Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq

EFFECT OF PEPPERMINT SUPPLEMENTATION AS POWDER OR

EXTRACT ON BROILER PERFORMANCE, SERUM BIOCHEMICAL
CONTENT AND GUT HEALTH UNDER E COLI CHALLENGE
H. I. Hasan S. A. M’Sadeq
Researcher Assist. Prof.
Dept.Anim Prod. Coll. Agric. Engin. Sci., University of Duhok, Iraq
[email protected]
ABSTRACT
The aim of this study was to evaluate the efficacy of different levels of peppermint (Mentha
piperita) on broiler performance, jejunum morphology, organs percentage and serum
biochemical content. A total of 576-day old ross 308 chickens was divided randomly into 12
treatments with 4 replicates pen per treatment and 12 birds each pen. Treatments were
arranged in a 2 × 6 factorial arrangement of treatments was employed. Factors were
challenge, - or +; control, antibiotic, 0.5 % peppermint in feed, 1% peppermint in feed, 0.5%
peppermint in water and 1% peppermint in water. The result of study was showed that the
peppermint supplementation was significant effect on feed intake, feed conversion ratio and it
has effect on body weight at day 24 and 35. The peppermint supplementation either in feed
and water at 24 day had a significant effect on jejunum muscle thickness and villus/ crypt
ratio while not effect on villus height and crypt depth. In conclusion, supplementation of
peppermint has positive effect on broiler performance and gut morphology.
Key words: meat chickens, productivity, gut morphology, Escherichia coli, peppermint

‫حسن وصديق‬ 310-299:)1)51: 2020- ‫مجلة العلوم الزراعية العراقية‬

‫تاثير اضافة النعناع على االداء االنتاجي و المحتوى الكيميائي لمصل الدم وصحة االمعاء لفروج اللحم تحت تاثير االصابة‬
‫االيكوالي‬
‫شوكت عبدالرزاق محمد صديق‬ ‫هلبين ادريس حسن‬
‫استاذ مساعد‬ ‫الباحث‬
‫كلية علوم الهندسة الزراعية – جامعة دهوك‬-‫قسم االنتاج الحيواني‬
‫المستخلص‬
‫ الصفات النسيجية‬،‫) على األداء االنتاجي‬Mentha piperita(‫اجريت هذه الدراسة لتقييم تاثير مستويات مختلفة من النعناع‬
‫ دجاج‬576 ‫ مجموع‬.‫ الوزن النسبي لالعضاء الداخلية ومحتوى الكيمياء الحيوية في مصل الدم لدجاج فروج اللحم‬،‫لالمعاء‬
. ‫ طائر لكل مكرر‬12 ‫ مكررات لكل معامله و‬4 ‫ معامالت مع‬12 ‫فروج اللحم بعمر يوم واحد تم تقسيمها بشكل عشوائي على‬
‫ ربيت الطيور في غرفتين مختلفتين بحيث في الغرفة االولى تم تربية الدجاج‬.‫ تجريبة عواملية‬6*2 ‫تم ترتيب المعامالت بشكل‬
‫ وزعت المعامالت في كل غرفة‬.‫تحت الظروف الطبيعية اما في الغرفة الثانية تعرضت الطيور الى االصابة ببكتريا االيكوالي‬
٪1 ‫ نعناع في الماء و‬٪0.5 ،‫ نعناع في األعالف‬٪1 ،‫ نعناع في األعالف‬٪0.5 ،‫ مضاد حيوي‬،‫بالشكل االتي (السيطرة‬
‫ وأظهرت نتائج الدراسة أن مكمالت النعناع كان لها تأثير كبير على تناول العلف وكفائة التحويل الغذائي ولها‬.)‫نعناع في الماء‬
‫ كانت‬24 ‫ اضافة مكمالت النعناع سواء الى الغذاء أو الى الماء في اليوم‬.35 ‫ و‬24 ‫ايضا تأثير على وزن الجسم في اليوم‬
‫ ) و كذلك نسبة طول الزغابات الى الخبايا بينما ال تؤثر على طول الزغابات‬Jejunum) ‫لها تأثير كبير على سماكة عضالت‬
‫ استنتجت هذة التجربة ان مكمالت النعناع لها تأثير إيجابي على االداء االنتاجي والصفات النسيجية المعاء‬.‫و عمق الخبايا‬
.‫فروج اللحم‬
‫ نعناع‬،‫ االيكوالي‬،‫ مجتمع مكروبى‬،‫ انتاج‬،‫ دجاج لحم‬: ‫الكلمات المفتاحية‬

*Received:11/7/2019, Accepted:8/10/2019

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
INTRODUCTION which main phenolic compound in oil
Commercial poultry egg and meat production peppermint. Some studies indicated that
dramatically increased all over the world essential oils of peppermint inhibit the growth
between 1970 and 2005, poultry production of Escherichia coli, Staphylococcus aureus,
was increased faster than other animal Salmonella enteritidis, and Candida albicans
production like beef (26). This due to (22). Thus this study aimed to determine the
efficiency of the feed conversion and lower effect of different levels of peppermint
production costs related with intensive poultry (Mentha piperita) in feed and water on broiler
production. There are many challenges related performance (weight gain, feed intake, FCR)
to food safety, animal well-being, production at 10,24,35 day and histology of jejunum
efficiency of the animal, housing environment, (villus height, crypt depth, villi/crypt ratio,
management (19). They face many challenges muscle thickness) and organs percentage and
of bacterial and viral infection. Colibacillosis serum biochemical at day 24.
and salmonella are considered major bacterial MATERIALS AND METHODS
diseases in the poultry industry worldwide The study was conducted at the animal house
(11). Escherichia coli belongs to the of animal production department, College of
enterobacteriacea family, gram-negative Agricultural Engineering Sciences, University
bacteria, growing aerobic and anaerobic with of Duhok, Kurdistan region of Iraq.
varying size and shape (20). They mainly Animal husbandry
affect intestine of birds which cause many A total of 576-day old Ross 308 chickens were
diverse diseases, in which Escherichia coli divided randomly into 12 treatments with 4
(APEC) is the main reason of colibacillosis in replicate pen per treatment and 12 birds each
broiler chickens (15). It has a negative pen (wire mesh partitioned at (120*75cm) will
economic impact on poultry production assign in two room to prevent birds from infect
worldwide. Antibiotics have been used in the E. coli. The rooms temperature at the first day
poultry industry to fight against infection was set on 33-34 °C and then decreased by 3
disease. However, the frequent use of °C until the end of week. Temperature and
antibiotics may increase the incidence of light program were adjusted according to the
antibiotic resistance. Which leads to economic Ross 308 guide (Aviagen 2012). The water
losses and treatment failure, and there are also and feed were provided ad libitum the feeding
concerns about human health about the program consist three periods the first is starter
presence of antimicrobial residues in meat (7, from 0 to 10 day and second is grower from
9). It is important to find an effective way to 10-24 day, third finisher from 24 to 35. The
replace antibiotics to control infectious ingredient and composition of basal diet was
diseases and reduce the spread of resistant showed at Table 1. The primary determinants
bacteria (14). Medicinal herbs have been used of performance, i.e., cumulative pen weight,
as alternatives to antibiotics. Medicinal plants feed intake (FI), and feed conversion ratio
and their products including plant extracts or (FCR) were measured at d 10, 24 and 35.
essential oils are introduced as candidates for Dietary treatment
use in broiler diets and they have beneficial In this study the negative control (no additive
effects on broiler health(6). The mechanism of and non-challenge) and negative antibiotic
action of those medical plants may be through (non-challenge and the diet 0.33 g/kg zinc
stabilization of normal gut microflora, bacitracin). 0.5% peppermint in feed (5 g/kg
prevention of pathogens colonization and play peppermint in feed and non-challenge),
important role of digestive enzymes 1%peppermint in feed (10g/kg peppermint in
production and activities improvement (13). feed and non-challenge) and (0.5% peppermint
Peppermint is considered therapeutic herbs in in water and non-challenge) (1%peppermint in
the world is member from the Labiate family. water and non-challenge). However, the
It is broadly utilized in herbal medicine(21). positive control (there is no additive and
The peppermint consists of many chemical challenge and positive antibiotic (challenge
compounds which play important role as and addition 0.33 g/kg zinc bacitracin in diet).
antibacterial activities, for instance, menthol in 0.5% peppermint in feed (5 g/kg peppermint in

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
feed and challenge), 1%peppermint in feed software and images was taken by using a
(10g/kg peppermint in feed and challenge) color video camera (Dino-eye 20). The height
(0.5% peppermint in water and challenge), of villi and depth of crypts and thickness of
(1%peppermint in water and challenge). muscles from each replication was measured
Preparation of powder peppermint ten times. The means of crypt depth, villus
Fresh peppermint plant was obtained from height was obtaining and also villus height per
Duhok during July and August month in 2018. crypt depth ratio was determined.
At the beginning the thick stem was removed Relative organ percentage
and spread on a clean cloth under the sun for 2 From each pen 1 birds was picked for
days to dried up. eviscerating to calculate the heart, spleen
Challenge of E. coli bursa, liver and gizzard weight at day 24.
These bacteria were isolated in a laboratory Blood biochemical
from local farms. Incubated E. coli was Blood samples were collected from bird at day
occurring at 37 °C overnight in the 24 old. Two birds from each cage were
MacConkey broth sterile (100 ml), Then 0.1 randomly selected and slaughtered. The blood
ml of the previous broth Subsequent was collected from jugular vein, centrifuged at
incubations in Eosin methylene blue agar for 3000 rpm for 15 min and serum were taken,
accounting colony. 1000 mL of MacConkey frozen at -20E°C. Determination of plasma
broth was used to inoculate a colony from total protein (TP), albumin, cholesterol,
Eosin methylene blue agar to get the vaccine glucose, creatinine (CR), alanine
challenge. Birds were inoculated with a 1.5 ml aminotransferase (ALAT), aspartate
of the suspension E. coli (3.8 × 108 CFU per aminotransferase (ASAT), triglyceride (TG)
mL) at 8 and 9 day of the bird’s age. and globulin were determined by automatic
Measurements studied traits analyses Biolis (Tokyo Boeki Medical system)
Jejunum morphology 24i by using respective biochemical kits of
The fixed samples were dried, disinfected and Cormay company the Cat. No 4-204 and 4-404
embedded in paraffin wax for histological and the application of the procedure are
analysis subsequently. Successive longitudinal available on (prestige 24i Biolis ).
Statistical analysis
Superfrost® slices (Thermo Scientific, The comparison between means were carried
Rockville, MD, USA) and were staining with out according to Duncan's multiple range test
eosin and hematoxylin. The Crypt depth and (P < 0.05) using a computerized program of
Villus height was measured by Dino-eye SAS (SAS, 2013).
Table 1. Composition of the basal starter, grower and finisher diets as percentage
Ingredients Starter Grower Finisher
Corn 47 49.9 51.5
Wheat 5 5 5
Wheat bran 5 3 5
Soybean meal 36.45 34 30
Vegetable oil 1.5 3.4 4.45
Limestone 1.8 1.69 1
Dicalcium 0.7 0.5 0.5
Salt 0.05 0.01 0.05
Vitamin premix 2.5 2.5 2.5
Nutrient composition%
ME(kcal/kg) 2878 3035 3116
Crude protein 22.86 21.33 19.58
Crude fiber 3.02 2.76 2.92
Fat 3.76 5.64 6.8
Linoleic acid 1.92 2.88 3.48
Lysine 1.58 1.47 1.37
Methionine 0.66 0.64 0.63
Tryptophan 0.37 0.36 0.34
Methionine +cysteine 1.05 1.01 0.96
Threonine 0.97 0.92 0.86
Arginine 1.59 1.5 1.37
Calcium 1.08 0.99 0.73
Phosphor 0.54 0.5 0.49
Sodium 0.19 0.18 0.19
Chloride 0.26 0.24 0.26

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
RESULTS AND DISCUSSION conversion ratio over all treatment. The non-
Broiler performance challenged antibiotic, non-challenged 1%
Performance results are presented in table 2 Peppermint in feed, challenged 0.5% and 1%
,3and 4. Broiler performance including body Peppermint in water significantly improved
weight gain, feed intake and feed conversion feed conversion ratio in compare to non-
ratio were measured at day 10, 24 and 35. challenged control. The performance results
The results of this study showed a significant from day 0-35 are present in table 4. There
effect of additives on broiler performance was no significant effect of challenge on body
(Table 2). From 0-10 day the inclusion of weight gain, feed intake and feed conversion
antibiotic significantly increased weight gain ratio. The bird’s performance was markedly
and feed intake over all treatments. However, different upon the feed additive treatments.
the birds fed 1%peppermint had lower feed The highest (P<0.001) body weight gain and
intake compared to the control and antibiotic feed intake was recorded for birds fed
(P<0.002). Moreover, birds fed peppermint antibiotics. No significant different were found
either in water or feed decreased feed intake between birds fed control and peppermint
when compared to birds fed antibiotic. A either in water or feed for body weight gain.
significant challenge ×additive interaction was Birds fed control diet had significantly higher
observed in feed intake (p<0.05). The non- feed intake than birds fed 0.5 and 0.1
challenge 1% peppermint in feed treatment peppermint in water. All additives, except 1%
significantly had decrease feed intake when peppermint in water, significantly improved
compared with challenged control and feed conversion ratio over control. Challenge
antibiotic. From 0-24 day the effect of × additive interactions (P<0.001) was
additives was visible (Table 3). Birds fed observed for body weight gain, feed intake and
Antibiotic had higher (P< 0.01) body weight feed conversion ratio. Body weight gain of
gain over other additives. All additives had challenged antibiotic birds were significantly
better (P<0.001) feed conversion ratio when higher overall treatments except non
compared to control. Significant challenge × challenged antibiotic. Challenged birds fed
additive interactions (P < 0.01) were observed control diet had lower body weight gain when
in body weight gain and feed conversion. the compared to non-challenge 1% peppermint in
non-challenged birds fed antibiotic feed, no challenged control and challenged
significantly increase body weight gain over control. Challenged birds fed antibiotic had
challenged control and non-challenged 0.5% significantly higher feed intake in compare to
peppermint in water. On the other hand, the all treatments, whereas challenged birds fed
challenged or no challenged birds fed 1% peppermint in water had lower feed intake,
peppermint either in water or feed (except except 0.5% peppermint in water, compared to
non-challenged 0.5% peppermint in water) other treatments. Poorer feed conversion ratio
were not different in body weight gain in was recorded for challenged birds fed control
compare to non-challenged birds fed diet. All treatments (challenged and non-
antibiotic. In term of feed conversion ratio, the challenged) significantly improved feed
challenged control had poorer(P<0.001) feed conversion ratio over challenged control.

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
Table 2. Effect the all treatments on bird performance at day 10
Initial
weight gain Feed intake
Treatment weight FCR
g/bird g/bird
g/birds
Non challenge control 40 212 242bc 1.143
Non challenge antibiotic 40 241 247bc 1.028
Non challenge 0.5% Peppermint in feed 40 223 238bc 1.071
Non challenge 1% Peppermint in feed 40 217 224c 1.030
Non challenge 0.5%Peppermint in water 40 212 243bc 1.155
Non challenge 1% Peppermint in water 40 211 236bc 1.124
Challenge control 40 218 250b 1.153
Challenge antibiotic 40 225 275a 1.223
Challenge 0.5% Peppermint in feed 40 210 234bc 1.121
Challenge 1% Peppermint in feed 40 208 228bc 1.108
Challenge 0.5% Peppermint in water 40 221 225bc 1.016
Challenge 1% Peppermint in water 40 211 236bc 1.118
SEM 2.1675 2.7209 0.0158
Main effects
Challenge
No 40 221 238 1.081
Yes 40 215 239 1.117
Additive
Control 40 215b 246ab 1.148
Antibiotic 40 233a 261a 1.125
0.5% Peppermint in feed 40 217b 236bc 1.096
1% Peppermint in feed 40 213b 226c 1.069
0.5% Peppermint in water 40 217b 234bc 1.085
1% Peppermint in water 40 211b 236bc 1.121
P-Value
Challenge 0.2325 0.7777 0.3319
Additive 0.0430 0.0020 0.7436
Challenge * additive 0.0964 0.0038 0.2300
a-c
Means the different column was superscript are significantly different (p<0.05)
Abbreviations: SEM, standard error of the mean., FCR, feed conversation ratio
Table 3. Effect the all treatments on bird performance at day 24
Weight gain Feed intake
Treatment FCR
g/bird g/bird
Non challenge control 1033ab 1401ab 1.358b
Non challenge antibiotic 1122a 1384ab 1.234cde
Non challenge 0.5% Peppermint in feed 1031ab 1332abc 1.292bcde
Non challenge 1% Peppermint in feed 1061ab 1275abc 1.202de
Non challenge 0.5%Peppermint in water 994b 1336abc 1.344bc
Non challenge 1% Peppermint in water 1074ab 1405a 1.310bcd
Challenge control 886c 1352abc 1.538a
Challenge antibiotic 1083ab 1304abc 1.204de
Challenge 0.5% Peppermint in feed 1047ab 1349abc 1.289bcde
Challenge 1% Peppermint in feed 1019ab 1306abc 1.285bcde
Challenge 0.5% Peppermint in water 1045ab 1237c 1.187e
Challenge 1% Peppermint in water 1041ab 1268bc 1.219de
SEM 11.9977 12.593 0.0161
Main effects
Challenge
No 1056 1346 1.276
Yes 1047 1293 1.237
Additive
Control 960b 1376 1.448a
Antibiotic 1103a 1344 1.219b
0.5% Peppermint in feed 1039ab 1340 1.290b
1% Peppermint in feed 1040ab 1290 1.244b
0.5% Peppermint in water 1019ab 1286 1.265b
1% Peppermint in water 1057a 1337 1.264b
P-Value
Challenge 0.6652 0.0567 0.076
Additive 0.017 0.2815 <.0001
Challenge * additive 0.0128 0.1007 <.0001

a-b
Means the different column was superscript are significantly different (p<0.05)
Abbreviations: SEM, standard error of the mean., FCR, feed conversation ratio

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
Table 4. Effect the all treatments on bird performance at day 35
weight gain Feed intake
Treatment FCR
g/bird g/ birds
Non challenge control 1910bc 2892b 1.514b
ab bc
Non challenge antibiotic 2015 2871 1.427b
Non challenge 0.5% Peppermint in feed 1921bc 2874bc 1.529b
Non challenge 1% Peppermint in feed 1959b 2826bc 1.476b
Non challenge 0.5% Peppermint in water 1856bc 2704bc 1.490b
bc bc
Non challenge 1% Peppermint in water 1914 2872 1.531b
Challenge control 1755cd 2861bc 1.637a
Challenge antibiotic 2159a 3261a 1.511b
Challenge 0.5% Peppermint in feed 1993b 2815bc 1.444b
bc bc
Challenge 1% Peppermint in feed 1896 2690 1.450b
Challenge 0.5% Peppermint in water 1867bc 2652cd 1.454b
Challenge 1% Peppermint in water 1678d 2474d 1.512b
SEM 21.9956 31.5643 0.0113
Main effects
Challenge
No 1933 2829 1.4906
Yes 1918 2778 1.4742
Additive
Control 1833bc 2876b 1.575a
a a
Antibiotic 2087 3066 1.469b
0.5% Peppermint in feed 1957b 2845bc 1.487b
1% Peppermint in feed 1927bc 2758bc 1.463b
0.5% Peppermint in water 1861bc 2678c 1.472b
c c
1% Peppermint in water 1796 2673 1.521ab
P-Value
Challenge 0.7565 0.4832 0.4392
Additive 0.0004 0.0006 0.0218
Challenge * additive <.0001 <.0001 0.0056
a-c
Means the different column was superscript are significantly different (p<0.05)
Abbreviations: SEM, standard error of the mean., FCR, feed conversation ratio
Jejunum morphology level of peppermint had significant effect on
The results of jejunal villus height, crypt villi crypt ratio that is increase villi crypt ratio
depth, villi/crypt ratio and muscle thickness at compared with control. Also, there is no
day 24 are present in table 5. Additives had a significant effect between challenge and non-
significant effect on jejunum muscle thickness challenge birds p>0.05 and there is non-
and villus/ crypt ratio. The birds fed 0.5% and significant challenge ×additive interactions
0.1% peppermint either in water or feed observed p<0.05 in villus height, crypt depth,
significantly reduced the muscle thickness villi/crypt ratio, muscle thickness.
compared to birds fed control diet. All Organs percentage
additives had higher (P<0.01) villus/ crypt Organ percentage from live body weight
ratio over control. There were significant results are presented in table 6. The inclusion
challenge*additive observed in crypt depth of all peppermint additives was not affect
and villus/ crypt ratio. The inclusion of organ percentage but only the antibiotic has
additive is non-significant effect on villus decrease intestine percentage. Also there is no
height, crypt depth, muscle thickness) at the 24 significant effect found between challenge and
day only the significant effect appears on non-challenge bird and also there is no
villi/crypt ratio p<0.05 in which the different

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
significant × additive interaction was observed albumin and globulin. Challenged control
in percentage of internal organ. birds had highest serum glucose concentration
Serum biochemical compare to non-challenged treatments. While
Serum biochemical results are present in Table the birds fed challenge 1% peppermint in
7 refer the effect of antibiotic, different levels water had significantly lower serum glucose
of peppermint (Mentha piperita) in feed or compared to challenged treatments. The serum
water on serum biochemical (glucose, total protein concentration of challenged birds
creatinine, total protein, albumin, cholesterol, fed 1% peppermint in feed or water and 0.5 %
triglyceride, GOT(ASAT), ALAT, globulin). peppermint in water were significantly lower
At day 24 the effect of challenge was clearly than birds fed challenged antibiotic and non-
visibly. Challenged birds have higher challenged 0.5 peppermint in water. The serum
(P<0.001) glucose, creatinine and albumin albumin concentration of challenged birds fed
than non-challenged birds. However, non- antibiotic and control was higher (P<0.001)
challenged birds had significantly higher total than all non-challenged treatments and
protein and globulin than challenged birds. No challenged 1% peppermint in feed or water.
significant effect of additives was observed in The peppermint treatment in water and feed
glucose, creatinine, total protein, albumin, and antibiotic under non-challenge condition
cholesterol, triglyceride, (ASAT), ALAT, and negative control had significantly
globulin. There were challenge*additive increased serum globulin compared with this
interaction observed for glucose, total protein, treatment under E. Coli challenge.
Table 5. Effect of all treatments on villi height, crypt depth, muscle thickness, villi/crypt
ratio at 24 day
Villi height Crypt depth Muscle thickness Villi / Crypt
Treatment means
um Um Um
Non challenge control 1194 224ab 209 5.449ab
Non challenge antibiotic 1155 157c 187 7.472a
Non challenge 0.5% Peppermint in feed 1693 239ab 191 7.093a
Non challenge 1% Peppermint in feed 1339 201bc 200 6.723a
Non challenge 0.5% Peppermint in water 1130 184bc 179 6.187a
Non challenge 1% Peppermint in water 1175 197bc 174 6.055a
Challenge control 1052 268a 237 4.038b
Challenge antibiotic 1316 241ab 216 5.532ab
bc
Challenge 0.5% Peppermint in feed 1335 199 185 6.826a
Challenge 1% Peppermint in feed 1356 202bc 205 6.683a
Challenge 0.5% Peppermint in water 1259 215abc 205 5.871ab
Challenge 1% Peppermint in water 1378 218ab 176 6.482a
SEM 38.4788 6.1522 4.5253 0.2015
Main effect
Challenge
No 1277 193 186 6.686
Yes 1329 215 197 6.279
Additive
Control 1123 246 223a 4.744b
Antibiotic 1235 199 201ab 6.502a
0.5% Peppermint in feed 1488 216 187b 6.940a
1% Peppermint in feed 1347 202 202ab 6.703a
0.5% Peppermint in water 1194 199 192ab 6.029a
1% Peppermint in water 1277 208 175b 6.268a
p-value
Challenge 0.5667 0.0606 0.1818 0.314
Additive 0.1103 0.2015 0.0442 0.0197
Challenge*additive 0.1665 0.0172 0.1277 0.0367
a-b
Means the different column was superscript are significantly different (p<0.05)
Abbreviations: SEM, standard error of the mean

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
Table 6. Effect of all treatments on organ percentage at day 24
Intestine Liver Gizzard Pancreas Spleen Bursa
Treatment means
% % % % % %
Non challenge control 6.536 2.534 3.422 0.359 0.073 0.202
Non challenge antibiotic 6.048 3.313 3.788 0.326 0.097 0.273
Non challenge 0.5% Peppermint in
feed 7.041 2.628 3.53 0.352 0.103 0.201
Non challenge 1% peppermint in feed 6.469 2.512 3.825 0.344 0.097 0.256
Non challenge 0.5%peppermint in
water 6.389 3.138 3.691 0.356 0.112 0.24
Non challenge 1%peppermint in water 7.282 2.811 3.728 0.375 0.093 0.238
Challenge control 6.669 2.781 3.774 0.345 0.088 0.198
Challenge antibiotic 4.961 3.569 3.896 0.288 0.124 0.208
Challenge 0.5% Peppermint in feed 6.96 3.014 3.845 0.336 0.077 0.258
Challenge 1% Peppermint in feed 7.245 2.995 3.562 0.382 0.086 0.243
Challenge 0.5%Peppermint water 6.927 2.964 3.843 0.385 0.095 0.229
Challenge 1%Peppermint in water 6.954 2.931 3.45 0.385 0.095 0.227
SEM 0.157 0.0795 0.0694 0.0071 0.0039 0.0073
Main effect
Challenge
No 6.625 2.88 3.712 0.351 0.1 0.242
Yes 6.609 3.094 3.719 0.355 0.095 0.233
Additive
Control 6.603a 2.658 3.598 0.352ab 0.08 0.2
b
Antibiotic 5.504 3.441 3.842 0.307b 0.111 0.24
0.5% Peppermint in feed 6.994a 2.821 3.687 0.344ab 0.09 0.229
a a
1% Peppermint in feed 6.857 2.753 3.694 0.363 0.091 0.25
a a
0.5% Peppermint in water 6.658 3.051 3.767 0.371 0.103 0.234
1% Peppermint in water 7.118a 2.871 3.589 0.380a 0.094 0.232
p-value
Challenge 0.9621 0.2311 0.966 0.7767 0.5447 0.5873
Additive 0.0313 0.053 0.9071 0.042 0.2654 0.5013
Challenge*additive 0.1437 0.2149 0.9454 0.1919 0.3625 0.5096
Abbreviations: SEM, standard error of the mean

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
Table 7. Effect all treatments on serum biochemical 24 day
Glu Cre Tp Alb Cho Tri Got ALAT Glo
Treatment means
mg/dl g/dl g/dl g/dl mg/dl Mg/dl U/L U/L g/dl
dc ab c
Non challenge control 239 0.163 3.040 1.153 116 72 591 3.55 1.888ab
Non challenge antibiotic 233dc 0.063 3.113ab 1.265bc 135 103 210 4.4 1.848abc
Non challenge 0.5% P.P
241dc 0.153 2.825abc 1.138c 89 58 234 3.3 1.688bcd
in feed
Non challenge 1% P.P in
234dc 0.078 3.185ab 1.285bc 117 57 281 4.05 1.900ab
feed
Non challenge 0.5% P.P
218d 0.143 3.470a 1.398bc 119 71 356 4.475 2.073a
in water
Non challenge 1% P.P in
228dc 0.108 3.165ab 1.305bc 138 106 235 4.25 1.860abc
water
Challenge control 335a 0.198 3.285ab 1.850a 170 79 233 6.8 1.435def
ab a a
Challenge antibiotic 300 0.183 3.370 1.863 150 89 268 6 1.508cde
Challenge 0.5% P.P in
312ab 0.153 2.850abc 1.613ab 144 89 260 4.2 1.238efg
feed
Challenge 1% P.P in
303ab 0.18 2.635bc 1.368bc 144 106 252 6.275 1.268efg
feed
Challenge 0.5% P.P in
298ab 0.148 2.663bc 1.540ab 117 70 220 7.825 1.123fg
water
Challenge 1%P.P in
272bc 0.145 2.243c 1.270bc 110 63 238 4.225 0.973g
water
SEM 6.7947 0.0109 0.0703 0.0436 4.9384 5.2993 27.156 0.4098 0.0581
Main effect
Challenge
No 231b 0.109b 3.152a 1.278b 119 79 263 4.095 1.874a
Yes 297a 0.162a 2.752b 1.531a 133 83 247 5.705 1.222b
Additive
Control 287 0.18 3.163 1.501 143 75 412 5.175 1.661
antibiotic 266 0.123 3.241 1.564 142 96 239 5.2 1.678
0.5% P.P in feed 276 0.153 2.838 1.375 117 74 247 3.75 1.463
1% P.P in feed 268 0.129 2.91 1.326 130 82 266 5.163 1.584
0.5% P.P in water 258 0.145 3.066 1.469 118 70 288 6.15 1.598
1% P.P in water 250 0.126 2.704 1.288 124 85 236 4.238 1.416
p-value
Challenge <.0001 0.0247 0.0088 0.0012 0.1201 0.7293 0.5378 0.07 <.0001
Additive 0.7248 0.6715 0.2129 0.4199 0.4889 0.7873 0.4247 0.6412 0.7562
Challenge*additive <.0001 0.2917 0.0037 <.0001 0.0612 0.5144 0.2466 0.4666 <.0001
Abbreviations: SEM, standard error of the mean., (p.p) peppermint (Glu)Glucose, (Cer)creatinine, (TP)total
protein, (Alb)albumin, (Cho)cholesterol, (Tri) triglyceride, (ALAT) alanine aminotransferase, (ASAT) aspartate
aminotransferase, (Glo)globulin
The peppermint supplementation in feed (Thymus vulgaris L.) leaves don’t have any
decreased feed intake at 10 days. The results considerable effect on feed intake. At the 24
were in agreement with the finding of Aroche day the peppermint supplementation either in
(5) who reported that the supplementation of feed and water had improve feed conversion
mixed leaves powder of medicinal plant ratio. Also, the all peppermint additive except
reduce feed intake. Also Amasaib (4) who 1% in water had significantly improved feed
showed that the addition of different levels conversion ratio at day 35. This result is
of spearmint (Mentha spicata) improved similar to Gurbuz (10) who found that the
feed intake. While, the results were in contrast peppermint supplementation had a
with finding of Ocak (18) who reported that significantly affect on FCR compared with
the dietary supplementation of dried control. Also Ahmed (2) they reported the
peppermint (Mentha piperita L.) and thyme addition 250 mg peppermint extract and

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
supplement 1 g flavomycin at the grower and mint groups(16). According to this study, the
Finisher periods had decrease the feed all peppermint additive in water or feed and
conversion ratio compared with other antibiotic don’t observed any considerable
treatment. Also the results were agreed with effect on internal organ percentage (intestine,
Mustafa (17) study who reported that (1% liver, gizzard, pancreas, spleen) at day 24. The
betony+ 1% peppermint) had significant effect results were similar to the finding of Toghyani
on feed conversion ratio in which have better (24) who reported that the internal organ
when compared with control at age 25- 42 weight are not affect by dietary addition of
day. In contrast, Witkowska (27) who peppermint . Also Ocak (18) who reported the
reported the essential oil mist in peppermint no dried peppermint (Mentha piperita L.) or
effect on feed conversion ratio from (1 to 42 thyme (thymus vulgaris L.) in diet supplements
days). Most of medical plants consist of was not have any significant effect on the
different essential oil in which stimulate weights of edible organs , pancreas and
digestive enzyme secretion and the effect on gizzard. Abdel-Wareth (1) found the addition
microbial that present in gut through the of menthol and leaves peppermint had no
balance of microbial (6). The peppermint effect on relative weight of pancreas ,gizzard,
supplementation either in water or feed have heart ,liver, spleen .While the result was in
effect on weight gain on day 24 and 35. The contrast with the finding Mustafa (17) who
result was agreement with finding showed the supplementation of dried
Witkowska (27) study who reported the peppermint 2% in broiler diet had significant
addition peppermint oil mist had increase the increase liver and heart and gizzard
mean BW and WG than control and birds percentage. According to this study,
exposed to thyme oil mist. While the result peppermint supplementation either in water or
was in disagreement with the finding of Ocak feed and antibiotic had not considerable effect
(18) who recorded the addition peppermint on serum biochemical at day 24. The results
had not any effect on body weight at 42 of age. were in agreement with finding of Khursheed
The reason the peppermint improve bird (12) who reported that the supplementation of
performance may be due to that the herbs like mint leaves with or without enzyme in both 1
peppermint contain different his was variety of or 2% levels were not observed any
active components which affect process of significant effect on serum glucose, total
digestion in which stimulate saliva secretion protein, cholesterol, SGPT and SGOT when
and enhance bile acid synthesis in liver and compared with control. Also the results in this
bile excretion which affect process digestion study were similar to the finding of Toghyani
and lipid absorption (8). The peppermint (24) who showed that the addition of different
supplementation either in feed or water at 24 levels of black seed (Nigellasativa) and
day had a significant effect on jejunum muscle peppermint (Mentha piperita) had no
thickness and villus/ crypt ratio while not significant effect on triglyceride , protein,
affect on villus height and crypt depth. The albumin, total cholesterol. In contrast, Ahmed
result were agreement with Yang (28) whom (2) indicated that peppermint had significant
reported that the addition of essential oils and effect in which they increase the cholesterol
organic acids in bird diet during finisher and total protein ,globulin and liver enzyme.
period were increased villus height to crypt According the study of Akbari (3)
depth ratio when compared to the bird fed diet supplementation of essential oil of peppermint
with 0.15 g per kg enramycin at grower period. and chromium picolinate under heat stress
While, the result disagree with finding of decrease the concentration of triglycerides and
Ahmed (2) whom recorded that the extract of glucose. Also Tayeb (23) in their study found
peppermint had desirable effect on crypt the medical plants had significant affect on
depth and villus height in ileal in small albumin, globulin, cholesterol and glucose
intestine. The increase of villus height: crypt concentration. The result of serum
depth ratio can indicate an increase in the biochemical, in this study were varied there
turnover of epithelial cells in the intestinal part are more factors affecting on serum
due to the increase of beneficial bacteria in biochemical. It can be nutritional situation and

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Iraqi Journal of Agricultural Sciences –2020:51(1):299-310 Hasan & Sadeq
compounds of herbs and also there are many extracts in animal nutrition. Acta Agric Slov,
other factor, like age, sex, type of bird, and 94(2): 95-102
environmental (25). 9. Furtula, V, E. Farrell, F. Diarrassouba, H.
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