Next Article in Journal
Test Studies on Geogrid–Soil Interface Behavior under Static and Dynamic Loads
Previous Article in Journal
Assessing Farmers’ Attitudes towards Rural Land Circulation Policy Changes in the Pearl River Delta, China
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Sustainability
Volume 14
Issue 7
10.3390/su14074298
sustainability-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Study on the Sustainability Potential of Thyme, Oregano, and Coriander Essential Oils Used as Vapours for Antifungal Protection of Wheat and Wheat Products

by
Voichita Bota
1,
Renata Maria Sumalan
2,*,
Diana Obistioiu
3,
Monica Negrea
1,
Ileana Cocan
1,
Iuliana Popescu
4 and
Ersilia Alexa
1
1
Faculty of Food Engineering, Banat’s University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania” from Timisoara, Calea Aradului No. 119, 300645 Timisoara, Romania
2
Faculty of Horticulture and Forestry, Banat’s University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania” from Timisoara, Calea Aradului No. 119, 300645 Timisoara, Romania
3
Faculty of Veterinary Medicine, Banat’s University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania” from Timisoara, Calea Aradului No. 119, 300645 Timisoara, Romania
4
Faculty of Agriculture, Banat’s University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania” from Timisoara, Calea Aradului No. 119, 300645 Timisoara, Romania
*
Author to whom correspondence should be addressed.
Sustainability 2022, 14(7), 4298; https://doi.org/10.3390/su14074298
Submission received: 4 March 2022 / Revised: 31 March 2022 / Accepted: 1 April 2022 / Published: 5 April 2022
(This article belongs to the Section Sustainable Food)
Browse Figures
Review Reports Versions Notes

Abstract

:
This study aims to highlight the antifungal, antimicotoxigenic potential and phytotoxic effect of three essential oils (EOs) of Origanum vulgare (OEO), Thymus vulgaris (TEO), and Coriandrum sativum (CEO) on wheat storage, but also the impact of EOs treatment on the sensory properties of bakery products obtained from the wheat seeds. The chemical composition of EOs was determined using GC-MS analysis; the fungal load was evaluated using the direct plating technique, while mycotoxin analyses were conducted using enzyme-linked immunosorbent assay (ELISA). A selective antifungal effect has been highlighted in terms of the action of EOs vapours. OEO and TEO are inhibited Alternaria, Fusarium and Drechslera, while Saccharomyces and Cladosporium have proven to be the most tolerant fungi. Drechslera is the most sensitive, the effect of all EOs being a fungicidal one. However, the fungicidal effect proved present in all EOs applied as vapours with values ranging between 0.2–0.4%. Regarding the phytotoxic effect of EOs vapours on the germination of the seeds, TEO and OEO had an inhibitory effect, especially at 0.4%. The effect is cumulative over time. The EOs inhibited deoxynivalenol (DON) occurrence; the maximum percentage of inhibition was obtained after 21 days of vapours exposure, being more effective in the case of 0.2%. EOs vapours treatment does not affect the quality of bread obtained from treated wheat seeds from a sensory point of view.

1. Introduction

The microflora of cereal seeds are diverse, determined by cultivation technology, variety, storage conditions, temperature and humidity levels during the vegetation period [1,2]. Cereal fungi are divided into two categories: (i) field fungi, which cause diseases during plant growth and (ii) storage fungi, involved in storage [3,4]. Often fungi involved in the mycotoxin infestation of cereal seeds are re-introduced by xerophilous species, metabolically adapted to humidity conditions around 14%. Field fungi have a chance to grow and multiply if the humidity is over 14%. At these humidity values, the activity of fungi affects the nutritional qualities of the seeds, being correlated with the increase of the temperature during the storage period [5]. Endophytic fungi, which survive during storage, can affect the quality of seeds and ensure their spreading in the following year.
Fusarium species fall into this category, fusarium blight being a disease that occurs for cereal plants during the growing season, affecting the quality of seeds during storage. Moreover, the genus is recognised, along with Aspergillus and Penicillium, as an essential producer of mycotoxins [4,5,6,7,8].
Great efforts have been made to control food deterioration by fungi, with synthetic fungicides being the most used storage strategy. However, despite the efficacy of these chemical substances, a significant number of them have led to the development of fungal resistance, proved to be toxic for the environment, and caused residual toxicity in grains [9].
Therefore, there is an increasing public demand for the development of new and safer antifungal agents for grain preservation. In recent years, many natural compounds have attracted the attention of scientists, such as plant extracts or essential oils (EOs) [10,11]. Given the positive impact of natural preparations on the environment and health, their use as antifungal agents in agriculture and food protection represents viable and sustainable alternatives to synthetic chemicals [11,12,13,14,15].
The establishment of natural alternatives with effective, efficient, and non-toxical preservative function derived from plants, whether extracts or oils, depends on several factors. Is discussed here about the chemical composition, concentration, possible synergies or antagonisms, forms of action (direct, vapours, nanoencapsulation), biological properties of the target microorganism and possible changes in the sensory properties of the preserved product [16,17,18,19,20,21].
EOs are hydrophobic substances of complex mixtures of volatile organic compounds obtained from different parts of aromatic plants. The antifungal activity of EOs has been extensively studied against Fusarium sp., Aspergillus sp., Penicillium sp., Alternaria sp., and Verticillium sp., among other economically important fungal genera [22,23,24,25]. However, the effectiveness of a particular EO as antifungal is determined not only by its fungitoxic effect but also by its ability to decrease or inhibit conidial production [26,27].
According to our previous studies, the treatment of wheat grains through direct contact with EOs from Coriandrum sativum, Thymus vulgaris, Mentha piperita determines lower mycotoxins occurrences belonging to Fusarium synthesis [28].
Considering the actual trends regarding the use of the three EOs: thyme (TEO), oregano (OEO) and coriander (CEO) in ecological agriculture, the present paper intends to clarify several aspects to assuring the sustainable use of EOs: (i) if the EOs can provide antifungal and antimicotoxigenic protection to wheat seeds during storage; (ii) if there is a phytotoxic effect of the essential oil vapours on the germination of the wheat seeds in the modified atmosphere; (iii) if the EOs provide antimicotoxigenic effect in wheat and if the mycotoxins are transferred in cereal products (flour, bran and bread); (iv) if the EOs treatment influences, from a sensory point of view, the bakery products obtained from EOs exposed wheat.

2. Materials and Methods

2.1. EOs Chemical Composition and the Working Variants

The essential oils used in the study come from the market (SOLARIS PLANT SRL, Bucharest, Romania).
The chemical characterisation of EOs was performed using GS/MS QP 2010 Plus (Shimadzu, Kyoto, Japan) equipped with AT WAX 30 m × 0.32 mm × 1 μm capillary column (Santa Clara, CA, United States). The program used to separate the compounds was: 40 °C for 1 min, a rate of 5 °C/min to 210 °C for 5 min. Helium was used as a carrier gas at a 1 mL/min flow rate. Injector and ion source temperatures were 250 °C and 220 °C, respectively. The injection volume was 1 μL at a split ratio of 1:50. The NIST 02, Wiley 275 spectra library has been used to identify the volatile compounds. The variants of the study are presented in Table 1.

2.2. Estimation of Antifungal Potential of EOs

42 hermetically sealed containers (800 mL vol) were used to determine the antifungal potential of EOs used in vapours form for the storage of wheat seeds. To remove opportunistic microorganisms the wheat seeds, obtained by ecological technology (Antille variety), were washed with 1:9 (v/v) hypochlorite solution then rinsed three times with sterile distilled water and dried in a sterile hood at 23 + 2 °C to a relative humidity of 14%.
The wheat seeds thus prepared were distributed in containers 300g each and the adjacent air volume of 395 mL (78% nitrogen and 21% oxygen, 0.03%carbon dioxide, 0.97% argon, 32% humidity) was used to calculate the concentration of the vapours from the air enriched with EOs. The amount of EOs (see Table 1) was added to a sterile filter paper attached to the lid. The containers were kept in the dark and periodically have been shaken for mixing and uniformly exposing the seeds to EOs vapours. At 7, 14 and 28 days from the experiment begin 14 containers were opened (7 variants in 2 repetitions) for evaluation of fungal colonisation, seeds germination test, and mycotoxin content.
To detect the fungal load of wheat seeds exposed to EOs vapour the direct plating technique was used. In this regard, dichloran rose bengal chloramphenicol (DRBC) (Oxoid, CM0727, Thermo Fisher Scientific Inc., Altrincham, UK) with 2 repetitions and 56 Petri plates for each moment of determination were used in the 7th, 14th, and 28th days. The plates were incubated in the dark, with the lid upside down at 22 + 2 °C for 3–4 days, after which the number of colonised seeds was noted. The fungi were isolated in pure culture on the sixth day for identification at genus and species levels. For identification and confirmation of Fusarium species, transfer was made to a specific medium, Dichlorane Chloramphenicol Peptone Agar (DCPA), with a photoperiod of 12 h light/night [29].
We established the frequency of occurrence of the fungal genus from the total fungal genera isolated per variant using the formula:
Fr (%) = (NOG/TNS) × 100
were,
  • NOG = number of occurrences of the genus
  • TNS = total number of fungi on sample

2.3. Estimation of the Phytotoxicity Effect of EOs on Wheat Seeds

As germination index used was the emergence of the radicles. The seedling (hypocotyl) and roots length (as a mean of 3 radicles) were recorded after 7 and 28 days of wheat exposure for each variant EOs vapours assay (Table 1). Briefly, 10 seeds were taken from each container and placed between 2 sheets of blotting paper in a Petri dish (Ø 90 mm) moistened with 10 mL of distilled water (2 Petri dishes were used per container). A total of 28 Petri dishes were used in each determination. After 4 days of incubating in the dark at temp 25 ± 2 °C, data were recorded.

2.4. Evaluation of Antimicotoxigenic Potential of EOs

Sample preparation and mycotoxin analyses were conducted using enzyme-linked immunosorbent assay (ELISA). The analysed mycotoxins were: deoxynivalenol (DON), fumonisin (FUMO), zearalenone (ZEA) and ochratoxin A (OTA).
For FUMO, OTA and ZEA the wheat samples (5 g) were extracted with 25 mL 70% methanol and for DON analysis 100 mL distilled water was used. The samples were extracted for 20 min using a high-speed shaker (IDL, Freising, Germany). The extracts were diluted at 1:13 for FUMO analysis and used directly for other mycotoxins analyses. R-Biopharm kits ELISA (Bio-Rad Laboratories, Redmond, WA, USA) were used to quantify mycotoxins. In this regard, 50 μL of filtrate and standard solutions were mixed with 50 μL of enzyme conjugate, then 50 μL antibody solution was added, mixed gently and incubated for 10 min. at room temperature. After washing three times with 250 μL distilled water, 100 μL substrate was added to each well and incubated for 5 min at room temperature. Finally, the resulting yellow colour’s intensity was measured at 450 nm using an ELISA 96-well plate reader (PR-1100, Bio-Rad Laboratories, USA) after adding 100 μL stop solution to each well. The results are expressed in ppm or ppb depending on the type of mycotoxin. Table 2 presents the regression coefficient of calibration curves and the Minimum Detection Limit (MDL) for each mycotoxin.
The mycotoxin content amplitude was evaluated using the formula:
AMy (%) = (IMC−TMC)/IMC × 100
were,
  • IMC—initial mycotoxin content (ppm)
  • TMC—mycotoxin content at time T (ppm)

2.5. The Obtaining of Flouring Derivative Products

The remaining wheat seeds were used to obtain flour, followed by the production of bread buns variants for mycotoxin analyses.
The wheat flour was obtained by grinding EOs fumigated wheat grains using the GM200 grinder (Retsch, Haan, Germany). The ground wheat was sieved to separate the flour from bran. The DON content was analysed for the flour and bran of each variant.
At 97 g flour obtained as above, 67 g of water and 2 g of yeast were added in granular form. The mixture was maintained for 20 min at room temperature and kneaded the dough again for another 8 min. The dough was left to ferment for 30 min at room temperature (23 °C), respectively, until the dough volume increased twice. This process was followed by modelling two buns weighing 83 g each and kept to the final fermentation (20 min). After baking, two bread buns weighing 62.95 g resulted for each EOs vapours treatment.
To highlight the transfer of mycotoxin occurring in endosperm or bran of seeds and the influence of heat treatment (baking) on the mycotoxin content in the bread, we experimented with wheat seeds contaminated with DON (6.45 ppm). From the 6 bread buns obtained with DON contaminated flour, 3 were baked at 180 °C and 3 at 250 °C, for 25 min.

2.6. Sensory Evaluation of Bread Derivates from EOs Fumigated Wheat

The sensory evaluation of bread buns was carried out only for wheat exposed 28 days to EOs vapours. The sensory evaluation of buns was evaluated by a panel of 38 assessors (males and females), non-smokers, without known cases of food allergies. A control sample (C) consisting of bread obtained from normal wheat flour was also evaluated with the other bread samples. Slices of bread with a thickness of 1 cm, with crust, were presented on sheet plates, coded with two digits, being served in random order, under normal lighting conditions and at room temperature. To evaluate the sensory attributes (appearance, texture, taste, flavour and overall acceptance), panellists used a five-point hedonic scale ISO 6658:2017 [30], with the following rates: 1 = strongly disliked; 2 = Slightly disliked; 3 = neither like nor dislike; 4 = Slightly liked; 5 = Strongly liked. The ranges of score and acceptability level were as follow: 1.00–1.49 = Not Acceptable (NA); 1.5–2.49 = Slightly Acceptable (SA); 2.50–3.49 = Moderately Acceptable (MA); 3.5–4.49 = Acceptable (A); 4.5–5.00 = highly acceptable (HA). All 38 panellists trained according to ISO 6658:2017 [30].

2.7. Statistical Analysis

All determinations were made in triplicate, and the results are reported as mean values ± standard deviation (SD). Differences between means were analysed with a one-way ANOVA, followed by multiple comparison analysis using the t-test (two-sample assuming equal variances). Differences were considered significant when p-values < 0.05. Data were processed with Statistica 10 (StatSoft, SAS Institute, Inc. Cary, NC, USA).

3. Results

3.1. GC-MS Composition of EOS

The chemical composition of the analysed EOs is presented in Table 3. The results show that TEO contains Thymol (35.862%) and o-Cymene (31.957%) as major components, followed in percentage over 4% by Linalool (6.301%). The commercial CEO contains 72.231% Linalool and in smaller percentages Limonene (4.766%) and α-Pinene (6.071%). In commercial oils, minor compounds have also been identified. In OEO, 17 volatile compounds were identified, of which Carvacrol represents 37.349% and o-Cymene 28.750% of the total components. Four volatile compounds are found in percentages between 4–7% (Caryophyllene, β-Myrcene, γ-Terpinene and β-Linalool).

3.2. Estimation of Antifungal Potential of EOs

To express the frequency of occurrence of fungi for each moment of determination, we used the sum of the values of the occurrence frequencies noted for each EOs variant (Total Frequency-TFr%, Figure 1). The structure of the seed’s microbiota has changed during 4-week. Thus, 5 dominant fungal genera have been identified, namely Saccharomyces (S. cerevisiae), Drechslera (D. tritici), Alternaria (A. alternata, A. infectoria), Fusarium (F. graminearum) and Cladosporium (C. cladosporioides).
After 7 days of exposure to EOs vapours, the frequency of occurrence of fungal genera has changed quantitatively but also qualitatively. Thus, all fungi were represented in the control version with values TFr between 5% and 30%. Fusarium and Cladosporium have appeared in all EOs treatments. The highest frequency for Fusarium (72%) was determined in the case of CEO2, followed by CEO1 with 46% and the lowest frequency was recorded for OEO1 and OEO2 with values of 20%. Fusarium is the most tolerant, with a total frequency of 237%.
After 14 days it is observed that the genus Drechslera is not found in any variant of EOs vapor treatment and Alternaria is totally inhibited at TEO1, registering the total frequency value of less than 34%. Fusarium appears with the highest frequency in the control version. The fungus with the highest TFr% is Saccharomyces with value of 379%. Fusarium was identified only in the control variant, the fungicidal effect of EOs concentrations in the vapor phase being responsible for this.
After 28 days the fungicidal effect of EOs is noticed for Drechslera and Fusarium which appear only in the control version. Alternaria reappears in all treatment variants alongside Saccharomyces and Cladosporium. Cladosporium is the genus with the highest frequency value of 268%.

3.3. Estimation of the Phytotoxicity Effect of EOs on Wheat Seeds

Due to the high volatilising properties of the oils and their persistence in closed environments, the evaluation of the impact on the germination capacity of wheat seeds is considered a fundamental analysis. Figure 2 shows the germination of wheat seeds stored in closed containers with a vapour-enriched atmosphere of EOs used in two concentrations. At first glance, the inhibitory effect of essential oil vapours can be observed. TEO1 and CEO1 applied in a concentration of 0.2% slightly affected the germination of seeds after 7 days of exposure, but after 28 days, the wheat germination is greatly affected. Seeds stored in atmospheres enriched with 0.2%/0.4% TEO and 0.4% OEO have lower germination values. In the case of CEO vapour, the highest values of all tested variants are noted for both concentrations at 70%.
Differences depending on concentrations of EOs-vapours are also noticeable at the length of the radicles and stem (Table 4). Overall, EOs vapours affect the growth of the radicles and stem in a differentiated way compared to the effect of the concentration. Thus, CEO1 stands out with appropriate control values for the length of the radicles, but significant differences occur for the size of the stems both at 7 days (3.00 ± 0.50 mm) and at 28 days (3.00 ± 0.58 mm) compared to the control sample (4.29 ± 0.41 mm). For CEO2, however, the negative effect on radicles growth occurs, both at 7 days and 28 days. The highest differences related to the radicle’s length were found in the OEO2 (2.97 ± 0.40) and TEO2 (2.84 ± 0.22) variants compared to the control (2.84 ± 0.22 mm). The negative effect remained even after 28 days of storage in the atmosphere enriched with EOS vapours.

3.4. The Effect of EOs Treatment on Mycotoxins Contamination in Wheat Samples and Derivative Products

The analysis of the mycotoxins content in the case of naturally contaminated samples showed that of the 4 mycotoxins analysed (DON, FUMO, ZEA and OTA), only DON was identified, both in the control sample and in the wheat samples treated with EOs (Table 5). The initial DON content in the contaminated natural wheat samples was 6.45 ppm. After 7 days of treatment, the highest content decrease is observed up to 4.863 ppm (inhibition percentage of 24.6%, Figure 3) in the case of TEO1, respectively, at 4.23 ppm (decrease of 34.42%) when TEO2 was used. The fumigation with OEO1 leads to a slight increase in DON content (9.77%), respectively, a decrease in OEO2 of 20.78%. The CEO did not show inhibition of DON after 7 days at any of the concentrations tested. The DON content increased 29.61% for CEO1 and 39.53% for CEO2. There is a slight decrease in DON content of 5.74% in control compared with the initial value, Figure 3.
After 14 days of fumigation, an increase of DON concentration in control (9.61%) compared to the initial concentration, respectively, of 9.92% in the case of fumigation with OEO1, were observed. For the other EOS analysed, regardless of the concentration tested, a decrease in DON content is observed, the highest inhibition being observed in the case of CEO2 (−57.05% compared to the initial concentration). A significant decrease is also noticed in the case of CEO1 (−37.83%), but also after fumigation with OEO2 (−34.57%) and TEO2 (−33.95%).
The effects of EOs fumigation after 28 days decrease compared to the effect after 14 days but show an essential percentage of inhibition related to the control for which the DON content increases by 28.99% compared to the initial one. For all EOs analysed, the inhibitory effect was resisted after 28 days. The values vary between −25.74% in the case of fumigation with CEO2 and −64.65% for TEO1, according to Figure 3.
Figure 4 presents the DON content in the derivative products. The experiment was performed on the control sample naturally contaminated with DON (6.45 ppm) to highlight the transfer of DON occurring naturally in wheat in the endosperm and bran and the influence of heat treatment (baking) on the DON content in bread.
The obtaining data show the content of DON in flour was 3.78 ppm, while in bran was 1.89 ppm of DON. In bread, after baking at 180 °C, 3.51 ppm of DON was recovered, and 3.22 ppm when the baking temperature was 250 °C.

3.5. Results of the Sensory Evaluation of Bread Derivates from EOs Fumigated Wheat

The samples obtained (Figure 5) were analysed from a sensory point of view to estimate the printing capacity of the EOS on taste and smell on the bread obtained from fumigated wheat.
Figure 6 shows the sensory evaluation results as mean and standard deviation differences. The most highly appreciated was TEO2, with 0.2% Thymus vulgaris oil, which obtained mean scores of 4.711 for overall acceptability, 4.842 for flavour and 4.816 for taste, falling within the 4.00–5.00 score range, which indicates high acceptability.
For the same attributes (overall acceptability, taste and flavour), the following high-scoring bread sample was TEO2 samples with 0.4% Thymus vulgaris oil, registering the following values: 4.632 (overall acceptability), 4.816 (flavour) and 4.789 (taste). However, in terms of appearance and texture, the scores were higher than in the TEO1 samples (Figure 6).

4. Discussion

4.1. Chemical Composition of Essential Oils

The content of volatile compounds and the dominant chemotype varies with the botanical species, geographical origin, and harvest time [28].
When TEO was used as an antifungal agent, thymol and o-cymene were the major components, with a sum of significant compounds over 60%. A smaller percentage of linalool was also detected (6.301%). These results agree with our previous study [11] regarding the chemical profile of TEO and which highlighted γ-terpinene and thymol as significant compounds. Other authors also reported thymol as a primary volatile compound [31,32,33,34], but o-cymene, γ-terpinene [35], camphor [35], carvacrol [32] and linalool [31] were found in significant quantities.
The chromatographic profile of CEO showed that linalool was the primary compound, followed by α-pinene and D-limonene. Our results are consistent with our previous investigations regarding the chemical composition of CEO obtained from Romania that reported the linalool as the principal constituent of CEO [36]. The content of linalool in CEO varied in the same range in other samples obtained from C. sativum seeds cultivated in Brazil (77.48% linalool) [37]. A high linalool content in CEO (60–80%) was also reported in other studies [38,39].
OEO analysed in our study was characterised by a high content of carvacrol and o-cymene (over 28%) and lower content (between 4–6.2%) of other compounds (caryophyllene, β-myrcene, γ-terpinene and β-linalool). Similar composition was reported by in the oregano leaf-flower oil, including carvacrol (30.73%), thymol (18.81%), p-cymene (10.88%), caryophyllene (7.73%), and 3-carene (4.06%) [40]. Carvacrol was the main compound (60%) in the fraction of OEO obtained after distillation at 140°C, while o-cymene represented the highest percentage (47.96%) of OEO obtained after distillation at 82 °C [41]. Higher content of carvacrol (70.2%) and similar minor compounds were reported by other authors [42].

4.2. Fungal Load of Wheat Seeds Kept in the Atmosphere Rich in Essential Oil Vapours

Research regarding the control of fungal contamination of cereals and cereal products is a significant focus; prevention of fungal infection during plant development, harvesting and storage of cereals are priorities in controlling the incidence of fungal infections and mycotoxins development [28]. Proper storage (humidity, temperature and insect control) and the addition of antifungal agents can reduce fungal growth and prevent the contamination of the sample. Boruga et al. [43] reported that the antimicrobial activity of EOs depends on their chemical components and is related to the presence of phenolic compounds and terpenic hydrocarbons, respectively [24,25,43].
The data resulting from the storage of wheat seeds in the atmosphere enriched with TEOs, CEOs and OEOs vapours show that the fungicidal or fungistatic effect is achieved depending on the type of oil and its concentration. The explanation lies in the antimicrobial compounds existing in each essential oil, whose diversity and weight vary depending on the plant, the plant part, the plant growth phase or the cultivation conditions [44].
Recent research has shown the different behaviour of essential oils applied in the form of vapours. The fact is that the antimicrobial activity of essential oils is more effective in the form of vapour-phase against bacteria [19,45,46,47,48,49].
This aspect was also highlighted in the case of our study. Therefore, when applying CEO1 and CEO2, Fusarium and Cladosporium recorded high occurrence frequency values after 7 days of seed fumigation. These high occurrence frequency values do not denote a weak antifungal effect of this EO for the concentrations taken in the study because in the weeks following, the frequency of occurrence for Fusarium was null. Based on these results, we can conclude that CEO acts over time, the fungicidal effect reaching the peak after 14 days of fumigation. The need for a more extended period to prove the irreversible antifungal effect in the case of EOs vapours has been reported by other authors [20].
The physical properties and chemical composition of significant compounds of EOs are responsible for the way of action and the achievement of the antimicrobial effect. Inouye et al. [50] highlighted that thymol and carvacrol are compounds with lower volatilisation and moderate solubility that deposit faster. With 35.862% thymol and OEO containing 37.349% carvacrol, our TEO study showed a more pronounced antifungal effect in the first week of fumigation. Linalool existing in the CEO in a high proportion (72.231%) considered very volatile, along with 4.766% limonene, has a low solubility in aqueous environments, demonstrating a poorly applied antimicrobial effect in liquid form but has high persistence and a strong antifungal effect if used in the form of vapours [49,51].
Regarding the mechanism of action of EOs, it consists in the loss of semipermeable properties of the cytoplasmic membrane, which leads to imbalances and loss of the integrity of the microbial cell [47]. On the other hand, the penetrating capacity of microbial membranes can be reduced due to the hydrophobic properties of oils used in the form of microemulsions or applicated in liquid form, producing mycelium-mycelium aggregate and thus reducing the affinity for microorganisms [47]. There are possible modes of antimicrobial action of phenolic compounds that could be explained by the denaturation of enzymes responsible for germinating spores or by interfering with amino acids that play a role in germination [52].
EOs obtained from Thymus vulgaris L. were evaluated for biological activity and chemical composition. Various studies [53,54] reported significant bacteriostatic activity. Tian et al. in 2019 [24] as well demonstrated the antifungal potential of Thymus essential oil by a total inhibition of Fusarium mycelium growth at all investigated concentrations and a synergistic/antagonistic effect when sage essential oil was added [11].
Oregano extracts and OEO affect all stages of the peroxidative process by neutralising free radicals, blocking catalysis-peroxidase, and interrupting the lipid radical [55].
A selective antifungal effect has been highlighted in terms of the action of EOs vapours. For example, in the case of OEO and TEO, Alternaria, Fusarium and Drechslera sp. are inhibited. At the same time, Saccharomyces and Cladosporium have proven to be the most tolerant species and Drechslera the most sensitive, the effect of all EOs being a fungicidal one. However, the fungicidal effect was demonstrated in all EOs applied as vapours in 0.2–0.4%.
For Alternaria the effect of EOs vapours is fungistatic, this being certified by the appearance of fungus at 28 days. Short-term studies have shown that TEO applied as vapour-phase at 66.7 μL/L has proven an antifungal effect in protecting artificially inoculated cherry tomatoes with Alternaria sp. kept for 5 days [16]. In our study, within 28 days, the concentration of 0.4% TEO provides a fungistatic effect for Alternaria.

4.3. About Phytotoxicity Effect of EOs Vapours on Wheat

As it is known, EOs are an effective, eco-friendly alternative in the management of weeds in ecological technologies; numerous studies initiated in recent years confirm the potential of bioherbicide attributed to them [56,57,58,59]. This potential is attributed to the phenomenon of allelopathy, which is not selective, only if the seeds or plants have enzymatic mechanisms of annihilation of the toxicogenic potential [36,60]. Recent studies have shown that the phytotoxic effect of EOs by allelopathy is valid for weed seeds and crop plant seeds [57,61]. Generally, reaching the phytotoxicity limit depends on the type of EO, concentration, and the plant’s sensitivity [62,63]. Using EOs vapours in wheat storage assay, it was demonstrated that exposure time is an essential factor in achieving the phytotoxic effect. As the seeds are exposed for a long time, in this case, 28 days, the germination capacity of the seeds is affected through direct action on the responsible enzymes, α-amylase being the main enzyme involved in initiating the germination process of monocots seeds [64]. However, even if germination has taken place, the negative effect of the essential oil vapours is evident in the seedling in the first step of growth.
Regarding the phytotoxic effect of EOs vapours on the germination of the seeds, TEO and OEO have an inhibitory effect, especially at 0.4%. The effect is cumulative over time.
According to the theory that the dose size makes the difference between good and bad, the effect on plants of EOs can be also stimulating or inhibiting. Phenomenon is characterised by a low-dose stimulation and a high-dose inhibition is known as hormesis [65]. Monoterpenes oxygenate from CEO determine the wheat germination release in a concentration-dependent and hormesis-type manner: high concentrations inhibit, and low concentrations enhance the induction of visible radicles. This could be due to the induction of α-amylases in the endosperm of germinated cereal grains [66]. In our study, 0.2% CEO with 72.231% linalool does not affect the radicles release, but the inhibitory effect is noticeable in the emission of the seedling, which is an involvement in the synthesis of cytokinins. This statement is in agreement with previous studies that affirm the repressive effect of EOs may be manifested in the first step of maise growth if the treatment is applied to the seeds [67] radicles growth of lettuce exposed to thyme, marjoram, vervain, and caraway EOs. In contrast, thyme and oregano EOs inhibited germination and radicle elongation for cress seeds [68].

4.4. The Effect of EOs Treatment on Mycotoxins Contamination in Wheat Samples and Derivative Products

In stored cereals, the application of natural preservatives and EOs generates the inhibition of mycotoxins production [69]. On the other hand, the combination of chemicals and natural products can lead to a 90% reduction in deoxynivalenol (DON) [70,71].
In our experiment, the initial DON content of the naturally contaminated wheat in control was 6.45 ppm. The analysed sample does not correspond to the food consumption requirements for wheat, considering the current legislation in force which provides for DON content a maximum admitted limit of 1.250 ppm for cereals other than durum wheat, oats and maise. In the European Union, the presence of Fusarium toxins in foodstuffs is regulated by Regulation (EC) No 1126/2007 [72] amending Regulation (EC) No 1881/2006 [73], and by Recommendation 2013/165/EU [74].
The treatment of cereals with EOs leads to a significant decrease in DON levels in wheat samples, depending on the type of EOs, concentration and time of fumigation. Our results show that the maximum percentage of inhibition is obtained after 21 days from fumigation, being more effective in the case of the concentration of 0.2% EOs (Figure 3). Regardless of the type of oil used and the concentration, after 21 days of fumigation, the inhibition is over 50%. Even after 7 days of fumigation, there is an inhibition of the development of DON in the case of TEO and OEO2 between 20–35%. After 14 days of fumigation, the inhibition capacity increases, being noticed in all experimental variants except OEO1. After 28 days of fumigation, an inhibitory capacity is observed compared to the control in the case of using EOs, but lower than the values obtained after 21 days.
A previous study regarding the reduction of DON contamination using EOs in the storage phase highlighted that after 22 days of treatment, DON was undetectable in all wheat samples [28]. A similar effect of EOs on the growth rate of DON produced by Fusarium species was reported [69].
Since mycotoxins are generally located in the outer shell of cereal seeds, their removal can reduce the content of mycotoxins in the seeds, and the effectiveness of decontamination depends on the degree of penetration of the seeds. Grinding does not directly affect the mycotoxin content but changes the distribution between different cereal fractions. In the case of foods derived from dried and ground cereals, the distribution of mycotoxins in different fractions resulting from grinding depends on the fungal penetration of the endosperm. Thus, cereals with surface contamination will have a lower content of mycotoxins in the flour and a higher concentration in germs, bran and fiber. fibre.
Our results showed that the content of DON is higher in flour than in bran (Figure 4), which means that contamination has occurred because the field or that conditions of temperature, humidity, water activity in storage deposition favoured the penetration of mycotoxins into the endosperm of the grain.
Regarding the reduction of DON content by heat treatments, studies have shown that DON is very stable in the temperature range of 170–350 °C, without indicating a decrease in its concentration after 30 min at 170 °C. This feature is hazardous for human and animal health [75]. Our results show a slight decrease in DON content during the bread processing process, from a concentration of 3.78 ppm in wheat flour to 3.51 ppm when the baking process took place at 180 °C (7.14% reduction), respectively, 3.22 ppm at 250 °C (14.81% reduction). A reduction in DON concentration up to 52% was achieved in cereals using hot steam at 185 °C for 6 min [76]. As the thermal decomposition products of the DON are not known, and the fact that no toxicity studies have been performed using these DON products, there is no clear evidence that these compounds resulting from the thermal instability of the DON lead to detoxification of food contaminated with DON.
Nowadays, different concentrations of OEO are used to control Penicillium sp. [77] and other fungal species such as Aspergilus niger, being stronger than rosemary or sage oil [78].

4.5. Sensorial Evaluation of Wheat Bread

Despite some positive studies on applying natural antimicrobials of plant origin, we have faced two significant problems in terms of food application: the change in smell occurring when using high concentrations and the additional costs that occur.
In this regard, we aimed to evaluate the effect of fumigation with EOs applied in wheat storage on the sensory properties of bread. Due to the low concentrations of EOs applied to fumigation, they do not have a negative effect on the smell or taste of the bread; on the contrary they were evaluated positively by the evaluators.
Regarding flavour attribute, the scores given by the evaluators increased in the order: TOE1 > TEO2 > CEO2 > CEO1 > OEO2 > OEO1 > Control suggesting that Thymus vulgaris, Origanum vulgare and Coriandrum sativum conferred to products an appreciated aroma by the evaluators. In terms of taste attribute, the scores increased in the following order TEO2 > TEO1 > CEO1 > CEO2 > OEO1 > OEO2 > Control and in terms of overall acceptability, the panellists ranked the bread samples as follows: TEO2 > TEO1 > CEO2 > CEO1 > OEO2 > OEO1 > Control.
If we refer to the difference in score between the two concentrations of essential oil used (0.4% and 0.2%), there are no notable differences between the evaluated attributes of the studied samples. Similar studies regarding consumer acceptability, in the case of the addition of essential oils in bakery products, have been carried out by Sikkhamondhol et al. [79] and Khalil et al. [80].
Previous studies highlighted that OEO could be used up to 2% in bread to improve nutritional and sensory qualities, specific volume and shelf life, having an inhibitory action on moulds [81]. In addition, natural antimicrobial compounds from OEO could influence the preservation of bread and other bakery products, being antimicrobial agents against fungal growth in many foods [55,82]. Oregano could be used successfully in producing graham bread, improving the taste and bread aroma [83].

5. Conclusions

In this study, we demonstrated the possibility of using essential oil of thyme, oregano, and coriander to control storage fungi and the prevention of DON mycotoxins occurring for two concentrations calculated at air volume, 0.2 and 0.4%, respectively. The fungicidal effect of EO vapours was achieved after 7 days for field fungi (e.g., Drechslera), while for Fusarium, the antifungal effect was proven after 14 days of EOs fumigation. The germination of the seeds was inhibited after 7 days of exposure; in the order, oregano > thymus > coriander, the effect of the EO vapours acting cumulatively over time and was affected both the emission of radicles and the seedling length.
There were no noticeable sensory differences between the two concentrations applied regarding the sensory evaluation of the bread obtained from seeds exposed for 28 days to EOs vapours. Concerning the overall acceptability, on the first place comes the variant of bread from seeds fumigated with TEO followed by the CEO and OEO.
TEO, OEO and CEO vapour-phase essential oils can be a sustainable alternative to preserving wheat seeds for baking, ensuring safety and protection against mycotoxin-producing fungi. Due to the negative effects on germination, it is not recommendable to apply them if the destination of the seeds is to obtain a field crop.

Author Contributions

Conceptualisation V.B., R.M.S. and E.A.; methodology, V.B., R.M.S., D.O., M.N., I.C. and I.P.; validation, E.A. and R.M.S.; writing—original draft preparation, V.B., R.M.S. and E.A.; writing—review and editing, V.B., R.M.S., D.O., M.N., I.C. and E.A.; visualisation, V.B., R.M.S., D.O., M.N., I.C., I.P. and E.A. All authors have read and agreed to the published version of the manuscript.

Funding

This research paper is supported by the project “Increasing the impact of excellence research on the capacity for innovation and technology transfer within USAMV.B. Timișoara” code 6PFE, submitted in the competition Program 1—Development of the national system of research—development, Subprogram 1.2—Institutional performance, Institutional development projects—Development projects of excellence in RDI.

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

The report of the analyses performed for the samples in the paper can be found at the Interdisciplinary Research Platform (PCI) belonging to the Banat University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania” from Timisoara.

Acknowledgments

We were able to carry out this research with the support of the Interdisciplinary Research Platform belonging to the Banat University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania” from Timisoara, where the analysis were made.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Alexa, E.; Poiana, M.-A.; Sumalan, R.-M. Mycoflora and Ochratoxin A Control in Wheat Grain Using Natural Extracts Obtained from Wine Industry By-Products. Int. J. Mol. Sci. 2012, 13, 4949–4967. [Google Scholar] [CrossRef] [PubMed]
  2. Matusinsky, P.; Zouhar, M.; Pavela, R.; Novy, P. Antifungal Effect of Five Essential Oils against Important Pathogenic Fungi of Cereals. Ind. Crops Prod. 2015, 67, 208–215. [Google Scholar] [CrossRef]
  3. Negrea, M.; Pop, G.; Sumalan, R.; Alexa, E.; Poiana, M.-A. Post-Harvest Fungi Production in Different Storage. In Proceedings of the 6th Central European Congress on Food-CEFood Congress, Novi Sad, Serbia, 23–26 May 2012. [Google Scholar]
  4. Fleurat-Lessard, F. Integrated Management of the Risks of Stored Grain Spoilage by Seedborne Fungi and Contamination by Storage Mould Mycotoxins—An Update. J. Stored Prod. Res. 2017, 71, 22–40. [Google Scholar] [CrossRef]
  5. Mohapatra, D.; Kumar, S.; Kotwaliwale, N.; Singh, K.K. Critical Factors Responsible for Fungi Growth in Stored Food Grains and Non-Chemical Approaches for Their Control. Ind. Crops Prod. 2017, 108, 162–182. [Google Scholar] [CrossRef]
  6. Dawlal, P.; Barros, E.; Marais, G.J. Evaluation of Maize Cultivars for Their Susceptibility towards Mycotoxigenic Fungi under Storage Conditions. J. Stored Prod. Res. 2012, 48, 114–119. [Google Scholar] [CrossRef]
  7. Streit, E.; Schatzmayr, G.; Tassis, P.; Tzika, E.; Marin, D.; Taranu, I.; Tabuc, C.; Nicolau, A.; Aprodu, I.; Puel, O.; et al. Current Situation of Mycotoxin Contamination and Co-Occurrence in Animal Feed—Focus on Europe. Toxins 2012, 4, 788–809. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  8. Enyiukwu, D.N.; Awurum, A.N.; Nwaneri, J.A. Mycotoxins in Stored Agricultural Products: Implications to Food Safety and Health and Prospects of Plant-Derived Pesticides as Novel Approach to Their Management. Gen. J. Microbiol. Antimicrob. 2014, 2, 32–48. [Google Scholar] [CrossRef]
  9. Margni, M.; Rossier, D.; Crettaz, P.; Jolliet, O. Life Cycle Impact Assessment of Pesticides on Human Health and Ecosystems. Agric. Ecosyst. Environ. 2002, 93, 379–392. [Google Scholar] [CrossRef]
  10. Pandey, P.K.; Kumar, P.; Singh, N.N.; Tripathi, N.N.; Bajpai, V.K. Essential Oils: Sources of Antimicrobials and Food Preservati. Front. Microbiol. 2017, 7, 2161. [Google Scholar] [CrossRef] [Green Version]
  11. Alexa, E.; Danciu, C.; Cocan, I.; Negrea, M.; Morar, A.; Obistioiu, D.; Dogaru, D.; Berbecea, A.; Radulov, I. Chemical Composition and Antimicrobial Potential of Satureja hortensis L. in Fresh Cow Cheese. J. Food Qual. 2018, 2018, 8424035. [Google Scholar] [CrossRef] [Green Version]
  12. Cocan, I.; Negrea, M.; Cozma, A.; Alexa, E.; Poiana, M.-A.; Raba, D.; Danciu, C.; Popescu, I.; Cadariu, A.I.; Obistioiu, D.; et al. Chili and Sweet Pepper Seed Oil Used as a Natural Antioxidant to Improve the Thermo-Oxidative Stability of Sunflower Oil. Agronomy 2021, 11, 2579. [Google Scholar] [CrossRef]
  13. Obistioiu, D.; Cocan, I.; Tîrziu, E.; Herman, V.; Negrea, M.; Cucerzan, A.; Neacsu, A.-G.; Cozma, A.L.; Nichita, I.; Hulea, A.; et al. Phytochemical Profile and Microbiological Activity of Some Plants Belonging to the Fabaceae Family. Antibiotics 2021, 10, 662. [Google Scholar] [CrossRef] [PubMed]
  14. Cocan, I.; Alexa, E.; Danciu, C.; Radulov, I.; Galuscan, A.; Obistioiu, D.; Morvay, A.; Sumalan, R.; Poiana, M.; Pop, G.; et al. Phytochemical Screening and Biological Activity of Lamiaceae Family Plant Extracts. Exp. Ther. Med. 2018, 15, 1863–1870. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  15. Alexa, V.T.; Szuhanek, C.; Cozma, A.; Galuscan, A.; Borcan, F.; Obistioiu, D.; Dehelean, C.A.; Jumanca, D. Natural Preparations Based on Orange, Bergamot and Clove Essential Oils and Their Chemical Compounds as Antimicrobial Agents. Molecules 2020, 25, 5502. [Google Scholar] [CrossRef] [PubMed]
  16. Feng, W.; Chen, J.; Zheng, X.; Liu, Q. Thyme Oil to Control Alternaria Alternata In Vitro and In Vivo as Fumigant and Contact Treatments. Food Control 2011, 22, 78–81. [Google Scholar] [CrossRef]
  17. Puškárová, A.; Bučková, M.; Kraková, L.; Pangallo, D.; Kozics, K. The Antibacterial and Antifungal Activity of Six Essential Oils and Their Cyto/Genotoxicity to Human HEL 12469 Cells. Sci. Rep. 2017, 7, 8211. [Google Scholar] [CrossRef] [Green Version]
  18. Teneva, D.; Denkova-Kostova, R.; Goranov, B.; Hristova-Ivanova, Y.; Slavchev, A.; Denkova, Z.; Kostov, G. Chemical Composition, Antioxidant Activity and Antimicrobial Activity of Essential Oil from Citrus aurantium L. Zest against Some Pathogenic Microorganisms. Z. Nat. C 2019, 74, 105–111. [Google Scholar] [CrossRef] [Green Version]
  19. Felšöciová, S.; Vukovic, N.; Jeżowski, P.; Kačániová, M. Antifungal Activity of Selected Volatile Essential Oils against Penicillium sp. Open Life Sci. 2020, 15, 511–521. [Google Scholar] [CrossRef]
  20. Shaaban, H.A. Essential Oil as Antimicrobial Agents: Efficacy, Stability, and Safety Issues for Food Application. In Essential Oils; de Oliveira, M.S., da Costa, W.A., Silva, S.G., Eds.; IntechOpen: Rijeka, Croatia, 2020. [Google Scholar]
  21. Sumalan, R.M.; Kuganov, R.; Obistioiu, D.; Popescu, I.; Radulov, I.; Alexa, E.; Negrea, M.; Salimzoda, A.F.; Sumalan, R.L.; Cocan, I. Assessment of Mint, Basil, and Lavender Essential Oil Vapor-Phase in Antifungal Protection and Lemon Fruit Quality. Molecules 2020, 25, 1831. [Google Scholar] [CrossRef]
  22. Harris, R. Synergism in the Essential Oil World. Int. J. Aromather. 2002, 12, 179–186. [Google Scholar] [CrossRef]
  23. Bakkali, F.; Averbeck, S.; Averbeck, D.; Idaomar, M. Biological Effects of Essential Oils—A Review. Food Chem. Toxicol. 2008, 46, 446–475. [Google Scholar] [CrossRef] [PubMed]
  24. Tian, X.; Liang, T.; Liu, Y.; Ding, G.; Zhang, F.; Ma, Z. Extraction, Structural Characterization, and Biological Functions of Lycium barbarum Polysaccharides: A Review. Biomolecules 2019, 9, 389. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  25. Tian, B.; Liu, M.; An, W.; Yu, L.; Zhang, J.; Liu, Y.; Zhao, J.; Li, J. Lycium Barbarum Relieves Gut Microbiota Dysbiosis and Improves Colonic Barrier Function in Mice Following Antibiotic Perturbation. J. Funct. Foods 2020, 71, 103973. [Google Scholar] [CrossRef]
  26. Nguefack, J.; Leth, V.; Amvam Zollo, P.H.; Mathur, S.B. Evaluation of Five Essential Oils from Aromatic Plants of Cameroon for Controlling Food Spoilage and Mycotoxin Producing Fungi. Int. J. Food Microbiol. 2004, 94, 329–334. [Google Scholar] [CrossRef] [PubMed]
  27. Bajpai, V.K.; Chul, S. In Vitro and In Vivo Inhibion of Plant Pathogenic Fungi by Essential Oil and Extracts of Magnolia liliflora Desr. J. Agric. Sci. Technol. 2012, 14, 845–856. [Google Scholar]
  28. Sumalan, R.-M.; Alexa, E.; Poiana, M.-A. Assessment of Inhibitory Potential of Essential Oils on Natural Mycoflora and Fusarium Mycotoxins Production in Wheat. Chem. Cent. J. 2013, 7, 32. [Google Scholar] [CrossRef] [Green Version]
  29. Pitt, J.I.; Hocking, A.D. Fungi and Food Spoilage; Springer: Berlin/Heidelberg, Germany, 2009; pp. 20–41. [Google Scholar]
  30. SR ISO 6658; 2017—Sensory Analysis—Methodology—General Guidance. ISO: Geneva, Switzerland, 2017.
  31. Schmidt, E.; Wanner, J.; Höferl, M.; Jirovetz, L.; Buchbauer, G.; Gochev, V.; Girova, T.; Stoyanova, A.; Geissler, M. Chemical Composition, Olfactory Analysis and Antibacterial Activity of Thymus vulgaris Chemotypes Geraniol, 4-Thujanol/Terpinen-4-Ol, Thymol and Linalool Cultivated in Southern France. Nat. Prod. Commun. 2012, 7, 1934578X1200700. [Google Scholar] [CrossRef] [Green Version]
  32. Hudaib, M.; Speroni, E.; Di Pietra, A.M.; Cavrini, V. GC/MS Evaluation of Thyme (Thymus vulgaris L.) Oil Composition and Variations during the Vegetative Cycle. J. Pharm. Biomed. Anal. 2002, 29, 691–700. [Google Scholar] [CrossRef]
  33. Baranauskienė, R.; Venskutonis, P.R.; Viškelis, P.; Dambrauskienė, E. Influence of Nitrogen Fertilisers on the Yield and Composition of Thyme (Thymus vulgaris). J. Agric. Food Chem. 2003, 51, 7751–7758. [Google Scholar] [CrossRef]
  34. Rus, C.; Rm, S.; Alexa, E.; Dm, C.; Pop, G.; Botau, D. Study on Chemical Composition and Antifungal Activity of Essential Oils Obtained from Representative Species Belonging to the Lamiaceae Family. Plant Soil Environ. 2016, 61, 297–302. [Google Scholar] [CrossRef] [Green Version]
  35. Imelouane, B.; El Bachiri, A.; Wathelet, J.-P.; Dubois, J.H.A. Chemical Composition, Cytotoxic and Antioxydant Activity of The Essential Oil of Lavandula dentata. World J. Chem. 2010, 5, 103–110. [Google Scholar]
  36. Sumalan, R.M.; Alexa, E.; Popescu, I.; Negrea, M.; Radulov, I.; Obistioiu, D.; Cocan, I. Exploring Ecological Alternatives for Crop Protection Using Coriandrum sativum Essential Oil. Molecules 2019, 24, 2040. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  37. Mandal, S.; Mandal, M. Coriander (Coriandrum sativum L.) Essential Oil: Chemistry and Biological Activity. Asian Pac. J. Trop. Biomed. 2015, 5, 421–428. [Google Scholar] [CrossRef] [Green Version]
  38. Zamindar, N.; Sadrarhami, M.; Doudi, M. Antifungal Activity of Coriander (Coriandrum sativum L.) Essential Oil in Tomato Sauce. Food Meas. 2016, 10, 589–594. [Google Scholar] [CrossRef]
  39. Momin, A.H.; Acharya, S.S.; Gajjar, A.V. Coriandrum Sativum-Review of Advances in Phytopharmacology. Int. J. Pharm. Sci. Res. 2012, 3, 1233. [Google Scholar]
  40. Han, F.; Ma, G.; Yang, M.; Yan, L.; Xiong, W.; Shu, J.; Zhao, Z.; Xu, H. Chemical Composition and Antioxidant Activities of Essential Oils from Different Parts of the Oregano. J. Zhejiang Univ. Sci. B 2017, 18, 79–84. [Google Scholar] [CrossRef] [Green Version]
  41. Leyva-López, N.; Gutiérrez-Grijalva, E.; Vazquez-Olivo, G.; Heredia, J. Essential Oils of Oregano: Biological Activity beyond Their Antimicrobial Properties. Molecules 2017, 22, 989. [Google Scholar] [CrossRef] [Green Version]
  42. Asensio, C.M.; Grosso, N.R.; Juliani, H.R. Quality Characters, Chemical Composition and Biological Activities of Oregano (Origanum Spp.) Essential Oils from Central and Southern Argentina. Ind. Crops Prod. 2015, 63, 203–213. [Google Scholar] [CrossRef]
  43. Borugă, O.; Jianu, C.; Mişcă, C.; Goleţ, I.; Gruia, A.T.; Horhat, F.G. Thymus vulgaris Essential Oil: Chemical Composition and Antimicrobial Activity. J. Med. Life 2014, 7, 56–60. [Google Scholar]
  44. Prakash, B.; Kedia, A.; Mishra, P.K.; Dubey, N.K. Plant Essential Oils as Food Preservatives to Control Moulds, Mycotoxin Contamination and Oxidative Deterioration of Agri-Food Commodities—Potentials and Challenges. Food Control 2015, 47, 381–391. [Google Scholar] [CrossRef]
  45. Antih, J.; Houdkova, M.; Urbanova, K.; Kokoska, L. Antibacterial Activity of Thymus vulgaris L. Essential Oil Vapours and Their GC/MS Analysis Using Solid-Phase Microextraction and Syringe Headspace Sampling Techniques. Molecules 2021, 26, 6553. [Google Scholar] [CrossRef] [PubMed]
  46. Houdkova, M.; Kokoska, L. Volatile Antimicrobial Agents and In Vitro Methods for Evaluating Their Activity in the Vapour Phase: A Review. Planta Med. 2020, 86, 822–857. [Google Scholar] [CrossRef] [PubMed]
  47. Brożyna, M.; Paleczny, J.; Kozłowska, W.; Chodaczek, G.; Dudek-Wicher, R.; Felińczak, A.; Gołębiewska, J.; Górniak, A.; Junka, A. The Antimicrobial and Antibiofilm In Vitro Activity of Liquid and Vapour Phases of Selected Essential Oils against Staphylococcus aureus. Pathogens 2021, 10, 1207. [Google Scholar] [CrossRef] [PubMed]
  48. Tullio, V.; Nostro, A.; Mandras, N.; Dugo, P.; Banche, G.; Cannatelli, M.A.; Cuffini, A.M.; Alonzo, V.; Carlone, N.A. Antifungal Activity of Essential Oils against Filamentous Fungi Determined by Broth Microdilution and Vapour Contact Methods. J. Appl. Microbiol. 2007, 102, 1544–1550. [Google Scholar] [CrossRef]
  49. Inouye, S.; Nishiyama, Y.; Hasumi, Y.; Yamaguchi, H.; Abe, S.; Uchida, K.; Yamaguchi, H. The Vapor Activity of Oregano, Perilla, Tea Tree, Lavender, Clove, and Geranium Oils against a Trichophyton Mentagrophytes in a Closed Box. J. Infect. Chemother. 2006, 12, 349–354. [Google Scholar] [CrossRef]
  50. Inouye, S.; Uchida, K.; Maruyama, N.; Yamaguchi, H.; Abe, S. A Novel Method to Estimate the Contribution of the Vapor Activity of Essential Oils in Agar Diffusion Assay. Nippon Ishinkin Gakkai Zasshi 2006, 47, 91–98. [Google Scholar] [CrossRef] [Green Version]
  51. Griffin, S.G.; Wyllie, S.G.; Markham, J.L.; Leach, D.N. The Role of Structure and Molecular Properties of Terpenoids in Determining Their Antimicrobial Activity. Flavour Fragr. J. 1999, 14, 322–332. [Google Scholar] [CrossRef]
  52. El Khetabi, A.; Ezrari, S.; El Ghadraoui, L.; Tahiri, A.; Ait Haddou, L.; Belabess, Z.; Merah, O.; Lahlali, R. In Vitro and In Vivo Antifungal Activities of Nine Commercial Essential Oils against Brown Rot in Apples. Horticulturae 2021, 7, 545. [Google Scholar] [CrossRef]
  53. Prashanth, N.S.; Marchal, B.; Devadasan, N.; Kegels, G.; Criel, B. Advancing the Application of Systems Thinking in Health: A Realist Evaluation of a Capacity Building Programme for District Managers in Tumkur, India. Health Res. Policy Syst. 2014, 12, 42. [Google Scholar] [CrossRef] [Green Version]
  54. Saleh, N.B.; Chambers, B.; Aich, N.; Plazas-Tuttle, J.; Phung-Ngoc, H.N.; Kirisits, M.J. Mechanistic Lessons Learned from Studies of Planktonic Bacteria with Metallic Nanomaterials: Implications for Interactions between Nanomaterials and Biofilm Bacteria. Front. Microbiol. 2015, 6, 677. [Google Scholar] [CrossRef] [Green Version]
  55. Chiș, M.S.; Muste, S.; Păucean, A.; Man, S.; Sturza, A.; Petruț, G.S.; Mureșan, A. A Comprehensive Review about Antimicrobial Effects of Herb and Oil Oregano. Hop Med. Plants 2017, 25, 17–27. [Google Scholar]
  56. Verdeguer, M.; Castañeda, L.G.; Torres-Pagan, N.; Llorens-Molina, J.A.; Carrubba, A. Control of Erigeron bonariensis with Thymbra capitata, Mentha piperita, Eucalyptus camaldulensis, and Santolina chamaecyparissus Essential Oils. Molecules 2020, 25, 562. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  57. Agnieszka, S.; Magdalena, R.; Jan, B.; Katarzyna, W.; Malgorzata, B.; Krzysztof, H.; Danuta, K. Phytotoxic Effect of Fiber Hemp Essential Oil on Germination of Some Weeds and Crops. J. Essent. Oil Bear. Plants 2016, 19, 262–276. [Google Scholar] [CrossRef]
  58. Koiou, K.; Vasilakoglou, I.; Dhima, K. Herbicidal Potential of Lavender (Lavandula angustifolia Mill.) Essential Oil Components on Bristly Foxtail (Setaria verticillata (L.) P. Beauv.): Comparison with Carvacrol, Carvone, Thymol and Eugenol. Arch. Biol. Sci. 2020, 72, 223–231. [Google Scholar] [CrossRef] [Green Version]
  59. Werrie, P.-Y.; Durenne, B.; Delaplace, P.; Fauconnier, M.-L. Phytotoxicity of Essential Oils: Opportunities and Constraints for the Development of Biopesticides. A Review. Foods 2020, 9, 1291. [Google Scholar] [CrossRef] [PubMed]
  60. Benarab, H.; Fenni, M.; Louadj, Y.; Boukhabti, H.; Ramdani, M. Allelopathic Activity of Essential Oil Extracts from Artemisia Herba-Alba Asso. on Seed and Seedling Germination of Weed and Wheat Crops. Acta Sci. Nat. 2020, 7, 86–97. [Google Scholar] [CrossRef]
  61. Synowiec, A.; Kalemba, D.; Drozdek, E.; Bocianowski, J. Phytotoxic Potential of Essential Oils from Temperate Climate Plants against the Germination of Selected Weeds and Crops. J. Pest Sci. 2017, 90, 407–419. [Google Scholar] [CrossRef]
  62. Angelini, L.G.; Carpanese, G.; Cioni, P.L.; Morelli, I.; Macchia, M.; Flamini, G. Essential Oils from Mediterranean Lamiaceae as Weed Germination Inhibitors. J. Agric. Food Chem. 2003, 51, 6158–6164. [Google Scholar] [CrossRef]
  63. Rahman, A. (Ed.) Studies in Natural Products Chemistry; Elsevier: Amsterdam, The Netherlands, 1988; ISBN 978-0-444-42970-4. [Google Scholar]
  64. Poonpaiboonpipat, T.; Pangnakorn, U.; Suvunnamek, U.; Teerarak, M.; Charoenying, P.; Laosinwattana, C. Phytotoxic Effects of Essential Oil from Cymbopogon citratus and Its Physiological Mechanisms on Barnyardgrass (Echinochloa crus-galli). Ind. Crops Prod. 2013, 41, 403–407. [Google Scholar] [CrossRef]
  65. Calabrese, E.J.; Blain, R.B. Hormesis and Plant Biology. Environ. Pollut. 2009, 157, 42–48. [Google Scholar] [CrossRef]
  66. Rentzsch, S.; Podzimska, D.; Voegele, A.; Imbeck, M.; Müller, K.; Linkies, A.; Leubner-Metzger, G. Dose- and Tissue-Specific Interaction of Monoterpenes with the Gibberellin-Mediated Release of Potato Tuber Bud Dormancy, Sprout Growth and Induction of α-Amylases and β-Amylases. Planta 2012, 235, 137–151. [Google Scholar] [CrossRef] [PubMed]
  67. Możdżeń, K.; Krajewska, A.; Bocianowski, J.; Jop, B.; Synowiec, A. Microencapsulated Caraway Essential Oil Affects Initial Growth of Maize Cultivars. Molecules 2021, 26, 5059. [Google Scholar] [CrossRef] [PubMed]
  68. De Almeida, L.F.R.; Frei, F.; Mancini, E.; De Martino, L.; De Feo, V. Phytotoxic Activities of Mediterranean Essential Oils. Molecules 2010, 15, 4309–4323. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  69. Jimenez-Garcia, S.N.; Garcia-Mier, L.; Garcia-Trejo, J.F.; Ramirez-Gomez, X.S.; Guevara-Gonzalez, R.G.; Feregrino-Perez, A.A. Fusarium Mycotoxins and Metabolites That Modulate Their Production. In Fusarium—Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers; Askun, T., Ed.; InTech: Rijeka, Croatia, 2018; ISBN 978-1-78923-318-6. [Google Scholar]
  70. Magan, N. Mycotoxin Contamination of Food in Europe: Early Detection and Prevention Strategies. Mycopathologia 2006, 162, 245–253. [Google Scholar] [CrossRef]
  71. Mshelia, L.P.; Selamat, J.; Iskandar Putra Samsudin, N.; Rafii, M.Y.; Abdul Mutalib, N.-A.; Nordin, N.; Berthiller, F. Effect of Temperature, Water Activity and Carbon Dioxide on Fungal Growth and Mycotoxin Production of Acclimatised Isolates of Fusarium verticillioides and F. graminearum. Toxins 2020, 12, 478. [Google Scholar] [CrossRef]
  72. Kyprianou, M. Commission Regulation (EC) No 1126/2007 of 28 September 2007. Amending Regulation (EC) No 1881/2006 Setting Maximum Levels for Certain Contaminants in Foodstuffs As Regards Fusarium Toxins in Maize and Maize Products; European Commission: Brussels, Belgium, 2007. [Google Scholar]
  73. European Commission. Commission Regulation (EC) No 1881/2006 of 19 December 2006 Setting Maximum Levels for Certain Contaminants in Foodstuffs; European Commission: Brussels, Belgium, 2006. [Google Scholar]
  74. European Commission. Commission Recommendation of 27 March 2013 on the Presence of T-2 and HT-2 Toxin in Cereals and Cereal Products; European Commission: Brussels, Belgium, 2013. [Google Scholar]
  75. Peng, Z.; Chen, L.; Nüssler, A.K.; Liu, L.; Yang, W. Current Sights for Mechanisms of Deoxynivalenol-Induced Hepatotoxicity and Prospective Views for Future Scientific Research: A Mini Review: DON-Induced Hepatotoxicity and Prospective Views. J. Appl. Toxicol. 2017, 37, 518–529. [Google Scholar] [CrossRef]
  76. Cenkowski, S.; Pronyk, C.; Zmidzinska, D.; Muir, W.E. Decontamination of Food Products with Superheated Steam. J. Food Eng. 2007, 83, 68–75. [Google Scholar] [CrossRef]
  77. Passarinho, A.T.P.; Dias, N.F.; Camilloto, G.P.; Cruz, R.S.; Otoni, C.G.; Moraes, A.R.F.; Soares, N.D.F.F. Sliced Bread Preservation through Oregano Essential Oil-Containing Sachet: Bread Preservation through Antimicrobial Sachet. J. Food Process Eng. 2014, 37, 53–62. [Google Scholar] [CrossRef]
  78. Borges, R.S.; Ortiz, B.L.S.; Pereira, A.C.M.; Keita, H.; Carvalho, J.C.T. Rosmarinus officinalis Essential Oil: A Review of Its Phytochemistry, Anti-Inflammatory Activity, and Mechanisms of Action Involved. J. Ethnopharmacol. 2019, 229, 29–45. [Google Scholar] [CrossRef]
  79. Sikkhamondhol, C.; Teanpook, C.; Boonbumrung, S.; Chittrepol, S. Quality of Bread with Added Turmeric (Curcuma longa): Powder, Essential Oil and Extracted Residues. Asian J. Food Agro-Ind. 2009, 2, 690–701. [Google Scholar]
  80. Khalil, P.; Masood, S.; Rehman, A.; Zafar, A.; Islam, Z.; Javaid, N.; Ilyas, A.; Qamar, S.; Zeb, A. Proximate and Sensory Analysis of Wheat Bread Supplemented with Nigella Sativa Oil and Nigella Sativa Extract. Pure Appl. Biol. 2021, 10, 1158–1165. [Google Scholar] [CrossRef]
  81. Dhillon, G.K.; Amarjeet, K. Quality Evaluation of Bread Incorporated with Different Levels Cinnamon Powder. Int. J. Food Sci. 2013, 2, 70–74. [Google Scholar]
  82. Ávila Sosa Sánchez, R.; Portillo-Ruiz, M.C.; Viramontes-Ramos, S.; Muñoz-Castellanos, L.N.; Nevárez-Moorillón, G.V. Effect of Mexican Oregano (Lippia berlandieri Schauer) Essential Oil Fractions on the Growth of Aspergillus Spp. in a Bread Model System: Modeling Aw and Mexican Oregano on Aspergillus. J. Food Process. Preserv. 2015, 39, 776–783. [Google Scholar] [CrossRef]
  83. Muresan, C.; Stan, L.; Man, S.; Scrob, S.; Muste, S. Sensory Evaluation of Bakery Products and Its Role in Determining of the consumer preferences. J. Agroaliment. Processes Technol. 2012, 18, 304–306. [Google Scholar]
Sustainability 14 04298 g001 550
Figure 1. The Total Frequency (TFr%) of fungi isolated at 7, 14 and 28 days from seeds wheat stored in an atmosphere enriched with essential oil vapours. EOs concentrations are calculated for the volume of air enriched with essential oil vapours (OEO1, TEO1, CEO1 = 0.2%, OEO2, TEO2, CEO2 = 0.4%).
Figure 1. The Total Frequency (TFr%) of fungi isolated at 7, 14 and 28 days from seeds wheat stored in an atmosphere enriched with essential oil vapours. EOs concentrations are calculated for the volume of air enriched with essential oil vapours (OEO1, TEO1, CEO1 = 0.2%, OEO2, TEO2, CEO2 = 0.4%).
Sustainability 14 04298 g001
Sustainability 14 04298 g002 550
Figure 2. Wheat germination test determined at 7 and 28 days of seeds exposure to EOs in vapours-form. (n = 4).
Figure 2. Wheat germination test determined at 7 and 28 days of seeds exposure to EOs in vapours-form. (n = 4).
Sustainability 14 04298 g002
Sustainability 14 04298 g003 550
Figure 3. Amplitude of DON (AMy, %) at 7, 14 and 28 days from seeds wheat storage in an atmosphere enriched with essential oil vapours. EOs concentrations are OEO1, TEO1, CEO1 = 0.2%, OEO2, TEO2, CEO2 = 0.4% calculated for the volume of air enriched with essential oil vapours.
Figure 3. Amplitude of DON (AMy, %) at 7, 14 and 28 days from seeds wheat storage in an atmosphere enriched with essential oil vapours. EOs concentrations are OEO1, TEO1, CEO1 = 0.2%, OEO2, TEO2, CEO2 = 0.4% calculated for the volume of air enriched with essential oil vapours.
Sustainability 14 04298 g003
Sustainability 14 04298 g004 550
Figure 4. DON content in derivative products (wheat, bran, flour and bread buns baked at 180 °C and 250 °C). The values are expressed as mean ± standard deviations (n = 3). t-test was used to compare the mean differences registered among samples. The error bars indicate standard deviation. Different letters among samples indicate significant differences (p < 0.05) among values according to the t-test.
Figure 4. DON content in derivative products (wheat, bran, flour and bread buns baked at 180 °C and 250 °C). The values are expressed as mean ± standard deviations (n = 3). t-test was used to compare the mean differences registered among samples. The error bars indicate standard deviation. Different letters among samples indicate significant differences (p < 0.05) among values according to the t-test.
Sustainability 14 04298 g004
Sustainability 14 04298 g005 550
Figure 5. Bread samples obtained after fumigation with Eos.
Figure 5. Bread samples obtained after fumigation with Eos.
Sustainability 14 04298 g005
Sustainability 14 04298 g006 550
Figure 6. Mean values of the sensory evaluation (consumer acceptance) of bread with different oils (OEO1, TEO1, CEO1 = 0.2%, OEO2, TEO2, CEO2 = 0.4%) using 5-point hedonic scale (n = 38).
Figure 6. Mean values of the sensory evaluation (consumer acceptance) of bread with different oils (OEO1, TEO1, CEO1 = 0.2%, OEO2, TEO2, CEO2 = 0.4%) using 5-point hedonic scale (n = 38).
Sustainability 14 04298 g006
Table
Table 1. Quantities and concentrations used in the study of the sustainability of essential oils.
Table 1. Quantities and concentrations used in the study of the sustainability of essential oils.
EOsµL% *
OEO1Oreganum sativum EOs8000.2
OEO2Oreganum sativum EOs16000.4
TEO1Thymus vulgaris EOs8000.2
TEO2Thymus vulgaris EOs16000.4
CEO1Coriandrum sativum EOs8000.2
CEO2Coriandrum sativum EOs16000.4
* The EO concentrations are calculated for the volume of air enriched with essential oil vapours.
Table
Table 2. Parameters of mycotoxins determination.
Table 2. Parameters of mycotoxins determination.
MycotoxinsCalibration Curve Parameters
(r2)		MDL *
DON0.99590.08 ppm
ZEA0.98845 ppm
FUMO0.99460.015 ppm
* Minimum detection limit.
Table
Table 3. Chemical composition (% of total) of EOs.
Table 3. Chemical composition (% of total) of EOs.
CompoundsTypeTREOs
OEOTEOCEO
α-PineneMH11.3142.4031.9186.071
CampheneMH12.641-1.6750.308
β-PineneMH13.4680.5943.4770.143
Β-MyrceneMH14.5494.2431.7730.213
α-PhellandreneMH14.8000.255--
α-TerpineneMH15.1592.1430.725-
D-LimoneneMH15.2453.7132.3884.766
γ-TerpineneMH16.5044.121-1.335
cis- β-OcimeneMH16.850--0.237
p-Mentha-1,4(8)-dieneMH17.224--0.281
TerpinoleneMH17.2251.048--
EucalyptolMO17.3380.1290.698-
o-CymeneMH18.49128.75031.957-
o-CymolMH18.551--5.386
1-Octen-3-ol * 22.141-0.239-
cis-Linalyl OxideMO23.520--1.375
Trans-Linalool oxideMO24.441--1.162
β- LinaloolMO24.5834.7866.30172.231
CaryophylleneSH25.1216.2101.425-
Thymol methyl etherMO25.701-1.041-
Benzene, 1-methoxy-4-methyl-2-(1-methylethyl)MO26.341-0.734-
Bornyl acetateMO26.484-0.513-
4-TerpinenolMO26.903-1.761-
CamphorMO27,9130.2060.3193.643
Geranyl acetateMO28.93--0.901
BorneolMO29.277-1.725-
trans-GeraniolMO30.96--0.077
p-PropenylanisoleMO32.462--0.948
p-Cymen-8-olMO32.887--0.091
3,7-Octadiene-2,6-diol, 2,6-dimethyl-MO35.316--0.679
Caryophyllene oxideSO36.090.2360.555-
CaryophylleneSH36.4193.5733.380-
ThymolMO37.378-35.862-
CarvacrolMO38.19837.3491.524-
1,7-Octadiene-3,6-diol, 2,6-dimethyl-MO38.275--0.1456
ViridiflorolSO40.6350.234--
Total major compounds ** 98.34396.21395.969
Monoterpene hidrocarbonates (MH) 46.42443.19317.559
Monoterpenes oxygenate (MO) 42.13648.21578.410
Sesquiterpene hidrocarbonates (SH) 9.7834.805-
Sesquiterpene oxygenate (SO) 0.4710.556-
* Other compounds; ** The difference up to 100% represents unidentified compounds (values not presented in the table).
Table
Table 4. Effects of EOs vapours-phase to radicles lengths and seedling lengths after 7 days and 28 days of seeds exposure.
Table 4. Effects of EOs vapours-phase to radicles lengths and seedling lengths after 7 days and 28 days of seeds exposure.
TreatmentRadicles (mm)Seedling (mm)
7 Days28 Days7 Days28 Days
Control11.50 ± 0.22 a10.94 ± 0.55 a4.29 ± 0.41 a4.29 ± 0.41 a
OEO19.64 ± 1.52 b5.52 ± 0.66 d3.64 ± 0.66 b3.65 ± 0.76 b
OEO22.97 ± 0.40 e3.33 ± 0.00 e2.17 ± 0.41 d2.00 ± 0.00 e
TEO19.14 ± 0.62 c9.07 ± 0.81 b4.00 ± 0.86 a3.84 ± 1.00 b
TEO22.84 ± 0.22 e3.00 ± 0.00 e2.75 ± 0.88 c2.67 ± 1.15 c
CEO111.09 ± 0.44 a11.03 ± 0.47 a3.00 ± 0.50 c3.00 ± 0.58 c
CEO26.19 ± 1.05 d5.99 ± 1.07 d2.38 ± 0.52 d2.43 ± 0.53 d
Results as mean ± SE. Different letters (a,b,c,d,e) in columns reveal significant differences according to Tukey test at p < 0.05 for EOs vapours concentration.
Table
Table 5. The DON content at 7, 14 and 28 days from seeds wheat storage in an atmosphere enriched with essential oil vapours.
Table 5. The DON content at 7, 14 and 28 days from seeds wheat storage in an atmosphere enriched with essential oil vapours.
DON Content (ppm)
EOs TreatmentInitial7 Days14 Days28 Days
CONTROL6.45 ± 0.04 a6.08 ± 0.06 f7.07 ± 0.05 c8.32 ± 0.10 g
OEO16.45 ± 0.07 a7.08 ± 0.08 c7.09 ± 0.08 c2.88 ± 0.05 c
OEO26.45 ± 0.06 a5.11 ± 0.09 a4.22 ± 0.04 bd3.62 ± 0.07 be
TEO16.45 ± 0.07 a4.86 ± 0.06 a3.76 ± 0.05 a2.28 ± 0.04 a
TEO26.45 ± 0.08 a4.23 ± 0.05 b4.26 ± 0.06 b3.88 ± 0.6 b
CEO16.45 ± 0.08 a8.36 ± 0.07 d4.01 ± 0.06 d3.61 ± 0.06 de
CEO26.45 ± 0.07 a9.00 ± 0.08 e2.77 ± 0.03 e4.79 ± 0.05 f
The values are expressed as mean ± SD (n = 2). A t-test was used to compare the mean differences among samples; data within the same column with different letters are significantly different (p < 0.05).
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Bota, V.; Sumalan, R.M.; Obistioiu, D.; Negrea, M.; Cocan, I.; Popescu, I.; Alexa, E. Study on the Sustainability Potential of Thyme, Oregano, and Coriander Essential Oils Used as Vapours for Antifungal Protection of Wheat and Wheat Products. Sustainability 2022, 14, 4298. https://doi.org/10.3390/su14074298

AMA Style

Bota V, Sumalan RM, Obistioiu D, Negrea M, Cocan I, Popescu I, Alexa E. Study on the Sustainability Potential of Thyme, Oregano, and Coriander Essential Oils Used as Vapours for Antifungal Protection of Wheat and Wheat Products. Sustainability. 2022; 14(7):4298. https://doi.org/10.3390/su14074298

Chicago/Turabian Style

Bota, Voichita, Renata Maria Sumalan, Diana Obistioiu, Monica Negrea, Ileana Cocan, Iuliana Popescu, and Ersilia Alexa. 2022. "Study on the Sustainability Potential of Thyme, Oregano, and Coriander Essential Oils Used as Vapours for Antifungal Protection of Wheat and Wheat Products" Sustainability 14, no. 7: 4298. https://doi.org/10.3390/su14074298

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop