The
utilization of essential oils (EO) as supplements for animals, particularly
dairy cows, has gained increasing attention in recent years due to their
perceived potential to enhance various aspects of animal health and
productivity (1). EO, extracted from aromatic plants,
are known for their diverse bioactive compounds, which possess a range of
biological and pharmacological activities such as antimicrobial, antioxidant,
antifungal, and antibacterial (2). In the context of animal husbandry,
these natural compounds are being explored as dietary supplements with the aim
of enhancing
overall well-being, mitigating stress, and potentially increasing products (3). Dairy cows, as vital contributors
to the agricultural sector, are subject to various stressors and health
challenges that can impact milk production. The incorporation of EO into their
diet as a supplement represents a novel avenue for addressing these concerns (4). Türkiye is one of the world's most
florally diverse countries. This distinction is attributed to the convergence
of the Mediterranean, Iran-Turan, and Euro-Siberian phytogeographic regions
within Anatolia, fostering dynamic interactions among plant communities (5). Furthermore, the considerable
variations in climate, topography, geology, geomorphology, and aquatic
environments play pivotal roles in shaping this botanical abundance. Sivas, a city in Türkiye,
emerges as a province characterized by a rich floral spectrum, influenced by the
distinctive geographical features and imprints of the Anatolian climate zone (6). Some of these plants analyzed in this study, collected from the Sivas
region, not only highlights
the local biodiversity but also unveiled the biotherapeutic potential inherent
in the flora of this region.
The
Umbelliferae (Apiaceae) family has a vast array of 300–455 genera and over 3000
species. Notably, anise (Pimpinella anisum) emerges as a crucial species
employed for its aromatic qualities in both the food and perfumery industries.
Beyond its industrial applications, P. flabellifolia holds significance
in folk medicine, particularly for its beneficial effects, notably during
lactation periods (7). Tepe et al. have documented that
the EO of P. flabellifolia contains key biotherapeutic compounds,
including limonene (47%), E-anethole (37.9%), and β-pinene (6%) (). Furthermore, studies have
identified E-anethole at 41% in the aerial parts and 63.5% in the fruits of (). Recent research has highlighted the
utilization of certain Pimpinella species as supplements in animal feed,
with a specific focus on enhancing milk secretion in dairy cows ().
Declarations
Conflict of Interest
The authors declare no conflicting interest.
Data Availability
The unpublished data is available upon request to the corresponding author.
The
Myrtaceae[Ma1] family includes 150 genera and
approximately 3300 species, exhibiting a global distribution. Within this plant
family, Myrtus communis is a key plant species renowned for its diverse
pharmacological effects, including antioxidant, antimicrobial, and
anti-inflammatory properties (11). EO composition of M. communis
species cultivated in Türkiye has been documented, revealing constituents such
as linalool (31.3%), linalyl acetate (17.8%), 1,8-cineole (14.7%), geranyl
acetate (9.1%), α-pinene (8.4%), and α-terpineol (6.5%) (12).
On the other hand, the genus Tanacetum encompasses over 200
species distributed across West Asia, North Africa, and Europe (13). Within Türkiye, there are 45
Tanacetum species and 18 of them are endemic. T. vulgare, a member of
the Tanacetum taxa, is a crucial plant because of its diverse EO compositions
and is recognized not only for its pharmacological effects but also for its
application as a repellent against insects (14). The EO composition of T. vulgare
includes camphor, borneol, β-thujone, and 1,8-cineole, albeit with varying
percentages attributed to the specific collection area (15).
In
this study, P.
flabellifolia, M. communis, and T. vulgare were
collected, and their EO compositions analyzed. Furthermore, diverse methods
were employed to investigate the antioxidant and antimicrobial activities of
these EOs. Lastly, in silico analyzes was performed to assess molecular
mechanisms to examining the effects of EO supplementation on dairy cows, with a
specific emphasis on its potential effects on milk production. Utilizing in
silico molecular docking methods, our objective is to delineate the intricate
interactions between EO compositions and crucial molecular targets associated
with lactation and metabolic pathways. Through this investigation, we aim to
provide valuable insights into the role of EO as dietary supplements, aiming to
optimize the performance and well-being of dairy cows, potentially influencing
both the quality and quantity of milk production.
Experimental Section
Plant Material Collections
The
plant species collection regions are listed, P. flabellifolia from Sivas
Gürün/Sivas, M. communis from Gürpınar/Sivas and T. vulgare from
Ulaş-Tecer/Sivas. Voucher plant specimens were identified by Dr. Erol DONMEZ
at the department of Biology, Cumhuriyet University, Sivas-Turkey and has been
deposited at the Herbarium of the Department of Biology, Cumhuriyet University,
Sivas-Turkey (CUFH-Voucher No: ED-11014 for P. flabellifolia, No:
ED-11015 for M. communis and No: ED11005 for T. vulgare)
Isolation of Essential Oils
Air-dried
and mechanically grinded aerial parts of all plant samples were distillated for
3 h using a Clevenger apparatus. Obtained oil samples were dried with anhydrous
sodium sulphate, filtrated, and stored at +4°C.
Gas Chromatography/Mass Spectrometry (GC/MS) Analysis
Chemical
compositions of the EO samples were analyzed using a Shimadzu GC2010 GC–MS,
equipped with a capillary column TC-5 (50 m× 0,32 mm i.d., 0.32 mm) and a 70 eV
EI Quadrapol detector. Helium was the carrier gas, at a flow rate of 2,1
ml/min., injector and MS transfer line temperatures were set at 265 and 280°C,
respectively. Column temperature was initially at 45°C held for 3 min, then
gradually increased to 150°C at a 2°C/min rate, held for 4 min, and finally
increased to 265°C at a 4°C/min and held for 4 min. Diluted samples (1:150
v/v, in hexane) of 0.5 µL were injected manually and splitless. The components
were identified by comparison with their lineer retention indices (LRIs) and MS
(NBS75K library data of the GC–MS system).
Antioxidant Activity Assays
Diphenylpicrylhydrazyl
(DPPH) Assays
The
methodology employed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) in the presence of
a hydrogen-donating antioxidant. DPPH solutions exhibit a robust absorption
band at 517 nm, manifesting as a deep violet color. As the absorption
diminishes, the resultant decolorization is stoichiometrically linked to the
extent of reduction. The residual DPPH, quantified after a specified duration,
serves as an inverse measure of the radical scavenging activity exerted by the
antioxidant. To execute this, 50 µl of various concentrations of the EOs were
introduced to 5 ml of a 0.004% methanol solution of DPPH. Following a 30-minute
incubation period at room temperature, absorbance readings were recorded at 517
nm against a blank. Inhibition free radical DPPH in percentage was calculated
in the Equation 1.
DPPH(
Equation 1
EO
concentrations providing 50% inhibition (IC50) were calculated from the graph
plotting inhibition percentage against varying concentrations. Synthetic
antioxidant reagent butylated hydroxytoluene (BHT) was used as a positive
control and all tests were carried out in triplicate.
β-Carotene-Linoleic
Acid Assays
Antioxidant
capacity was also determined by assessing the inhibition of volatile organic
compounds and conjugated diene hydroperoxides resulting from the oxidation of
linoleic acid. This method relies on the alteration of the yellow color of
β-carotene, attributed to its reaction with radicals generated during linoleic
acid oxidation within an emulsion (16). The velocity of β-carotene
bleaching is attenuated in the presence of antioxidants. This alteration serves
as the basis for evaluating the antioxidant activity of the EO in comparison to
well-established synthetic and natural antioxidants, such as BHT. The
antioxidative capacities of the EOs were measured against BHT at equivalent
concentrations, with a blank comprising only 350 µl of ethanol. All experiments
were replicated three times for robustness and reliability.
Antimicrobial Activity Assays
The
antimicrobial and antifungal activities of the EOs were systematically assessed
against three Gram-positive bacteria, five Gram-negative bacteria, and one
fungus. The bacterial and fungal strains utilized in this study were sourced
from the Refik Saydam Hygiene Institute in Ankara, Turkey. Bacterial cultures
were appropriately suspended in Mueller Hinton Agar (MHA-Oxoid-CM337), while
yeasts were suspended in Sabouraud Dextrose Agar (OxoidCM41). To evaluate
antimicrobial activity, a disc diffusion assay was performed (17). Initially, suspensions of the
tested microorganisms (0.1 ml containing 108 cells per ml) were evenly spread
on solid media plates. Subsequently, filter paper disks (6 mm in diameter),
treated with 10 μl of EO, were placed on these plates. Then plates were incubated
at 4°C for 2 hours and at 37°C for 24 hours for bacteria, and at 30°C for 48
hours for yeasts. The diameters of the resulting inhibition zones were measured
in millimeters. All assays were performed triplicate.
In Silico Analysis
Molecular Docking
The
use of computer-aided methodologies for drug and supplement development within
the veterinary medicine domain has become a prominent focus in recent times. In
silico experiments have been instrumental in elucidating specific molecular
interactions between target protein structures and bioactive compounds within
herbal mixtures. Prolactin, a hormone secreted by the pituitary gland, plays a
pivotal role in milk production during pregnancy and postpartum in mammals (18). Increasing prolactin secretion in
mammals has been identified as a strategy to enhance milk production,
particularly in dairy cows (19). Various approaches in the
literature decrease prolactin inhibitors, with dopamine being a notable
example. Extensive studies have reported that the dopamine molecule and related
processes act as prolactin inhibitors, thereby diminishing lactation (19). Similarly, literature suggests that
elevated progesterone levels during pregnancy in mammals serve to prevent
prenatal milk loss (20, 21).
From
this knowledge, structures of dopamine (Uniprot ID: P20288) and progesterone
(Uniprot ID: Q8MIL9) of the Bos taurus were selected as receptors in a
semi-flexible molecular docking procedure (22). Molecular docking studies were
conducted for ligands common in EO compositions of plants with the highest
quantities. The in silico investigation employed Autodock Vina (23) and Discovery Studio Visualizer (24) programs for comprehensive
assessment.
PASS Prediction
The
PASS (Prediction of Activity Spectra for Substances) online web tool enables
prediction of the expected biological function profile of a chemical compound
with similarities to a drug (25). The compounds showing the lowest
binding affinity and highest inhibitory potential from molecular docking
analyses were further subjected to PASS analysis to reveal their other
pharmacological effects. The PASS tool prediction results 2 category labels of
“probability to be active” (Pa) or “probability to be inactive” (Pi) as
biological activity.
Mutagenicity Prediction
Lazar,
a web-based computational tool helps in the prediction of complicated
toxicological properties such as toxicity, carcinogenicity, and blood brain
barrier (BBB) permeation (26). Lazar employs data mining
algorithms to input experimental data and generate predictions for unknown
chemical ligands. For the analysis, the chemical SMILES data of phytochemicals
were introduced to the system, and results were obtained by selecting the
desired options for analysis.
Carcinogenicity
Prediction
The
potential of any chemical to induce carcinogenicity in humans or animals can be
predicted computationally by using Carcinogenicity Prediction using Ensembled
Learning Methods (CarcinoPred-EL). This web tool is created by combining
different algorithms (RF, SVM and XGBoost), resulting in values with high
sensitivity, accuracy, and specificity rates (27).
ADMET Analysis
ADMET
(absorption, distribution, metabolism, excretion, and toxicity) analysis was
performed by using the pkCSM (https://biosig.lab.uq.edu.au/pkcsm) web tool (28). For analysis, the SMILES chemical
data format of the bioactive compounds was retrieved from the Pubchem database.
Statistical Analysis
The
data were statistically analyzed by ANOVA (n=3) and statistical significance
was accepted at a level of p<0.05.
Table 1. Chemical components of Tanacetum vulgare L., Pimpinella flabellifolia and Myrutus communis L.subsp .communis Essential Oils.
No.
LRI
(cal)
LRI
(lit)
Compound
Tanacetum
vulgare L (%)
Pimpinella
flabellifolia (%)
Myrutus
communis L. subsp. communis (%)
IM
1
930
931
(-)-α-Pinene
3.02±0.03
5.1±0.03
14.94±0.05
a,b
2
953
954
camphene
1.48±0.02
nd
0.57±0.00
a
3
976
977
sabinene
2.35±0.04
nd
nd
a
4
977
979
β-pinene
nd
3.7±0.05
2.87±0.02
a
5
992
994
myrcene
nd
nd
0.4±0.02
a
6
1012
1010
β-Cymene
12.77±0.13
nd
nd
a
7
1013
1011
p-cymene
nd
1.6±0.03
nd
a,b
8
1014
1013
m-cymene
nd
nd
4.57±0.06
a
9
1031
1030
4-carene
nd
nd
1.35±0.01
a
10
1042
1041
ocimene
nd
nd
4.75±0.05
a,b
11
1050
1051
Thujan-4-ol
2.90±0.02
nd
nd
a
12
1062
1062
α-Terpinyl
acetate
nd
nd
1.61±0.01
a
13
1065
1068
1,8-Cineole
5.15±0.04
nd
27.06±0.15
a,b
14
1081
1080
linalool
0.05±0.00
0.30±0.01
6.45±0.24
a,b
15
1086
1088
terpinolene
nd
0.80±0.03
2.53±0.03
a
16
1090
1091
β-Thujone
32.62±0.35
nd
nd
a,b
17
1117
1120
fenchol
nd
nd
0.35±0.02
a
18
1134
1133
(+)-cis-sabinol
0.23±0.01
nd
0.33±0.02
a
19
1145
1144
camphor
5.1±0.03
nd
nd
a,b
20
1161
1164
4-terpineol
1.49±0.01
1.45±0.03
5.28±0.04
a
21
1168
1165
thujanol
nd
nd
0.22±0.03
a
22
1182
1183
α -terpinene
nd
0.9±0.04
nd
a
23
1184
1185
myrtenol
nd
nd
3.14±0.05
a
24
1185
1186
α-terpineol
1.90±0.11
nd
nd
a
25
1202
1203
d-limonene
nd
34.2±0.23
5.25±0.08
a,b
26
1216
1218
β-phellandrene
nd
0.3±0.01
nd
a
27
1229
1227
anisaldehyde
nd
0.1±0.00
nd
a
28
1236
1235
linalyl
acetate
nd
nd
0.93±0.10
a
29
1275
1276
sabinyl
acetate
2.97±0.03
nd
nd
a
30
1284
1283
bornyl
acetate
5.47±0.02
nd
nd
a
31
1286
1285
(E)-anethol
nd
48.30±0.24
nd
a,b
32
1297
1298
carvacrol
nd
0.30±0.00
2.42±0.01
a,b
33
1301
1299
myrtenyl
acetate
nd
nd
3.25±0.06
a
34
1302
1306
thymol
nd
nd
3.85±0.03
a
35
1358
1360
geranyl
acetate
nd
nd
1.07±0.01
a
36
1580
1578
spatulenol
nd
0.4±0.02
nd
a
Note: (a) Compounds listed in order of elution from a DB-5 column. (b) Identification of components based on standard compounds; All values are mean ± standard deviation of triplicates; LRI(cal): Linear retention indices (DB-5 column) calculated against n-alkanes. % calculated from FID data with standart; LRI(lit):https://pubchem. ncbi.nlm.nih.gov; IM: Identification Method; nd: not detected.
Results and Discussion
GCMS Analysis Results
The results of the volatile composition analysis of the EOs are presented in Table 1 and Figure 1. First,
the EO samples extracted from T. vulgare, composed of mostly beta-thujone
(32.62%), beta-cymene (12.77%), and bornyl-acetate (5.47%).
Figure 1. Venn diagram for the EO composition.
Table 2. Antioxidant properties of all three plants.
Sample
Inhibition IC50 (mg/mL) (DPPH)
Inhibition %
(β-carotene- Linoleic acid)
Tanacetum
vulgare
0.200±0.01
18.5±0.09
Pimpinella
flabellifolia
0.083±0.003
57.2±2.18
Myrtus communis
0.500±0.085
37.07±1.25
BHT
0.0145±0.002
98.8±1.75
In an EO analysis study on Canadian varieties, the highest component was camphor at 30.48%, with the beta-thujone amount reported as 3.66% (29). In a similar study conducted on Poland species, the highest component was trans-chrysanthenyl acetate at 18.39%, while the beta-thujone amount was found to be 14.28% (15). Although the obtained content analysis results show regional variations, a common observation is that T. vulgare EO contains a high amount of beta-thujone. On the other hand, EO of P. flabellifolia, consisted of of (E)- anethol 48.30%, D-limonene 34.2% and (-)-α-Pinene 5.1% volatile compounds. Similar studies in the literature have consistently identified anethol and derivatives as the predominant component in the EO composition of this species (8). In the EO content of M. communis, a total of 93.19% has been identified, with high levels of volatile components, including 1,8-Cineole at 27.06%, (-)-α-Pinene at 14.94%, and linalool at 6.45%. A similar chemical profile was reported by Maharik et al., with the highest content being 1,8-Cineole at 31.98% and linalool at 21.94% (30). Considering the analysis of three essential oil samples, it was concluded that the chemical profile of the essential oil can vary significantly depending on the region from which it is collected, with T. vulgare identified as one such component.
Venn diagram analysis, conducted to uncover common component profiles and determine chemical synergy, revealed that the common component, alpha-pinene, was found in all three EO formulations, with the highest content determined in M. communis at 14.94%. Alpha-pinene, recognized as a high biotherapeutic product, has been reported to exhibit positive effects on the motor nervous system, in addition to its antimicrobial, antioxidant, and anti-inflammatory properties, particularly when exposed during the prenatal period in mice (31). In addition, regarding the commonality of chemical components, it has been concluded that M. communis and P. flabellifolia may exhibit volatile compound synergies with 7 common compounds (Figure 1).
Antioxidant Assays
Free radical DPPH and β-carotene-linoleic acid assay results for each EO samples were shown in Table 2. The
antioxidant activities of EOs were analyzed using both DPPH and β-carotene
assays. From the results, P. flabellifolia showed the highest
antioxidant activity for both DPPH and β-carotene tests. Compared with the
control groups, the plants exhibited a significantly high antioxidant profile,
especially in the DPPH assays. The antioxidant properties obtained were found
to be slightly lower than those of the T. vulgare species studied in
Latvia (0.032-0.181 mg/mL depending on the plant parts) (32), and similar to those of the
Romanian samples (33). On the other hand, antioxidant
analyses for P. flabellifolia have been reported 0.00849 mg/mL for DPPH
and 59.5% for β-carotene (8). It is possible that the species
collected from the Erzincan/Turkey region exhibited antioxidant activity
similar to the Sivas region. The antioxidant properties of M. communis
were higher than those observed in previous studies conducted on M. communis
EOs in Algeria, Tunisia, and Morocco, demonstrating IC50 values ranging between
0.80 and 4.50 mg/mL (34-36).
Table 3. Antimicrobial activities of the EOs using agar disc diffusion method.
Microorganisms
Disc Diffusiona
Results
P. flabellifolia
M. communis
T. vulgare
Gentamicin
Staphylococcus aureus
18±1.01
29±1.00
16.5 ±0.50
23±0.54
Escherichia coli
11±0.26
18±0.45
3±0.04
16±0.20
Proteus vulgaris
11±1.93
10±1.70
-
22±1.45
Pseudomonas aeruginosa
6±1.01
6±1.01
84±2.04
20±0.28
Salmonella typhi
10±1.00
17±1.05
21±1.45
10±1.45
Bacillus subtilis
90±172
20±1.55
21±1.45
29±1.45
Klebsiella pneumoniae
60±1.81
23±1.62
16±0.88
20±1.45
Corynebacterium diphteriae
11±1.44
23±1.14
-
23±1.45
Candida albcans
38±1.47
15±1.47
12±1.45
-
Antimicrobial Assays
The antimicrobial properties of all EO samples were determined using the disc diffusion method, and the results are presented in Table 3.
The
application of essential oils due to their antimicrobial activities is known in
both animal husbandry and ethnoveterinary practices (37, 38) because bacterial and fungal
infections in dairy cows can directly impact milk yield (39). Some findings have shown that these
infections can be transmitted to humans if proper pasteurization processes are
not ensured, such as E. coli contamination (40). On average, the antimicrobial
activities were found to be in the following order, P. flabellifolia > M.
communis > T. vulgare. In case of P. flabellifolia, results from
the disc diffusion method indicate that Bacillus subtilis is the most
sensitive microorganism, showing the highest inhibition zone, which is
approximately three times larger than that of gentamicin. This followed by
antimicrobial activity on Pseudomonas aeruginosa with T. vulgare
EO. P. aeruginosa infections
leading to mastitis are frequently observed in livestock, including animals
raised in Türkiye (41) . The high activity of T. vulgare
EO observed in the species could be considered a scientific basis for the
traditional application of farmers administering the plant to cattle (38). Similarly, studies conducted on Candida
albicans, a species causing mastitis in dairy cows, also confirm the high
efficacy of Pimpinella EOs (42).
Molecular Docking Results
According
to the results of in silico molecular docking analysis (Figure 2),
alpha-terpinyl acetate (Pubchem ID: 111037) from M. communis and
spathulenol (Pubchem ID: 92231) from P. flabellifolia exhibited the
highest scores for both receptors.
Binding
affinities for the dopamine receptor were -5.4 kcal/mol for alpha-terpinyl
acetate and -5 kcal/mol for spathulenol (Figure 3). Likewise, for the
progesterone receptor, the binding affinities were -5.3 kcal/mol for
alpha-terpinyl acetate and -5 kcal/mol for spathulenol (Figure 4). For dopamine
receptors, alpha-terpinyl acetate exhibited pi-sigma interactions at TYR71,
formed a carbon-hydrogen bond at THR153, and engaged in alkyl interactions at
VAL78, ILE156, and TRP160 residues. Additionally, Van der Waals interactions
were observed at VAL74, SER75, and VAL152 residues. Conversely, spathulenol
formed a carbon-hydrogen bond at THR153 and an alkyl interaction at ILE156.
Furthermore, Van der Waals interactions occurred at TYR71, VAL74, SER75,
VAL152, ALA157, and TRP160 residues.
Table 4. Carcinogenicity and mutagenicity prediction results.
CarcinoPred
-EL Method
Average
Predicted
Result
Mutagenicity
Prediction
T. vulgare Volatiles
β-Thujone
XGBoost
0.59
Non-Carcinogen
non-mutagenic
beta-Cymene
XGBoost
0.36
Non-Carcinogen
non-mutagenic
bornyl
acetate
XGBoost
0.49
Non-Carcinogen
non-mutagenic
P. flabellifolia Volatiles
(E)-
anethol
XGBoost
0.38
Non-Carcinogen
non-mutagenic
d-limonene
XGBoost
0.47
Non-Carcinogen
non-mutagenic
(-)-α-Pinene
XGBoost
0.48
Non-Carcinogen
non-mutagenic
M. communis Volatiles
1,8-Cineole
XGBoost
0.48
Non-Carcinogen
non-mutagenic
linalool
XGBoost
0.39
Non-Carcinogen
non-mutagenic
thymol
XGBoost
0.46
Non-Carcinogen
non-mutagenic
On the other hand, when examining the interactions with progesterone receptors, it was observed that alpha-terpinyl acetate established conventional hydrogen bonds specifically at the HIS71 residue, engaging in alkyl interactions at both LYS12 and LEU15 residues. Additionally, this compound exhibited Van der Waals interactions with residues such as LEU16, ASN18, LEU70, VAL74, and HIS78. In stark contrast, spathulenol, another compound under scrutiny, demonstrated its ability to form conventional hydrogen bonds, primarily at the LEU15 residue, and also showed Van der Waals interactions with residues such as LEU16, ASN18, LEU70, VAL74, HIS71, and HIS78.
As depicted in Figures 3 and 4, the Bos taurus dopamine receptor demonstrates relative hydrophilicity at the binding pocket, whereas the progesterone receptor is even more hydrophilic than the dopamine receptor. This discrepancy may contribute to differences in the residue interaction types and distances for the same ligands. In addition, reference molecular docking analyses were conducted using dopamine for the dopamine receptor and progesterone for the progesterone receptor, yielding binding affinities of -4 kcal/mol and -6.8 kcal/mol, respectively. This comparative study highlights higher values obtained for the dopamine receptor, whereas the values obtained for the progesterone receptor are considered acceptable compared with those obtained for the progesterone molecule, which serves as the receptor’s natural ligand. The results obtained from our analyses support the use of various parts of the M. communis plant as veterinary supplements for ewes, goats, and dairy cows for different purposes, which is consistent with the literature (43, 44). Considering the limited literature available due to its endemic nature, it is possible to establish the consistency of our results for P. flabellifolia by drawing parallels with other plant species within the Pimpinella genus used for similar purposes as veterinary supplements (10, 45).
Mutagenicity and Carcinogenicity Predictions
The computational analysis results regarding the probability of 3 highly abundant volatiles from the Eos being mutagenic and carcinogenic are presented in Table 4.
The
average values calculated using the XGBoost method for highly carcinogenic
compounds are close to 1, whereas the values obtained for these volatile compounds
remain below the average carcinogenicity (0.5) (26). Most of these bioactive compounds,
currently utilized in the pharmaceutical and cosmetic industries, as evident
from the results, do not possess carcinogenic or mutagenic effects at moderate
concentrations (46). In addition, based on predictions
conducted on alpha-terpinyl acetate and spathulenol using data obtained from
molecular docking results, no carcinogenic or mutagenic properties were
detected.
ADMET Predictions
Estimation
of various ADMET properties of the volatiles were given in the Table 5.
LogP values measure the hydrophilicity or lipophilicity of a compound.
Compounds with high logP values are categorized as lipophilic (hydrophobic),
whereas those with low values are considered hydrophilic (47). Because all samples in the results
are EOs, the volatile components are found moderately hydrophobic. These
findings support the hypothesis that these components exhibit antimicrobial
properties by disrupting the structure of bacterial cell membrane lipids (48). Upon examining the average logP
values, it is evident that the values obtained for the three compounds, which
constitute over 50% of the EO structure, demonstrate the highest hydrophobicity
for P. flabellifolia. In addition, antimicrobial analysis results
indicated that P. flabellifolia exhibited the highest activity. When
evaluated in terms of BBB permeability, once again, P. flabellifolia has
been found to exhibit high content and potency in terms of both impact speed
and strength. Research on other Pimpinella species regarding BBB permeability
also emphasizes their rapid and potent efficacy (49). CNS permeability refers to the
ability of a compound to penetrate the BBB and reach the tissues of the central
nervous system. The average value of -2 obtained in the results indicates low
CNS permeability (50). From this, it is understood that
the application of essential oils carries a low probability of causing
significant neurological effects or complications. Finally, it was observed
that P. flabellifolia has the shortest clearance time among the applied
essential oils. This suggests that the EO may remain in circulation for a
longer period, potentially leading to an extended bioterapeutic effect.
Table 5. ADMET prediction results for three tested plants.
LogP
Water solubility
(log mol/L)
Intestinal absorption (%)
BBB permeability
CYP1A2 inhibitor
CNS permeability
Total Clearance
(log ml/min/kg)
Hepatotoxicity
T. vulgare Volatiles
β-Thujone
2.2
-2.9
98.1
0.7
Yes
-1.9
0.13
No
beta-Cymene
3.1
-4
93.6
0.4
Yes
-1.3
0.2
No
bornyl
acetate
2.7
-3.03
95.3
0.5
Yes
-2.3
1
No
P. flabellifolia Volatiles
(E)-
anethol
2.7
-2.9
95.5
0.5
Yes
-1.6
0.2
No
d-limonene
3.3
-3.5
95.8
0.7
Yes
-2.3
0.2
No
(-)-α-Pinene
2.9
-3.7
96
0.7
Yes
-2.2
0.04
No
M. communis Volatiles
1,8-Cineole
2.7
-2.6
96.5
0.3
Yes
-2.9
1
No
linalool
2.6
-2.6
93.1
0.6
Yes
-2.3
0.5
No
thymol
2.8
-2.7
90.8
0.4
Yes
-1.6
0.2
Yes
Conclusion
This
study aimed to assess the biological and biotherapeutic activities of the EOs
from Tanacetum vulgare L., Myrtus communis L. subsp. communis L.,
and Pimpinella flabellifolia (Boiss.) Benth. Et Hook. ex Drude medicinal
plants. GC/MS analysis results indicate that all three plants are rich in
bioactive volatile components. According to computational prediction results,
none of the components exhibit carcinogenicity or mutagenicity properties.
Additionally, the antimicrobial results obtained, along with the ADMET
properties, suggest that Pimpinella flabellifolia is the strongest plant
in terms of biotherapeutic effect. Additionally, in silico analysis suggested
that these plant species could be utilized as veterinary supplements for dairy
cows, potentially enhancing lactation and overall well-being. Nevertheless,
further studies are necessary to investigate in vitro and in vivo activity of
lactating performance and other behaviors.
This study aimed to assess the biological and biotherapeutic activities of essential oils derived from the medicinal plants Tanacetum vulgare L., Myrtus communis L. subsp. communis L., and Pimpinella flabellifolia (Boiss.) Benth. Et Hook. ex Drude. Plant samples were systematically collected from the Sivas region of Türkiye. Subsequently, essential oils were extracted using a Clevenger-type apparatus, and their compositions were assessed by gas chromatography–mass spectrometry (GC-MS) analysis. Then, antioxidant activities of the essential oil samples were investigated using β-carotene-linoleic acid and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Furthermore, the antimicrobial activity of these species was assessed via the disc diffusion assay. Finally, the potential effects of the essential oil compositions from these plants on milk production in dairy cows were analyzed through in-silico methods.
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