Environmental Science and Pollution Research

https://doi.org/10.1007/s11356-018-1685-z

RESEARCH ARTICLE

Effect of dietary phytogenics (herbal mixture) supplementation

on growth performance, nutrient utilization, antioxidative properties,
and immune response in broilers
Ahmed A. Saleh 1 & Tarek A. Ebeid 1,2 & Alaeldein M. Abudabos 3

Received: 5 October 2017 / Accepted: 4 March 2018

# Springer-Verlag GmbH Germany, part of Springer Nature 2018

Abstract
This experiment was conducted to study that effect of a phytogenics (herbal mixture) supplementation as alternative to antibiotics
on the growth performance and immune response in broilers. Three thousand broiler male chicks at 1 day of age were housed in
floor pens and divided into five experimental groups with five replicates of 120 birds each. The first group was fed the basal diet
without any additive in feed or water; the second group was fed the basal diet, and the drinking water was supplemented with
mixture of antibiotics (Doxycycline 1 g/5 L + Colistine sulfate 1 g/5 L) for the first 10 days of life. The other three treatments of
chickens were fed the control diet supplemented with herbal mixture at a level of 0.5, 1.0, and 1.5 kg/ton, respectively. Dietary
herb mixture supplementation improved growth performance significantly in comparison with control group. In addition, herb
mixture supplementation reduced plasma total cholesterol and triglycerides concentrations, whereas, high density lipoprotein
HDL-cholesterol and glutathione peroxidase (GPX) were increased significantly. Furthermore, herb mixture supplementation
increased plasma levels of total protein and antibodies titers for Newcastle disease virus before and after the infection. It could be
concluded that dietary herb mixture supplementation had positive effects on growth performance, antioxidative properties, and
humoral immunity in broiler chickens.

Keywords Herb mixture . Broilers . Growth performance . Antioxidative status . Immunity

Introduction animal diets due to the efflorescence of microbe resistance,

Apart from significant role of phytogenics, pharmaceutical
plants, herbs, and spices in daily human food for raise of taste,
flavor, and color, the herb-feed additives have also been effi-
ciently applied in poultry and animal nutrition for perfection
of their health and comfort. With the ban of antibiotics use in
Responsible editor: Philippe Garrigues
* Ahmed A. Saleh
ahmed.saleh1@agr.kfs.edu.eg
1
Department of Poultry Production, Faculty of Agriculture,
Kafrelsheikh University, Kafr El-Sheikh 33516, Egypt
2
Department of Animal Production and Breeding, College of
Agriculture and Veterinary Medicine, Qassim University,
Buraydah 51452, Saudi Arabia
3 action in the intestine against different E. coli, Salmonella,
Department of Animal Production, College of Food and Agriculture
Sciences, King Saud University, Riyadh, Saudi Arabia
alternative growth promoters must be found (Steiner 2009).
Antimicrobial additives such as different types of antibiotics
are used to reduce harmful microorganisms in the bird’s intes-
tine and to develop their growth, feed conversion ratio, and
immunity; however, habit antibiotic employment as feed ad-
ditive has been forbid in different countries worldwide in the
last 10 years (Leeson 2007). Administering antibiotics to
poultry and animal in their feed or drinking water has had a
high impact on the commercial production of meat and egg for
human consumption since the early 1950s (Jukes 1972).
Bettenay et al. (2003) reported that doxycycline is a second-
generation tetracycline mainly active against gram-positive
and gram-negative aerobic and anaerobic bacteria as well as
mycoplasma, chlamydia, Rickettsia, and spirochetes.
Furthermore, Brander et al. (1993) reported that colistin is
acting mainly against gram-negative bacteria. As it is not
digested from the gastrointestinal tract, it develops its main
and Enterobacter strains.

Numerous phytogenics have pharmacological effects and
widely used in human traditional medicine. In poultry and
animal nutrition, it has been studied the effects of a wide range
of herbs on feed efficiency, antimicrobial, coccidiostatic, an-
thelmintic, and immune-stimulating (Panda et al. 2006).
Several medicinal plants [e.g., garlic, onion, thyme, spear-
mint, pepper, yucca, black cumin (black seed), ginger] have
been used in animal and poultry production due to their health
benefits, e.g., spearmint leaves is used as carminative, anti-
spasmodic, and choleretic in human reported by Al-Kassie
(2010). It has been reported that onion has pharmaceutical
activities for antioxidant, antimicrobial, and antiallergic; and
a combination of garlic and ginger prevents high blood pres-
sure, high cholesterol level, and cholesterol oxidation which
are the primary causes of atherosclerosis, the precursor of
cardiovascular diseases (El-Meleigy et al. 2010). Also,
Bamidele and Adejumo (2012) found that the combined effect
of garlic and ginger mixture has greater influence as an anti-
hypercholesterolemic agent and improved growth perfor-
mance in broilers.
Pepper is one of the traditional herbs used in human food.
Yoshioka et al. (2001) reported that pepper plays an important
role in analyzing fat and cholesterol in the animal body and
shares in reducing levels of plasma triglycerides and works in
backing the vascular system in the human and animal bodies.
Moreover, pepper is wealthy in vitamin C which has a major
impact on improving production out of claiming the reduction
of heat (Yoshioka et al. 2001). Yucca schidigera is one of the
good sources of saponins that prevent the growing of protozoa
by reactive with the total cholesterol existent on the parasite
cell membrane, therefore accomplishing parasite death (Wang
et al. 1998). Sundry experiments with saponins have ex-
plained their good effects on meliorative nutrient absorption
by rising intestinal permeability through cell membrane depo-
larization; however, saponins can also affect the absorption of
different vitamins and minerals. Cheeke (1999) reported that
oral rule of saponins is also reconditioned with improving the
animal impedance to field infections overing an immunomod-
ulatory effect will happen. Thyme is used traditionally for
several medicinal purposes: respiratory disease, antimicrobial,
antinociceptive, and etc. (Demir et al. 2008). Moreover,
thymol and carvacrol are the main antibacterial active
substances, so they could be used instead of different
antibiotics. Aktug and Karapinar (1986) found that the bene-
ficial value of thyme in poultry industry has been shown to
inhibit the growth of Salmonella typhimurium and E. coli
when added to media. It could be hypothesized that herb mix-
ture (garlic, onion, thyme, spearmint, pepper, yucca, black
cumin seed, and ginger) might have several vital benefits of
such as antioxidant, antimicrobial, and anti-inflammatory,
which raise its use as feed additives as alternative to antibiotics
in poultry diets. Thus, the purpose of the present experiment
was to evaluate the influence of a phytogenics (herb mixture)
Environ Sci Pollut Res
supplementation as alternative to antibiotics on the growth
performance, plasma lipids, nutrient utilization profile, antiox-
idative properties, lipid peroxidation, and immune restraint in
broilers.
Materials and methods
The present study was done in the research unit of Al-Sabeel
Al-Gadidah Company for poultry production, Tanta, Al-
Gharbia, Egypt and affirmed by the Committee on the Ethics
of Animal Experiments of Kaferelsheikh University, Egypt.
All procedures and experiments complied with the guidelines
and were approved by the Local Ethic Commission of the
Animal Experiments of the Department of Poultry
Production, Faculty of Agriculture, Kaferelsheikh
University, with respect to animal experimentation and care
of animals under study. All efforts were made to minimize
suffering. Three thousand male broiler chicks (Cobb avian
48), at 1 day of age were housed in floor pens and divided
into five experimental groups with five replicates of 120 birds
each (20 bird/m2) in floor pens. The experiment was conduct-
ed in close system farm with 21-h light: 3-h dark cycle, and the
chickens were housed in floor pens; each pen area was 2 ×
3 m. Pens were provided with litter (pine wood shavings) to a
depth of 5–6 cm. Birds were provided with water and five
experimental diets starter diet from 1 to 10 days (granular
form), grower diets from 11 to 22 days (pellet form), and
finisher from 23 to 35 days (pellet form). The compositions
of the basal diets are shown in Table 1. The first treatment was
fed a basal diet without any additives and served as control;
the second group was fed on the basal diet, and the drinking
water was supplemented with mixture of antibiotic
(Doxycycline 1 g/5 L + Colistine sulfate 1 g/ 5 L) for the first
10 days of life. The other three groups of chicks were fed the
control diet supplemented with herb mixture at a level of 0.5,
1.0, and 1.5 kg/ton, respectively, until 35 days of age. The
diets, herb mixture, and antibiotics were kindly provided by
Fat-Hens CO., LTD (Egypt). After brooding period, room
temperature was kept as 24 °C with relative humidity from
50 to 70% throughout the experiment. The birds were fed each
experimental diet from 1 to 35 days of age, while, the second
group was given the antibiotics in water from 1 to 10 days of
age and completed until 35 days without any additives.
Herb mixture preparation
The herb mixture contains eight different herbs (stems and
leaves of spearmint, thyme, and yucca; seeds of pepper and
black cumin; roots of ginger; and developing bulb part of
onion and garlic). Each raw herb (Ragab El-Attar Co,
Egypt) was dried separately in oven by 40 °C for 24 h then
the herb was grinded. Powder of each herb was weighed and
Environ Sci Pollut Res

Table 1 Composition and nutrient analysis of the basal diets Table 2 Phytogenics (herbal mixture) composition

Ingredients Starter Grower Finisher Botanical name (English common name) Content (%)
(g/kg) (g/kg) (g/kg)
Mentha spictata (spearmint) 25.0
Maize 574 607.7 626 Allium sativum (garlic) 25.0
Soybean meal, 44% 310 276 246 Allium cepa (onion) 15.0
Corn gluten meal, 44 30 34 Thymus (thyme) 10.0
62%
Capsicum annuum (pepper) 5.0
Soy oil 30 35 43
Yucca elephantipes (yucca) 10.0
Limestone 11 10 11
Nigella sativa (black cumin) 5.0
Dicalcium phosphate 17 17 17
Zingiber purpureum (ginger) 5.0
Premixa 3 3 3
Sodium bicarbonate 2 2 2
Salt 3.5 3.5 3.5
L-Lys HCl 3 3 1.7
dried samples were then homogenized after drying. Samples
DL-Met 2.5 2.5 2.5
were randomly taken and ground to analysis according to
L-threonine 0.3 0.3 0.3 AOAC (2000) for crude protein (CP, Method 968.06), ether
Nutrient compositionb extract (EE, Method 920.39), and crude fiber (CF, Method
CP, % 21.5 19.3 18.1 932.09). The calculations were as follows: nitrogen retention
ME, Mcal/Kg 12.15 12.45 12.80 (%) = (total nitrogen intake − total nitrogen excreted)/total ni-
Ca, % 1.1 0.95 0.93 trogen intake × 100.
P, % 0.46 0.45 0.44 At 35 days, after the decapitation, blood samples (12 sam-
Na, % 0.22 0.22 0.22 ple/group) were collected in heparinized test tubes and quickly
CL, % 0.25 0.25 50.2 centrifuged (3000 rpm for 20 min at 5 °C) to separate the
a plasma. Plasma was stored at − 20 °C until further analysis.
Each 3 kg of vit and min in Premix contains 6000000 IU vit A,
900000 IU vit D3, 40,000 mg vit E, 2000 mg vit K, 2000 mg vit. B1, Plasma total cholesterol, triglycerides (TG), HDL-cholesterol,
4000 mg vit B2, 2000 mg vit B6, 10 mg vit B12, 50,000 mg niacin, AST, ALT, glucose, GSH-Px, total protein, albumin, and glob-
10,000 mg pantothenic acid, 50 mg biotin, 3000 mg folic acid, ulin were measured calorimetrically by using commercial kits
250,000 mg choline, 50,000 mg Zn, 8500 mg Mn, 50,000 mg Fe,
(Diamond Diagnostics, Egypt) according to the procedure
50,000 mg Cu, 200 mg I, 100 mg Se, and 100 mg Co
b outlined by the manufacturer. Muscle total cholesterol was
According to NRC (1994)
measured by the method of Saleh (2013). Muscle and liver
TBARS were measured according to Ohkawa et al. (1979).
its supplementation ratio was indicated in Table 2. All the
herbs were mixed by small mixer for 5 min. The final herbal
mixture was stored in a cool and dark glass jars with tight-
Assay of serum antibody titers
fitting lids until used.
At 28 days, 25 birds from each experimental group were ran-
domly chosen and grown in a fortified room to Newcastle
Growth performance and carcass parts infection, measuring the hemagglutination inhibition (HI) test
before and after the infection, and the birds were infected by
Bird’s body weight was measured weekly, and feed consump- live vaccines containing LaSota Newcastle disease virus
tion was measured daily during the experiment. At the end of (NDV) strain which is used in the vaccination procedures.
the experimental period, 75 chicks (15male/group) were ran- This strain is commercially used in the immunization of
domly chosen, weighed and slaughtered by hand using a con- chickens in Egyptian farms. Serum antibody titer against
ventional unilateral neck cut to sever the carotid artery and Newcastle disease (ND) was determined by means of HI test
jugular vein, and bled for 120 s. Carcasses were dissected to using standard methods qualified in OIE (OIE 2009).
measure the weights of breast muscle, liver, and abdominal
fat. Statistical analysis

Nutrient digestibility
At the last 3 days of the trial, excreta and feed were collected
and weighted by 20 birds from each group. Then, the samples
were dried by the drying oven at 60 °C for 24 h. The whole
The differences between the treatment groups and the control
group were analyzed with a general liner model using SPSS
Statistics 17.0 (Statistical Packages for the Social Sciences,
released 23 August 2008). Tukey s multiple comparison test
was used to identify which treatments conditions were
 Environ Sci Pollut Res

Table 3 Effect of feeding phytogenics (herb mixture) on growth performance in broilers

Treatmentsd

Control Antibiotics Herb mixture (kg/ton)

0.5 1.0 1.5

Initial body weight (g) 43.2 ± 0.001 43.1 ± 0.002 42.9 ± 0.001 43.0 ± 0.002 43.4 ± 0.001
Body weight gain (g/34 day) 2005 ± 48b 2095 ± 58ab 2080 ± 43ab 2140 ± 57a 2155 ± 66a
Feed intake (g/34 day) 3290 ± 88b 3415 ± 77a 3390 ± 85a 3455 ± 98a 3470 ± 91a
Feed conversion ratio (kg/kg) 1.641 ± 0.03a 1.630 ± 0.02a 1.629 ± 0.05ab 1.614 ± 0.03b 1.610 ± 0.04b
Carcass weight (g/100 g body weight) 65.98 ± 2.78 66.73 ± 1.89 67.43 ± 1.55 68.93 ± 2.72 68.51 ± 2.81
Breast muscle weight (g/100 g body weight) 23.93 ± 0.92b 23.71 ± 1.11b 24.98 ± 1.55a 25.27 ± 1.72a 25.84 ± 1.81a
Liver weight (g/100 g body weight) 2.99 ± 0.13 3.24 ± 0.22 3.18 ± 0.30 3.23 ± 0.23 3.39 ± 0.32
Abdominal fat (g/100 g body weight) 2.26 ± 0.09a 2.18 ± 0.14b 1.92 ± 0.13b 1.68 ± 0.09c 1.74 ± 0.13b

Values are expressed as means ± standard error. Data were analyzed by one-way analysis of variance and Tukey’s multiple tests.
a–c
Means within the same row with different superscripts differ (P < 0.05)
d
Treatments: control (no supplementation), antibiotics (Doxycycline 1 g/5 L + Colistine sulfate 1 g/5 L) for the first 10 days of life, and the other three
experimental groups of chicks were fed the basal diet supplemented with herb mixture at a concentration of 0.5, 1.0, and 1.5 kg/ton, respectively, until
35 days of age

significantly different from each other at a significance level
of P < 0.05.
Results
The effect of dietary phytogenics (herb mixture) on growth
performance and organs weight were summarized in Table 3.
Dietary herb mixture supplementation increased feed con-
sumption, chickens body weight gain, and breast muscle
weight significantly. Feed conversion ratio was improved by
herb mixture supplementation (P < 0.05). A significant reduc-
tion of abdominal fat weight was observed in chicken fed herb
mixture-supplemented diets, whereas, carcass and liver
weights were not significantly influenced.
Table 4 shows the nutrient retention of broiler chickens fed
the herb mixture-supplemented diet. Dietary 1.0 and 1.5 kg/
ton herb mixture supplementation resulted in a significant in-
crease in crude protein and ether extract retentions compared
with control. However, no significant differences were found
between control group and chickens drank antibiotics in crude
protein and crude fat retentions. Crude fiber retention was not
affected by feeding the herb mixture-supplemented diet or
adding antibiotic in water.
The influences of dietary herb mixture supplementation on
plasma concentrations of total cholesterol, HDL-cholesterol,
triglyceride, GOT, GPT, and glucose are presented in Table 5.
Plasma total cholesterol, glucose, and triglyceride were signif-
icantly reduced by adding the herb mixture-supplemented diet
compared to the control diet, while, plasma high-density lipo-
protein (HDL-cholesterol) level was increased (P < 0.05).

Table 4 Effect of feeding

phytogenics (herb mixture) on Treatmentsd
coefficients nutrient retention
Control Antibiotics Herb mixture (kg/ton)

0.5 1.0 1.5

Crude protein retention, % 69.87 ± 6.1b 71.21 ± 5.8 ab 71.54 ± 4.3ab 73.47 ± 8.8a 73.97 ± 7.7a
Crude fat retention, % 39.48 ± 5.1b 39.21 ± 4.1b 40.97 ± 5.4a 41.24 ± 3.1a 42.01 ± 3.8a
Crude fiber retention, % 30.38 ± 5.1 38.41 ± 3.1 38.97 ± 4.4 39.10 ± 4.4 39.87 ± 3.4

Values are expressed as means ± standard error. Data were analyzed by one-way analysis of variance and Tukey’s
multiple tests
a–c
Means within the same row with different superscripts differ (P < 0.05)
d
Treatments: control (no supplementation), antibiotics (Doxycycline 1 g/5 L + Colistine sulfate 1 g/5 L) for the
first 10 days of life, and the other three experimental groups of chicks were fed the basal diet supplemented with
herb mixture at a concentration of 0.5, 1.0, and 1.5 kg/ton, respectively, until 35 days of age
Environ Sci Pollut Res

Table 5 Effect of feeding phytogenics (herb mixture) on blood traits in broilers

Treatmentsd

Control Antibiotics Herb mixture (kg/ton)

0.5 1.0 1.5

TG (mg/dL) 27.42 ± 1.23a 25.2 ± 0.88a 21.27 ± 1.04b 21.02 ± 1.07b 20.78 ± 0.86b
Total cholesterol (mg/dL) 152.6 ± 5.1a 148.94 ± 6.3ab 134.75 ± 6.1b 137.67 ± 5.1b 128.92 ± 5.3b
HDL-cholesterol (mg/dL) 73.2 ± 21.8b 75.2 ± 3.3b 82.9 ± 1.2a 83.2 ± 2.7a 85.2 ± 3.3a
GOT (U/I) 190.0 ± 22 189.2 ± 26 188.3 ± 33 187.0 ± 32 187.7 ± 26
GPT (U/I) 23.4 ± 5.1 21.8 ± 3.4 21.3 ± 2.8 21.8 ± 5.7 22. ± 3.4
Glucose (mg/dL) 194 ± 22a 195 ± 19a 173 ± 17b 179 ± 21b 175 ± 14b

Values are expressed as means ± standard error. Data were analyzed by one-way analysis of variance and Tukey’s multiple tests
TG, triglycerides; HDL, high-density lipoprotein; GOT, glutamic oxalacetic transaminase; GPT, glutamate pyruvate transaminase)
a–c
Means within the same row with different superscripts differ (P < 0.05).
d
Treatments: control (no supplementation), antibiotics (Doxycycline 1 g/5 L + Colistine sulfate 1 g /5 L) for the first 10 days of life, and the other three
experimental groups of chicks were fed the basal diet supplemented with herb mixture at a concentration of 0.5, 1.0, and 1.5 kg/ton, respectively, until
35 days of age

Neither plasma GOT nor GPT level was affected by feeding
herb mixture.
Interestingly, in Table 6, it is clear that plasma total protein,
globulin, and ND titers at 28 days and after infection were
significantly improved in treatments fed the herb mixture-
supplemented diet compared with the control treatment.
With respect to the effect of dietary herb mixture on muscle
total cholesterol, muscle MDA, liver MDA, and plasma GSH-
Px (Fig. 1a–d), it could be spotted that dietary 0.5, 1.0, and
2.5 kg/ton herb-mixture significantly decreased muscle total
cholesterol, muscle MDA, and liver MDA, while, plasma
GSH-Px concentration was significantly increased in compar-
ison with control.
Discussion
Results of the present study illustrated that feeding the herb
mixture-supplemented diet increased body weight gain and
breast muscle weight, while, it decreased abdominal fat
weight and improved feed conversion ratio (Table 3). These
results might be attributed to the growth promoting effect of

Table 6 Effect of feeding phytogenics (herbal mixture) on plasma proteins and HI test for Newcastle disease virus (ND)

Treatmentsd

Control Antibiotics Herb mixture (kg/ton)

0.5 1.0 1.5

Plasma total protein (mg/dL) 3.11 ± 0.31b 3.58 ± 0.34ab 3.78 ± 0.41ab 3.94 ± 0.32a 4.04 ± 0.44a
Plasma albumin (mg/dL) 1.19 ± 0.05b 1.56 ± 0.08a 1.52 ± 0.04a 1.1 ± 0.06b 1.21 ± 0.08b
Plasma globulin (mg/dL) 1.92 ± 0.18b 2.02 ± 0.35ab 2.26 ± 0.23a 2.84 ± 0.23a 2.82 ± 0.25a
ND titers, 14 days 1.98 ± 0.21 2.36 ± 0.31 2.62 ± 0.32 2.79 ± 0.21 2.76 ± 0.30
ND titers, 28 days 1.94 ± 0.23b 2.52 ± 0.33a 2.78 ± 0.25a 2.95 ± 0.34a 2.92 ± 0.33a
ND titers, 35 days 3.14 ± 0.19 3.42 ± 0.27 3.73 ± 0.31 3.93 ± 0.29 3.52 ± 0.34
ND titers, after infection 7.13 ± 0.27b 7.92 ± 0.36ab 8.66 ± 0.42a 9.03 ± 0.29a 8.74 ± 0.31a

Values are expressed as means ± standard error. Data were analyzed by one-way analysis of variance and Tukey’s multiple tests
a–c
Means within the same row with different superscripts differ (P < 0.05)
d
Treatments: control (no supplementation), antibiotics (Doxycycline 1 g/5 L + Colistine sulfate 1 g/5 L) for the first 10 days of life, and the other three
experimental groups of chicks were fed the basal diet supplemented with herb mixture at a concentration of 0.5, 1.0, and 1.5 kg/ton, respectively, until
35 days of age

Fig. 1 Effect of feeding phytogenics (herbal mixture) on muscle total
cholesterol (a), thiobarbituric acid reactive substance (mg MDA/kg meat)
(b), thiobarbituric acid reactive substance (mg MDA/kg liver) (c), and
plasma glutathione peroxidase (GSH-Px, mg/dL) (d). Values are means
represented by vertical bar. a,b,c,dMean values with unlike letters were
significantly different (P < 0.05)
some ingredients of herb mixture. This assumption is in agree-
ment with previous studies (Saleh et al. 2014) which noted
that an enhanced in body weight, weight gain, and organ
weights were observed in birds fed a diet adding onion or
black seed. In a similar way, feeding spearmint increased feed
composition and thus improved growth of birds reported by
Environ Sci Pollut Res
Amasaib et al. (2013), and dietary herb mixture supplementa-
tion increased feed composition in the present experiment.
One tolerable explanation is that the difference between the
feeding single herb and the herb mixture might be due to some
antimicrobial components involved in the ingredient of
phytogenics of the present study. Since it has been familiar
that microflora condition in the gastrointestinal tract enhanced
digestibilities of diets and consequently influences the growth
performance of broiler chickens (Metzler et al. 2005).
Essential oils extracted from thyme negatively affected both
gram-positive and gram-negative bacteria (Delaquis et al.
2002) and improved growth performance via increasing the
digestibility of feed (Demir et al. 2008). Similarly, Aktug and
Karapinar (1986) reported that carvacrol, which is involved in
the essential oils from thyme, negatively affected Salmonella
typhimurium, Escherichia coli, Staphylococcus aureus, and
Clostridium perfingers. These results suggested that antimi-
crobial combination involved in the ingredient of herb mixture
such as thyme, garlic, and ginger controlled the microflora in
the gastrointestinal tract which consequently involved in en-
hancing the digestibility of feed and growth performance in
broiler chickens fed the herb-mixture-supplemented diet
might be enhanced.
Wenk (2003) found that medical plants and their ex-
tracts can improve appetite and endogenous extraction
such as enzymes or have coccidiostat and improved di-
gestibility. Thus, growth performance and digestibility
were improved in the present study by herb mixture prob-
ably due to the antimicrobial and antifungal effects of
herb mixture ingredients (Kamel 2000).
In the present study, dietary herb mixture supplementation
decreased plasma levels of TG, total cholesterol, and glucose,
while, plasma HDL-cholesterol concentration was significant-
ly increased (Table 5). The underlying mechanisms for im-
provement of plasma lipid profiles might be caused by pepper,
garlic, and ginger. It has been reported that either pepper or
garlic has a hypolipidemic capability (Bamidele and Adejumo
2012; Saleh et al. 2017, 2018). Feeding either garlic or onion
meal significantly decreased serum total cholesterol, LDL-
cholesterol, GOT, GPT, and glucose levels of broiler chickens
(Al-Kassie 2010). Garlic enhances bile acid synthesis and
degradation of cholesterol to fecal bile acid and natural sterols
also; ginger has a vital role in decreasing total cholesterol by
inhibiting the enzymes such as hydroxyl-methyl-glutaryl-
CoA reductase, which plays an important role in cholesterol
biosynthesis (Bamidele and Adejumo 2012). In addition, Al-
Mashhadani et al. (2011) reported that the inclusion of carva-
crol resulted in a decrease in serum glucose. These results
might suggest that feeding the herb-mixture-supplemented di-
et might improve plasma lipid profiles including lower glu-
cose concentration. Furthermore, dietary herb mixture supple-
mentation did not affect the levels of plasma GOT and GPT.
These results are in agreement with the previous study of Al-
Environ Sci Pollut Res
Jaff (2011) which documented that feeding herb mixture did
not affect the level of GOT. In general, plasma levels of AST
and ALT were considered as liver indexes for liver cells dam-
age, and the values acquired in the present experiment are
within the normal physiological value results. Therefore, these
results may provide evidences for occurrence of no toxicity of
feeding herb-mixture-supplemented diet in broiler chickens.
One of the important results in the current experiment
was the perfection of the antioxidative properties ex-
plained by the significant increase in serum GSH-Px and
the decrease of lipid peroxidation index (liver MDA and
muscle MDA) in broilers due to dietary herb mixture sup-
plementation (Fig. 1). Actually, the antioxidant properties
of herb mixture have been largely described and among
the different of herbs with antioxidant effects. Mahmoud
et al. (2002) found that black seed and mint have known
to possess strong antioxidant activities. The antioxidative
activities in black seed and mint are by their contents of
phenolic and terpenes (Cuppett and Hall 1998). Other
medical plants with antioxidative merit such as oregano
and thyme contain high levels of carvacrol and thymol.
Also, Nakatani (1994) reported that some herbs rich with
flavonoids such as green tea have been described as nat-
ural antioxidant, and the black pepper and red pepper
contain also different antioxidative substances. The
antioxidant properties of different herbs can share in to
dietary unsaturated fatty acid protection from oxidation.
Cuppett and Hall (1998) found that using Labiatae family
plants as antioxidants in feed diet of poultry is very effec-
tive. Moreover, the antioxidant and free-radical scaveng-
ing properties of different plants have been found to play
a great role in their hepatoprotective activity (Shertzer
et al. 1994; Thabrew et al. 1995). Therefore, it could be
documented that dietary herb mixture supplementation
would enhance antioxidative properties and reduce lipid
peroxidation in broilers. From another opinion, the use of
herb mixture as an antioxidant is not only effective for the
animal health, but also for the oxidative stability of the
meat and eggs. Botsoglou et al. (2003) found that feeding
of turkey feed with 200 mg extract of oregano/kg diets
significantly reduces lipid per-oxidation of meat.
Interestingly, results of the present study illustrated that
dietary herb mixture supplementation enhanced plasma pa-
rameters related to health and immunity (i.e., total protein,
albumin, globulin, and antibodies titers for Newcastle disease
virus before and after the infection, respectively (Table 6).
These results suggested that immune-defense system might
be enhanced due to dietary herb mixture supplementation. It
has been found that either ginger or peppermint has a role for
anti-inflammatory (Al-Kassie 2010). In addition, Huang et al.
(2004) reported that the improvement of immune response
condition to microbial and/or fungi is positively related to
performance of broilers. Furthermore, increased immune
response to microbial and/or fungi resulted in enhance in ox-
idants level in the gastrointestinal tract of broilers and laying
hens. Moreover, black seed (Nigella sativa) has abilities of
antimicrobial effects (Saleh 2014) and antifungal effects
(Salem and Hossain 2000), and either garlic or onion has high
antioxidative properties, which cancel occurrence of oxidants
or counter the negative effect of oxidants (Al-Mamary 2002).
Also, either black seed or onion might also be contributed to
the increase in immune-related parameters. Therefore, it could
be assumed that dietary herb mixture supplementation would
influence the viability and immune function in broiler in the
present study. Moreover, by taking into account our outcome
in antioxidative properties and lipid per-oxidation, it appears
more likely that dietary herb mixture supplementation may
promote the immune responsiveness and the protection versus
diseases in broilers.
In summary, the results obtained in the present study might
suggest that feeding herb-mixture-supplemented diet im-
proved growth performance, nutrient digestibility, antioxida-
tive properties, and humoral immunity and could be used as
alternative to antibiotic in broiler chickens.
Acknowledgements Authors would like to extend their sincere appreci-
ation to the Deanship of Scientific Research at King Saud University for
its funding of this research through Research Group Project No. RGP-
VPP-273. The authors wish to acknowledge the helpful suggestions of
members of department of poultry production, faculty of agriculture,
Kafrelsheikh University, Egypt. The authors wish to thank Al-Sabeel
Al-Gadidah Company for poultry production for theirs helps in this
experiment.
Compliance with ethical standards
Conflict of interest The authors declare that they have no conflict of
interest.
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