**11.2 Essential oils of T. broussonetii and T. maroccanus**

*Thymus* (thyme) is an aromatic and medicinal plant belonging to the *Lamiaceae* family. This plant is native to the Mediterranean region. (Duke, 1989; Zargari, 1990; Newall et al., 1996). *Thymus broussonetii* Boiss and *Thymus maroccanus*. are an endemic plants of Morocco. Its species are used for traditional medicine for the treatment of various illnesses (Bellakhdar, 1997; Sijelmassi, 1993). The studies of essential oil of *T. broussonetii* have been published indicating the antimicrobial, anti-inflammatory, the immunological, behavioural effects and antitumor properties (Lattaoui et al., 1994; Lattaoui and Tantaoui-Elaraki, 1994, Ismaili et al. 2001; 2002; 2004, Elhabazi et al., 2006, Jaafari et al, 2007).

*T. broussonetii* and *T. maroccanus* were collected at flowering stage in July 2006 in a High Atlas mountain respectively, in "Ait Ourir" and in "Essaouirra" from the region, Centre and Southwest of Morocco. The inflorescences, leaves and stem were separated by hand. Samples were dried at the shade.

#### **11.3 Chemical composition of essential oils of T. broussonetii and T. maroccanus**

The identification of 93.1% (36 compounds) of the chemical composition of the essential oil of some leaf of *T. broussonetii* (figure 7) is shown in Table 2. This part produces a yield of 1.6% essential oils. The chemical identity of this oil shows that it consists mainly of monoterpene hydrocarbons which represent more than half of this oil (53.3%). In this class of monoterpenes, p-cymene (21.0%), the α-pinene (11.8%) and camphene (8.5%) are the most important compounds. The class of oxygenated monoterpenes is second with 33%, represented mainly by borneol (16.5%) and thymol (11.3%). The sesquiterpene hydrocarbon represent 6.6% and consist mainly leden (3.2%). The spathulénol is the only oxygenated sesquiterpene identified in this part of the plant with a percentage of 0.2%.

The identification of 96.7% of the chemical composition of the essential oil of some leaf of T. maroccanus (figure 8) is shown in Table 3. The yield of essential oils of this part is 1%. The monoterpene hydrocarbon species represent 49.5% of the leaves of this species. The αpinene (11.6%) and p-cymene (25.3%) are the most important compounds in this class. The oxygenated monoterpenes represent 37.9%, they consist mainly of carvacrol (33%). The sesquiterpene hydrocarbon represents 6.6% and consists of 2% β-caryophyllene. The oxygenated sesquiterpenes represented only 0.4% of this oil.

Fig. 7. *T. broussonetii*

cultures by 18 h, diluted with saline so as to contain about 108 cells / ml, at a density of between 0.08 and 0.1 at 625 nm (Careaga and al. 2003; Joffin and Leyral,2001). We evaluated the sensitivity of this strain against the essential oils of *thymus broussonetii* and *thymus maroccanus* from the region of Marrakech (Morocco). We used the Mueller Hinton medium for our study. The antimicrobial activity of essential oils, we used the method of diffusion from antibiotic susceptibility discs of pure essential oil (Jacob et al., 1979). The media poured in Petri dishes are inoculated with 1 ml of bacterial suspension of 108 cells / ml and excess inoculum was removed by aspiration (Shunying et al., 2005). The essential oil is deposited in the volume of 10 ml on Whatman sterile paper discs of 6 mm in diameter. In parallel, we use cookies to check the growth of the strain tested. The petri dishes were left 30 min at room temperature to allow complete diffusion of the product (CA-SFM, 1993). The antibacterial activity was determined in terms of diameter of inhibition zone around the discs recorded after 24 h of incubation at 37 ° C. The test is

*Thymus* (thyme) is an aromatic and medicinal plant belonging to the *Lamiaceae* family. This plant is native to the Mediterranean region. (Duke, 1989; Zargari, 1990; Newall et al., 1996). *Thymus broussonetii* Boiss and *Thymus maroccanus*. are an endemic plants of Morocco. Its species are used for traditional medicine for the treatment of various illnesses (Bellakhdar, 1997; Sijelmassi, 1993). The studies of essential oil of *T. broussonetii* have been published indicating the antimicrobial, anti-inflammatory, the immunological, behavioural effects and antitumor properties (Lattaoui et al., 1994; Lattaoui and Tantaoui-Elaraki, 1994, Ismaili et al.

*T. broussonetii* and *T. maroccanus* were collected at flowering stage in July 2006 in a High Atlas mountain respectively, in "Ait Ourir" and in "Essaouirra" from the region, Centre and Southwest of Morocco. The inflorescences, leaves and stem were separated by hand.

**11.3 Chemical composition of essential oils of T. broussonetii and T. maroccanus**  The identification of 93.1% (36 compounds) of the chemical composition of the essential oil of some leaf of *T. broussonetii* (figure 7) is shown in Table 2. This part produces a yield of 1.6% essential oils. The chemical identity of this oil shows that it consists mainly of monoterpene hydrocarbons which represent more than half of this oil (53.3%). In this class of monoterpenes, p-cymene (21.0%), the α-pinene (11.8%) and camphene (8.5%) are the most important compounds. The class of oxygenated monoterpenes is second with 33%, represented mainly by borneol (16.5%) and thymol (11.3%). The sesquiterpene hydrocarbon represent 6.6% and consist mainly leden (3.2%). The spathulénol is the only oxygenated

The identification of 96.7% of the chemical composition of the essential oil of some leaf of T. maroccanus (figure 8) is shown in Table 3. The yield of essential oils of this part is 1%. The monoterpene hydrocarbon species represent 49.5% of the leaves of this species. The αpinene (11.6%) and p-cymene (25.3%) are the most important compounds in this class. The oxygenated monoterpenes represent 37.9%, they consist mainly of carvacrol (33%). The sesquiterpene hydrocarbon represents 6.6% and consists of 2% β-caryophyllene. The

sesquiterpene identified in this part of the plant with a percentage of 0.2%.

oxygenated sesquiterpenes represented only 0.4% of this oil.

performed in three repetitions in the same experimental conditions.

**11.2 Essential oils of T. broussonetii and T. maroccanus** 

2001; 2002; 2004, Elhabazi et al., 2006, Jaafari et al, 2007).

Samples were dried at the shade.

Fig. 8. *T. maroccanus*

Use Thyme Essential Oils for the Prevention of Salmonellosis 321

**Monoterpenes hydrocarbon 49.5**  Tricyclene 921 1001 tr α-Thujene 924 - 0.8 α-Pinene 934 1012 11.6 Thuja-2,4(10)-diene 937 1112 0.1 Camphene 944 1051 0.8 β-Pinene 971 1093 0.4 Myrcene 984 1141 1.4 α-phellandrene 998 1185 0.3 α-terpinene 1010 1158 1.1 *p*-cymene 1017 1244 25.3 Limonene 1024 1176 2.7 γ-Terpinene 1052 1220 4.6 Terpinolene 1079 1254 0.1 **Oxygenated Monoterpenes 37.9**  (E)-Sabinene hydrate 1054 1423 0.2 Linalol 1085 1510 2.3 Camphre 1122 1466 0.2 Borneol 1148 1651 0.8 Terpinen-4-ol 1161 1554 0.5 Dihydrocarvone 1 1171 1557 0.3 Dihydrocarvone 2 1179 1579 0.2 Carvone 1233 1676 0.2 Thymol 1268 2124 0.4 Carvacrol 1280 2151 33.0 Eugenol 1328 - tr **Sesquiterpenes hydrocarbon 6.6**  α-Cubebene 1348 1431 tr α-ylangene 1370 1461 tr Copaene 1375 1463 0.1 β-Bourbonene 1382 1488 0.1 β-Patchoulene 1386 - tr β-caryophyllene 1416 1558 2.0 β-Cubebene 1425 1667 tr Aromadendrene 1437 1570 1.4 α-Humulene 1449 - 0.1 allo-Aromadendrene 1456 1605 0.2 D-Germacrene 1474 1667 tr β-Gurjunene 1484 1681 0.2 Ledene 1491 1655 1.0 γ-Cadinene 1504 - 1.3 Calamenène 1508 1784 0.1 (Z)-α-Bisabolène 1532 - tr Guaiazulène 1652 - tr

IRa IRb %


IRa: Index of retention in non-polar chromatographic column HP-1 IRb: Index of retention in the polar chromatographic column HP-20 tr: trace

Table 2. Percentage and chemical composition of the EO part of the leaf of *T. broussonetii* 

**Monoterpenes hydrocarbon 53.3**  Tricyclene 921 1001 0.3 α-Thujene 924 - 0.3 α-Pinene 934 1012 11.8 Thuja-2,4(10)-diene 937 1112 0.1 Camphene 944 1051 8.5 Sabinene 966 1105 0.2 β-Pinene 971 1093 1.9 Myrcene 984 1141 2.1 α-phellandrene 998 1185 0.2 3-δ-carene 1006 1128 0.1 α-terpinene 1010 1158 1.0 *p*-cymene 1017 1244 21.0 Limonene 1024 1176 2.2 γ-Terpinene 1052 1220 2.5 Terpinolene 1079 1254 0.2 **Oxygenated Monoterpenes 33.0**  (E)-Sabinene hydrate 1054 1423 0.3 Linalol 1085 1510 0.2 Camphre 1122 1466 0.1 Isoborneol 1140 1651 tr Borneol 1148 1651 16.5 Terpinen-4-ol 1161 1554 0.4 Dihydrocarvone 1 1171 1557 0.4 Dihydrocarvone 2 1179 1579 0.1 Carvenone 1233 1676 tr Thymol 1268 2124 11.3 Carvacrol 1280 2151 3.7 **Sesquiterpenes hydrocarbon 6.5**  α-Cubebene 1348 1431 tr β-Bourbonene 1382 1488 0.1 α-Gurjunene 1408 1497 0.1 β-caryophyllene 1416 1558 0.3 Aromadendrene 1437 1570 2.1 allo-Aromadendrene 1456 1605 0.4 γ-Muurolene 1469 1683 0.2 Ledene 1491 1655 3.2 Calamenene 1508 1784 tr **Oxygenated Sesquiterpènes 0.2**  Spathulenol 1561 2059 0.2

IRa: Index of retention in non-polar chromatographic column HP-1 IRb: Index of retention in the polar chromatographic column HP-20

Table 2. Percentage and chemical composition of the EO part of the leaf of *T. broussonetii* 

tr: trace

IRa IRb %


Use Thyme Essential Oils for the Prevention of Salmonellosis 323

(Shunying et al., 2005). They are able to disintegrate the bacterial cell membrane (Ultee et al., 1999). The membrane loses its structure and becomes more permeable to ions (Lambert et al., 2001). Damages to the cell membrane may also allow the dissipation of pH gradient and

The study by (Imelouane & ElBachiri, 2010) of the antibiotic susceptibility shows that the strain used is relatively sensitive to Florenfinicol, Spectinomycin, Enrofloxacin, Cotrimoxazole, Flumequine Tiafen, Tetracycline and Gentamycin. This sensitivity varies

*Salmonella sp* 0 0 0 30 0

*Salmonella sp* 28 25 0 32 18 34

UB: Fluméquine ,SXT: Cotrimoxazole; MY:Lyncomycine ; ENR:Enrofloxacine ; SH : Spectinomycine;

Table 5. Inhibitory effect of different antibiotic discs: Diameter of inhibition zones in mm

The study of antibiotic susceptibility shows that the strain used is sensitive to Florenfinicol, Spectinomycin, Enrofloxacin, Flumequine, Tetracycline and Gentamycin. This sensitivity varies from one antibiotic to another (Table), while the strain is resistant to Cephalothin,

Quantitative comparison of the results of EO to antibiotics is difficult because the nature of the activity and composition of the molecules are not comparable. Unlike EO, complex mixtures of volatile compounds that evaporate, but also that diffuse into the agar at different speeds, antibiotics are large non-volatile molecules. There Dissemination takes

microorganisms Thymus pubescens

*Salmonella sp* 08

Table 6. Antibioaromatogramme of the essential oil of *Thymus pubenscens*. Diameter of inhibition zone of *Salmonella sp* determined using the diffusion method on solid medium.

Inhibition Zone (mm)

Kf: Cephalothin; AMP: Ampiciline; AML:Amoxycilline; TPC:Tiafen; E:Erythromycine

place probably at the surface and / or volume in the mass of the agar.

Inhibition Zone (mm) antibiotics KF30 AMP10 AML25 TPC30 E15

Inhibition Zone (mm) antibiotics UB30 SXT25 MY ENR5 SH FFC

decreased membrane potential (Ultee et al., 1999).

from one antibiotic to another and from one strain to another

**11.5 The antibacterial activity of antibiotics** 

FFC: Florenfinicol

among Salmonella sp.

Ampicillin, Amoxicillin and Lyncomycine.


IRa: Index of retention in non-polar chromatographic column HP-1

IRb: Index of retention in the polar chromatographic column HP-20, tr: trace

Table 3. Percentage and chemical composition of the HE part of the leaf of *T. maroccanus*
