

Recently the high concern of the eco-friendly and healthy means used in the improvement of animal production resulted in banning the use of antibiotics in animal feeding in 2006 by the European Union1 due to their potential residues in milk and meat, meanwhile, their effects consequently on human health. So, there has been an increasing interest to search for natural alternatives for growth promoters and feeding antibiotics in ruminant nutrition such as medicinal plants and their essential oils2-4. Essential Oils (EO) are a mixture of volatile aromatic secondary metabolites that are extracted from plants by steam and/or water distillation5. Santos et al.6 recorded an increase in milk fat content with feeding EO mixture to dairy cows. However, the inclusion of EO increased milk production decreased somatic cell count and did not affect the milk composition of dairy ewes7.
Thyme is planted worldwide for many uses such as culinary, medical and cosmetic perennial purposes. Thyme is considered as antispasmodic, expectorant, antiseptic, antimicrobial and antioxidant8. Thymol, a phenolic monoterpene [(5-methyl-2-(1-methyl ethyl) phenol)] is one of the major compounds of thyme (Thymus vulgaris) and organo (Origanum vulgaris) essential oils9. The strong antimicrobial activity of thymol against a wide range of gram-positive and negative bacteria10 is attributed to its ability in plasma membrane disruption of bacteria and the reduction in glucose uptake5,11. Salamataza et al.12 suggested that thyme methanolic extract can affect fermentation efficiency in the rumen and could be used as a mitigating agent of methane. Ebrahimi et al.13 found that the addition of thyme (Thymus vulgaris) oil to the diet of Brown Swiss Neonatal calves improved nutrient digestibility of DM and OM, with no significant effect on feed intake and feed conversion ratio. Also, no significant effects on DMI and feed efficiency with thyme oil supplementation were observed with feedlot Holstein calves14 or with sheep15. While El-Naggar et al.16 indicated an improvement in digestibility and feed efficiency of growing lambs with thyme oil supplementation. In contrast, there were no significant effects of thyme leaves addition at 3% of DM on nutrient digestibility of lambs17. Boutoial et al.18 reported that using thyme leaves in rations of dairy goats significantly increased the milk contents of fat, protein and PUFA. In the same trend, El-Essawy et al.19 recorded increases in milk TS, the total antioxidant capacity and concentration of USFA profile in the milk of Barki ewes fed the diet supplemented with thyme EO.
The objectives of this study were to investigate the effects of thyme addition as leaves or essential oil on digestibility, nutritive value, milk production and its composition, growth performance of kids, ruminal fermentation characteristics and blood metabolites of Damascus lactating goats.
Study area: The field trial of the current study was carried out at Sheep and Goats Unit, Agriculture Experimental and Research Station, Faculty of Agriculture, Cairo University through the period from January to Mach, 2019. While the lab analysis extended to August 2020.
Ethical approval: The protocol of the present study was accepted by the Institutional Animal Care and Use Committee, Cairo University (IACUC) (Approval No. CU/II/F/22/21).
Thyme leaves and essential oils: Thyme (Thymus vulgaris) leaves were obtained from Beni Mazar districts, Minia Governorate, Egypt. The fresh leaves were dried at shadow till drying, then it was saved in polyethylene bags. A hydro distillation method by Clevenger type apparatus was used to extract the essential oil from fresh thyme leaves as described by Vakili et al.14. Then the collected oil was dehydrated by using anhydrous sodium sulfate. Oil percentage was calculated (1.65%) and stored in the fridge until using. The bioactive components in the thyme oil were determined by Gas Chromatography-Mass Spectrometry (GC-MS) instrument (a trace GC Ultra Gas Chromatographs (THERMO Scientific Corp., USA), coupled with a THERMO mass spectrometer detector (ISQ™EC Single Quadrupole Mass Spectrometer) at the Department of Medicinal and Aromatic Plants Research, National Research Center.
Thyme in the current study contained 42.70% Thymol, 2.93% Carvacrol, 17.41% p-Cymene, 15.62% γ-Terpinene, 4.25% α-Terpinene, 1.65% Myrcene, 2.01% Linalool, 3.30% α-Pinene, 1.91% β-Pinene, 3.72% β-Caryophyllene and 4.50% Limonene.
Experimental animals and rations: Thirty-six multiparous lactating Damascus goats, 4 years old and an average weight 35 kg, after 7 days of parturition were divided into three groups (12 animals each) according to the milk production then, the experimental groups were randomly assigned to receive one of the experimental rations. The experiment lasted for 80 days (30 days adaptation period, milk yield was recorded 4 times with 2 weeks interval, then followed by 4 days for digestion trial and collection of rumen liquor and blood samples). Kids were separated from their mothers and changes in body weight of mothers and kids were determined biweekly.
Table 1: | Chemical composition of the concentrate feed mixture, clover hay and experimental ration |
Feedstuffs | |||
Items | CH | CFM | *Experimental ration |
DM | 91.75 | 90.97 | 91.28 |
Chemical composition (%) (DM basis) | |||
OM | 88.47 | 94.83 | 92.29 |
Ash | 11.53 | 5.17 | 7.71 |
CP | 13.62 | 14.57 | 14.19 |
EE | 1.49 | 2.58 | 2.14 |
CF | 27.37 | 5.98 | 14.54 |
NFE | 45.99 | 71.70 | 61.42 |
Fibre fraction (%) | |||
NDF | 43.83 | 29.87 | 35.45 |
ADF | 31.99 | 11.23 | 19.53 |
ADL | 5.53 | 2.99 | 4.01 |
Cellulose | 26.46 | 8.24 | 15.53 |
Hemi-cellulose | 11.84 | 18.64 | 15.92 |
CH: Clover hay, CFM: Concentrate feed mixture and *Calculated |
Animals in the control group (C) were fed Egyptian clover (Trifolium alexandrinum) plus Concentrate Feed Mixture (CFM) without additives, while the other experimental animals were fed control ration plus 1% thyme leaves of total DM (G1) or the equivalent as thyme essential oil (G2). The equivalent amount of thyme essential oil was based on oil% in thyme which was 1.65%. All animal groups were offered rations at 3% of LBW (60% CFM: 40% Egyptian clover) to cover the total requirements recommended by NRC20. Both thyme leaves (G1) and EO (G2) were mixed with a small amount of CFM daily just before feeding to ensure that animals consumed all amounts. The concentrate feed mixture consists of 68% yellow corn, 13% wheat bran, 17% soybean meal, 0.5% common salt and 1.5% limestone. The chemical compositions of the experimental ratio are shown in Table 1.
Feeding procedures: The concentrate feed mixtures and Egyptian clover were divided into two equal portions fed twice daily at 0800 and 1600 hrs. Freshwater was always available. Minerals blocks were available for all animals.
Milk sampling: Milk yield was recorded and milk samples were collected four times during the collection period for all animals by hand milking twice daily at 7 am and 5 pm. Whereas, one-tenth of the morning and the evening milk yields were mixed for each animal and stored at (-18°C) for further analysis.
Digestion trial and faeces sampling: After the collection period of milk, the digestion trial was performed using six animals from each group for three successive days using the bag technique to determine nutrient digestibility via the Acid Insoluble Ash (AIA) method as described by Lee and Hristov21. Nutrient’s digestibility was calculated according to the following formula:
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A subsample (10%) of total collected faeces was sprayed with 10% sulfuric acid, then the drying process was done at 70°C for 24 hrs. The dried faeces were ground and stored for chemical analysis.
Rumen liquor sampling: Rumen liquor samples were collected at 0, 3 and 6 hrs. post-feeding by stomach tube from the six animals at the end of the faeces collection period. Samples were strained via two layers of cheesecloth, the ruminal liquor pH was determined immediately using a digital pH meter. Rumen fluid samples, after adding ortho-phosphoric acid, were stored using glass bottles at deep freeze (-18°C), then used for the analysis of Total Volatile Fatty Acids (TVFA’s) and Ammonia Nitrogen (NH3-N).
Blood sampling: Blood samples were taken from the jugular vein from the six animals on the last day of the experimental period before the morning feeding in tubes containing Ethylene Diamine Tetra Acetic Acid (EDTA) as an anticoagulant. Blood plasma was obtained by centrifugation of the blood samples at 4000 rpm for 20 min and kept at deep freeze (-18°C) for further analysis.
Analytical procedures
Feeds and faeces analysis: Chemical analysis of feedstuffs and faeces samples was carried out according to AOAC22. The Nitrogen Free Extract (NFE) was calculated by difference. Fibre fractions were determined in feeds and faeces according to Van Soest et al.23.
Rumen liquor analysis: Values of rumen pH were determined using Hanna digital pH meter. The concentration of Ammonia-Nitrogen (NH3-N) in the rumen liquor was determined by the Kjeldahl distillation method22. Rumen total volatile fatty acids were estimated by steam distillation method as recommended by Wang et al.24.
Blood plasma analysis: Blood plasma total protein and creatinine were measured as mentioned by Tietz25. Blood plasma albumin was determined by the method of Doumas et al.26. Blood plasma urea, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total triglycerides, total cholesterol and glucose were estimated according to Reed27.
Milk analysis: Total solids, fat, protein and lactose of milk samples were analyzed by infrared spectrophotometer (Foss Matic 120 Milko-Scan, Foss Q3 183 Electric, Hillerød, Denmark) as described by AOAC22. Solids not fat content of milk was calculated by the difference between total solids and fat content. Milk fatty acids profile was determined via High-Pressure Liquid Chromatography (HPLC) according to AOAC28.
Statistical analysis: The general linear model procedure of SAS29 was used for data analysis. Data of nutrients digestibility, blood and milk were analyzed using one way ANOVA procedure by following the next model:
Yij = μ+Rij+Eij |
Where:
μ | = | Overall mean of Yij |
Rij | = | Treatment effect |
Eij | = | Experimental error |
While two-way ANOVA procedure was used to analyse the data of pH, NH3-N and TVFA according to the following model:
Yij = μ+Li+Tj+[L×T]ij+Eij |
Where:
Yij | = | Experimental observation |
μ | = | General mean |
Lj | = | Effect of time |
Tj | = | Effect of experimental rations |
[L×T] | = | Interaction effect due to time and experimental rations |
The differences among means were separated according to Duncan’s New Multiple Range Test.
Nutrient’s digestibility and nutritive value: Data concerns nutrients digestibility and nutritive value in Table 2 indicated significant increases (p<0.05) in digestibility of DM (73.09 and 72.14%), OM (76.10 and 74.62%) and CP (72.59 and 73.39%) with G1 and G2, respectively compared with C (70.48, 73.75 and 70.69%), in the same order. Mirzaei-Aghsaghali et al.8 explained the digestion improvement with thyme EO addition by the positive effect of herbs and spices and their active compounds on activation of ruminal bacteria that are involved in the digestion process or by stimulation the secretion of bile salt, saliva or other digestive enzymes19.
Table 2: | Effect of thyme leaves or essential oil addition on nutrients digestibility and nutritive value |
Experimental groups | |||||
Items | C | G1 | G2 | ±SE | p-value |
Apparent digestibility (%) | |||||
DM | 70.48b | 73.09a | 72.14ab | 0.48 | 0.026 |
OM | 73.75b | 76.10a | 74.62ab | 0.75 | 0.037 |
CP | 70.69b | 72.59a | 73.39a | 0.47 | 0.018 |
CF | 65.89a | 63.23b | 61.56c | 0.67 | 0.002 |
EE | 72.76 | 72.48 | 71.22 | 0.38 | 0.227 |
NFE | 76.35b | 80.09a | 78.11b | 0.89 | 0.01 |
Fiber fractions | |||||
NDF | 62.43a | 60.16b | 59.48b | 0.53 | 0.027 |
ADF | 63.01 | 62.57 | 61.12 | 0.41 | 0.136 |
Nutritive values (%) | |||||
TDN | 70.01b | 72.18a | 70.77ab | 0.71 | 0.099 |
DCP | 10.03 | 10.3 | 10.41 | 0.29 | 0.891 |
Different alphabetic letters means in the same row with various superscripts are significantly different (p<0.05), C: (Control group) goats fed CFM+clover hay, G1 and G2: Control group supplemented with thyme leaves or essential oil, respectively, TDN: Total digestible nutrients and DCP: Digestible crude protein |
Also, this improvement could be due to the anti-microbial properties of the main active components identified in the thyme EO such as thymol, carvacrol, pinene, p-cymene and limonene, which inhibit some microbial activity and stimulate the digestion process30,31.
In the same trend, El-Naggar et al.16 and Ebrahimi et al.13 reported increases in DM and OM digestibility when thyme EO was added to lambs or calves’ rations, respectively. The same result was obtained by Mehrabadi et al.32 with in vitro DM and OM degradability when thyme leaves were added by 5, 10 and 15% of concentrate DM. However, in other studies, neither addition of thyme leaves33 nor thyme EO15,19 had a significant effect on DM and OM digestibility. In the same context, no significant effect of thyme EO was detected on in vitro DM34 or OM degradability35. The variations in the digestibility results with EO addition could be attributed to many factors such as dose, chemical composition, source of plant, extraction procedures of EO14.
The improvement in CP digestibility may be due to the anti-microbial effect of some phenolic compounds in EO such as thymol, which can modulate the microbial population in the rumen, which reduce the degradation of dietary protein, increases N escape and increase the protein flow to the lower gut, which result in improvement of nitrogen utilization33,36.
In the same trend, significant16,33 or insignificant increases19 were recorded in CP digestibility either with thyme leaves or EO addition.
In contrast, a significant reduction in CF digestibility was observed with thyme addition especially with EO being 61.56% compared with control being 65.89%. The same result was obtained with NDF digestibility, with no significant effect on ADF digestibility because of thyme addition. The NDF digestibility was reduced by 3.6 and 4.7% with G1 and G2, respectively. The decrease in CF and fibre fraction digestibility may be attributed to the sensitivity of fibrolytic bacteria to phenolic compounds as thymol found in thyme EO34. Also, Cieslak et al.37 indicated that, thymol may inhibit the growth of certain microorganisms in the rumen, which affects proteolysis and reduce fibre degradation.
In agreement with the previous results, Benchaar et al.34 noted a significant decrease in vitro disappearance of NDF with thyme EO supply. The same results in CF and NDF digestibility were conducted by El-Essawy et al.19 when ewes’ ration supplemented with thyme EO, with no significant effect on ADF digestibility. On the other hand, increases in digestibility of CF16, NDF and ADF13 were reported with thyme EO supply. While Khattab et al.38 observed no significant effect of thyme leaves addition on in vitro disappearance of NDF and ADF.
The highest TDN value (72.18%) was recorded with thyme leaves addition (G1), with no significant difference between G1 and G2 (70.77%). This improvement may be due to the increase in DM and OM digestibility with G1 and G2. Although DCP value was not significantly affected by thyme addition, there was an improvement especially with EO supply because of digestibility improvement of CP. The DCP values were 10.03, 10.30 and 10.41% for C, G1 and G2, respectively.
In agreement with our findings, TDN and DCP values were improved either with thyme leaves33 or EO addition16,19.
Rumen liquor parameters: Table 3 indicated no significant (p<0.05) effect of sampling time or thyme supplementation either in form of leaves or essential oil on rumen fermentation parameters. The lack of significant effect of essential oils on fermentation characteristics may be explained by the results of McIntosh et al.39 that the active EO dose that can affect ruminal bacteria ranges from 35-360 mg L1, which cannot be achieved by in vivo trial. Also, Cardozo et al.40 and Busquet et al.41 reported many factors that may eliminate the effect of EO on rumen parameters such as inadequate dose, concentrate to roughage ratio and adaption time, as longer adaption time will result in more adaption of rumen microorganisms to EO.
The mean values of ruminal pH ranged from 6.25 for C and G1 to 6.28 for G2. Matching with our results, neither thyme leaves addition17,36,38, nor essential oil14,16,19 had a significant effect on ruminal pH value. However, Benchaar et al.34 recorded a significant increase in vitro pH value with thyme EO supply.
Current data showed insignificant decreases in NH3-N concentrations either at 3 (26.73 and 26.12 mg/100 mL) and 6 hrs (24.65 and 24.04 mg/100 mL) post-feeding or the mean value (24.96 and 24.63 mg/100 mL) with thyme addition (G1 and G2), respectively compared with control (29.66, 25.28 and 26.17, in the same order). Molero et al.42 indicated that, the active components in EO including thymol may reduce proteolysis and degradation of amino acids in the rumen because of reduction either inactivity of ammonia producing bacteria or preventing their growth39. The major mode of action of these active components in herbal plants was suggested by their reduction effect on adhesion sites of ruminal bacteria to feed particles, which reduce the deamination process and NH3 production5,36. In the same context, Evans and Martin43 noted that thymol may affect the energy metabolism of Selenomonas ruminantium and Streptococcus bovis, the major deamination bacteria in the rumen.
Table 3: | Effect of thyme leaves or essential oil addition on rumen parameters of lactating goats |
Experimental groups | ||||||
Items | Sampling time (hrs) | C | G1 | G2 | ±SE | p-value |
pH | 0 | 6.73 | 6.6 | 6.68 | 0.15 | 0.953 |
3 | 5.68 | 5.37 | 5.48 | 0.25 | 0.903 | |
6 | 6.35 | 6.77 | 6.69 | 0.11 | 0.267 | |
Mean | 6.25 | 6.25 | 6.28 | 0.19 | 0.997 | |
NH3-N, mg/100 ml rumen liquor | 0 | 23.58 | 23.5 | 23.72 | 0.41 | 0.982 |
3 | 29.66 | 26.73 | 26.12 | 0.77 | 0.127 | |
6 | 25.28 | 24.65 | 24.04 | 0.45 | 0.601 | |
Mean | 26.17 | 24.96 | 24.63 | 0.67 | 0.675 | |
TVFA, mleq/100 ml rumen liquor | 0 | 9.19 | 9.04 | 8.97 | 0.41 | 0.981 |
3 | 12.28 | 10.89 | 10.54 | 0.63 | 0.561 | |
6 | 8.1 | 7.19 | 7.67 | 0.51 | 0.813 | |
Mean | 9.86 | 9.04 | 9.06 | 0.55 | 0.83 | |
a,b,cMeans in the same row with various superscripts are significantly different (p<0.05), C: (Control group) goats fed CFM+clover hay, G1 and G2: Control group supplemented with thyme leaves or essential oil, respectively |
Patra and Yu44 showed that reducing the deamination of amino acids in the rumen may alter the digestion site of protein to the intestine. In the same line, Bodas et al.45 reported that the decreases in vitro CP disappearance and NH3-N concentration may lead to an increase in the passage of dietary protein to the intestine and improve the nitrogen utilization by ruminants.
In the same trend, significant16,17,35,46 and insignificant19,32,38 decreases in the ruminal concentration of NH3-N were reported with thyme addition. However, Benchaar et al.34, Vakili et al.14, Ahmed et al.47 and Ribeiro et al.15 recorded no significant effect.
Although the ruminal concentration of Total Volatile Fatty Acids (TVFA’s) was not affected by thyme supplementation, there were insignificant decreases with G1 and G2 being 9.04 and 9.06 mL eq/100 mL compared with control being 9.68 mL eq/100 mL. Castillejos et al.48 suggested that, the reduction in TVFAʼs concentration with EO supply may be due to the anti-microbial effect of thymol, which inhibits rumen microorganisms and fermentation process. This anti-microbial activity of thymol may be attributed to the hydroxyl group in its structure, which reduce the membrane integrity of bacterial cells and reduce glucose uptake49-51.
Also, a reduction in vitro TVFAʼs concentration was observed when thyme EO was added by 30, 300 and 600 ppm35 or 100 mL L1 46. On the other hand, TVFA’s concentrations were not significantly changed with thyme leaves or EO supply in many studies14,19,32,34.
Blood parameters: However, Table 4 showed no significant differences among groups in blood concentrations of Total Protein (TP), Albumin (ALB) and Globulin (GLO), there were enhancement in TP (7.35 and 7.39 g dL1) and ALB (4.27 and 4.36 g dL1) values with thyme addition (G1 and G2), respectively compared to 6.93 and 3.81 g dL1 in control. This result may be attributed to the improvement in CP digestibility with thyme groups.
In the same trend, no significant effect was detected in blood TP, ALB with the addition of either thyme EO19 or thyme leaves17. Also, Ebrahimi et al.13 did not find a significant difference in serum TP among dairy calves supplemented either with 1- or 2-ml thyme oil and control group. In contrast, Soltan52 recorded increases in blood TP concentration with EO supplementation at a level of 187 or 281 mg/day.
In the current study, there were decreases (p<0.05) in concentrations of Blood Urea Nitrogen (BUN) with thyme leaves (G1) being 22.36 mg dL1 or EO (G2) supply being 21.09 mg dL1 compared with control being 24.25 mg dL1. The high correlation between BUN concentration and NH3-N concentration in the rumen53,54, may explain the reduction in BUN concentration in the current study by the inhibition effect of EO on ruminal bacteria that is responsible for ammonia production such as Peptostreptococcus anaerobius and Clostridium astyklandy 11.
This reduction in BUN concentration is in accordance with results obtained by El-Essawy et al.19, who recorded insignificant decreases in BUN concentration with the addition of thyme EO to ewes’ ration. On the other hand, Vakili et al.14, Biricik et al.55 and Khamisabadi et al.17 found no significant effect. While Seirafy and Sobhanirad56 reported a significant increase in BUN concentration when 5 mL/day thyme oil was added to Holstein suckling calves.
The present data indicated that blood creatinine and glucose concentrations were not affected by thyme addition. Also, Vakili et al.14 and Khamisabadi et al.17 agree with the previous result. However, a significant increase in glucose concentration was observed by Ebrahimi et al.13 with thyme EO supply.
Table 4: | Effect of thyme leaves or essential oil addition on blood parameters of lactating goats |
Experimental groups | |||||
Items | C | G1 | G2 | ±SE | p-value |
Total proteins (g dL1) | 6.93 | 7.35 | 7.39 | 0.1 | 0.119 |
Albumin (g dL1) | 3.81 | 4.27 | 4.36 | 0.12 | 0.13 |
Globulin (g dL1) | 3.12 | 3.08 | 3.03 | 0.02 | 0.131 |
Urea-N (mg dL1) | 24.25a | 22.36ab | 21.09b | 0.96 | 0.106 |
Creatinine (mg dL1) | 0.82 | 0.89 | 0.91 | 0.03 | 0.492 |
AST (IU L1) | 86.18b | 87.05ab | 88.24a | 0.39 | 0.065 |
ALT (IU L1) | 12.41b | 13.15ab | 13.73a | 0.29 | 0.018 |
Glucose (mg dL1) | 52.19 | 53.54 | 52.87 | 0.34 | 0.305 |
Total cholesterol (mg dL1) | 105.72a | 92.17c | 94.51b | 2.09 | 0.000 |
Total triglycerides (mg dL1) | 19.08a | 15.14c | 15.65b | 0.24 | 0.000 |
Different alphabetic letters means in the same row with various superscripts are significantly different (p<0.05), C: (Control group) goats fed CFM+clover hay, G1 and G2: Control group supplemented with thyme leaves or essential oil, respectively, ALT: Alanine aminotransferase and AST: Aspartate aminotransferase |
Current results revealed that the highest (p<0.05) concentrations of AST and ALT were recoded with thyme EO addition (G2) being 88.24 and13.73 IU L1, with no significant difference between G1 (87.05 and 13.15 IU L1) and G2 compared with control (86.18 and 12.41 IU L1), in the same order.
In the same context, Seirafy and Sobhanirad56 noted increases in AST and ALT concentrations with thyme EO supply. While in the opposite to current findings, no significant effect was detected by Vakili et al.14, Khamisabadi et al.17 and El-Essawy et al.19.
The lowest (p<0.05) Concentrations of Total Cholesterol (CHOL) being 92.17 mg dL1 and Total Triglycerides (TRIG) being 15.14 mg dL1 were observed with thyme leaves-supplemented goats (G1), followed by those supplemented with thyme EO (G2) being 94.51 and 15.65 mg dL1 compared with the control group being 105.72 and 19.08 mg dL1, respectively. Michel et al.57 explained these reductions in blood CHOL and TRIG concentrations by the hypolipidemic and hypercholesterolemia properties of aromatic plants and their EO. Also, it was indicated that the product of herbal plants metabolism may inhibit the production of the enzyme (3-hydroxy-3-methylglutaryl CO. A) that controls the cholesterol synthesis58.
In matching with our results, Morsy et al.59, Khamisabadi et al.17, Seirafy and Sobhanirad56 and El-Essawy et al.19 reported decreases in blood CHOL and TRIG concentrations with thyme leaves or EO addition. In contrast, concentrations of CHOL and TRIG were not affected by the addition of thyme EO in the study conducted by Vakili et al.14.
Milk yield, composition and feed efficiency: Data of Dry Matter Intake (DMI), milk yield and composition, feed efficiency in Table 5 showed an insignificant (p<0.05) increase in DMI with thyme leaves supplementation (1215 g/day), while DMI tended to be decreased insignificantly (p<0.05) with thyme EO supply (1135 g/day) compared with control (1175 g/day). The depression in total DMI with EO supply may have resulted from palatability problems, which require an encapsulation of EO to overcome that problem5. Moreover, Ribeiro et al.15 cleared that mainly composed of EO may be affected by sensory properties such as odour, flavour and acceptability.
In the same trend, Khamisabadi et al.17 recorded an increase in DMI when lambsʼ ration was supplemented with thyme leaves. While decreases in DMI were observed with blend EO addition to dairy cows’ ration6,60,61. In contrast, Seirafy and Sobhanirad56 reported an increase in total DMI with thyme EO addition to Holstein suckling calves. However, no effect of thyme leaves or EO addition on DMI was detected by Vakili et al.14, El-Naggar et al.16, Mohamed et al.33, Ebrahimi et al.13, El-Essawy et al.19 and Ribeiro et al.15. The variation in DMI with EO supply may be attributed to many factors such as EO source, diet type, the adaption of rumen microbes to EO3,62. Furthermore, the chemical and physical characteristics of the diet can affect DMI3,63.
In the current study, the highest (p<0.05) milk and 4% Fat Corrected Milk (FCM) yields were observed with thyme leaves addition (G1) being 1170 and 1133 g/h/day, respectively, with no significant difference between G1 and G2 (1125 and 1059 g/h/day) compared with control (935 and 939 g/h/day), in the same order. The increase in milk yield with thyme addition maybe because of the improvement in DM and OM digestibility. In this context, many theories explain the increase in milk production with thyme addition. One of these theories indicated that the increase in milk production with EO supply may be attributed to the change of ruminal fermentation pattern, through reducing acetate: Propionate ratio5,64 or due to reduction in methane production11.
Table 5: | Effect of thyme leaves or essential oil addition on dry matter intake, milk yield and composition and feed efficiency |
Experimental groups | |||||
Items | C | G1 | G2 | ±SE | p-value |
DMI (g) | 1175 | 1215 | 1135 | - | - |
Milk yield (g/h/day) | 935b | 1170a | 1125a | 38.32 | 0.01 |
4% FCM (g/h/day) | 939b | 1133a | 1059ab | 38.75 | 0.03 |
Milk composition (%) | |||||
Fat | 4.03 | 3.79 | 3.61 | 0.13 | 0.494 |
Protein | 3.26 | 3.51 | 3.6 | 0.12 | 0.533 |
Lactose | 4.77 | 4.68 | 4.78 | 0.15 | 0.965 |
TS | 12.98 | 12.93 | 12.93 | 0.18 | 0.992 |
SNF | 8.95 | 9.14 | 9.32 | 0.11 | 0.49 |
Ash | 0.92 | 0.95 | 0.94 | 0.03 | 0.901 |
Feed efficiency | |||||
Milk yield/DMI (g g1) | 0.80b | 0.96a | 0.99a | 0.03 | 0.009 |
Different alphabetic letters means in the same row with various superscripts are significantly different (p<0.05), C: (Control group) goats fed CFM+clover hay, G1 and G2: Control group supplemented with thyme leaves or essential oil, respectively |
Also, it may be because of stimulating endocrine glands in animals to release some hormones such as growth hormone65, which stimulate milk production by elevating the conversion rate of feed nutrients into milk66. Furthermore, the antioxidant property of the thymol may inhibit the lipid peroxidation of the free radicals, which protect the mammary secretory cells and keep them healthy67 which lead to more milk production.
In parallel to the previous results, Mohamed et al.33 recorded significant increases in milk and FCM yields when dairy goats fed ration supplemented with thyme leaves. Also, higher milk yield was recorded with EO addition to rations of dairy cows61,68 or Chios dairy ewes7. Furthermore, Smeti et al.69 demonstrated higher milk yield with goats supplemented with rosemary leaves than those supplemented with rosemary EO. On the other hand, EO addition had no significant effect on milk yield of lactating goats59, lactating ewes19 and dairy cows6,60,70,71.
Although milk composition was not affected either by thyme leaves or EO supplementation, there were insignificant decreases in fat contents (3.79 and 3.61%) and increases in protein contents (3.51 and 3.60%) with thyme (G1 and G2), respectively compared to 4.03 and 3.26% for control, in the same order. The previous result could be a reflection of the decrease in CF and the increase in CP digestibility (Table 2) and the improvement in blood total protein (Table 4) with thyme addition. Moreover, the reduction in NH3-N concentration in the rumen (Table 3) may be an indicator of improving the synthesis of microbial protein, which reflects on higher milk protein content63.
In matching with current findings, Mohamed et al.33 and El-Essawy et al.19 found that milk fat content was decreased with thyme leaves or EO supply. Also, in coincidence with present results, Morsy et al.59 and El-Essawy et al.19 reported an increase in milk protein with EO addition. On contrary, no change in milk fat content60,61 and protein content6,71 was detected with EO supply. However, Giannenas et al.7 and Flores et al.70 did not record any effect of EO addition on milk composition.
Feed efficiency (milk yield, g/DMI, g) was significantly (p<0.05) improved with thyme leaves and EO addition by 20 and 24%, respectively compared with control.
In agreement with our data, feed efficiency was improved by 16.2% when dairy goats fed ration supplemented with thyme leaves33. Also, it was reported that the supplementation of blend EO improved feed efficiency of Chios dairy ewes7 and dairy cows61,68,71. However, El-Essawy et al.19 found that the addition of thyme EO to ration of lactating ewes did not affect feed efficiency.
Milk fatty acids profile: The milk fatty acids profile in Table 6 indicated significant and insignificant decreases in C6:0 to C15:1, C17:0, C18:0 and 20:0 with thyme addition. However, C16:0, C16:1 and C18:1 to C18:3 were increased with both thyme leaves and EO supply. As a coincidence to the previous results, concentrations of Total Saturated Fatty Acids (TSFA) were decreased (69.55 and 69.52%), while concentrations of Total Unsaturated Fatty Acids (TUFA) being (30.45 and 30.48%) and Total Poly Unsaturated Fatty Acids (TPUFA) being (2.99 and 2.98%) were increased either with thyme leaves or EO addition, respectively compared with control being 71.62, 28.38 and 2.70%, in the same order.
Table 6: | Effect of thyme leaves or essential oil addition on milk fatty acids profile |
Experimental groups | |||||
Items | C | G1 | G2 | ±SE | p-value |
Caproic acid (C6:0) | 1.29 | 1.21 | 1.15 | 0.04 | 0.110 |
Caprylic acid (C8:0) | 3.04a | 2.67b | 2.88ab | 0.16 | 0.029 |
Capric acid (C10:0) | 11.24a | 10.14b | 10.02b | 0.20 | 0.001 |
Lauric acid (C12:0) | 5.96 | 5.78 | 5.82 | 0.14 | 0.879 |
Myristic acid (C14:0) | 12.13a | 11.32b | 11.51ab | 0.32 | 0.069 |
Myristoleic acid (C14:1) | 0.57 | 0.35 | 0.43 | 0.08 | 0.625 |
Pentadecanic acid (C15:0) | 1.35 | 1.24 | 1.21 | 0.07 | 0.770 |
Ginkgolic acid (C15:1) | 0.14 | 0.11 | 0.10 | 0.02 | 0.623 |
Palmitic acid (C16:0) | 24.53 | 27.08 | 26.50 | 0.71 | 0.350 |
Palmitoleic acid (C16:1) | 1.82b | 2.19ab | 2.35a | 0.19 | 0.035 |
Heptadecanoic acid (C17:0) | 0.22 | 0.19 | 0.15 | 0.04 | 0.841 |
Stearic acid (C18:0) | 11.43 | 9.76 | 10.02 | 0.47 | 0.331 |
Oleic acid (C18:1) | 23.15 | 24.81 | 24.62 | 0.37 | 0.124 |
Linoleic acid (C18:2) | 1.49 | 1.71 | 1.68 | 0.05 | 0.214 |
Linolenic acid (C18:3) | 1.21 | 1.28 | 1.30 | 0.05 | 0.788 |
Arachidic acid (C20:0) | 0.43 | 0.25 | 0.26 | 0.07 | 0.528 |
TSFA | 71.62 | 69.55 | 69.52 | 0.82 | 0.547 |
TUFA | 28.38b | 30.45a | 30.48a | 0.45 | 0.065 |
TPUFA | 2.70b | 2.99a | 2.98a | 0.06 | 0.024 |
Different alphabetic letters means in the same row with various superscripts are significantly different (p<0.05), C: (Control group) goats fed CFM+clover hay, G1 and G2: Control group supplemented with thyme leaves or essential oil, respectively, TSFA: Total saturated fatty acids, TUFA: Total unsaturated fatty acids and TPUFA: Total polyunsaturated fatty acids |
Table 7: | Effect of thyme leaves or essential oil addition on kids performance |
Experimental groups | |||||
Items | C | G1 | G2 | ±SE | p-value |
Birth BW (kg) | 3.07 | 3.22 | 3.6 | 0.15 | 0.365 |
Final BW (kg) | 9.32b | 11.58a | 11.22a | 0.41 | 0.03 |
Total BW gain (kg) | 6.25b | 8.36a | 7.62a | 0.38 | 0.022 |
Average daily gain (g) | 83.33b | 111.47a | 101.61a | 4.95 | 0.008 |
Different alphabetic letters means in the same row with various superscripts are significantly different (p<0.05), C: (Control group) goats fed CFM+clover hay, G1 and G2: Control group supplemented with thyme leaves or essential oil, respectively, average daily gain: Total BW gain, g/75 days |
The decrease in TSFA and the increase in TUFA of milk with thyme supplementation may be attributed to the high anti-microbial properties of some phenolic compounds in thyme such as thymol, eugenol and carvacrol, because of the hydroxyl group in their structure49,72 which negatively affect both gram-negative and gram-positive bacteria73 via disturbance in the cytoplasmic membrane10. These compounds can inhibit the rumen microorganisms such as bacteria74, which are involved in the biohydrogenation process of unsaturated fatty acids in the rumen33. Furthermore, Tholstrup et al.75 indicated that, the reduction in saturated fatty acids is correlated with less blood cholesterol, as reported in the present study (Table 4), which is considered as an indicator for more quality and healthy milk to consumers.
In the same trend, milk fatty acids profile either of lactating goats or ewes was improved because of EO supply19,59 or due to thyme leaves addition33.
Kids’ performance: Table 7 revealed no significant difference in birth Body Weight (BW) of kids among different groups. The average values of birth BW were 3.07 kg for C, 3.22 kg for G1 and 3.60 kg for G2. However, final BW (11.58 and 11.22 kg), total BW gain (8.36 and 7.62 kg) and Average Daily Gain (ADG) (111.47 and 101.61 g) were significantly (p<0.05) increased for kids of G1 and G2, respectively compared with those of control group being 9.32 kg, 6.25 kg and 83.33 g, in the same order. These results may be attributed to the improvement in energy metabolism with thyme addition because of the thymol effect on decreasing acetate and decreasing propionate production without affecting total VFA33. Furthermore, residues of EO in milk may improve the appetite and weight of kids due to the digestion stimulant, antioxidant and antiseptic properties of thymol19. Moreover, a positive correlation between the growth rate of kids and milk production of mothers was reported by Smeti et al.69, which matches with higher milk production with thyme addition in the current study.
In the same context, significant increases in final BW and ADG of kids and lambs were observed either with dairy goats supplemented with thyme leaves33 or with lactating ewes supplemented with thyme EO19, in the same order. Moreover, Smeti et al.69 demonstrated that kids of dairy goats who received rosemary leaves had higher final BW and ADG than those who received rosemary EO.
It could be concluded that both thyme forms (leaves or EO) improved digestibility and lactation performance of Damascus goats with no significant difference between the two forms. However, the supplementation of lactating goat's ration especially with thyme leaves recorded the best values of DM and OM digestibility, TDN and kids’ growth rate, the lowest concentrations of blood total cholesterol and triglycerides and the highest milk production with more healthy aspects for consumers.
This study discovered that incorporating thyme (Thymus vulgaris) especially in form of leaves could be used successfully as a natural feed additive that can be beneficial for lactating goats’ performance. Furthermore, it has the potential to produce functional milk with a low concentration of total saturated fatty acids and a high concentration of total unsaturated fatty acids. This study will help the researchers to investigate more natural alternatives for antibiotics.
Wafaa Ghoneem Reply
Is the International Journal of Dairy Science still indexed in Scopus?
Salah El-Din Abd El-moniem Hassan Abo El-Nor Reply
This study included all possible measurements. It is noted that this study is extracted from a master’s or doctoral thesis, and the effort expended in it is valued by the researcher. However, in the methods and materials, the researcher used a statistical analysis to measure the morale of milk and blood measurements as a false scale because the use of One way clacesification was not taken into consideration as a stage Milk and the effect of the interaction between the treatment and the effect of the stage of milk, therefore, this error needs to be corrected, which could change the results obtained.