Determination of sensitivity of some food pathogens to spice extracts

Gökhan Akarca; Oktay Tomar; İlknur Güney; Sena Erdur; Veli Gök

doi:10.1007/s13197-019-03994-1

. 2019 Sep 16;56(12):5253–5261. doi: 10.1007/s13197-019-03994-1

Determination of sensitivity of some food pathogens to spice extracts

Gökhan Akarca ^1,^✉, Oktay Tomar ², İlknur Güney ¹, Sena Erdur ¹, Veli Gök ³

PMCID: PMC6838422 PMID: 31749472

Abstract

Spices are primarily used as flavor enhancers and have attracted attention as natural food preservatives since their antimicrobial effects were determined. In the present study, the antimicrobial effects, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values on 5 important food-borne pathogenic bacteria were investigated in 20 different types of spices that are not commonly used. The results indicated that Hibiscus (Hibiscus sabdariffa) was the most effective against Listeria monocytogenes (26.37 mm zone diameter) and Staphylococcus aureus (24.15 mm zone diameter) (P < 0.05) followed by the chebulic myrobalan (Terminalia chebula) (21.34 ± 0.35 and 23.85 ± 1.69 mm diameter zone respectively) (P < 0.05). Likewise, Hibiscus (H. sabdariffa) showed the lowest MICs and MBCs concentration values on five important food-borne pathogens (L. monocytogenes) MIC; 0.187 mg/L, MBC; 0. Thus, this study determined that spices with antimicrobial activities can be used as natural preservatives.

Keywords: Antimicrobial activity, MICs, MBCs, Spices, Food-borne pathogen

Introduction

Pathogenic microorganisms found in foods can lead to food spoiling, as well as the emergence of new foodborne epidemics. In recent years there has been a severe increase in the rate of emergence of antibiotic-resistant bacteria, thus, causing an increase in the rates of morbidity and mortality of infections and food poisonings caused by these bacteria (Miladi et al. 2016). Every year in the world today, thousands of people are affected by pathogenic microorganisms and their food poisoning. Moreover, tons of food is rendered unconsumable due to the presence of saprophytic organisms. Therefore, manufacturers use a variety of chemical preservatives to prolong the shelf life of food and prevent the development of pathogens in food (De Souza et al. 2005). Most preservatives include synthetic chemical products. Moreover, these substances can neither prevent the development of all pathogens that may be present in food nor extend the shelf life of the product due to their inability to curb the growth of saprophytic microorganisms. They may also cause microbiological resistance to growth (Tajkarimi et al. 2010; Silva and Domingues 2017).

Recent studies have determined that many spices have significant antimicrobial effects, medicinal value, and antioxidant properties (Peter 2001; Kustes 2013). Consumers prefer foods, which have a longer shelf life and do not bear the risk of foodborne illness. Thus, consumers pressurize the producers to produce microbiologically reliable food using natural alternative preservatives and minimal chemical preservatives (Brul and Coote 1999).

In this study, 20 different types of spices (not well-known by consumers and producers) were extracted with ethyl alcohol. The antibacterial effects of these extracts on some important foodborne pathogens were then determined by disk diffusion method. Moreover, minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of spice extracts on these foodborne pathogens were identified.

Materials and methods

Preparation of spice extracts

The spices used in the research were obtained from the Afyonkarahisar market as packaged in bags of 50 g each. Each spice was then pulverized with the aid of a grinder mill. From the powdered spices, 100 g were weighed, and 400 mL of 80% ethyl alcohol was added. This mixture was stirred at 120 rpm using a shaker (Wise Shake^® SHO-2D) for 24 h, after which the mixture was filtered through a sterilized 22 mm filter paper. The filtered solution was transferred to a rotary evaporator (Heidolph Hei-VAP value) to separate the alcohol and the extracts at 100 rpm and 60 °C.

Bacteria

Escherichia coli ATCC 25922, Listeria monocytogenes ATCC 51774, Staphylococcus aureus ATCC 6538, Enterococcus aerogenes ATCC 13048 and Salmonella Typhimurium ATCC 14028 were used in the study.

Antibiotic discs

The present study utilized susceptibility test discs of the antibiotics Cefalexin, Lincomycin, Fusidic Acid, Triple Sulfonamides, Gentamicin, Tetracycline and Amoxicillin–Clavulanic Acid (Oxoid) antibiotic susceptibility test discs were used.

Preparation of extract disks

The obtained extracts were taken into 5 mL sterile Petri dishes. Empty antibiotic discs (oxoid) were placed on them. Petri dishes were placed in a refrigerator (4 °C) for 1 h to absorb the extracts of the discs.

Preparation of inoculums

Overnight cultures reproduced in a non-selective medium were taken from single colonies with the aid of a sterile loop. These colonies were suspended in a physiological saline solution to form homogeneous turbidity. The density of the obtained inoculum suspension was adjusted to be equal to 0.5 McFarland standard. Density control was checked with the McFarland turbidity standard (Bauer et al. 1959, 1966).

Placement of discs and incubation

The resulting inoculum was inoculated with Muller Hinton agar (Merck 1.05437, Germany) (MHA) at room temperature without any additives within 15 min. A sterile swab was immersed in the prepared suspension of bacteria and rotated several times within. Outside, the suspension was pressed against the inner wall of the tube to remove the excess fluid on the swab. Then, the inoculation was spread evenly across the MHA surface in three directions using a sterile cotton swab.

The inoculated medium was incubated for 10 min. Then, the discs impregnated with antibiotics and discs impregnated with plant extracts were placed on separate plaques at a distance from each other such that the zones were well-separated (Bauer et al. 1959, 1966). The Petri dishes were incubated at 35 °C for 18–20 h (Anonymous 2018). The zones formed at the end of the incubation period were measured with a digital caliper under sufficient light.

Determination of the minimum inhibitory concentrations (MICs)

Empty sterile test tubes were labeled A, 2, 3, 4, 5. The + and − control tubes were also maintained. Starting from tube A (except + and −), each tube was supplemented with 2 mL of nutrient broth (Merck 1.05443, Germany), and 4 mL of plant extract was added to tube A. Then, 2 mL of this extract was transferred to tube number 2. After mixing the contents of tube two thoroughly, 2 mL of the extract and the broth mixture were transferred to tube number 3. This process was repeated until tube 5. Finally, 2 mL of the mixture of extract and the broth mixture from tube number 5 was taken and discarded, thus, achieving equal concentrations in each tube, but half-reduced concentrations compared to the previous tube. Only 2 mL of extract was added to the “+ control” tube, and only 2 mL of nutrient broth was added to the “− control” tube.

Subsequently, cultures containing 10⁵ CFU/mL of bacteria were added to all tubes except 0.1 mL of “− control” tube. All tubes were incubated at 37 °C for 24 h. At the end of the incubation, the bacterial growth was controlled in the tubes. The tubes, which showed turbidity and membrane formation on the surface, were evaluated as those with positive development. Accordingly, the first tube in which (+) development was seen and the tube before that were observed. The MIC values were determined according to the following formula.

M I C = LC + HC / 2

LC: The lowest concentration of antimicrobial’s for microorganism inhibition, HC: The highest concentration of microorganism that can be developed.

Moreover, no growth was observed in the “+ control” tube, and there was growth in the “− control” tube (By Aamer et al. 2015; Chikezie 2017).

Determination of minimum bactericidal concentrations (MBCs)

A volume of 0.1 mL was taken from all tubes (except for the one used for MIC determination) using a sterile pipette. A 0.1 mL volume (with the aid of a sterile pipette) was inoculated on the specific medium on which the relevant bacterium could grow, using the spread plate method (By Aamer et al. 2015; Chikezie 2017). The plated cultures were incubated at the appropriate temperature, time and conditions (ISO 1999, 2001a, b, 2015, 2017a, b, c, d, e).

Statistical analyses

All microbiological analyses were performed in three replicates. Statistical analysis of the data was made by using the analysis of one-way ANOVA of SPSS program, version 23.0.0.0 (Anonymous 2015a). Mean values with a significant difference (P < 0.05) were compared with Duncan’s multiple range tests.

Results and discussion

The results obtained by the disc diffusion method to determine the antimicrobial effects of spices on some food pathogens are shown in Table 1 and details the zone diameters as a result of the effect of the antibiotics on the pathogens (Table 2). Eucast and CLSI (Table 3), provide the standards of resistance and susceptibility of some antibiotics on pathogens used in the present study.

Table 1.

Antimicrobial effect of spices on some food pathogens (mm zone diameter)

Spice	Escherichia coli	Staphylococcus aureus	Salmonella Typhimurium	Enterobacter aerogenes	Listeria monocytogenes
Hibiscus sabdariffa	23.43 ± 1.82^a	24.15 ± 1.26^a	23.85 ± 1.54^a	8.49 ± 1.22^b	26.37 ± 1.36^a
Punica granatum (flos)	20.02 ± 0.11^b	22.65 ± 0.20^a	18.03 ± 0.17^d	16.37 ± 0.27^e	19.24 ± 0.15^c
Terminalia chebula	13.49 ± 0.88^c	23.85 ± 1.69^a	20.50 ± 1.51^b	20.04 ± 0.58^b	21.34 ± 0.35^ab
Cinnamomum verum	12.54 ± 0.25^d	20.75 ± 0.09^a	13.28 ± 0.23^c	10.01 ± 0.25^e	17.43 ± 0.09^b
Artemisia dracunculus	15.63 ± 0.29^b	18.66 ± 1.15^a	14.84 ± 0.33^b	14.54 ± 0.25^b	10.68 ± 0.28^c
Helichrysum arenarium	14.32 ± 0.23^b	13.46 ± 0.27^c	14.04 ± 0.17^bc	16.40 ± 0.33^a	12.47 ± 0.16^d
Verbascum thapsus (flos)	13.36 ± 0.11^b	16.20 ± 0.17^a	8.24 ± 0.25^e	10.38 ± 0.20^d	12.53 ± 0.23^c
Ocimum basilicum	13.26 ± 0.12^a	7.68 ± 0.06^c	10.40 ± 0.17^b	10.38 ± 0.32^b	12.89 ± 0.11^a
Asphodelus albus	13.58 ± 0.23^c	15.14 ± 0.16^a	9.06 ± 0.24^e	14.32 ± 0.29^b	10.17 ± 0.42^d
Verbascum thapsus (flos)	13.36 ± 0.11^b	16.20 ± 0.17^a	8.24 ± 0.25^e	10.38 ± 0.20^d	12.53 ± 0.23^c
Ocimum basilicum	13.26 ± 0.12^a	7.68 ± 0.06^c	10.40 ± 0.17^b	10.38 ± 0.32^b	12.89 ± 0.11^a
Ocimum basilicum	13.26 ± 0.12^a	7.68 ± 0.06^c	10.40 ± 0.17^b	10.38 ± 0.32^b	12.89 ± 0.11^a
Hypericum perforatum	12.51 ± 0.55^c	14.35 ± 0.34^b	9.49 ± 0.31^d	10.32 ± 0.19^d	17.04 ± 0.35^a
Lamium Album	11.46 ± 0.40^e	19.43 ± 0.22^a	12.50 ± 0.20^d	13.55 ± 0.41^c	15.63 ± 0.24^b
Veronica chamaedrys	11.50 ± 0.37^a	7.33 ± 0.27^c	9.01 ± 0.25^b	7.49 ± 0.18^c	7.40 ± 0.32^c
Rosmarinus officinalis	9.72 ± 0.20^c	23.14 ± 0.39^a	11.01 ± 0.17^b	9.37 ± 0.28^c	8.10 ± 0.45^d
Matricaria chamomilla	8.30 ± 0.31^c	11.44 ± 0.33^b	13.42 ± 0.05^a	8.0 ± 0.30^c	13.62 ± 0.20^a
Olea europaea (Leaf)	8.01 ± 0.79^b	11.89 ± 1.07^a	8.52 ± 0.22^b	8.48 ± 0.33^b	7.35 ± 0.17^b
Glycyrrhiza glabra	7.99 ± 0.15^c	13.93 ± 0.40^a	8.10 ± 0.36^c	8.01 ± 0.22^c	11.47 ± 0.21^b
Capsella bursa-pastoris	7.41 ± 0.08^c	9.56 ± 0.06^b	7.39 ± 0.17^c	7.55 ± 0.37^c	10.31 ± 0.24^a
Sorghum bicolor	7.40 ± 031^bc	8.03 ± 0.17^ab	7.99 ± 0.21^ab	7.28 ± 0.23^c	8.20 ± 0.25^a
Laurus nobilis	7.36 ± 0.13^c	15.57 ± 1.61^a	10.25 ± 1.29^b	8.02 ± 0.52^bc	7.21 ± 0.06^c
Achillea millefolium	7.21 ± 0.03^d	14.74 ± 0.12^a	7.12 ± 0.09^d	8.24 ± 0.11^c	11.59 ± 0.18^b

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^a–eValues with the same capital letters in the same column for each analysis differ significantly (P < 0.05)

Table 2.

Antimicrobial effect of antibiotics on some food pathogens (mm zone diameter)

Antibiotics	Escherichia coli	Staphylococcus aureus	Salmonella Typhimurium	Enterobacter aerogenes	Listeria monocytogenes
Chloramphenicol	17.43 ± 0.27^b	18.48 ± 0.13^a	17.24 ± 0.08^b	17.21 ± 0.18^b	12.38 ± 0.10^c
Ampicilin	15.27 ± 0.16^c	NE	12.31 ± 0.25^d	16.45 ± 0.23^b	17.58 ± 0.33^a
Cepholexin	13.34 ± 0.11^e	23.00 ± 0.08^b	16.05 ± 0.14^c	27.05 ± 0.05^a	15.05 ± 0.18^d
Erythromycin	14.44 ± 0.25^c	25.02 ± 0.20^a	8.00 ± 0.13^e	16.31 ± 0.04^b	12.32 ± 0.34^d
Penicilin	16.32 ± 0.19^b	16.33 ± 0.08^b	15.13 ± 0.33^c	18.25 ± 0.16^a	14.67 ± 0.17^c
Streptomycin	13.29 ± 0.18^d	24.04 ± 0.28^a	13.20 ± 0.16^d	18.31 ± 0.08^b	14.10 ± 0.11^c
Lincomycin	14.32 ± 0.12^c	10.12 ± 0.04^d	10.03 ± 0.16^d	16.36 ± 0.08^b	18.30 ± 0.09^a
Amoxicillin–clavulanic acid	20.32 ± 0.24^b	NE	18.10 ± 0.06^c	27.01 ± 0.25^a	14.17 ± 0.06^d
Fusidic Acid	9.01 ± 0.11^e	25.50 ± 0.09^a	11.12 ± 0.06^d	17.26 ± 0.08^b	13.31 ± 0.24^c
Triple Sulfanamids S3 30	8.08 ± 0.06^d	32.37 ± 1.01^a	10.37 ± 1.02^c	8.01 ± 0.14^d	23.03 ± 1.20^b
Gentamicin GN10	24.03 ± 0.26^c	26.27 ± 0.88^b	22.28 ± 0.12^d	19.01 ± 0.38^e	28.06 ± 0.10^a
TetracydineTE 30	12.36 ± 0.43^c	27.14 ± 0.44^a	13.05 ± 0.40^c	12.08 ± 0.49^c	22.48 ± 0.19^b
Cefalexin	15.24 ± 0.60^a	7.36 ± 0.17^c	14.89 ± 0.32^a	14.01 ± 0.74^a	10.09 ± 0.46^b
Ceftizoxime	32.68 ± 0.26^b	23.08 ± 0.33^c	34.99 ± 0.28^a	35.55 ± 1.26^a	14.60 ± 0.69^d
Cefepime	35.37 ± 1.00^b	36.55 ± 0.45^b	36.42 ± 1.07^b	46.55 ± 0.62^a	27.01 ± 0.20^c
Kanamycin	16.18 ± 0.06^d	31.05 ± 1.99^a	24.11 ± 0.40^c	23.27 ± 0.22^c	27.02 ± 0.27^b

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NE not effective

^a–eValues with the same capital letters in the same column for each analysis differ significantly (P < 0.05)

Table 3.

CLSI and Eucast clinical microbiological zone diameter standards (mm) (Anonymous 2015b, 2018)

Antibiotics	Escherichia coli			Staphylococcus aureus			Salmonella Typhimurium			Enterobacter aerogenes			Listeria monocytogenes
Antibiotics	S	I	R	S	I	R	S	I	R	S	I	R	S	I	R
Ampicilin	≥ 17	14–16	≤ 13	NT	NT	NT	≥ 17	14–16	≤ 13	≥ 17	14–16	≤ 13	NT	NT	NT
Benzylpenicillin	≥ 14	–	≤ 14	≥ 26	–	≤ 26	≥ 14	–	≤ 14	≥ 14	–	≤ 14	≥ 13	–	≤ 13
Amoxicillin–clavulanic acid	≥ 18	14–17	≤ 13	NT	NT	NT	≥ 18	14–17	≤ 13	≥ 18	14–17	≤ 13	NT	NT	NT
Gentamicin GN10	≥ 15	13–14	≤ 12	≥ 15	13–14	≤ 12	≥ 15	13–14	≤ 12	≥ 15	13–14	≤ 12	NT	NT	NT
Penicilin	NT	NT	NT	≥ 29	–	≤ 28	NT	NT	NT	NT	NT	NT	NT	NT	NT
Netilmicin	≥ 15	13–14	≤ 12	≥ 18	–	≤ 18	≥ 15	13–14	≤ 12	≥ 15	13–14	≤ 12	NT	NT	NT
Erythromycin	NT	NT	NT	≥ 23	14–22	≤ 13	NT	NT	NT	NT	NT	NT	≥ 25	–	≤ 25
Streptomycin	≥ 15	12–14	≤ 11	NT	NT	NT	≥ 15	12–14	≤ 11	≥ 15	12–14	≤ 11	NT	NT	NT
Chloramphenicol	≥ 18	13–17	≤ 12	≥ 18	13–17	≤ 12	≥ 18	13–17	≤ 12	≥ 18	13–17	≤ 12	NT	NT	NT
Sulfonamides	≥ 17	13–16	≤ 12	≥ 17	13–16	≤ 12	≥ 17	13–16	≤ 12	≥ 17	13–16	≤ 12	NT	NT	NT
Kanamycin	≥ 18	14–17	≤ 13	≥ 18	14–17	≤ 12	≥ 18	14–17	≤ 13	≥ 18	14–17	≤ 13	NT	NT	NT
Tetracycline	NT	NT	NT	≥ 19	15–18	≤ 14	NT	NT	NT	NT	NT	NT	NT	NT	NT
Clindamycin	NT	NT	NT	≥ 21	15–20	≤ 14	NT	NT	NT	NT	NT	NT	NT	NT	NT
Fusidic Acid	NT	NT	NT	≥ 24	–	≤ 44	NT	NT	NT	NT	NT	NT	NT	NT	NT
Penicilin	NT	NT	NT	≥ 29	–	≤ 28	NT	NT	NT	NT	NT	NT	NT	NT	NT

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NT no test record

It was determined that Hibiscus sabdariffa and Punica granatum (flos) show highest antibacterial effect on Escherichia coli with a diameter zone of 22.93 ± 1.95 and 20.02 ± 0.11 mm (P < 0.05).

The values of ethanol extracts of spices used in our study as zone diameters of antimicrobial activities on Escherichia coli according to disc diffusion method were compared with the standard values (Table 2). Eucast and CLSI (Table 3) hibiscus (Hibiscus sabdariffa), and pomegranate (Punica granatum) (flos) were found to be more effective than all other antibiotics (Table 4; P < 0.05).

Table 4.

Comparison of the antimicrobial effect of spices on some food pathogens according to Eucast and CLSI standards

Spice	Escherichia coli	Staphylococcus aureus	Salmonella Typhimurium	Enterobacter aerogenes	Listeria monocytogenes
Hibiscus sabdariffa	S	S	S	S	S
Laurus nobilis	R	I	R	R	R
Terminalia chebula	R	S	S	S	I
Olea europaea (Leaf)	R	R	R	R	R
Veronica chamaedrys	R	R	R	R	R
Artemisia dracunculus	R	R	I	R	R
Rosmarinus officinalis	R	S	R	R	R
Achillea millefolium	R	R	I	R	R
Glycyrrhiza glabra	R	R	R	R	R
Sorghum bicolor	R	R	R	R	R
Punica granatum (flos)	I	R	R	R	R
Lamium Album	S	R	I	S	I
Asphodelus albus	I	I	R	I	R
Verbascum thapsus (flos)	I	I	R	R	R
Ocimum basilicum	R	R	R	R	R
Capsella bursa-pastoris	R	R	R	R	R
Hypericum perforatum	R	I	R	I	R
Helichrysum arenarium	I	I	R	I	R
Matricaria chamomilla	R	R	I	R	R
Cinnamomum verum	I	R	R	R	R

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S sensitive, R resistant, I intermediate

Cerit (2008) determined that the antimicrobial effects of 100% concentrations of bay and rosemary essential oils on Escherichia coli were 10.66 and 6.66 mm, respectively, which are similar to the results obtained in our study. Similarly, Sagdic et al. (2002) found that bay essential oils did not inhibit Escherichia coli O: 157 H: 7. Likewise, Borrás-Linares et al. (2015) reported that 25 different Hibiscus sabdariffa L. variants created 10–16 mm zone on Escherichia coli.

Salmonella Typhimurium growth was most significantly inhibited by hibiscus (Hibiscus sabdariffa), (23.85 ± 1.54 mm diameter zone) followed by chebulic myrobalan (Terminalia chebula) (20.50 ± 1.51 mm diameter zone) and tarragon (Artemisia dracunculus) (14.84 ± 0.33 mm diameter zone) (Table 1; P < 0.05). The extracts of these three spices were more effective than the other standard antibiotics (Table 2; P < 0.05).

Similarly, chebulic myrobalan (Terminalia chebula) was more effective than standard antibiotics on Enterobacter aerogenes activity (20.04 ± 0.58 mm zone diameter). The chebulic myrobalan effect was followed by asplenium ceterach (Helichrysum arenarium) (16.40 ± 0.33 mm zone diameter) and pomegranate (Punica granatum) (16.37 ± 0.27 mm diameter zone) (Table 1; P < 0.05)

Hibiscus (Hibiscus sabdariffa) (24.15 ± 1.26 mm zone diameter), the chebulic myrobalan (Terminalia chebula) (23.85 ± 1.69 mm zone diameter) and rosemary (Rosmarinus officinalis), (23.14 ± 0.39 mm diameter zone) were the most effective on Staphylococcus aureus growth (Table 1; P < 0.05) Kanamycin (31.05 ± 1.99 mm diameter zone) is the most effective among standard antibiotics on Staphylococcus aureus activity (Table 2; P < 0.05).

The antimicrobial effects of the bay and rosemary essential oils on Staphylococcus aureus, determined regarding zone diameter, were 12.96 and 13.66 mm, respectively (Cerit 2008).

Hibiscus (Hibiscus sabdariffa) showed the strongest antimicrobial activity (26.37 ± 1.36 mm zone diameter⁾ on Listeria monocytogenes, followed by the chebulic myrobalan (Terminalia chebula) (21.34 ± 0.35 36 mm zone diameter) (P < 0.05). Although there is not much data on the effect of standard antibiotics on Listeria monocytogenes, Gentamicin GN10 is determined as the most effective antibiotic eliciting a 28.06 ± 0.10 mm zone diameter (Table 2; P < 0.05).

Our results corroborated the study by Cerit (2008), which determined the antimicrobial effects (10.33 and 9.66 mm) of 100% concentrations of bay and rosemary essential oils on Listeria monocytogenes. Nakahara and Alzoreky (2003) demonstrated that the bay (Laurus nobilis) essential oil had bactericidal activity on Listeria monocytogenes.

Reason for strong antibacterial properties of spices can be attributed to the presence of rich phytochemical metabolites. The presence and quantities of alkaloids, phenolic compounds, flavonoids, cyanidins and saponins found in spices were demonstrated with previously performed studies. These compounds found in plants are generally thought to be the main groups showing antibacterial effect on pathogenic bacteria. These compounds are generally produced by plants as a defense mode against microbial infections (Fernández-Arroyo et al. 2012; Edwards-Jones 2013; Abdallah 2016; Akarca 2019).

Table 5 shows the MICs and MBCs values of the spices in our study and Table 6 shows the MIC values of the different antibiotics on the pathogens using Eucast.

Table 5.

Effect of MICs and MBCs on spices, some food pathogens (mg/L)

Spice	Esherichia coli		Staphylococcus aureus		Salmonella Typhimurium		Enterobacter aerogenes		Listeria monocytogenes
Spice	MIC	MBC	MIC	MBC	MIC	MBC	MIC	MBC	MIC	MBC
Hibiscus sabdariffa	0.046	0.015	0.046	0.015	0.093	0.015	0.015	0.015	0.187	0.015
Laurus nobilis	> 2	–	> 2	–	> 2	–	> 2	–	> 2	–
Terminalia chebula	0.375	0.015	0.375	0.015	0.75	0.25	0.375	0.015	0.187	0.25
Olea europaea (flos)	> 2	–	> 2	–	> 2	–	> 2	–	> 2	–
Veronica chamaedrys	> 2	–	> 2	–	> 2	–	> 2	–	> 2	–
Artemisia dracunculus	0.75	0.062	0.75	0.031	0.375	0.062	0.187	0.031	0.375	0.062
Rosmarinus officinalis	> 2	–	> 2	–	> 2	–	> 2	–	> 2	–
Achillea millefolium	> 2	–	> 2	–	> 2	–	> 2	–	> 2	–
Glycyrrhiza glabra	> 2	–	> 2	–	> 2	–	> 2	–	> 2	–
Sorghum bicolor	0.75	0.5	> 2	–	> 2	–	> 2	–	> 2	–
Punica granatum (flos)	> 2	–	> 2	–	> 2	–	> 2	–	0.75	0.062
Lamium album	0.75	0.5	0.375	0.25	0.75	0.5	0.375	0.25	0.375	–
Asphodelus albus	0.375	0.031	0.375	0.125	0.375	0.031	0.75	0.5	1.5	0.1
Verbascum thapsus	1.5	0.25	0.75	0.062	0.75	0.5	0.375	0.125	0.750	0.25
Ocimum basilicum	> 2	–	> 2	–	> 2	–	1.5	–	> 2	–
Capsella bursa-pastoris	> 2	–	> 2	–	> 2	–	1.5	–	> 2	–
Hypericum perforatum	0.75	0.031	0.75	0.015	0.375	0.125	0.375	0.031	0.75	0.062
Helichrysum arenarium	0.375	0.25	0.375	0.25	0.375	0.25	0.75	0.5	0.375	0.062
Matricaria chamomilla	0.75	0.5	0.75	0.5	0.75	0.5	0.75	0.5	0.75	0.5
Cinnamomum verum	0.75	0.031	0.75	0.031	0.75	0.031	0.75	0.031	1.5	1

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MBCs value not detected

Table 6.

Eucast clinical MIC breakpoint tables (mg/L) (Anonymous 2018)

Antibiotics	Escherichia coli		Staphylococcus aureus		Salmonella Typhimurium		Enterobacter aerogenes		Listeria monocytogenes
Antibiotics	S≤	R>	S≤	R>	S≤	R>	S≤	R>	S≤	R>
Benzylpenicillin	NT	NT	0.125	0.125	NT	NT	NT	NT	1	1
Ampicilin	8	8	NT	NT	8	8	8	8	1	1
Gentamicin	2	4	1	1	2	4	2	4	NT	NT
Netilmicin,	2	4	1	1	2	4	2	4	NT	NT
Erythromycin	NT	NT	1	2	NT	NT	NT	NT	1	1
Clindamycin	NT	NT	0.25	0.5	NT	NT	NT	NT	NT	NT
Fusidic Acid	NT	NT	1	1	NT	NT	NT	NT	NT	NT
Chloramphenicol	8	8	8	8	8	8	8	8	NT	NT
Tetracycline	NT	NT	1	2	NT	NT	NT	NT	NT	NT
Cefalexin	16	16	NT	NT	16	16	16	16	NT	NT
Amoxicillin–clavulanic acid	8	8	NT	NT	8	8	8	8	NT	NT

Open in a new tab

NT no test record

The main objective of the MIC test is to control the absorbance of samples containing microbial activity, inoculations, antimicrobial agents, and disinfectants. The MIC test describes the efficacy of a given antimicrobial agent.

The MICs values of the ethanol extracts of spices used in our study on Escherichia coli (Table 5) were determined. Compared to the standard values determined by Eucast (Table 6), the Hibiscus (Hibiscus sabdariffa) MICs value was much lower compared with the other antibiotics. Hibiscus MIC values were followed by chebulic myrobalan (Terminalia chebula), asphodel (Asphodelus albus) and the ceterach officinarum (Helichrysum arenarium).

The lowest MICs for Staphylococcus aureus was demonstrated by hibiscus (Hibiscus sabdariffa) (0.046 mg/L) followed by chebulic myrobalan (Terminalia chebula), Henbit (Lamium Album), Asphodel (Asphodelus albus), and ceterach officinarum (Helichrysum arenarium) (0.375 mg/L). For this pathogen, the lowest MICs from standard antibiotics were determined to be 1 mg/L.

The lowest MIC value on Salmonella Typhimurium was shown by hibiscus, tarragon, Asphodel, tutsan, and Ceterach officinarum, respectively. Similarly, hibiscus was effective at concentrations much lower than the MICs values on Enterobacter aerogenes of the standard antibiotics determined by Eucast. Hibiscus was followed by tarragon with a value of 0.187 mg/L.

The lowest MIC values of 0.187 mg/L (for hibiscus and ceterach officinarum) were obtained for Listeria monocytogenes. Our study showed only the lowest concentration of MICs for the three antibiotics (Benzylpenicillin, Ampicillin, and Erythromycin) on this bacterium (1 mg/L) (Table 6).

The minimum bactericidal concentration (MBC) is the lowest concentration of an antibacterial agent required to kill a particular bacterium (Amyes et al. 1996). It can be determined from broth dilution minimum inhibitory concentration (MIC) tests by subculturing on agar plates that do not contain the test agent. The MBC is identified by determining the lowest concentration of an antibacterial agent that reduces the viability of the initial bacterial inoculum by ≥ 99.9% (French 2006).

The MBC of the ethanolic extract of Hibiscus and ceterach officinarum on E. coli was found to be 0.015 mg/L. MBCs effect at the lowest concentration on Staphylococcus aureus; was similarly demonstrated by hibiscus and ceterach officinarum with 0.015 mg/L. tarragon and cinnamon watched these two plants with a value of 0.031 mg/L.

The MBC of Hibiscus was 0.015 mg/L on Salmonella Typhimurium, and that of Cinnamon and tutsan was 0.31 mg/L.

Hibiscus was tested on all other pathogens used in this study and showed an MBC effect at a low concentration of 0.015 mg/L on Listeria monocytogenes followed by tarragon and tutsan at 0.062 mg/L (Table 5).

Kim et al. (1995) demonstrated the antimicrobial activity of 11 different plant essential oils on five different food pathogens and determined that carvacrol was the most effective on Salmonella Typhimurium and Vibrio vulnificus. They found the MBCs value of carvacrol on these bacteria to be 250 μg/mL.

Hammer et al. (1999) investigated the antimicrobial activities of 52 plant extracts on ten different microorganisms and found that 20 plant extracts had antimicrobial activity, particularly on Escherichia coli and Staphylococcus aureus. Among these, thymus essential oil was the most effective with the lowest MICs of 0.03% (v/v) for Escherichia coli and 0.88% (v/v) for Staphylococcus aureus.

Bonyadian and Moshtaghi (2008) reported that Tarragon had MICs of 15.00 and 4.00 on Salmonella Typhimurium and Listeria monocytogenes respectively, whereas the MBCs were 17.00 and 5.00, respectively. Similarly, Raeisi et al. (2012) found that the MICs values of Tarragon extract on Escherichia coli and Staphylococcus aureus were 2500 and 1250 μg/mL, while the MBC values were 5000 and 2500 μg/mL, respectively.

Although many of the spices used in this research have high antibacterial properties, microbiological analysis shows that generally spices have a significantly high microbial load. Furthermore, the rate of contamination caused by process errors along the production chain is also very high. For this reason, in every stage of the production of spices which have a wide usage area in the food industry; Good production practices (GMPs) and hazard analysis and good implementation of the critical control point (HACCP) system will avoid potential difficulties (Cusato et al. 2014; Gomez et al. 2014).

Conclusion

Turkey holds a unique position in the World regarding plant diversity. A significant part of Turkey’s flora consists of aromatic plants that have been consumed for years by the Turkish people as spices or in alternative medicine. The possibility of using spices as an alternative to food preservation has gained momentum in recent years primarily due to increase in antibiotic resistance and the high cost and side-effects of antibiotics. Studies have revealed the antimicrobial activities of many spices used by the Turkish people for many years; however, these studies are not well-known internationally.

As a result of these studies, spices determined to have antimicrobial activities will be used as natural preservatives in addition to their use as antimicrobials. This will ease the anxiety associated with the use of artificial preservatives and enhance the nutrient values of foods.

Footnotes

Publisher's Note

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References

Abdallah EM. Antibacterial efficiency of the Sudanese roselle (Hibiscus sabdariffa L.), a famous beverage from Sudanese folk medicine. J Intercult Ethnopharmacol. 2016;5(2):186–190. doi: 10.5455/jice.20160320022623. [DOI] [PMC free article] [PubMed] [Google Scholar]
Akarca G. Composition and antibacterial effect on food borne pathogens of Hibiscus surattensis L. calyces essential oil. Ind Crops Prod. 2019;137:285–289. doi: 10.1016/j.indcrop.2019.05.043. [DOI] [Google Scholar]
Amyes S, Miles SR, Thomson CJ, Tillotson G. Antimicrobial chemotherapy: pocketbook. London: CRC Press; 1996. [Google Scholar]
Anonymous (2015a) SPSS Version 23.0.0.0 for Windows SPSS Inc. Shicago IL,USA
Anonymous (2015b) CLSI (Clinical and Laboratory Standards Institute), Zone diameter and minimal inhibitory concentration (MIC) Standards
Anonymous (2018) Eucast, European Committee on antimicrobial susceptibility testing. http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_8.0_Breakpoint_Tables.pdf. Accessed 28 Apr 2018
Bauer AW, Perry DM, Kirby WMM. Single disc antibiotic sensitivity testing of Staphylococci. AMA Arch Intern Med. 1959;104:208–216. doi: 10.1001/archinte.1959.00270080034004. [DOI] [PubMed] [Google Scholar]
Bauer AW, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966;36:493–496. doi: 10.1093/ajcp/45.4_ts.493. [DOI] [PubMed] [Google Scholar]
Bonyadian M, Moshtaghi H. Bactericidal activity of some plants essential oils against Bacillus cereus, Salmonella typhimurium, Listeria monocytogenes and Yersinia enterocolitica. Res J Microbiol. 2008;3(11):648–653. doi: 10.1016/j.indcrop.2012.02.042. [DOI] [Google Scholar]
Borrás-Linares I, Fernández-Arroyoa S, Arráez-Romana D, Palmeros-Suárezc PA, Del Val-Díazc R, Andrade-Gonzálesc I, Fernández-Gutiérreza A, Gómez-Leyvac JF, Segura-Carreteroa A. Characterization of phenolic compounds, anthocyanidin, antioxidant and antimicrobial activity of 25 varieties of Mexican Roselle (Hibiscus sabdariffa) Ind Crop Prod. 2015;69:385–394. doi: 10.1016/j.indcrop.2015.02.053. [DOI] [Google Scholar]
Brul S, Coote P. Preservative agents in foods. Mode of action and microbial resistance mechanisms. Int J Food Microbiol. 1999;50:1–17. doi: 10.1016/S0168-1605(99)00072-0. [DOI] [PubMed] [Google Scholar]
By Aamer AA, Abdul-Hafeez MM, Sayed SM. Minimum inhibitory and bactericidal concentrations (MIC and MBC) of honey and bee propolis against multidrug resistant (MDR) Staphylococcus Spp. isolated from bovine clinical mastitis. Glob J Sci Front Res D Agric Vet. 2015;15(2):1–9. [Google Scholar]
Cerit LS (2008) Antimicrobial properties of some spice volatile oils. Msc thesis. Pamukkale university graduate school of natural and applied science. Denizli, Turkey
Chikezie IO. Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using a novel dilution tube method. Afr J Microbiol Res. 2017;11(23):977–980. doi: 10.5897/AJMR2017.8545. [DOI] [Google Scholar]
Cusato S, Gamerioz AH, Sant’Ana AS, Corrassin CH, Cruz AG, de Oliveira CAF. Assessing the costs involved in the implementation of GMP and HACCP in a small dairy factory. Qual Assur Saf Crop. 2014;6(2):135–139. doi: 10.3920/QAS2012.0195. [DOI] [Google Scholar]
De Souza EL, Stamford T, Lima ED, Trajano VN, Barbosa JM. Antimicrobial effectiveness of spices: an approach for use in food conservation systems. Braz Arch Biol Technol. 2005;48:549–558. doi: 10.1590/S1516-89132005000500007. [DOI] [Google Scholar]
Edwards-Jones V. Alternative antimicrobial approaches to fighting multidrug-resistant ınfections. In: Rai M, Kon K, editors. Fighting multidrug resistance with herbal extracts, essential oils and their components. Amsterdam: Elsevier; 2013. pp. 1–9. [Google Scholar]
Fernández-Arroyo S, Herranz-López M, Beltrán-Debón R, Borrás-Linares I, Barrajón-Catalán E, Joven J, Fernández-Gutiérrez A, Segura-Carretero A, Micol V. Bioavailability study of a polyphenol-enriched extract from Hibiscus sabdariffa in rats and associated antioxidant status. Mol Nutr Food Res. 2012;56:1590–1595. doi: 10.1002/mnfr.201200091. [DOI] [PubMed] [Google Scholar]
French GL. Bactericidal agents in the treatment of MRSA infections the potential role of daptomycin. J Antimicrob Chemother. 2006;58(6):1107–1117. doi: 10.1093/jac/dkl393. [DOI] [PubMed] [Google Scholar]
Gomez CCB, Lemos GFC, Silva MC, Hora IMC, Cruz AG. Training of food handlers in a hotel: tool for promotion of the food safety. J Food Saf. 2014;34:218–223. doi: 10.1111/jfs.12116. [DOI] [Google Scholar]
Hammer KA, Carson CF, Riley TV. Antimicrobial activity of essential oils and other plant extracts. J Appl Microbiol. 1999;86:985–990. doi: 10.1046/j.1365-2672.1999.00780.x. [DOI] [PubMed] [Google Scholar]
ISO (1999) International Standard Organization, 6888-1, Horizontal method for the enumeration of coagulase-positive Staphylococci technique using baird parker agar medium
ISO (2001a) International Standard Organization, 16649-1:2001, Microbiology of food and animal feeding stuffs—horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 1: colony-count technique at 44 degrees C using membranes and 5-bromo-4-chloro-3-indolyl beta-d-glucuronide
ISO (2001b) International Standard Organization, 16649-2:2001, Microbiology of food and animal feeding stuffs—horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 2: colony-count technique at 44 degrees C using 5-bromo-4-chloro-3-indolyl beta-d-glucuronide
ISO (2015) International Standard Organization, 16649-3:2015, Microbiology of the food chain—horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 3: Detection and most probable number technique using 5-bromo-4-chloro-3-indolyl-β-d-glucuronide
ISO (2017a) International Standard Organization, 6579-1:2017, Horizontal method for the detection, enumeration and serotyping of Salmonella
ISO (2017b) International Standard Organization, 21528-1:2017, Microbiology of the food chain—horizontal method for the detection and enumeration of Enterobacteriaceae—Part 1: detection of Enterobacteriaceae
ISO (2017c) International Standard Organization, 21528-2:2017, Microbiology of the food chain—horizontal method for the detection and enumeration of Enterobacteriaceae—part 2: colony-count technique
ISO (2017d) International Standard Organization, 11290-1:2017, Microbiology of the food chain, horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp.—Part 1: detection method
ISO (2017e) International Standard Organization, 11290-2:2017, Microbiology of the food chain, horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp.—Part 2: enumeration method
Kim J, Marshall MR, Wei CI. Antibacterial activity of some essential oil components against five foodborne pathogens. J Agric Food Chem. 1995;43(11):2839–2845. doi: 10.1021/jf00059a013. [DOI] [Google Scholar]
Kustes S (2013) The spice of life. Realfood University Cook Book, USA
Miladi H, Zmantar T, Chaabouni Y, Fedhila K, Bakhrouf A, Mandouani K, Chaieb K. Antibacterial and efflux pump inhibitors of thymol and carvacrol against food-borne pathogens. Microb Pathog. 2016;99:95–100. doi: 10.1016/j.micpath.2016.08.008. [DOI] [PubMed] [Google Scholar]
Nakahara K, Alzoreky NS. Antibacterial activity of extracts from some edible plants commonly consumed in Asia. Food Microbiol. 2003;80:223–230. doi: 10.1016/S0168-1605(02)00169-1. [DOI] [PubMed] [Google Scholar]
Peter KV. Introduction. In: Peter KV, editor. Handbook of herbs and spices. Washington: CRC Press; 2001. pp. 1–6. [Google Scholar]
Raeisi M, Tajik H, Razavi Roohani SM, Maham M, Moradi M, Hajimohammadi B, Naghili H, Hashemi M, Mehdizadeh T. Essential oil of tarragon (Artemisis dracunculus) antibacterial activity on Staphylococcus aureus and Escherichia coli in culture media and Iranian white cheese. Iran J Microbiol. 2012;4(1):3–34. [PMC free article] [PubMed] [Google Scholar]
Sagdic O, Kuşçu A, Özcan M, Özçelik S. Effects of Turkish spice extracts at various concentrations on the growth E. coli O157:H7. Food Microbiol. 2002;1:473–480. doi: 10.1006/fmic.2002.0494. [DOI] [Google Scholar]
Silva F, Domingues FC. Antimicrobial activity of coriander oil and its effectiveness as food preservative. Crit Rev Food Sci Nutr. 2017;57:35–47. doi: 10.1080/10408398.2013.847818. [DOI] [PubMed] [Google Scholar]
Tajkarimi MM, Ibrahim SA, Cliver DO. Antimicrobial herb and spice compounds in food. Food Control. 2010;21:1199–1218. doi: 10.1016/j.foodcont.2010.02.003. [DOI] [Google Scholar]

[CR1] Abdallah EM. Antibacterial efficiency of the Sudanese roselle (Hibiscus sabdariffa L.), a famous beverage from Sudanese folk medicine. J Intercult Ethnopharmacol. 2016;5(2):186–190. doi: 10.5455/jice.20160320022623. [DOI] [PMC free article] [PubMed] [Google Scholar]

[CR2] Akarca G. Composition and antibacterial effect on food borne pathogens of Hibiscus surattensis L. calyces essential oil. Ind Crops Prod. 2019;137:285–289. doi: 10.1016/j.indcrop.2019.05.043. [DOI] [Google Scholar]

[CR3] Amyes S, Miles SR, Thomson CJ, Tillotson G. Antimicrobial chemotherapy: pocketbook. London: CRC Press; 1996. [Google Scholar]

[CR4] Anonymous (2015a) SPSS Version 23.0.0.0 for Windows SPSS Inc. Shicago IL,USA

[CR5] Anonymous (2015b) CLSI (Clinical and Laboratory Standards Institute), Zone diameter and minimal inhibitory concentration (MIC) Standards

[CR6] Anonymous (2018) Eucast, European Committee on antimicrobial susceptibility testing. http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v_8.0_Breakpoint_Tables.pdf. Accessed 28 Apr 2018

[CR7] Bauer AW, Perry DM, Kirby WMM. Single disc antibiotic sensitivity testing of Staphylococci. AMA Arch Intern Med. 1959;104:208–216. doi: 10.1001/archinte.1959.00270080034004. [DOI] [PubMed] [Google Scholar]

[CR8] Bauer AW, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966;36:493–496. doi: 10.1093/ajcp/45.4_ts.493. [DOI] [PubMed] [Google Scholar]

[CR9] Bonyadian M, Moshtaghi H. Bactericidal activity of some plants essential oils against Bacillus cereus, Salmonella typhimurium, Listeria monocytogenes and Yersinia enterocolitica. Res J Microbiol. 2008;3(11):648–653. doi: 10.1016/j.indcrop.2012.02.042. [DOI] [Google Scholar]

[CR10] Borrás-Linares I, Fernández-Arroyoa S, Arráez-Romana D, Palmeros-Suárezc PA, Del Val-Díazc R, Andrade-Gonzálesc I, Fernández-Gutiérreza A, Gómez-Leyvac JF, Segura-Carreteroa A. Characterization of phenolic compounds, anthocyanidin, antioxidant and antimicrobial activity of 25 varieties of Mexican Roselle (Hibiscus sabdariffa) Ind Crop Prod. 2015;69:385–394. doi: 10.1016/j.indcrop.2015.02.053. [DOI] [Google Scholar]

[CR11] Brul S, Coote P. Preservative agents in foods. Mode of action and microbial resistance mechanisms. Int J Food Microbiol. 1999;50:1–17. doi: 10.1016/S0168-1605(99)00072-0. [DOI] [PubMed] [Google Scholar]

[CR12] By Aamer AA, Abdul-Hafeez MM, Sayed SM. Minimum inhibitory and bactericidal concentrations (MIC and MBC) of honey and bee propolis against multidrug resistant (MDR) Staphylococcus Spp. isolated from bovine clinical mastitis. Glob J Sci Front Res D Agric Vet. 2015;15(2):1–9. [Google Scholar]

[CR14] Cerit LS (2008) Antimicrobial properties of some spice volatile oils. Msc thesis. Pamukkale university graduate school of natural and applied science. Denizli, Turkey

[CR15] Chikezie IO. Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using a novel dilution tube method. Afr J Microbiol Res. 2017;11(23):977–980. doi: 10.5897/AJMR2017.8545. [DOI] [Google Scholar]

[CR16] Cusato S, Gamerioz AH, Sant’Ana AS, Corrassin CH, Cruz AG, de Oliveira CAF. Assessing the costs involved in the implementation of GMP and HACCP in a small dairy factory. Qual Assur Saf Crop. 2014;6(2):135–139. doi: 10.3920/QAS2012.0195. [DOI] [Google Scholar]

[CR17] De Souza EL, Stamford T, Lima ED, Trajano VN, Barbosa JM. Antimicrobial effectiveness of spices: an approach for use in food conservation systems. Braz Arch Biol Technol. 2005;48:549–558. doi: 10.1590/S1516-89132005000500007. [DOI] [Google Scholar]

[CR18] Edwards-Jones V. Alternative antimicrobial approaches to fighting multidrug-resistant ınfections. In: Rai M, Kon K, editors. Fighting multidrug resistance with herbal extracts, essential oils and their components. Amsterdam: Elsevier; 2013. pp. 1–9. [Google Scholar]

[CR19] Fernández-Arroyo S, Herranz-López M, Beltrán-Debón R, Borrás-Linares I, Barrajón-Catalán E, Joven J, Fernández-Gutiérrez A, Segura-Carretero A, Micol V. Bioavailability study of a polyphenol-enriched extract from Hibiscus sabdariffa in rats and associated antioxidant status. Mol Nutr Food Res. 2012;56:1590–1595. doi: 10.1002/mnfr.201200091. [DOI] [PubMed] [Google Scholar]

[CR20] French GL. Bactericidal agents in the treatment of MRSA infections the potential role of daptomycin. J Antimicrob Chemother. 2006;58(6):1107–1117. doi: 10.1093/jac/dkl393. [DOI] [PubMed] [Google Scholar]

[CR21] Gomez CCB, Lemos GFC, Silva MC, Hora IMC, Cruz AG. Training of food handlers in a hotel: tool for promotion of the food safety. J Food Saf. 2014;34:218–223. doi: 10.1111/jfs.12116. [DOI] [Google Scholar]

[CR22] Hammer KA, Carson CF, Riley TV. Antimicrobial activity of essential oils and other plant extracts. J Appl Microbiol. 1999;86:985–990. doi: 10.1046/j.1365-2672.1999.00780.x. [DOI] [PubMed] [Google Scholar]

[CR23] ISO (1999) International Standard Organization, 6888-1, Horizontal method for the enumeration of coagulase-positive Staphylococci technique using baird parker agar medium

[CR24] ISO (2001a) International Standard Organization, 16649-1:2001, Microbiology of food and animal feeding stuffs—horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 1: colony-count technique at 44 degrees C using membranes and 5-bromo-4-chloro-3-indolyl beta-d-glucuronide

[CR25] ISO (2001b) International Standard Organization, 16649-2:2001, Microbiology of food and animal feeding stuffs—horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 2: colony-count technique at 44 degrees C using 5-bromo-4-chloro-3-indolyl beta-d-glucuronide

[CR26] ISO (2015) International Standard Organization, 16649-3:2015, Microbiology of the food chain—horizontal method for the enumeration of beta-glucuronidase-positive Escherichia coli—Part 3: Detection and most probable number technique using 5-bromo-4-chloro-3-indolyl-β-d-glucuronide

[CR27] ISO (2017a) International Standard Organization, 6579-1:2017, Horizontal method for the detection, enumeration and serotyping of Salmonella

[CR28] ISO (2017b) International Standard Organization, 21528-1:2017, Microbiology of the food chain—horizontal method for the detection and enumeration of Enterobacteriaceae—Part 1: detection of Enterobacteriaceae

[CR29] ISO (2017c) International Standard Organization, 21528-2:2017, Microbiology of the food chain—horizontal method for the detection and enumeration of Enterobacteriaceae—part 2: colony-count technique

[CR30] ISO (2017d) International Standard Organization, 11290-1:2017, Microbiology of the food chain, horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp.—Part 1: detection method

[CR31] ISO (2017e) International Standard Organization, 11290-2:2017, Microbiology of the food chain, horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp.—Part 2: enumeration method

[CR32] Kim J, Marshall MR, Wei CI. Antibacterial activity of some essential oil components against five foodborne pathogens. J Agric Food Chem. 1995;43(11):2839–2845. doi: 10.1021/jf00059a013. [DOI] [Google Scholar]

[CR33] Kustes S (2013) The spice of life. Realfood University Cook Book, USA

[CR34] Miladi H, Zmantar T, Chaabouni Y, Fedhila K, Bakhrouf A, Mandouani K, Chaieb K. Antibacterial and efflux pump inhibitors of thymol and carvacrol against food-borne pathogens. Microb Pathog. 2016;99:95–100. doi: 10.1016/j.micpath.2016.08.008. [DOI] [PubMed] [Google Scholar]

[CR35] Nakahara K, Alzoreky NS. Antibacterial activity of extracts from some edible plants commonly consumed in Asia. Food Microbiol. 2003;80:223–230. doi: 10.1016/S0168-1605(02)00169-1. [DOI] [PubMed] [Google Scholar]

[CR36] Peter KV. Introduction. In: Peter KV, editor. Handbook of herbs and spices. Washington: CRC Press; 2001. pp. 1–6. [Google Scholar]

[CR37] Raeisi M, Tajik H, Razavi Roohani SM, Maham M, Moradi M, Hajimohammadi B, Naghili H, Hashemi M, Mehdizadeh T. Essential oil of tarragon (Artemisis dracunculus) antibacterial activity on Staphylococcus aureus and Escherichia coli in culture media and Iranian white cheese. Iran J Microbiol. 2012;4(1):3–34. [PMC free article] [PubMed] [Google Scholar]

[CR38] Sagdic O, Kuşçu A, Özcan M, Özçelik S. Effects of Turkish spice extracts at various concentrations on the growth E. coli O157:H7. Food Microbiol. 2002;1:473–480. doi: 10.1006/fmic.2002.0494. [DOI] [Google Scholar]

[CR39] Silva F, Domingues FC. Antimicrobial activity of coriander oil and its effectiveness as food preservative. Crit Rev Food Sci Nutr. 2017;57:35–47. doi: 10.1080/10408398.2013.847818. [DOI] [PubMed] [Google Scholar]

[CR40] Tajkarimi MM, Ibrahim SA, Cliver DO. Antimicrobial herb and spice compounds in food. Food Control. 2010;21:1199–1218. doi: 10.1016/j.foodcont.2010.02.003. [DOI] [Google Scholar]

PERMALINK

Determination of sensitivity of some food pathogens to spice extracts

Gökhan Akarca

Oktay Tomar

İlknur Güney

Sena Erdur

Veli Gök

Abstract

Introduction

Materials and methods

Preparation of spice extracts

Bacteria

Antibiotic discs

Preparation of extract disks

Preparation of inoculums

Placement of discs and incubation

Determination of the minimum inhibitory concentrations (MICs)

Determination of minimum bactericidal concentrations (MBCs)

Statistical analyses

Results and discussion

Table 1.

Table 2.

Table 3.

Table 4.

Table 5.

Table 6.

Conclusion

Footnotes

References

ACTIONS

PERMALINK

RESOURCES

Cite

Add to Collections

PERMALINK

Determination of sensitivity of some food pathogens to spice extracts

Gökhan Akarca

Oktay Tomar

İlknur Güney

Sena Erdur

Veli Gök

Abstract

Introduction

Materials and methods

Preparation of spice extracts

Bacteria

Antibiotic discs

Preparation of extract disks

Preparation of inoculums

Placement of discs and incubation

Determination of the minimum inhibitory concentrations (MICs)

Determination of minimum bactericidal concentrations (MBCs)

Statistical analyses

Results and discussion

Table 1.

Table 2.

Table 3.

Table 4.

Table 5.

Table 6.

Conclusion

Footnotes

References

ACTIONS

PERMALINK

RESOURCES

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