (which are ucually guite severe), diarrhoea, sweating, headache, prostration and sometimes a fall in body temperature, generally lasting from 24 to 48 h [20 ]. In the study, four samples (1, 2, 3 and 7) had antibacterial activity against Staphylococcus aureus ATCC 29213. Sample 2 was more effective than the other samples against it (Table 3). Bacillus cereus is another important pathogenic foodborn bacteria. It is an aerobic, spore-forming rod, normally present in soil, dust and water. This bacterium produces a wide variety of extracellular toxins and ezymes which cause diarrhoeal and emetic syndromes. In the study, four samples (1, 2, 4 and 5) had antibacterial activity against Bacillus cereus. The effective samples displayed similar antibacterial activity [20 ]. Proteus vulgaris, Pseudomonas pseudoalkaligenes, Streptococcus pyogenes and Streptococcus thermophilus were other sensitive bacteria. But the samples had not antimicrobial effect against important foodborne pathogenic bacteria as Salmonella Typhimurium and Listeria monocytogenes strains (Table 3). Conclusion. The study indicated that PSS had high phenolic content, antioxidant and antimicrobial activity. These results, therefore, indicate that pomegranate sour sauce is a potent antioxidant food and might have potential value as a functional food ingredient.

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[7 ] Kaur C., H. C. Kapoor. International Journal of Food Science and Technol., 37, 2002, 153–161. [8 ] AOAC, “Official Methods of Analysis. Association of Official Chemists” 14th Edition, Association of Official Analytical Chemists, Arlington, 1984. ˙ sleme End¨ ˇ lu B. Meyve ve Sebze I¸ [9 ] Cemerog ustrisinde Temel Analiz Metodları. Biltav Yayınları, Ankara, T¨ urkiye, 1992. [10 ] Holt J. G., N. R. Krieg, P. H. A. Sneath, J. T. Staley, S. T. Williams. Bergey’s Manual of Determinative Bacteriology. Lippincott, 1994. [11 ] Kim J., M. R. Marshall, C. Wie. Journal of Agriculture and Food Chemistry, 43, 1995, 2839–2845. ¨ u ¨ ¸ elik, F. B. Erim. Food ¨ltekin-Ozg ¨ven, T. Diken, B. Ozc [12 ] Tezcan F., M. Gu Chemistry, 115, 2009, 873–877. [13 ] Eksi A., I. Ozhamamci. Gıda, 34, 2009, 265–270. [14 ] Guo C., J. Wei, J. Yang, J. Xu, W. Pang, Y. Jiang. Nutrition Research, 28, 2008, 72–77. [15 ] Cano A., M. B. Arnao. International Journal of Food Properties, 8, 2005, 521–528. [16 ] Mousavinejad G., Z. Emam-Djomeh, K. Rezaei, M. H. H. Khodaparast. Food Chemistry, 115, 2009, 1274–1278. [17 ] Lu J., Y. Wei, O. Yuan. Journal of Chromatography B, 857, 2007, 175–179. [18 ] De Nigris F., M. L. Balestrieri, S. Williams-Ignarro, F. P. D’Armiento, C. Fiorito, L. J. Ignarro, C. Napoli. Nitric Oxide, 17, 2007, 50–54. [19 ] Jay J. M. Modern food microbiology. An AVI Book, New York, 1992. [20 ] Jay J. M., M. J. Loessner, D. A. Golden. Modern Food Microbiology. Springer Science-Business Media, New York, 2005. ∗

Department of Food Engineering Faculty of Engineering and Natural Science Gumushane University Gumushane, Turkey ∗∗

Department of Horticulture Faculty of Agriculture Ataturk University Erzurum, Turkey

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Department of Food Engineering Faculty of Agriculture Ataturk University Erzurum, Turkey e-mail: [email protected]

Dzemal Bijedic University Agromediterranean Faculty 88104 Mostar, Bosnia and Herzegovina ∗∗∗
