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Sudan Academy of Sciences (SAS)
Atomic Energy Council
Determination of Trace Elements in Chicken Feeds in Khartoum state
A thesis Submitted for partial Fulfillment of Requirements for the Degree of
M.Sc. in Nuclear Sciences and Technology
By
Salma Yahya Mohammed Hassan
Supervisor
Dr. Ashraf Mohammed Khair
March, 2017
Sudan Academy of Sciences (SAS)
Atomic Energy Council
Determination of Trace Elements in chicken feeds in Khartoum State
By
Salma Yahya Mohammed Hassan
Examination Committee:
Name Title Signature
Prof. Mohammed Ahmed External Examiner - ٧ر
Dr. Ashraf Mohammed
Khair
Supervisor'.'١ .
Examination Date: March,2017
؛
اا
Lأ
حرج،rI
اا،
I
ق؛
DEDICATION
To My Family
To My Teaches
To My Fends
ا
1
S،' ١.١١LEDGE١1E()ACK١
I would like to express my deepest appreciation to all those who provided me the
possibility to complete tliis research. A special gratitude to Dr. Ashraf Mohammed
Khair, my research supervisor, for Iris patierrt guidairce, encouragement arrd usel'ul
critiqrres of this research work.
I am grateful for Mr. Mgzoob Alamien who Irelped rrre to collect my sanrples. And
I would like also to extend my tharrks to Ms. Nwaal Mgzoob for her help m samples
preparation.
Abstract
Trace elements are very important for chicken because they contribute m biochemica:
processes repaired tor normal growth, development and formation 0؛ the eggshell. The
deficiency 01* the elevation of these trace elements may affect the normal physiological
activity and biochemical process of their bodies. In this study the concentrations of trace
elements in chicken feed samples were determined by XRF spectrometry.
The results showed that there were 9 trace elements in sample (R, Ca. Mn, Fe, Cu. Zn. Pb.
Rb, Sr) the mean concentrations were R (2.12^10^), Ca (2.22x0), Mn (48.1 )١ Fe (191).
Cu (9.81), Zn (24.6), Pb (1.63), Rb (7.79), Sr (4.09) ppm.
Comparing the trace elements concentrations obtained in this study with the recommended
values showed that some of them e.g., Mn, Cu, Zn, Fe were found in higher concentration
tlian the recommended values, which stipulated by national research council NRC Europin
uonion. While, lead concentration was in the permissible limit (5mg/kg). However, the
statistical analysis revealed that there wei'e no significant difference between all
concentrations of trace elements in the two types of chicken feeds l.e.. layer and broiler.
The Pearson correlation test displayed a strong correlation between R-Rb (().885), Mn-Zn
(O.874).The negative correlations between Mn-Rb (-0.680), R-Mn (-0.600), R-Ca ( -0.565).
Zn-Rb (-0.541), Ca-Rb (-0.458) were found. Further investigations were performed using
the principal component analysis (PCA), which involved the exraction of principal factoi-
to study the total variance in the feed samples in terms of the trace elemeirts concentrations
in each. The obtained results revealed that the first principal component have a positive
correlation with the elements Ca, Mn, Pb and Zn, while R and Rb showed negative
correlation with it. Similarly, the second principal component showed positive correlation
vrith the elements Mn, Pb and Zn, while Ca and Sr slrowed negative, on the other hand the
third component was found to have a positive correlation with Cu and negative with Fe,
and these !'esults explained the loading of each element in the components. As a conclusion,
the PCA factors were found to explain approximately 71% of the total variance in the
chicken feed samples.
،المستخلصى
همية بالغة ائنذرة العناصر قشرة وتكوين الطبيعي للنمو ومطلوبة الحيوية العمليات في لمساهمتها وذلك للدواجن ا
اجسام في الحيوية والعمليات الطبيعي النمو في يوثر ان يمكن العناصر هذه تراكيز في اوالنقصان الزيادة" البيض.
شعة مطيافية جهاز بواسطة الدواجن اعلاف في النذرة العناصر تراكيز علي التعرف تم الدراسة هذه في الدواجن. ا
المفغورة. السينية
1
Ii
نإا
4
،النحاس ،المنجنيز،الحديد ،الكالسيوم ،البوتاسيوم ) وهي العينات كل في عناصر 9 وجود أظهرت النتائج
سترانشيوم( و الرصاص،الربيديوم الزينك، تراكيز بمتوسط ا
10 *2٠12 ,0 *4.09,7.79,1.63,24.6,9.81,191,48.1,2.22 ppm تراكز قورنت فدما الزار. طي بحاث هينة من بها الموصي المستويات مع الدراسه هذه في المتحصله العناصر وربيه ا مطم وجدت ،ا
الحد ضمن بتركيز وجد فقد الرصاص اما به، الموصي المستوي من اعلي بتركيز (Mn, Cu, Zn, العذاصر)ح^
به. المسموح
حصائى التحليل الدجاج غذاء ) العينات من المجموعتين في العناصر تراكيز بين معنوي اختلاف وجود عدم أظهر ا
العناصر تركيز بين قويه ارتباط علاقة وجود بيرسون ارتباط معامل اختبار أظهر اللاحم(. الدجاج وغذاء البياض
(0.874) K-Rb (0.885), Mn-Zn (0.600,) بين عكسية ارتباط علاقة وجدت ٠- MR (-0.680), K-Mn
(0.458)- K-Ca (-0.565), ZRb (-0.541), Ca-Rb ٠
ساسيه المكونات تحليل اجراء تم ايضا ساسيه المكونات معامل استحلاس يتضمن الذي (PCA) ا ختلاف لدراسة ا ا
المكون ان توضح عليها المتحصل النتائج ٠ عينه لكل النذره العناصر تركيز ناحية من الدواجن غذاء عينات في الكلي
ساسي ول ا K, مع عكسية ارتباط علاقة وجدت بينما Ca, Mn, Pb, Zn العناصر مع موجبه ارتباط علاقة له ا
Rb ساسي المكون مماثل نحو .علي اظهرعلاقة بينما Mn, Pb, Zn العناصر مع موجبة ارتباط علاقة أظهر الثاني ا
ساسي المكون اخري ناحية من . Ca,S٢ مع عكسية ارتباط وعلاقة ء مع موجبه ارتباط علاقة أظهر الثالث ا
ساسيه المكونات في العناصر تأثير توضح النتائج هذه Fe مع عكسيه ارتباط ساسيه المكونات تحليل أخيرا . ا وجد ا
ختلاف يفسر انه %.71 بنسبة الدواجن علف عينات في الكلي ا
ا—
'***
ا
IV
Tabic ot Contents
DEDICATION........................................................................................................................1ACKNOWLEDGEMENTS................................................................................................. iiABSTRACT........................................................................................................................... iii.........................................................................................................................المستخلص iv
LIST OF TABLES............................................................................................................. viiLIST OF FIGURES.......................................................................................................... viilLIST OF ABBREVIATIONS............................................................................................ixChapter 1
INTRODUCTION.............................................................................................................. 10
1.1 Background....................................................................................................... 101.2 Trace Elements...................................................................................................... 1 11.3 Objectives.............................................................................................................. 12
Chapter 2
LITERATURE REVIEW.................................................................................................132.1 Trace Element....................................................................................................... 132.2 Functions of Minerals............................................................................................132.3 Poultry Nutrition................................................................................................... 142.4 Trace elements in poultry feeds........................................................................... 142.5 .Effects of some trace elements and heavy metals in chickens........................... 18
2.5.1 Chromium Cr.................................................................................................182.5.2Zinc..............................................................................................................182.5.3 Calcium.......................................................................................................... 192٠5.4Lead.................................................................................................................192.5.5Cadmium.........................................................................................................202٠5.6Iron.................................................................................................... ......202.5.7Manganese..................................................................................................... 21
2٠5.8Nickel..............................................................................................................212.5.9Iodine..............................................................................................................21
.......................................................................................................Selenium؛■ 2.5.10 222.5.1 IPhosphorus...................................................................................................23ح٠٠t
١٠
؛
Chapter 3
MATERIAL AND METHOD..........................................................................................243.1 Sample collection...................................................................................................24
3.2 Sample preparation and analysis........................................... 24
3.2.1 Radioactive soui'ce.......................................................................... . 24
3.2.2X-ray emission............................................................................ -.24
3.2.3 Semiconductor Si(Li) detector.......................................................................25
3.3 Statistical Analysis.................................................................................................26
Chapter 4
RESULTS AND DISCUSSION 27
4.1 Validation of the x-ray Fluorescence...................................................................274.2 Trace Elements Concentrations in Chicken Feeds...............................................28
4.3 The Difference of Concentrations of Heavy Metals in Layer and BroilerChicken Feeds................................................................................................................... 304.4 The Correlation between Heavy Metals in two Types of Feed........................... 304.5 Principal Component Analysis..............................................................................3 1
4.5.1 Principal Correlation Matrix..........................................................................334.6 Comparison of the Trace Elements Concentrations in Chicken Feeds and thePrevious Studies and Recommended Level.....................................................................34
CONCLUSION................................................................................................................... 36RECOMMENDATIONS......................................................... 37REFERENCES.................................................................................................................... 38APPENDIX A...................................................................................................................... 43
VI
١
LIST OF TABLES
Table 4.1. Trace elements concentration in the hey powder standard.............................. 2S
Table 4.2 The concentrations (ppm) of the trace elements in chicken feeds .... ٠...٠..,,,٠٠ 20
Table 4.3. The correlation between the trace elements concentration in chicken feeds .٠. 3 1
Table 4.4. The total variance explained by PCA................................................................32
Table 4.5. Principal correlation matrix (n = 20).................................................................33
vn
LIST OF FIGURES
Figure 3٠ 1. Working principle of a Si(Li) detector............................................................. 26
Figure 4.1. XRF spectrum of the Hey standard...................................................................27
Figure 4.2. XRF spectrum of the chicken feeds samples.................................................... 30
Figure 4.3. The scree plot of the principal components...................................................... 32
Figure 4.5. The score plot of the first component (PCI) and the second component (PC2).
..............................................................................................................................................34
vm
Chapter 1
INTRODUCTION
1.1 Background
World Health Organization define food is any substance consumed to provide nutritional
support for the body( It is usually of plant or animal origin. Some food is obtained directly
from plants; but even animals that are used as food sources are raised by feeding them food
derived from plants (Cereal grain. Com (maize), wheat, and rice) Most of the gran) that is
produced worldwide is fed to livestock.
Poultry is a category of domesticated birds kept by humans for the purpose of collecting
their eggs, or killing for their meat and/or feathers) Poultry IS the second most widely eaten
meat in the world accountiirg for about 30% of meat pi'oduction worldwide, after bee!'at
38%. Poultry is basically a source of economic, palatable and healthy food protein. Poultry
feed is food for farm poultry, including chickens, ducks, geese and other domestic birds.
Feed for poultry mostly consists of grains (Mahesar ct al٠, 2010).
Chickens are one of the main sources of protein. It have genetically been selected for
improve feed conversion and rapid growth or production of eggs. This has led to two
different types of chickens: broiler- and layer-type chickens as a consequence of this
selection foi' economically important production traits. These chickens differ in body
١veight gain, duration of life and, recently data ai'e becoming available, also in immune
system caused by genetic differences. Therefore concern has risen with respect to their
ability to a mount an immune response to the whole array of pathogens as encountered in
poultry breeding (Roenen et al., 2002).
Minerals are the inorganic part of feeds or tissues. They are often divided into two
categories, based on the amount that is required in the diet. Requirements for major, or
macro, minerals usually are stated as a percentage of the diet, whereas requirements for
minor, or ti'ace, minerals are stated as milligrams pel' kilogram of diet or as parts per
million. Minerals are required for the formation of the skeleton, as c٢ mponents of various
10
compounds with particular functions within the body, as couictors of enzymes, and 01؛' the
maintenance of osmotic balance within the body of the bird. Calcium and phosphorus are
essential for the formation and maintenance of the skeleton. Sodium, potassium,
magnesium, and chloride function with phosphates and bicarbonate to maintain
homeostasis of osmotic !'elationships and pH throughout the body (Dale, 1994).
Most of the calcium in the diet of the growing bird is used for bone formation, whereas in
the mature laying fowl most dietary calcium is used for eggshell formation. Other functions
of calcium include roles iir blood clotting and as a second messenger in intracellular
communications (Dale, 1994).
1.2 Trace Elements
The elements found in body in small amounts such as cobalt (Co), copper (Cu), manganese
(Mn), molybdenum (Mo) and selenium (Se) are elements important for maintaining the life
processes in plairts and/or animals including humans. Low uptake/intake causes deficiency
and high cause’s toxicity (Johnsson, 2005).
Trace elements in nutrition depends on total content of elements in soil which effects on
plants, airimals and humans of the 21 most environmentally important heavy metals and
metalloids, deficiencies of heavy metals, such as zinc, copper . manganese and metalloids
such as selenium in agricultural soil are affecting agricultural productivity and human
health (Alloway, 2013).
Heavy metals can be classified as potentially toxic (aluminium, arsenic, cadmium, lead,
mercury, etc.), pi'obably essential (nickel, vanadium and cobalt) and essential (iron,
manganese, copper, zinc and selenium). Toxic elements can be very harmful even at low'
concentration when ingested over a long time period. The essential nretals can also produce
toxic effects when the metal intake is excessively elevated (Uluozlu et al., 2009).
WH0/FA0 set peianissible level to metals in chicken feed, metals are added to chicken
feed. Excessive or deficiency use of tlrese metals may catise problems in chicken body
11
rI'الو
health, some of these elements may cause severe toxicity ٠. ٠٠ iieh اأ١٠اًئ؛١ m death. la .Sudan
there is lack of information and researches. The concentrations of metals in chicken feeds
were stndied in Khartoum state 2016٠ Trace elements in chicken feeds are significant for
normal growth, development, and egg shell formation. All metals are toxic at higher
concentration aird accumulation over time in chicken body can cause serious illness.
1.3 Objectives
Generally, this work aims to identify and estimate the trace elements concentrations m
chicken feeds, specifically to:
1. Identify the trace elements and determination theii' concentrations in chicken feeds.
2. Compai'e the trace elements concentrations in layer and broiler chicken feeds.
3. Measui'e the correlation between concentrations of trace elements in the chicken feeds.
12
Chapter 2
LITERATURE RE١IE١٠V
2.1 Trace Element
Trace elements is significant of both human beings and animal’s life although the content
of each trace element is less than 0.01%. Trace elements are contributory in metabolism
and maintain the dynamic balance in the body with variety of forms (Macharia et al., 2005).
2٠2 Functions of Minerals
Minerals have four broad kinds of functions for exist structural, physiological, catalytic
and regulatory- although they are not exclusive to particular element in the same individual.
1. Structural: the structural components of body organs and tisstics can be form by
minerals, represent by calcium, phosphorus and sulphur in muscle proteins,
Miirerals such as zinc and phosphorus can also conti'ibute structural stability to the
molecules and membranes of which they are part.
2. Physiological: minerals act as electrolytes in body fluids and tissue, interested to
maintenance of osmotic pressure, acid- base balance, membrane permeability and
tissue irritability: sodium, potassium, chloride, calcium and magnesium in blood,
cerebrospinal fluid and gastric juice provide examples of such functions.
3. Catalytic: minerals can act as catalysts in enzyme and hormone systems, as integral
and specific components of the structure of metalloenzymes or as less specific
activators with in those systems.
4. Regulatory: in recent years, minerals have been found to regulate cell replication
and differentiation; calcitim, for example influences signal transduction and zinc
influences transcription adding to long-established regulatory roles, such as that of
the element iodine as a constituent of thyroxin (Underwood. 1999). Basically
essential trace elements are needed for' tissue repair, for growth, for metabolism and
for slowing down some of the degenerative processes (Macharia et ah, 2005).
The deficiency or elevation of these trace elements can affect the normal physiological
activity and biochemical process of the body, thereby incurring abnor nal cell metabolism.
13
)
growth and reproductive disorders, and sever oxidative 'damage. Therefore, the trace
elements that exist in their different organs might significantly maintain in balanceLu et
al.,2O15).
People need varying amount of trace element iron, cobalt, copper, manganese,
molybdenum and zinc for living organisms. Excessive levels of which can damage an
organism. Other toxic elements including mercury and lead have no vital and beneficial
effects on organisms and their accumulation in the body over time leads to very serious
diseases. Some elements which are normally toxic are useful for some organism in certain
cases (Altundag et ah, 2015).
2.3 Poultry Nutrition
Poultry feeds are known as a complete feed, since it is prepared in such a way to contain
all the vitamins, minerals, energy, protein and other nutrients essential for proper health of
the birds, egg production and growth (Bukar and Saeed, 2014). Feed ingredients and
additives in poultry diets contain different sources: Com and soybean meal sources of
energy, fish meals and meat meals good sources of protein and amino acids Bale et al.
(2015) reported also that the feed consists of gains such as com, wheat or barley, oil seeds,
cake meal (originating mainly from oil producing seeds such as soybeans) sunflower seeds,
peanuts, cotton seed and protein products of animal origin such as fish meal, meat and bone
meal.
2.4 Trace elements in poultry feeds
Trace minerals are important in poultry feeds because they contribute in biochemical
processes required for normal growth, development and eggshell formation (Yenice
et al., 2015). Heavy metals residues may concentrate in poultry meat and eggs which take
up from different sources (Nisianakis et al., 2009). Some trace elements such as Iron (Fe)
and copper (Cu) are necessary for good human health which are common found in the diet
nevertheless, all metals are toxic at higher concentrations (Todd w Lane and Morel, 2000).
Other heavy metals as arsenic (As), Cadmium (Cd) lead (Pb) are toxic and their
accumulation over time inside chicken can cause serious illness. Certain elements that
14
normally toxic are beneficial for certain organism'؟ or under certain conditions (Al-
Ashmawy, 2013; Todd w. Lane et al٠. 2005).
Toxic mineral snch as arsenic, cadmium, lead and mercury are harmful to animals’ health
which are occur naturally in the environment as a result of natural carrses, as well as
industrial and agricultural practices (.Bampidis et ah, 2013). It found in feed pose serious
health hazards to consumers and secondary consumers due to bio magnifications. These
toxic elements are also normally present in natural environment. Due to many reasons tlrese
elements can easily entered in feed ingredients of different animals and complexes of
nature these elements are not digested at all. Many studies showed that elements used in
feed accumulated in body and cause harmful impacts. In case of poultry industry deposition
of heavy metals in body of broiler were result of their* excessive rrse iir poultry feed (K.
Rehman et ah, 2012). some trace elements emitted to the environment from industrial
activities, are harmfirl contaminates to humans which important element of pollution and
briirg about considerable damage to live ecosystems and the environment through natural
and artificial means, easily accumulated and create complex structures particularly in soil,
and they are defined as hazardous pollutants (Altundag et al., 2015).
Essential elements such as Cu, Zn, and As arc often added to commercial feeds to promote
optimum growth rate and antimicrobial properties (Moral et al., 2008). Other nonessential
elements such as Cd, Pb, Cr are found in feeds due to their presence iir concentrates aird
supplements, abrasion with iron cages or tools, and environmental pollution (Sager and
Martin-Gil, 2009; Wang et al., 2013).
Recent trend in poultry indttstry various metals are added in poultry diets to enhance the
growth rate, increase weight and prevention of diseases (Alkhalaf et al., 2010) but
sometimes these heavy metals are much higher in poultry feed than required value. On the
other hands, deficiency of heavy metals is another reason for their addition in feed because
it causes different problems in broiler as they perform many functions as trace elements.
Many studies are conducted to assess the heavy metals from poultry feed and results
showed the mixing of high concentration of heavy metals. The -use of heavy metals in
excess are accumulated in body tissues of broiler (Iwegbue et al., 2008; K. Rehman et al.,
2012).
15
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<١.^
*٠
.....
١ ٠٠.
•• •٠٠
٧٩٠٠
I
eggshell thickness, ylk colour, albumen height) and perk-، jancc <-٢'iaymg hen. ١. ٠ \i ١٦ '٠ rhc
effect of increased microelement doses in biological form on egg quality parameters and
hens performance was investigated. Their results showed that in the groups fed with tile
new biological supplement, egg quality parameters improved, including eggshell strength,
eggshell thickness, albumen height and yolk colour. The biological form of microelement
also improved the feed conversion rate, especially in the group fed with a biological form
of Cr (III). K. Rehman et al. (2012)fcund the concentration of heavy metals (copper,
manganese and iron) in feed samples is greater too times than WHO/FAO permissible
limits.
Bukai' and Saeed (201.4) analyzed poultry feed samples to determine heavy metalsI ,concentration. The results showed that different concentrations of cadmium, cobalt إ
copper, iron, manganese, nickel, lead, chromium and zine were detected in all samples.
They range from 0.53-03.19mg/kg for Cd, O.13-3.33mg/kg for Co,2.03-5.41 mg kg for Cu,' 08.79-19.74mg/kg foi' Fe, 12.5O-37.5Omg/kg for Mn, l.O3-2.O6mg/kg for Ni, 0.27-
.mg/kg for Pb, o.47-2.36mg/kg for Cr and 16.3O-38.O4mg/kg for Zn respectivelyإ 0.80
Suleiman et al. (2015) used atomic absorption spectrophotometry to determine six metals
(Cu, Pb, Zn, Cd, Mn and Ni) in chicken feeds in Sokoto-Nigiria. The concentration in
pg/ml of the metals ranged between 0.04 and 1.21 for Cu, 0.01 and 0.55 for Pb, 1.43 and
l 11.65 for Zn, 0.10 and 0.12 Cd, 0.94 and 3.12 for Mn and 0.004 and 0.25 for Ni. In mostof the analyzed samples, the concentration of Cu, Zn, Mn and Ni was found to be lower ؛
than the nutritional requirement of broiler chicken at a level which coLild be harmful for إ
I the poultry. Also the study showed the presence of heavy metals (Pb and Cd) in all the feed
I samples analyzed, but none exceeded permissible levels as set by European Union and
National Research Council.
Malu et al. (2015) determined the level of toxic minerals (Pb, Cd, Ni, Fe, Cu, JMn, Zn, Cr)
in poultry feeds using Atomic Absorption Spectrophotometer. The result showed that the
concentration in mg/kg obtained in broilei' feed brand was 4.378:,:0.0001, 1.952 00006.
1.803±0.0004, 0.260 0.0002, 0.095 ±0.0002. 0.043±0.0004, 0.009 ±0.0002, 0.043i-0.0000
for Mn, Zn, Fe, Cu, Ni, Cr, Cd and Pb. While the concentration, of toxic minerals in the
layer brand were 4.146 ± 0.0003, 2.051 ± 0.0003, 1 (147 ± 0.0003, 0.499 ± 0.0002. 0٠ 102 =
17
لج 0.002, 0.074 لج 0.0002 0.081 ,0002)0 and 0.008 0.0002 لج ior Mn, Zn٠ Fe, Cu٠ Cr. Pb and
Cd respectively. This implies that the concentration of Mn, Zn and Fe ١vere the highest in
all the feed samples with Cd and Pd being the lowest however comparison of these results
with other researchers and those acceptable standard values for food nutrition as stipulated
by European Union (EU) and World Health Organization (WHO).
2.5 Effects of some trace elements and heavy metals in chickens
2.5.1 Chromium
Chromium (Cr) is an important trace element in human and animal bodies although it’s
toxic metal at high exposure. Cr actively involved in tire metabolism of carbohydrates,
lipids and protein. In particular, Cr poteirtiates insulin action, resulting in tire increased
cellular uptake of glucose and improved intracellular carbohydrate and lipid metabolism.
Studies also showed that Cr 3 Can-induced histopathological changes and oxidative stress
in the liver and kidney in chicken. In low content, trivalent chromium can promote tire
growtlr and development of chickens thereby improving the quality of their nreat. For tlris
reason Cr is added into animal feed, sometimes people may put in excess trivalent
chromium to gain more benefits, adversely causing such element to be poisonous and
dangerous for chicken (Liu et al., 2015).
1٠5٠1
Zinc has important role in biological process, including imnrune function, growth,
development and reproduction. It is component of many enzynnes contributing in the
energy metabolism, protein synthesis and degradation biosynthesis of nucleic acids, CC>2
transport and many other important problems in poultry industry influencing economic
profitability of egg production. Zinc participating on formation calcite crystal and
modifying crystallographic structure of egg shell (Mabe et al., 2003; Torki et al., 2014).
Zinc (Zn) is necessary in poultry diets and its deficiency can causes various disorders, in
addition to depressed growth and performance. It is now recognized that Zn has a major
role in antioxidation, growth and developnnent, productior, innnnunity and stress related
.18
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issues. Supplementation of Zn ean improve growth, augment immunity, enhance
antioxidant capacity, increase endocrine secretion and interact with other minerals in the
gut (Abdelrahman et ah, 2016).Torki et al. (2014) zinc effects on laying hens under cold
condition, alleviates cold stress-induced depression in productive parameters and increases
zinc plasma concentration. Increased eggshell thickness was seen in the laying hens fed the
Zn included diet.Sunder et al. (2008) Reported that zinc deficiency in poultry diets end in
reduction irr weight gain, skeletal malformations, poor bone mineralization, and
immunological dysfunctions. Therefore, broiler chickens are often supplemented by zinc
during early age of it to elicit a positive response in broiler chickens. Because Zn
supplementation was found to be essential, the recommended 40 ppm for broiler chickens,
which appeared to be based on the results that considered growth performance as the only
criterion. However, higher Zn levels (60 to 180 ppm) produced better immune status in
broiler chickens.
2.5.3 Calcium
Dietary calcium concentration in hen feed has a significant effects on some of hen and egg
characteristics. For instance, low dietary calcium leads to weaker eggshells and hen bone
quality, comparing to hen fed by control diet or high calcium one, on the other hands, its
higher concentration leads to increase serum concentration, osteoprotegerin mRNA and
osteocalcin protein and inhibited serum alkaline phosphatase activity and decreased its
mRNA. It leads also to reduce egg production, similarly to the high-energy diet. They also
increase fat deposition, but had negative effects on bone metabolic homeostasis (Jiang et
al., 2013).
2.5.4 Lead
Lead (Pb) causes acute and chronic poisoning and induces a broad range of physiological,
biochemical and behavioral dysfunctions in man, animals and aquatic life (Erdogan et a٠l.,
2005).
Bakalli et al. (1995) studied the effects of supplemental dietary lead on broiler chickens.
They found lead supplementation caused linear decreases in body weight gain, delta-
aminolevulinic acid dehydratase activity. Lead additions to the diet resulted in a dose-
related increase of Pb in blood, kidney, liver and tibia. Higher dietary calcium caused
19
reduced Pb in blood and liver. Lead is toxic to chickens at much lower levels than
previously recognized.
Mahesar et al. (2010) found the amount of lead in most analyzed poultry feed samples was
greater than maximum tolerable level which could be harmful for the poultry. Ebrahimi et
al٠ (2015) stated that Pb-induced oxidative stress suppresses growth performance and feed
efficiency in broiler chickens. Lead supplement to chickens diets decreased feed intake,
body weight gain, and feed efficiency. Moreover the expression of all sugar, peptide, and
amino acid transporters was significantly down regulated in the birds under Pb induced
oxidative stress. Exposure to Pb also upregulated the antioxidant enzymes gene expression
together with the down regulation of glutathione S-transferase.
2.5.5 Cadmium
Cd toxicity has been related to decrease in birds body health (Pollock and Machin, 2009).
Several studies have shown that in most chickens cadmium accumulate in liver and kidney
(Marettova et al., 2012). Cd suppress the viability of the chicken splenic lymphocyte and
induces the oxidative stress and subsequently DNA damage and apoptosis (Li et al., 2010).
Cd exposure change the activities of antioxidant enzymes of erythrocytes and produce
oxidative stress by disturbing the oxidative and antioxidative balance of the adult poultry
birds (Kant et al., 2011; Li et al., 2013).
Cadmium (Cd) is a widespread contaminants element to environment associated with
many industrial processes (Marettova et al., 2015). Cd pollutants found in the air, drinking
water, and food are a potential health threat to humans, animals, and plants. Previous
studies indicated that exposure to Cd !*esults in severe damage to multiple organs and
tissues, including the brain (Zhang et al., 2016).
2.5.6 Iron
Iron is significant trace element for chicks, and it is required for fonctions of numerous Fe-
containing enzymes and protein systems. Fe prevent clinical deficiencies in chickens or
ensure to reach their optimal growth, for this reason Fe have been supplemented into their
20
k
؛١٠٠
إ٠
diets. Traditionally, Fe is supplemented into diets in the ;.om of :norganic salts, such a.١
sulfates and oxides. However, inorganic Fe additives have many disadvantages. sued as
low bioavailability, high hydroscopicity and oxidation, high excretion, and so on (Ma el
al., 2014).
Iron is an important tr'ace element that plays a vital 1'ole in many physiologie proeesses.
European Union and National Research Council (Dale, 1994) has recommended 80 mg
Fe/kg diet for broiler chicken. Iron fortification is critical for broiler chickens because of
increasing demand for' Fe because of the rapid increases in red blood cell volume and body
mass (Sun etal., 2015).
2.5.7 Manganese
Mn is significant element for bone and tendon development as its deficiency causes tibial
twisting and bending and tibiometatarsal joint enlargement and malformation, resulting in
slippage of the gastrocnemius tendon fi'om its condyles. Mn is also associated with shorter
and thicker long bones in poultry (Underwood, 1999).
2.5.8 Nickel
Nickel is an essential element for the chick. It is required for activity of vitamin B12 and
biotin during metabolism of odd-chain fatty acid in animals (Nielsen. 1991). And is
involved in methionine-folate metabolism (Uthus and Pocllot 1997). And during the past
decade, nickel has been shown to participate in several enzyme reactions. In
chemoautotrophic hydrogen bacteria, nickel is incorporated into hydrogenases (Friedrich
et al., 1982) and carbon monoxide dehydrogenase from elostridia lias also been shown to
be a nickel enzyme (Ragsdale et al., 1983). Nickel, high concentrations of which can affect
human health badly, can accumulate on plants, animals, and soil (Demil' et al., 2005).
2.5.9 Iodine
Slupczyhska et al. (2014) found that Used of KI as source of iodine enhanced the egg
weight. The increased I level in the diet had an equivocal influence on egg weight'
improved the feed convei'sion per 1 kg of eggs, and deci'eased the proportion of damaged
21
ا
- eggs.When used KI as to enhance diet, the Hematic gical -؛؛diee.١ ot hen bkmd
demonstrated significantly higher led blood cells numbers and hemoglobin concentrations
Opaliriski et al. (2012) studied the effect of iodine yeast (!-yeast) supplementation on the
performance, egg traits, and iodine content of eggs of laying hens. Their results suggested
that iodine yeast supplementation in the diet of laying hens is an effective method for
increasing iodine concentration in eggs and thus could contribute to elimination of iodine
deficiency disorders in humans consuming iodine-enriched eggs.
2.5.10 Selenium
In poultry. Selenium is an important micronutrient for normal growth and maintenance.
The recommended selenium concentration in broiler diet is from 0.1 n٦g/kg to (). 1 5 mg/kg
(Sevcikova et al., 2006).
Puerto et al. (2014) studied the effect of selenium supplementation in diet broilers on the
lipid and protein oxidation, GPX activity and drip loss in pectoralis and Gastrocnemius
muscles. Diet supplemented with organic (seleniomethionine'; and inorganic (sodium
selenite) selenium at 0.30 ppm from thirty-five until fifty-six days. Their results showed
that In Pectoralis and in Gastrocnemius fresh during 5 days, Se supplemented in diet did
not affect lipid or protein oxidation. However, GPx activity was enhanced in both muscles
and the drip loss was reduced significantly at 24 hours post mortem. Selenium
supplementation could be a strategy to reduce the di'ip loss in poultry meat and the effect
could be mediated by the GPx activity in muscle.
Tufarelli et al. (2016) studied the effect of organic Se (2-hydroxy-4methylselenobutanoic
acid) (HMSeBA) supplementation on selected performance criteria and Sc deposition in
egg of laying hens. They found (HM Se BA) as Sc supplement affected positively egg yolk
quality without effecting hen's productive traits aird it offers an efficient alternative to
fortify eggs with Se that lead to greater supply of Se for humans. Utterback et al. (2005)
investigated the effect of Se supplement hens diets on egg selenium content. The Results
indicate that use of Se yeast in laying hens diets is very effective for increasing the Sc
content of eggs.
٠riإ
22
2.5.11 Phosphorus
In addition to its function in bone formation, is also required in the utilization o! energ\
and in structured components of cells. Examples of phosphorus-containing compounds i١
phospholipids(Dale, 1994).
These forms of phosphorus, if present in plants, can be digested by poultry; howeeer. such
digestible forms usually account for only 30 to 40 percent of the total phosphorus. The
remaining phosphorus is present as phytatc phosphorus and is poorly digested. Only about
10 percent of the phytate phosphorus in corn and wheat is digested by poultry. The
phosphorus from animal products and phosphorus supplements is generally considered to
be well utilized (Dale, 1994).
23
Chapter3
MATERIAL A^D METHOD
used.
A total of 20 of poultry feed simple random samples (broiler, and layer chickens) were
obtained from the local market of Khartoum state. Two replicates for each sample were
3.2 Sample Preparation and Analysis
Samples was divided in to two groups (broiler and layer) and processed by dry-ashing
method for analysis. Samples were placed in crucibles then ashed gradually in furnace at
5OO٥C. Ashes samples grinded into a completely homogeneous powder. The hey powder
standard and samples was weighed accurately (1.00) gram and then fused and pressed into
a pellet that will adequately absorb the primary x-rays.
3.2.1 Radioactive Source
30 radioactive source was used, which is a source of an initial nominal activity of ؟؛؛Cd
by electron captut'e transformation ؟؛؛decays to Ag ؟؛؛mCi. In this type of decay the Cd
and produces characteristic x-rays of the daughter isotope. In the isotopic vacancies are
created in the K-shell, electroirs fall from L, M and N shells to fill the created vacancies.
9 are emitted of؛؛In the process Ka and Kp charactei'istic x-1'ays of the daughtei' isotope Ag
energy.
3٠2٠2 X-ray Emission
The x-ray from source used to excite element in spiceman when the inner shell electron
ejected from atom. The emission of characteristic x-ray photon occurs when a vacancy in
an inner electron state is formed, and an outer Ol'bital electron makes a transition to that
vacant state. The energy of the emitted photon is equal to the differen e in electron energy
24
Iج
levels state. The energy of the emitted photon is equal to till aiffererce m electron energy
levels of the transition. As the electron energy levels are characteristic of the atom, the
energy of the emitted photon is characteristic of the atom. The fluorescence from the
sample was detected by Si (Li) semiconductor detector.
3.2.3 Semiconductor Si (Li) Detector
To compensate and bind all free electrons, lithium ions are drifted (allowed to diffuse at
elevated temperature) into the silicon crystal to neutralize the Si crystal defects in a
particular zone. Afterwards. Au contacts are evaporated onto the crystal and a reverse
voltage applied. In the crystal, the energy difference ع (band gap) bctvccn the valence and
conduction band is 3.8 eV. At room temperature, the conduction band is partially populated
so that the crystal is a (semi)conductor. To keep the leakage current as low as possible, the
crystal is cooled with LN2 by placing it a vacuum cryostat. At -I96٠c almost all electrons
remain in the valence band. The radiation to be measured needs to enter the cryostat
through a thin entrance window, usually made of Be. By applying a reverse voltage to the
charge carrier free intrinsic zone, an absorbed x-ray photon is converted into charge by
ionization. Electrons are promoted from the valence to the conduction band, leaving
"positive holes” in the valence band؛ thus the crystal temporarily becomes conducting of
electron- hole pairs are created.
The electrons and holes are quickly swept to the contact layers by the electric, field created
by the applied reverse bias on the Cl'ystal. Figure 3.1 shows the operation principle
schematically. The charge induces a signal at the gate of a cooled field effect transistor
(FET) that is the input stage of a charge sensitive preamplifier.
The output signal is fed to a pulse processor that shapes the pulse and amplifiers it further.
This signal is in the range up to 10 V and is proportional to the energy of the absorbed
photon. The pulse height is digitized by means of an analog-to-digital converter (ABC)
and the resulting digital value stored in a multichannel analyzer (MCA). This is an array
of memory cells, called channels; by using the digital value associated with a single event
؛
25
as address offset into the memory array, the content of the appropriate channel is
incremented with one count.
I Thus, all detector events having the same pulse height are stored in tire same chanirel. After
-furtlrer amplification and shaping, tlris is converted into a bell-shaped pulse: the pulse ا
1 height is then digitized by an ADC, resulting for instance in adigital number of 420.
Ultimately, this causes the content of channel 420 to be incremented with one count. After ا
readout, tire MCA memory (typically 1024 or 2048 chamrels in size, each corresponding ا
to al0-20eVwide energy range) yields a pulse-height distribution of the detected events or ا
.an energy dispersive x-ray spectrum ا
Figure 3.1. Working principle of a Si(Li) detector
3.3 Statistical Analysis
The whole data were subjected to a statistical analysis using SPSS statistical software
version 20. Data were checked for normal distribution and significant differences between
feed types for different trace and heavy metals were assessed by independent t-test. The
Pearson correlation between trace and heavy elements in feeds was performed by
nonparametric test. The principal component analysis (PCA) was performed usiirg Minitab
version 14 in order to provide further identification and examine the correlation between
the trace elements concentrations in feed.
26
Chapter 4
RESULTS AND DISCUSSION
4.1 Validation of the X-ray Fluorescence
The concentrations of trace elements in chicken feeds were determined using x-ray
Fluorescence (XRF). The validation of' XRF technique was performed using Hey powder
standard. Figure 4.1 shows the typical XRF spectrum of the standard. It was observed
clearly the XRF peaks of the detected elements. The typical concentrations on elements in
the standard are shown by Table 4.1.
90 -
SO -
70 -
60 -
0 to 00^ 600 500 400 00 3 00 ن sooChannel Number
Figure 4.1. XRF spectrum of the Hey standard
The quality control of XRF analysis was performed using hey powder standard, and the
results of the trace elements concentration (in part pel' million (ppm)), are shown in T'able
4.1. In order to check the accuracy of the method, standard hey powder was analyzed to
determinate the trace elements concentrations.
Accuracy and precision of analysis were further established by simultaneous analysis of
the standard refei'ence hey powder samples. By applying the relative comparison method
27
for material content of the samples with other elements identified in samples, The tabulated
values in Table 4.1 showed good agreement between the international values of the trace
elements in the standard and concentrations obtained from OUI* experimental results*
therefore, the concentrations of the trace elements in the standards were comparable to their
reference values.
Table 4.1. Trace elements concentration in the hey powder standard
Element Mean STD cv% Certified Value Accuracy®/(* Error Recovery %K 2.12E+04 461.8802 2.18E+00 2.1OE+O4 9.52E-01 9.52E-01 99.05
Ca 2.22EO4 519.6152 2.34E+OO O42.16؛ E 2.78E+OO 2.78E+OO 97.22
Mn 4.81E+01 1.385641 2.88E-+00 4.70 EOl 2.34E+OO 2.34E+OOFe 1.91E+02 5.773503 3.O3EO 1.86 E+O2 2.51E+OO 2.51E+OO 97.49
Cu 9.81EOO 0.011547 1.18E-01 9.4 E+00 4.33E+OO 4.33E+OO 95.6٩Zn 2.46E + 01 0.69282 2.82EOO 2.4 E+00 2.5OE+OO 2.5OE+OOPb 1.63 E+00 0.046188 2.83E + OO 1.60 E+00 2.O8E+OO 2.O8E+OO 97.92
Rb 7.79E+OO 0.005774 7.41E-02 2.54EOO 2.54E+OO 97.46
Sr 4.09E01 0.057735 1 4 IE-01 4.00 Etoo 2.33E؛00 2.33E+OO 6٦.9٦
4.2 Trace Elements Concentrations in Chicken Feeds
In the present study, 9 trace elements and heavy metals (K, Ca, Mn, Fe, Cu, Zn, Pb, Rb,
Sr) were determined in layer and broiler chicken feeds by XRF spectrometer and the results
are shown in Table 4.2. It was observed that the highest concentration of trace element in
chicken feeds was Ca followed by K, and Fe. The concentration of Ca in the layer chicken
feed was higher than broiler chicken feeds about (2.57x0 4-6.75 xio٠4) ppm in layer feeds
and (2.9xio4-3.6xio4) ppm in broiler feeds.
Potassium (K) concentrations were about (1.61x0-4.904) ppm in the layer chickens
feed and nearly equal in the broiler feeds (1.23x0-4.8x104) ppm, and Fe concentrations
were found to be about (1.75x 0-4.99x0) and (1.38x0-6.27x10ذ) ppm layer and broiler
feeds, respectively. On the other hand, Pb showed the lowest concentrations among these
trace elements in the feed samples with concentrations about (1.24-3.36) and (1.45-9.71)
ppm in layer and broiler feeds, respectively. Moreover, Mn exhibited approximately equal
concentrations in both layer and broiler chicken feed, which were exactly about (5.1x0-
28
ا؛ا
إا
3.7xlO٦) ppm in layer feeds and (5.6xlfo-2.67xicb) ppm 11. lclci feeds. Similarly, Zinc
concentrations were approximately equal in both feeds samples, about (2٠36x 1-1.15x 10٦)
and (1.84x10^-1.01x10^) ppm in layei' and broiler feeds, respectively. In addition, layer
feeds showed lower concentrations of Cu compared to the broiler feeds, exactly about
(13.3-49.1) and (15.9-1.7x0) ppm. Similarly, the concentrations of Rb in broiler feeds
(9.41-240) ppm were slightly, higher than its concentration in layer feeds (11.6-179) ppm.
The concentrations range of Sr in layer feeds was between 53 to 270 ppm, while, m the
broilei' were found to be about (44-173) ppm (Table 4.2.))
Table 4.2 The concentrations (ppm) of the trace elements in chicken feeds
Statistical ParametersK Ca Mn Fe Cu Zn Pb Rb Sr
1١١
أ\
١
Layer Feeds
Mean 29600 134400 1706 2808 11 544 1.83 82.4 115
Std. Error of Mean 3811 161 425 296 4.00 13 0.28 19.2 19.6
Median 161 111150 852 2525 22.3 6\١٦ 1.92 70.4 111؛
Std. Deviation 12052 65523 1346 937 12.66 151 .890 61.0 62.2
Minimum 16100 67500 514 1750 13.30 116 0.00 11.60 53.3 '
Maximum 49000 151666 3740 4990 49.10 1150 3.36 \16.6 273
Broiler Feeds
Mean 30250 107320 1475 3400 147 628 2.87 106 94.14
Std. Error or Mean 4863 23938 116 560 113 106.8 0.78 29.4 11.03
Median ٦1566 ^6166 1139 2610 25 616 2.04 661 82.85 1
Std. Deviation 15380 15666 854 111 360 111.1 2.48 93.1 44.38
Minimum 12300 36900 5%6 1380 15.90 184 1.45 9.41 44.00
Maximum 54100 291000 1616 61 1170 1010 9.71 241 173.0
29
لi
Figure 4.2. XRF spectrum of the chicken feeds samples
4.3 The Difference of Concentrations of Heavy Metals in Layer and Broiler
Chicken Feeds
The differences between the mean concentrations of trace elements in the two different
groups of feeds (layer-broiler) was tested by the independent t-test and the results are
shown in Appendix (A), there were no significant difference between all concentrations of
trace elements in two groups. National research council NRC reported that the Mineral
requirements of egg-type chickens in production are similar to mineral requirements of
other poultry, with the exception of calcium (Dale, 1994).
4.4 The Correlation between Heavy Metals in two Types of Feed
Pearson’s Correlation test was performed to investigate the correlations between ti'ace
elements concentrations. There were sti'ong COI elation between K-Rb (0.885), Mn-Zn
(0.874), while, negative correlations were observed between Mn-Rb (-0.680), K-Mn (-
0.600), K-Ca (-0.565), Zn-Rb (-0.541), Ca-Rb (-0.458). In contrast, these results showed
no correlation between the Fe and Zn, which has been reported in previous study (Skfivan
et al., 2005), this could be attributed to the supplementary addition of these elements in the
chicken feeds, and suggested that the quality of the feed from the local market need some
30
ا
اا
attention. However, more information about the association between the trace elements in
chicken feeds will be discussed later in the results of the principal component analysis.
Table 4.3. The correlation between the trace elements concentration in chicken feeds
Elements K Ca Mn Fe Cu Zn Pb Rb Sr
K 1
Ca -0.565** 1
Mn -0.600** -0.037 1
Fe 0.005 0.337 0-.086 1
Cu -0.251 -0.179 0.240 -0.246 1
Zn -0.407 -0.280 0.874** -0.103 اددا 5 1
Pb -0.312 0.106 0.037 0.022 -0.113 0.079 1
Rb 0.885** -0.458* -0.680** 0.096 -0.231 -0.541* -0.245 1
Sr 0.104 0.304 -0.225 -0.028 .0.171 -0.385 -0.184 0.098 1
** correlation is significant at the 0.01 level (2-tailed),
*correlation is significant at the 0.05 level (2-tailed).
4.5 Principal Component Analysis
Principal component analysis was perform to identify which of the elements contribute
mostly of the total variance between the feeds samples. The first three components were
extract because their eigenvalues were higher than one, and therefore, they could be
regarded as the components which have the significant impacts in the total variance, this
can be seen clearly in the scree plot (Figure 4.3). The Scree plot show the eigenvalues of
the variable against the principal components and suggested that the most significant
components are the first (PCI), second (PC2) and the thii’d (PC3); the variance percentages
explained by each component can be seen in Table 4.3, and the cumulative variance
i percentage revealed that the first three components explained 71.5% of the total variance.٠
31
Table 4.4. The total variance explained h PCA
Component Initial Eigenvalues Extraction Sums of Squarcd Eoadings
Total % of Variance Cumulative 0/0 Total 0 of Variance Cumulative؛(-؟
1 3.290 36.551 36.551 ١!.١ 36.551 36.551
2 1.962 21.804 58.355 1.962 21.804 ذقلإ3 1.185 13.168 91.524 1.185 13.168 71.524
4 0.983 10.926 82.450
5 0.802 8.915 ١65\.9
6 0.504 5.601 ٩%.%٦ 0.114 1.269 98.236
8 ا آ 1.038 99.274
9 0.065 100.000 آ٦٦6
123456789 Component Number
Figure 4.3. The scree plot of the principal components
32
4.5.1 Principal Correlation Matrix
The correlation between the extracted factoi's and the trace elements in feeds was
investigated rising the correlation matrix and the resrilts are shown in Table 4.3. The PCA
results revealed that the most contributed elements in PCI are the elements Ca, Mn. Zn
and Pb, which is correlated strongly with the element with coefficients (,around 0.4 to 0٠8 )٠
١vhile the elements Rb and K were found to be negatively (coefficient about -0.9)
correlated with PCI. These results suggested that these elements vary together, thus ifone
increases, the other tend to as well, except Mn and Zn which negatively correlated with
fil'st component.
Moreover, the second component PC2 was found to be significantly correlated with the
elements Mn, Zir and Pb, positively, while Ca and Sr, showed negative correlation with the
second component. The third component showed positive correlation (0.6) with Cu, and
on the other hand, Fe showed negatively correlated with it. These results l'evealed that the
feed sample with higher concentrations of Mn Zn and Pb, will show a lower concentration
of Ca, Sr, and similar conclusion was observed to Cu and Fe.
Table 4.5. Principal correlation matrix (n = 20)
Variable Factorl Factor2 Factor3 Communality
K -0.954 0.043 -0.036 0.914
Ca 0.584 -0.619 -0.439 0.917
Mn 0.708 0.519 0.202 0.812
Fe 0.055 0.084 -0.876 11%.٠Cu 0.222 0.155 0.627 0.466
0.520 0.728 0.236 0.855
Pb 0.870 0.481 0.005 0.988
Rb -0.949 -0.039 -0.125 0.918
Sr -0.048 -0.780 0.085 0.618
33
ا
اااً
The score plot of the first two components was obtained am.؛ showed by figure 4.4. it was
observed that the PCA results could not discriminate between the two groups of feed
samples (layer and broiler). These results are in agreement with the basic statistical results
(test), and suggested that there was no significant difference between the layer and the
broiler samples in terms of trace elements concentration.
A
Group broiler د1 layer
-
-3-2-1012 3First Component
Figure 4٠4. The score plot of the first component (PCI) and the second component
(PC2).
4.6 Comparison of the Trace Elements Concentrations in Chicken Feeds and the
Previous Studies and Recommended Level
In this study, trace elements were found in chicken feeds with diffei'ent concentrations
indicating the differences in use of feed additives and environmental pollution. Sagei' and
Martin-Gil (2009) stated that the presence of the essential elements such as Cd, Pb, Cr in
feeds, is due to their presence in concentrates and supplements, and environmental
pollution.
For instance, national l'esearch council NRC reported that the mineral requirements of egg-
type chickens in production are similar to mineral requirements of other poultry, with the
exception of calcium. The onset of egg production creates a need for more calcium to make
34
i
ا5؟إ
Iاا
؛
t١٦e eggs١٦eU. Concentration 0؛' Ca ؛١٦ layer chicken feed ٠\as higher ؛١١٦١٦ broke؛’ feed becatise it supplemented to layer chicken feed to improve quality of eggs Jiang et ah (2() 13)
reported that calcium ireeded for eggslrell and for production of eggs)
Potassium K concentrations nearly equal in layer and broiler chickens feeds. Manganese
in all samples is higher than recommended value which recommended by NRC (1994) (60
ppm) and value gotteir from this study is also higher than value obtained by (Suleiman et
ah, 2015) and nearly equal to (K٠ u. Rehman et al., 2012) were found Mn in broilei' feeds
5425 mg /kg . the concentration of Cu in layer feeds was (49.1-13.3.) mg/kg and broiler
feeds (1.7x1015.9) mg/kg is higher than NRC recommended value for layer feeds
was(2.5)mg/kg and for broiler feeds was 8mg/kg (SANCO, 2002) and is lower value than
study (Wang et al., 2013) was found in layer feeds (3.8-132.1) mg/kg and in broiler feeds
(5.2-198.7) mg/kg.
Zinc level in layer chicken feeds is (2.36x0- 1.15x0) ppm and in broiler feeds is
(1.84x10 1.01x0) ppm is very higher than NRC recommended level (40ppm) for broiler
feeds and 30٠40 mg/kg in layer feeds. And higher than permissible limit 500 mg/kg as
stipulated by European Union(Bukar and Saeed, 2014). The present study concentrations
is also higher than (Wang et ah, 2013) study who found Zn concentration in layer feeds
about 28.6 to 235.7 mg/kg and broiler feeds was 5.6 to2.963 mg/kg and higher than(Malu
et al., 2015) study, the concentration were found in layer feeds 4.14 mg/kg and 1.95 mg/kg
in broiler feeds. Iron concentrations in layer feeds were found 1.75x0 -4.99x0 and in
broiler* feeds 1.38xO-4.99xlO٩ and in broiler feeds were found 1.3810^-6.2710^ is too
higher than permissible limit 45-80 mg/kg as stipulated by FA0//WH0. And too higher
than (Bukar and Saeed, 2014) stdy, were fourrd concentration of iron (8.79-19.74) mg/kg
in chicken feeds. Lead concentrations were found in layer* feeds 1.24-3.36 ppm and in
broiler feedsl.45-9.71 ppm is lower* than (Wang et al., 2013) concentrations 0.7-34.5 for'؛
layer* feeds and 0.3-13.lmg/kg in broiler feeds. But higher* than concentrations which found
■ ir٦ (Malu et al., 2015) found concentration in layer chicken foeds 0.043mg/kg and in br*oiler*
feeds were 0.008 mg/kg.
i
35
جاإإ
؛ا
اا
ا
iIا؛
؛
اإ!
CONCLUSION
II) this study determined the trace element concentrations in the layer and broiler feeds
results revealed that the highest concentration in all samples was Ca (134400) ppm in layer
feeds (107320) ppm in broiler feeds. While Pb the lowest concentration (1.83) ppm.
Calcium concentration was found in layer feeds (134400) ppm higher than broiler feeds
(107320) ppm. However, there was no significant difference between trace elements
concentration in layer and broiler feeds. The concentrations of most of the trace element
were found to be higher than recommended level which stipulated by national research
council. Lead was found in layer feeds with concentration of about 1.8 ppm and 2.87 ppm
broiler feed, these values were lower concentration thair permissible level (5mg/kg).
The principal component analysis (PCA) results showed that the elements Ca, Mn, Pb and
Zn and they have a positive correlation with the first principal component, and K and Rb
are associated and showed negative correlation with it. The second principal component
correlation values confirmed these results and showed clearly the association between these
elements Mn, Pb and Zn, while Ca and Sr were found to have a negative correlation with
it. In addition, the third component was found to have a positive correlation with Cu and
negative with Fe. Nevertheless, these results could not cluster the feed groups (layer and
broiler), indicating the similar composition of the feeds and confirming the results obtained
from the t-test results.
!
iI؛
iIأ1؛ا
36
ا
RECOMMENDATIONS
1٠ Trace elements should be supplement to chickens feed in international
recommended level.
2. Some ti'ace elements should be add to chickens feeds such as Se. I, Ni, and p
because it’s very important for chickens.
3. More attention should be paid foi' chickens feeds industry to assure the productions
quality.
4. Chickens feeds industry should be localized far away from industrial complexes
and pollution from general roads.
5. Availability of devices and substances which are essential for applying this type of
scientific researches to facilitate and encourage researcher.
Iأ
1
REFERENCES
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Alkhalaf, N. A., Osman, A. K., and Salama, K. A. (2010). Monitoring of aflatoxinsand heavy metals in some poultry feeds. African Journal of Food Science, 4(4)' 192- 199.
Alloway, B. (2013). Introduction. In B. J. Alloway (Ed.), Heavy Metals in Soils (Vol. 22, pp. 3-9): Springer Netherlands.
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42
Appendix A
The student t-test results
Levene's Test for Equality of Variances
t-test for Equality of Means
95% Confidence Inter al of tile DifferenceLower
Std. Error Difference
MeanDifference
Sig.(2- tailed)
dftSig.
12332.09088-13632.090886179.23494.650.000000.91718-0.1050.3301.001K
Ca
Mn
Fe
Cu
Zn
Ph
Equal variances assumedEqual variances not assumed Equal variances assumedEqual variances not assumed Equal variances assumedEqual variances not assumed Equal variances assumedEqual variances not assumed Equal variances assumedEqual variances not assumed Equal variances assumedEqual variances not assumed Equal variances assumedEqual variances not assumed
-0.105 17.027 0.917 -650.00000 6179.23494 -13685.49509 12385.495(19
0.028
8.524
5.637
4.511
0.082
1.832
0.869
0.009
0.029
0.048
١٥٠٦٦٦
0.193
0.855
0.855
0.458
0.458
-0.933
-0.933
-1.056
-1.056
-0.544
-0.544
-1.254
-1.254
18 0.404 mu 31660.14600 -39435.49852 92٦IAH
62١٦ .٦ 0.404 لا1١د١1١لم١ 31660.14600 -39533.63320 93693.63320
18 0.652 231.20000 504.51500 -828.74668 1291.14668
15.239 9.652 231.20000 5()4.51500 -1(.١٦ع1 \١٩ل١٦9لم١٩
18 0.363 -59211 634.5511 1 -1925.14241 741.14241
\2i! 0.2471 -592.00000 634.55111 -1956.09122 1٦٦ذ\!١
18
9.02
18
17.940
18
11.279
0.305
0.318
0.592
0.593
0.226
0.235
-120.48000
-120.48000
-84.60000
-84.60000
-1.04500
-1.04500
114.06637
114.06637
155.64443
155.64443
().83338
0.83338
-360.12455
-378.41917
-411,59682
-411.67530
ز٦٩1١
-2.87372
119.1645
137.4591
.42.39682
.42.47539
0.70586
لم١.٩جلإ١7ح
43
I
Rb
Equal variances assumed
3.094 0.096 ام 18 0.506 .23.89100 35)20297 -97.84970 5؟١؟.١6٦79
Equal variances not assumed م6٦9 ١)٦5.5 ،ه5٩ل٦ -23.89100 35.20297 -98.70335 50.92.135
Sr
Equal variances assumed ؟15 0.817 0.895 18 0.382 21.65000 24.18424 -29.15921 HA592A
Equal variances not assumed 0.895 0.384 21.65000 24.18424 -1٩.5ه٩٦٦ I٦.n٦
44
