Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.

55

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Evaluation of Digestarom and thyme as phytogenic feed additives for broiler

chickens
Prfung von Digestarom und Thymian als phytogene Futterzusatzstoffe bei Broilern
1

K.M. Sadek , H.A. Ahmed , M. Ayoub and M. Elsabagh

Dept. of Biochemistry, Nutrition and Vet. Clinical Nutrition, Hyiegene, Faculty of Veterinary Medicine, Damanhour University

Dept. of Nutrition and Vet. Clinical Nutrition, Faculty of Veterinary Medicine, Kafer El-sheikh University, Egypt

Correspondence: ksaadek@yahoo.com
Manuscript received 11 April 2014, accepted 29 May 2014

Introduction
The development and use of growth-promoting additives from natural sources is currently needed due to the growing
consumer awareness about food safety. Phytogenic feed additives or plant extracts have been gaining increased
popularity within the feed industry as possible natural alternatives to antibiotic growth promoters because they are
believed to be safer and healthier with fewer hazards. These phytogenic additives are an extremely heterogeneous
group of feed additives originating from the leaves, roots, tubers or fruits of herbs, spices or other plants. They are
available in either a solid, dried and ground form or as extracts or essential oils. This class of feed additives has
recently gained interest, especially for use in swine and poultry, with promising results regarding the growth
performance and immune response (JAMROZ et al., 2006). The growth performance, feed intake and feed conversion
ratio have been shown to be improved in broilers fed phytogenic additives either separately or in a mixture (ERTAS et
al., 2005). Additionally, the active components of herbs may improve digestion and the antioxidant status and
stimulate immune function in broilers (OUWEHAND et al., 2010). Moreover, numerous reports have demonstrated
the antioxidative, antimicrobial and immune-stimulating efficacy of phytogens in vitro. However, the experimental
in vivo data are still quite limited. The objective of this study was to evaluate the possible effects of a blend of
phytogenic additives (essential oils) and herbal plants on the performance, oxidative status, immunity, carcass
characteristics and intestinal microflora of broilers. Furthermore, a comparison between the effects of Digestarom
1317 and thyme on different nutritional and biochemical parameters was performed.

17.07.2014

Seite 1 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Materials and methods

Birds and dietary treatments

The experiment was conducted in accordance with the animal welfare guidelines of the Faculty of Veterinary

Medicine, Damanhour University, Egypt. One hundred and twenty 1-d-old chicks (Cobb 500) of mixed gender
obtained from a local broiler chicken hatchery were randomly assigned to four treatment groups consisting of three
replicates of ten birds each kept in wire-floored cages. The birds were treated as follows: (1) basal diet without

phytogenic additives (CON), (2) basal diet supplemented with Digestarom 1317 (D1.5) at 1.5g/10kg diet, (3) basal
diet supplemented with Digestarom 1317 at 3.0g/10kg diet (D3) and (4) diet contained thyme (Thymus vulgaris
leaves powder purchased commercially as dried herb supplements) at 100g/10kg diet (T10). Digestarom 1317 (a

blend of 8% peppermint, 2% eugenol or clove, 3.4% anethole or anise and thyme and sodium chloride as a carrier up
to 100%; Micro-Plus Konzentrate GmbH, Germany). The initial temperature of 32C was reduced sequentially
according to the age of the birds until reaching 26C at 21 d. The chicks were kept on a 23-h light program, with free
access to feed and water throughout the experiment. The birds were fed a starter diet until 21 d of age followed by a
finishing diet from d 21 to 35. The ingredients, the chemical composition and essential oils of thyme (Thymus
vulgaris) leaves powder and the basal diet are shown in Tables1 and2. The basal diets were formulated using the
National Research Council (1994) guidelines. The body weight development, feed intake, feed conversion ratio and
relative growth rate (RGR) of the broilers in different groups were recorded weekly.
Table 1. Composition, content of nutrients and essential oils of the used thyme (Thymus vulgaris)
Zusammensetzung und Gehalte an Nhrstoffen sowie essentiellen len des verwendeten Thymians (Thymus vulgaris)
Nutrients

g/kg

DM
Protein

880.0
92.8

Total Fat
Ash
Dietary Fiber

28.1
117.0
233.8

ME (MJ ME/kg diet)

12.9

Essential oils
Carvacrol
Thymol

%
29.9
14.4

-Terpinyl acetate
p-Cymene
Linalool
-Terpinene

10.4
10.1
6.8
3.3

-Caryophyllene
-Terpineol
Geraniol

3.3
3.1
2.9

Calculated according to NRC, 1994

17.07.2014

Seite 2 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Table 2. Composition and nutrient content of experimental diets (g/kg)

Zusammensetzung und Nhrstoffgehalte der Versuchsrationen (g/kg)
Ingredients

Starter

Thyme contained diet

Finisher

Thyme contained diet

Ground yellow corn

SBM (CP 44%)
Fish meal (CP 72.5%)
Soybean oil
Lime stone
Dicalcium phosphate

499.9
368.3
30.0
71.6
12.5
12.7

492.9
366.3
30.0
70.6
12.5
12.7

619.9
277.4
30.0
44.7
10.0
13.0

612.9
275.4
30.0
43.7
10.0
13.0

Salt (Nacl)
DL-Methionine
L-Lysine HCl

2.0
46.0
12.0

2.0
46.0
12.0

2.0
39.0
90.0

2.0
39.0
90.0

2.0

2.0

2.0

2.0

1000.0

10.0
1000.0

1000.0

10.0
1000.0

88.0

90.0

93.0

90.5

Vitamin-mineral premix
Thyme (dried leaves)
Total
2

Moisture

Dry matter

Crude protein

912.0

910.0

907.0

909.5

232.0

230.0

199.0

197.0

45.5

44.3

47.2

46.0

Ash

68.8

70.0

82.7

83.5

12.8

12.8

13.6

13.6

Ether extract

ME (MJ/kg diet)
1

Vitamin-mineral mixture was composed of: Each 3kg contain: retinol 3600 mg; cholecalciferol 50 mg; DL-alpha-tocopheryl acetate 10000 mg; menadione
sodium bisulphite 2000mg, thiamine mononitrate 1000mg, riboflavin 5000mg, pyridoxine 1500mg, cyanocobalamin 10mg, Biotin 50mg, calcium
pantothenate,10000mg, Nicotinic acid 30000mg, Folic acid 1000mg, Manganese 60000mg, Zinc 50000mg, Iron 30000mg, Copper 10000mg, Iodine
1000mg, Selenium 100mg, Cobalt 100mg, carrier (CaCo3) add to 3kg. Vitamin-mineral mixture produced by AGRI-VET 10th of Ramadan city A2, Egypt.
2
Calculated according to the chemical composition of feed components
3
Calculated according to NRC (1994)

Sampling and analyses

Thyme and basal diet sample were analyzed according to A.O.A.C. (1990) methods for dry matter (oven drying at
105C for 24h), crude protein (N-Kjeldahl 6.25) by Vapodest 10, fat (solvent extraction with petroleum ether, by
Soxtherm), ash (muffle furnace incineration at 550C) and crude fiber by fibertherm. Essential oils of thyme were
determined by GC-MS (Agilent Technologies, GS-sampler 80).
Blood samples were collected from the wing vein of experimental birds of each group at 35 days of age without an
anticoagulant for the separation of sera to detect antibody titer against Newcastle disease vaccine using the
Hemagglutination Inhibition (HI) test as an indicator of the birds immune responses in each of the different
experimental groups. The microtechnique of the HI test was performed according to TAKATASY (1955). The geometric
mean titer (GMT) was calculated according to BRUGH (1978).
Another blood sample with anticoagulants (0.1ml of 4% sodium citrate solution/1ml blood) was obtained for the
determination of the total red blood corpuscles (RBCS) and total leukocyte counts (WBCS) by hemocytometer
according to MAXINE and BENJAMIN (1985) and the hemoglobin content (Hb) by Sahlis method (LUCKY, 1977). In
addition, serum samples were evaluated to determine the serum total protein level (PETERS, 1968) and for the serum
albumin level (DOUMAS et al., 1971) using colorimetry. Serum globulin was determined as the difference between the
total protein and albumin (COLES, 1974). The serum ALT and AST activities were determined photometrically
according to the method described by REITMAN and FRANKEL (1957). The ALP activity in serum was determined
according to the method of BELFIELD and GOLDBERG (1971). At the end of the experimental period and after night
fasting, the birds were slaughtered, and the liver, pancreas and RBC hemolysate were collected, perfused with
phosphate-buffered saline solution, pH 7.4 and used for the determination of the following: glutathione peroxidase
activity using cumene hydroperoxide as a substrate (CHIU et al., 1976), glutathione S-transferase activity using a
spectrophotometer at room temperature as the rate of the GSH conjugation of CDNB (HABIG et al., 1974), ascorbic
acid level according to HARRIS and RAYS (1935), nitric oxide level according to MONTGOMERY and DYMOCK (1961) and
the phagocytic activity (PA) and index (PI) according to KAWAHARA et al. (1991). At the end of the experimental

17.07.2014

Seite 3 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

period, five chicks from each group were randomly selected, fasted for 12 hours and weighed. The chicks were then
slaughtered to complete bleeding and weighed to determine the dressing percentage. The liver, spleen, thymus and
bursa were weighed, and the relative weight to body weight of each organ was calculated. The gizzards, heart and
visible fat from each quail chick were also weighed.

At the ages of 14 and 18 days, birds from each replicate (four birds per treatment) were randomly selected and
slaughtered. After separation of the cecum, one gram of cecum content was removed from the bird. To determine the
colony forming units (CFU), one gram of the cecum content in the vicinity of a hot flame was added to 9ml of peptone

water (tube number one), and the solution was shaken. Then, 1ml of solution was added to the next tube (tube
number two) by a sampler containing 9ml of sterile peptone water. This operation was conducted until tube number
eight, and a dilution series was prepared. Dilutions of 10:2, 10:4, 10:6 and 10:8 of the samples were performed
according to MOUNTZOURIS et al. (2007), and the samples were placed on a plate containing eosin methylene blue
agar, McConkey agar and Rogasa medium for the growth of Escherichia coli, coliforms, lactobacillus bacteria, and

lactic acid bacteria; however, nutrient agar medium was prepared for the counting of the total cultured bacteria. This
process was repeated for each sample. Culture McConkey, eosin methylene blue and nutrient agar at 37C for 24h
and Rogasa media. Finally, the samples were evaluated when they reached between 25 and 300 colonies, which was
selected as an appropriate dilution. After counting, the number of colonies was multiplied by the inverse of the
dilution to obtain the number of bacteria.

Statistical analysis
Data were statistically analyzed by GLM fixed model procedures. Differences between mean values were compared by
Duncan's multiple rang test. The analysis of variance for the obtained data was performed using SAS (2002). The
data are expressed as the mean SE. A significant level of 0.05 was used.

Results and Discussion

Growth performance and carcass traits

The effect of essential oils and herb on the body weight gain, feed intake and feed conversion ratio of the broiler
chickens is presented in Table3. Analysis of variance of the obtained data indicated that there is no significant
difference between different experimental groups at the start of the experiment, while at the end of the experiment
chickens receiving the diet containing the Digestarom 1317 at 1.5g/10kg diet (D1.5) had a higher (P<0.05) final
body weight and body weight gain than those of the control and other treated groups. It was observed an
improvement about 8.1% of final body weight in chickens receiving the diet containing the Digestarom 1317 at
1.5g/10kg diet (D1.5) when compared to the control group. While chickens of groups T10 and D3 showed 1.4 and
1% improvement in final body weight when compared to the control group, respectively.
Table 3. Effect of experimental diets on performance of broilers
Einfluss der Versuchsrationen auf die Leistung der Broiler
Treatments
Item
Initial weight (g)
Final weight (g)
Gain (g)
RGR
Feed intake
FCR

CON

D1.5

D3

T10

46.90.16a
173928.9b
169228.8b
1890.16b
2976
1.760.03a

47.20.17a
187930.9a
183230.8a
1920.12a
2852
1.570.03c

47.20.15a
175529.8b
170829.7b
1890.16b
2832
1.670.03b

47.10.17a
176334.7b
171534.6b
1900.18b
2913
1.720.04ab

CON=control, D1.5=basal diet supplemented with Digestarom 1317 at 1.5g/10kg diet, D3=basal diet supplemented with Digestarom 1317 at 3g/10kg
diet, T10=diet contained Thyme at 100g/10kg diet.
Means within the same raw under the same category carry different superscripts are significantly different.

17.07.2014

Seite 4 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

These results are in agreement with those of (ERTAS et al., 2005), who found that EO mix derived from oregano, anise
and clove increased BW gain significantly when compared to the control group. The mixture was tested at different

levels, whereas 200mg/kg of the composition gave best results after 5 weeks of trial.

Additionally, these results are in agreement with those of MUKHTAR (2011), who observed that the addition of

600mg/kg clove oil to the broiler diet improved the weight gain compared with that of the control group by
approximately 2.24% and improved feed intake by 54.8%. Digestarom 1317 contains 2% eugenol (clove), which
has a stimulatory effect on the digestive system.
The active principles of phytogenic feed additives include promoting digestion, balancing the gut microbial
ecosystem and stimulating the secretion of endogenous digestive enzymes, thereby improving the growth

performance of poultry (CROSS et al., 2007). Digestarom 1317 contains 8% peppermint, which may improve the
digestibility of the diet as a digestion stimulant and hence increase the nutrient entry rate at an early stage of the
birds life (OCAK et al., 2008). Additionally, the improvement in the feed intake could be due to the positive effects of

clove oil on the digestive tract, and the high concentration of essential oil in addition to clove oil is super-rich in the
trace mineral manganese, which is necessary for nutrient metabolism and the synthesis of fatty acids and cholesterol
(ERTAS et al., 2005). The absence of an effect of thyme is consistent with the results of CROSS et al. (2003), who
observed no significant difference in the body weight gain between broilers fed a wheat-soybean meal based diet with
or without thyme.

Significant improvement in chicks fed at 1.5g Digestarom 1317/10kg diet (D1.5) when compared to high level of
Digestarom 1317 (D3) may be due to high levels negatively affecting the digestive system. (ERTAS et al., 2005).
Digestarom 1317 and thyme supplementation (groups D1.5, D3 and T10) significantly (P<0.05) improved average
FCR throughout the whole experimental period by about 10.8, 5.1 and 2.3% respectively, when compared to the
control. These results are in agreement with those of ERTAS et al. (2005) who observed a significant improvement
after feeding 200mg/kg of a mixture containing EO from oregano, clove and anise. Also, TOGHYANI et al. (2010)
observed that thyme herb at an inclusion level of 10g/kg decline FCR by approximately 4% when compared with the
control one.

Hematology and immune response

Table4 shows the comparative effects of the Digestarom 1317 supplementations at 1.5 and 3g/10kg diet and
thyme at 100g/10kg diet on HI compared with the ND, phagocytosis, WBC, RBC and Hb of the broiler chickens. The
Digestarom 1317 supplementations at 1.5 and 3g/10kg showed significant differences (P<0.05) compared with
the control diet. Additionally, all treated groups showed a significant difference (P<0.05) in phagocytosis compared
with the control group. These results agreed with those of DURRANI et al. (2008), who found that the hemoglobin
concentration, WBC and serum antibody titer for IB were significantly higher (P<0.05) in the broilers in group fed
mint-supplemented diets compared with the control diet. The higher antibody titer of the treated broilers in the
Digestarom 1317 -supplemented groups could possibly be due to the immuno-stimulant properties of the mint and
eugenol.

17.07.2014

Seite 5 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Table 4. Effect of experimental diets on some haematological and biochemical parameters of broilers at the end of the experiment
Einfluss der Versuchsrationen auf einige hmatologische und biochemische Parameter der Broiler am Versuchsende
Treatments
Item

CON

D1.5

D3

T10

HI
PA
PI
Wbcs
Rbcs
Hb
Total protein(g/dl)
Albumin (g/dl)
Globulin(g/dl)

4.1+0.17b
18.9+0.17c
3.4+0.07a
21.2+0.28
1.8+0.02
8.2+0.16
8.00.03b
4.40.03b
3.30.03b

5.10+0.22a
21.9+0.19a
1.6+0.13b
22.0+0.14
1.8+0.03
8.6+0.18
7.50.02c
4.40.03b
3.30.03b

4.9+0.21a
20.6+0.69b
2.7+0.39a
21.6+0.27
1.8+0.03
8.4+0.26
8.00.03a
4.50.03a
3.40.03ab

4.6+0.33ab
20.3+0.34b
2.9+0.38a
21.3+0.37
1.8+0.02
8.2+0.16
8.00.0.03a
4.60.03a
3.50.03a

GPT
GOT
ALP

59.15.23a
88.55.42a
93.14.12b

51.16.34b
90.57.42a
107.26.12ab

53.35.14b
75.06.57b
118.54.46a

51.27.84b
81.48.40ab
96.85.12b

CON=control, D1.5=basal diet supplemented with Digestarom 1317 at 1.5g/10kg diet, D3=basal diet supplemented with Digestarom 1317 at 3g/10kg
diet, T10=diet contained Thyme at 100g/10kg diet.
HI=haemaglutination inhibition titer, PA=phagocytic activity, PI=phagocytic index, Rbcs =red blood cells, Wbcs =white blood cells
Means within the same raw under the same category carry different superscripts are significantly different.

Biochemical parameters and oxidative status

The present data revealed that the supplementation of broilers with Digestarom 1317 and/or thyme has beneficial
effects, as represented by a significant improvement in the oxidative status in many tissues, including the liver, RBC
hemolysates and pancreas, without any pathological effects on the liver, as reflected by normal parameters related to
liver functions, such as normal protein profiles and enzymatic activities (Tables4,5 and6). The essential oils
considered to be components of Digestarom 1317 have been shown to have antioxidant activity (DEANS et al., 1993)
and to increase the plasma total protein, albumin and immunoglobulin G levels in pigs (YAN et al., 2010).
Table 5. NO (m/l) and ascorbic acid (mg/l) in liver, RBCs' hemolysate and pancreas in chickens fed diet supplemented with phytogenic feed
additives at the end of the experiment
Gehalte an NO (g/l) und Ascorbinsure (mg/l) in Leber, Erythrozyten-Hmolysat und Bauchspeicheldrse der Broiler am Versuchsende in
Abhngigkeit von der Zulage der Zusatzstoffe zum Futter
Treatments
Item

CON

D1.5

D3

T10

1502.88 a
1134.91a
1005.78a

1273.78b
87.25.81b
96.14.35a

1123.78c
86.84.35b
78.74.61b

1402.90a
93.02.30b
1034.35a

Liver
RBCs hemolysate

83.33.46c
45.32.90a

78.63.17c
47.63.17a

1033.48b
51.82.90a

1343.17a
49.43.75a

Pancreas

65.32.90c

68.63.17c

82.82.60b

1132.60a

NO (um/l)
Liver
RBCs hemolysate
Pancreas
Ascorbic acid (mg/l)

CON=control, D1.5=basal diet supplemented with Digestarom 1317at 1.5g/10kg diet, D3=basal diet supplemented with Digestarom 1317at 3g/10kg diet,
T10=diet contained Thyme at 100g/10kg diet.
Means within the same raw under the same category carry different superscripts are significantly different.

17.07.2014

Seite 6 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Table 6. GPX activity (IU/g wet tissue) and GST activity (mol CDNB/min/g wet tissue) of liver, RBCs hemolysate and pancreas in chickens fed
diet supplemented with phytogenic feed additives at the end of the experiment
GPX-Aktivitt (IU/g Frischsubstanz) und GST-Aktivitt (mol CDNB/min/g Frischsubstanz) in Leber, Erythrozyten-Hmolysat und
Bauchspeicheldrse der Broiler am Versuchsende in Abhngigkeit von der Zulage der Zusatzstoffe zum Futter
Treatments
Item

CON

D1.5

D3

T10

GPX Activity IU/g Wet tissue

Liver
RBCs hemolysate
Pancreas

59.22.30b
25.12.02c
46.22.60b

60.33.17b
40.02.88b
50.32.64b

77.32.90a
52.72.88a
63.52.88a

80.53.17a
42.82.60b
54.53.17ab

GST Activity mol CDNB/min/g Wet tissue

Liver
RBCs hemolysate
Pancreas

6266.06b
3266.65b
4487.81c

6326.96b
4748.11a
4536.93c

7559.24a
4855.50a
6697.51b

7427.53a
4968.08a
5866.35a

CON=control, D1.5=basal diet supplemented with Digestarom 1317 at 1.5g/10kg diet, D3=basal diet supplemented with Digestarom 1317 at 3g/10kg
diet, T10=diet contained Thyme at 100g/10kg diet.
Means within the same raw under the same category carry different superscripts are significantly different.

Thyme supplementation has been found to affect the serum antioxidant status in broilers (HOFFMAN-PENNESI and
WU, 2011). Likewise, the antioxidant effect of thyme has been shown, as indicated by lower TBARS levels, in tuna
flesh after treatment with dried thyme during chilled storage (SALAH and SALOUA, 2008). The antioxidative effect of
thyme is based essentially on polyphenolic compounds, such as flavonoids (Luteolin) (JUSTESEN and KNUTHSEN,
2001), and occurs via various mechanisms, such as the inhibition of lipid peroxidation and the scavenging of
radicals. Chicks fed with rations containing thyme and cinnamon have been shown to have reduced (P<0.05) serum
cholesterol but significantly increased total proteins, albumin, total WBCs, RBCs, Hb and heterophil to lymphocyte
ratio (P<0.05) compared with the control group (AL-KASSIE, 2009). The plasma total cholesterol, triglyceride and
HDL levels are not affected by the dietary inclusion of thyme in broilers, but there are increases in the albumin and
globulin levels (NAJAFI and TORKI, 2010). SARICA et al. (2005) found no difference in the performance, total plasma
cholesterol concentration, and relative weights of some internal organs of broilers when thyme and garlic powder
were incorporated at the 1g/kg level into wheat-based diets, with normal ALT and AST activities.

Carcass traits
Concerning the carcass traits (Table7), there were no significant differences in the slaughter performance (dressing
%, liver %, heart %, gizzard %, bursa %, thymus %) between different experimental groups when compared with the
control one except Digestarom 1317 -supplemented group (D1.5) showed increases in the dressing %, liver % and
gizzard %.

17.07.2014

Seite 7 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Table 7. Dressing %, liver, heart, gizzard, spleen, bursa and thymus (%) of chickens fed diet supplemented with phytogenic feed additives at
the end of the experiment
Schlachtausbeute (%) und relative Gewichte (%) von Leber, Herz, Muskelmagen, Milz, Bursa und Thymus der Broiler am Versuchsende in
Abhngigkeit von der Zulage der Zusatzstoffe zum Futter
Treatments
Item

CON

D1.5

D3

T10

Dressing %
Liver %
Heart %
Gizzard %

66.00.81b
2.30.30a
0.60.04a
1.80.19a

68.60.95a
2.50.11a
0.60.01a
2.00.03a

66.10.43b
2.20.12a
0.60.01a
1.70.08a

66.20.34ab
2.50.04a
0.60.01a
1.80.07a

Spleen %
Bursa %
Thymus %

0.10.00
0.120.00a
0.20.01a

0.10.00
0.120.00a
0.20.01a

0.10.00
0.120.00a
0.20.01a

0.10.00
0.120.00a
0.20.01a

CON=control, D1.5=basal diet supplemented with Digestarom 1317 at 1.5g/10kg diet, D3=basal diet supplemented with Digestarom 1317at 3g/10kg
diet,T10=basal diet supplemented with Thyme at 100g/10kg diet.
Means within the same raw under the same category carry different superscripts are significantly different.

These results are in agreement with those of ABUK et al. (2006) who found that carcass yield and some internal
organ weights such as the liver, pancreas, gizzard and small intestine were not affected by the addition of the
essential oil mixture to the diet. Their results disagree with those of OCAK et al. (2008), who found that a peppermintor thyme-supplemented diet had a disadvantage in the slaughter performance but caused no significant differences
in carcass weight, dressing percentage and edible inner organs.

Gastrointestinal tract health status

In this work, phytogens were shown to modulate the cecal flora composition via the reduction of coliforms at the
ages of both 14 and 28 days and the fortification of the gut microflora with beneficial members, such as lactobacilli
(Table8). The beneficial effects of phytogens on the gut flora composition have also been reported by other studies
(JAMROZ et al., 2005). The addition of Digestarom 1317 at a level of 1.5g/10kg diet, as prescribed by the
manufacturer, provides beneficial results, with a significant reduction in the counts of total bacterial, Escherichia coli
and coliform bacteria and an increase in the beneficial lactobacillus and lactic acid bacteria compared with the
control group. There was a non-significant difference in the number of harmful bacteria between the group treated
with 1.5g/10kg Digestarom 1317 and either the groups treated with 3g/10kg Digestarom 1317 or 100g/10kg
thyme but a significant difference in the increase in the beneficial bacteria with 1.5g/10kg Digestarom 1317. These
results support those reported by MOUNTZOURIS et al. (2011).

17.07.2014

Seite 8 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Table 8. Cecum microflora count of chickens fed diet supplemented with phytogenic feed additives at the end of the experiment
Zusammensetzung der Mikroflora des Blinddarms der Broiler am Versuchsende in Abhngigkeit von der Zulage der Zusatzstoffe zum Futter
Treatments
Item

CON

D1.5

Total bacteria (cfu/g)

7.2110 1.80410

Escherichia coli (cfu/g)

4.2210 2.38110

Coliforms bacteria (cfu/g)

7.5710 1.01010

Lactobacillus bacteria (cfu/g)

1.2210 2.39210

Lactic acid bacteria (cfu/g)

D3

4.9510 6.17810

2.1310 6.45110

4.4310 2.00910

1.3410 2.95910

3.2610 1.41510 a

2.1810 0.69310 b

T10

5.2810 5.84610

2.2110 1.63110

4.4510 1.09910

1.2110 3.48310

5.2510 1.50110

3.4610 1.44310

3.3010 2.85710

1.9910 2.64610

3.3110 3.43510 a

3.3510 2.02110 a

CON=control, D1.5=basal diet supplemented with Digestarom 1317 at 1.5g/10kg diet, D3=basal diet supplemented with Digestarom 1317at 3g/10kg diet,
T10=basal diet supplemented with Thyme at 100g/10kg diet.
Means within the same column under the same category carry different superscripts are significantly different.

The microbial population of the gastrointestinal tract plays an important role in the performance of bird health
(JAVED et al., 2002). The change in the number or types of bacteria can affect the immune system or the physiology of
the digestive system; bacteria may compete for nutrients or the release of nutrients with the host. Most absorption of
nutrients in birds occurs in the small intestine. Due to the increase in bacteria, a competition between the microbes
and host occurs during feed intake. Bacteria in the colon were affected by the fermentation of undigested nutrients
producing volatile fatty acids (NAHAVANDINEJAD et al., 2012). The action of both Digestarom 1317 and thyme on the
gut micro-flora depends mainly on their content of essential oils. Essential oils and their components have healthpromoting potential as feed additives in poultry (HOFFMAN-PENNESI and WU, 2011).
This study concludes that Digestarom 1317 at 1.5g/10 k g had a growth promoter effect and that Digestarom 1317
and thyme had antioxidant, immuno-stimulant and gastrointestinal health effects. The beneficial effects of the
addition of Digestarom 1317 and/or thyme to the broiler diets resulted in the economical production of chicken
meat.

Summary
The objective of this study was to investigate the effects of dietary supplementation with phytogenic feed additives,
Digestarom 1317 and thyme, on the growth performance, oxidative stress and intestinal microflora in broiler
chickens. Approach: A total of 120 1-day-old broiler chicks were randomly allocated to four groups with three
replicates per treatment and 10 birds per replicate. The dietary treatments were the basal diet without phytogenic
additives (control), basal diet supplemented with Digestarom 1317 at 1.5g/10kg diet, basal diet supplemented
with Digestarom 1317 at 3.0g/10kg diet and basal diet supplemented with thyme at 100g/10kg diet.
The results revealed that 1.5g/10kg Digestarom 1317 increased the body weight significantly compared with
either the higher dose of Digestarom 1317 or thyme. Digestarom 1317 and thyme significantly increased the
phagocytic activity and index. The addition of Digestarom 1317 (at two dosing levels) or thyme significantly
increased the levels of antioxidant enzymes and significantly decreased protein nitrosylation, as reflected by
decreased nitric oxide levels in the examined tissues. Thyme significantly increased the serum protein pattern,
whereas both phytogenic additives non-significantly increased the level of liver-specific enzyme. The addition of
phytogenic additives ameliorated the intestinal microflora via the reduction of coliforms at the ages of 14 and 28 days
and via the fortification of beneficial gut flora, such as lactobacilli. This study concludes that Digestarom 1317 at
1.5g/10kg had a growth promotion effect and that Digestarom 1317 and thyme had antioxidant, immunostimulant and gastrointestinal health effects. The beneficial effects of the addition of Digestarom 1317 and/or
thyme to the broiler diets resulted in the economical production of chicken meat.

17.07.2014

Seite 9 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

Key words
Broiler, phytogenics, Digestarom 1317, thyme, growth promoter, antioxidant, gut flora.

Zusammenfassung
Prfung von Digestarom und Thymian als phytogene Futterzusatzstoffe bei Broilern
Das Ziel der Studie war die Untersuchung des Einflusses eines Zusatzes der phytogenen Substanzen Digestarom
1317 und Thymian zum Futter auf die Wachstumsleistung, den oxidativen Stress und die Zusammensetzung der

Darm-Mikroflora von Broilern. Hierzu wurden insgesamt 120 nicht nach Geschlecht sortierte Eintagskken zufllig
auf die vier Behandlungsgruppen mit je drei Wiederholungen a 10 Tieren verteilt. Als Versuchsrationen wurden
eingesetzt: Basisration ohne Zusatzstoffe (Kontrolle), Basisration mit dem Zusatz von 1,5g Digestarom/10kg,
Basisration mit dem Zusatz von 3,0g Digestarom/10kg, Basisration mit dem Zusatz von 100g Thymian/10kg.
Der Zusatz von 1,5g Digestarom 1317 fhrte zu einer signifikant hheren Lebendmasse im Vergleich zur hheren
Dosierung von Digestarom und zu Thymian. Sowohl Digestarom als auch Thymian erhhten die
Phagozytenaktivitt und den -index signifikant. In hnlicher Weise wurden durch den Zusatz an Digestarom in
beiden Zulagehhen und von Thymian die Spiegel der antioxidativen Enzyme signifikant erhht und die ProteinNitrosylierung signifikant verringert. Dies wurde durch die verminderten Gehalte an Stickstoffmonoxid in den
Geweben angezeigt. Whrend Thymian zu einer signifikanten Zunahme des Serum-Protein-Spiegels fhrte, erhhte
der Zusatz an Digestarom in beiden Zulagehhen den Spiegel an Leber-spezifischen Enzymen nur nicht-signifikant.
Der Zusatz der Phytobiotika verbesserte die Darmflora ber eine Verminderung der coliformen Keime im Alter von
14 und 28 Tagen und ber die Frderung von gnstigen Darm-Keimen, wie Laktobazillen. Die Untersuchung hat
gezeigt, dass sowohl Digestarom als auch Thymian antioxidative, immuno-stimulierende und die Darmflora
verbessernde Effekte aufweisen. Die gnstigen Effekte des Zusatzes von Digestarom 1317 bzw. von Thymian zum
Broilerfutter ermglichten eine konomischere Broilerfleischproduktion.

Stichworte
Broiler, Ftterung, Phytobiotica, Digestarom, Thymian, Wachstumsfrderer, antioxidativ, Darm-Mikroflora

Acknowledgements
We are grateful for the assistance of the Faculty of Veterinary Medicine, Damanhur University.

References
AOAC, Association of Official Analytical Chemist, 1990: Official Methods of Analysis 15th ed. Arlington, Virginia,
22201, USA.
AL-ANKARI, A.S., M.M. ZAKI, S.I. AL-SULTAN, 2004: Use of Habek Mint (Mentha longifolia) in Broiler Chicken Diets.
International Journal of Poultry science 3(10), 629-634.

AL-KASSIE, G.A.M., 2009: influence of two plant extracts derived from thyme and cinnamon on broiler performance.
Pakistan Veterinary Journal 29(4), 169-173.
BELFIELD, A., D.M. GOLDBERG, 1971: Revised assay for serum phenyl phosphatase activity using 4-amino-antipyrine.
Enzyme 12, 561-573.
BRUGH, M.J., 1978: A simple method for recording and analyzing serological data. Journal of Avian disease 22, 362365.

ABUK, M., M. BOZKURT, A. ALIEK, Y. AKBA, K. KKYILMAZ, 2006: Effect of a herbal essential oil mixture on
growth and internal organ weight of broilers from young and old breeder flocks. South African Journal of Animal
Science 36(2), 135-141.
CHIU, D., H. FREDRICK, A.L. TAPPEL, 1976: Purification and properties of rat lung soluble glutathione peroxidase.
Biochemica et Biophysica Acta 445, 558-566.

17.07.2014

Seite 10 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

st

COLES, E.H., 1974: Veterinary Clinical Pathology. 1 Ed. W. B Saunders Co. Philadelphia, London, Toronto, 212213.

CROSS, D.E., R.M. MCDEVITT, K. HILLMAN, T. ACAMOVIC, 2007: The effects of herbs and their associated essential oils
on performance, dietary digestibility and gut microflora in chickens from 7 to 28 days of age. British Poultry
Science 48,496-506.
CROSS, D.E., K. SVOBODA, R.M. MCDEVITT, T. ACAMOVIC, 2003: The performance of chickens fed diets with or
without thyme oil and enzymes. British Poultry Science 44, 18-19.

DEANS, S.G., R.C. NOBLE, L. PENZES, S.G. IMRE, 1993: Promotional effects of plant volatile oils on the polyunsaturated
fatty acid status during aging. Age 16, 71-74.
DOUMAS, B.T., W.A. WATSON, H.G. BIGGS, 1971: Determination of serum albumin. Journal of Clinical Chemistry 31,
87-89.
DURRANI, F.R., N. ABIDULLAH, Z. CHAND, S. AKHTAR, 2008: Hematological, biochemical, immunomodulatory and
growth promoting effect of feed added wild mint (Mentha longifolia) in broiler chicks. Journal of Agriculture 24(4),
135-141.
ERTAS, O.N., T. GLER, M. IFTI, B. DALKILIC, U.G. SIMSEK, 2005: The effect of an essential oil mix derived from
oregano, clove and anise on broiler performance. International Journal of Poultry Science 4, 879-884.
HABIG, W.H., M.I. PABST, W.B. JAKOBY, 1974: Glutathione S-transferase: The first enzymatic step in mercapturic
acid formation. Journal of Biological Chemistry 249, 1730-1739.
HARRIS, L.J., S.N. RAYS, 1935: Determination of plasma ascorbic acid by 2, 6 dicholrophenol indophenol titration.
Lancet 1, 462.
HOFFMAN-PENNESI, D., C. WU, 2011: The effect of thymol and thyme oil feed supplementation on growth
performance, serum antioxidant levels, and cecal Salmonella population in broilers. Journal of Applied Poultry
Research 19, 432-443.
JAMROZ, D., T. WERTELECKI, M. HOUSZKA, C. KAMEL, 2006: Influence of diet type on the inclusion of plant origin
active substance on morphological and histochemical characteristics of the stomach and jejunum walls in chicken.
Journal of Animal Physiology and Animal Nutrition 90, 255-268.
JAMROZ, D., A. WILICZKIEWICZ, T. WERTELECKI, J. ORDA, J. SCORUPINSKA, 2005: Use of active substances of plant origin
in chicken diets based on maize and domestic grains. British Poultry Science 46, 485-493.

JAVED, M.T., M.A. SARWAR, R. KAUSAR, J. AHMAD, 2002: Effects of feeding different levels of formalin (37%
formaldehyde) and urea on broiler health and performance. Veterinarski Archives 72, 285-302.
JUSTESEN, U., P. KNUTHSEN, 2001: Composition of flavonoids in fresh herbs and calculation of flavonoid intake by use
of herbs in traditional Danish dishes. Food Chemistry 73, 245-250.
KAWAHARA, E., T. UEDA, S. NOMURA, 1991: In vitro phagocytic activity of white spotted shark cells after injection
with Aeromoas salmonicida extraceular products. Gyobyo Kenkyu Japan 26(4), 213-214.
LUCKY, Z., 1977: Methods for diagnosis of fish disease. Ameruno publishing Co, PVT, Ltd New Delhi, Bomby, New
York.
rd

MAXINE, M., B.S. BENJAMIN, 1985: Outline of veterinary clinical pathology. 3 Ed., Colorado State University.
NAHAVANDINEJAD, M., A. SEIDAVI, L. ASADPOUR, 2012: Temperature treatment of soybean meal on intestinal
microbial flora in broilers. African Journal of Microbiological Research 6(26), 5464-5471.
MONTGOMERY, H.A.C., J.F. DYMOCK, 1961: the determination of nitrite in water. Analyst 86, 414.
MOUNTZOURIS, K.C., P.T. SIRTSIKOS, E. KALAMARA, H. NITSCH, G. SCHATZMAYR, K. FEGEROS, 2007: Evaluation of the
efficacy of a probiotic containing Lactobacillus, Bifidobacterium, Enterococcus, and Pediococcus strains

17.07.2014

Seite 11 von 12

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.55

inpromoting broiler performance and modulating cecal microflora composition andmetabolic activities. Poultry
Science 86, 309-317.
MOUNTZOURIS, K.C., V. PARASKEVASA, P. TSIRTSIKOS, I. PALAMIDIA, T. STEINERB, G. SCHATZMAYRC, K. FEGEROSA, 2011:

Assessment of a phytogenic feed additive effect on broiler growth performance, nutrient digestibility and caecal
microflora composition. Animal Feed Science and Technology 6, 37-43.

MUKHTAR, A.M., 2011: The Effect of Dietary Clove Oil on Broiler Performance.Australian Journal of Basic and
Applied Science 5(7), 49-51.
NAJAFI, P., M. TORKI, 2010: Performance, Blood Metabolites and Immunocompetaence of Broiler Chicks Fed Diets
Included Essentioal Oils of Medicinal Herbs. Journal of Animal and Veterinary Advances 9(7), 1164-1168.
th

NRC, 1994: National Research Council: Nutrient requirements of poultry, 9

Washington, DC.

Ed National Academy press,

OCAK, N., G. ERENER, F. BURAK, M. SUNGU, A. ALTOP, A. OZMEN, 2008: Performance of broilers fed diets
supplemented with dry peppermint (Mentha piperita L.) or thyme (Thymus vulgaris L.) leaves as growth promoter
source. Czech Journal of Animal Science 53(4), 169-175.
OUWEHAND, A.C., K. TIIHONEN, H. KETTUNEN, S. PEURANEN, H. SCHULZE, N. RAUTONEN, 2010: In vitro effects of
essential oils on potential pathogens and beneficial members of the normal microbiota. Veterinarni Medicina 55,
71-78.
PETERS, T.J., 1968: Proposals for standardization of total protein assays. Journal of Clinical Chemistry 14, 1147.
REITMAN, S., S. FRANKEL, 1957: A colorimetric method for determination of glutamic pyruvic transaminase.
American Journal of Clinical Pathology 25, 56. Cited in pamphlet of Diamond Diagnostics.
SALAH, S., S. SALOUA, 2008: The effect of natural antioxidant (Thymus vulgaris Linnaeus) on flesh quality of tuna
(Thunnus thynnus (Linnaeus)) during chilled storage. Pan-American Journal of Aquatic Sciences 3(1), 36-45.
SARICA, S., A. CIFTCI, E. DEMIR, K. KILIN, Y. YILDIRIM, 2005: Use of an antibioticgrowth prometer and two herbal
natural feed additives with and without exogenous enzymes in wheat based broiler diets. South African Journal of
Animal Science 35, 61-72.
th

SAS, 2002: User's Guide statistical Analysis system, version 6, 4 Edition, SAS Institute, Cary, NC. USA.
TAKATASY, G.Y., 1955: The use of spiral 100 Min Serological and Virological micromethods. Acta Microbial
Academic Science 3, 191.

TOGHYANI, M., M. TOHIDI, A.A. GHEISARI, S.A. TABEIDIAN, 2010: Performance, immunity, serum biochemical and
hematological parameters in broiler chicks fed dietary thyme as alternative for an antibiotic growth promoter.
African Journal of Biotechnology 9, 6819-6825.

YAN, L., J.P. WANG, H.J. KIM, Q.W. MENG, X. AO, S.M. HONG, I.H. KIM, 2010: Influence of essential oil
supplementation and diets with different nutrient densities on growth performance, nutrient digestibility, blood
characteristics, meat quality and fecal noxious gas content in grower-finisher pigs. Livestock Science 128, 115122.

Correspondence: Kadry M. Sadek, Dept. of Biochemistry, Nutrition and Vet. Clinical Nutrition, Hyiegene, Faculty of Veterinary Medicine, Damanhour
University, Egypt; E-mail: ksaadek@yahoo.com

17.07.2014

Seite 12 von 12