Assessing feed effects on serum biochemical parameters of poultry chickens

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Assessing feed effects on serum biochemical parameters of poultry chickens
M. Alam 1, M. Ohid Ullah2 , Mirajul Islam2 and M. Shahidul Islam2*
1Department of Agricultural Statistics, Sylhet Agricultural University, Tilaghor, Sylhet-3100,
Email: malamstat@gmail.com,
2Department of Statistics, Shahjalal University of Science & Technology, Sylhet-3114,
Email: ohidullah@gmail.com, miraj.isrt10@gmail.com and shahed.stat@gmail.com2*
2*Corresponding author
Abstract: Accelerating body weight with feed efficiency of broiler chicken is one of the primary objectives in
poultry industry. Any adverse effect in chicken’s body through feed might produce health hazard to the consumers.
There are large varieties of feed available in the poultry industry. However, there could be trade-off between easy
weight gain and the accumulation of toxic biochemical component in chicken’s body. A study was conducted on
seventy randomly selected chickens which were randomly allocated to provide two types of feeds (so called
balanced study). Blood samples were taken at the end of experiment (35 days) and readings for total cholesterol
(TC), triglycerides (TG), high density lipoprotein (HDL), low density lipoprotein (LDL), calcium (Ca), creatinine
along with body weight were determined through lab experiment. Moreover, liver enzyme parameters aspartate
amino transferase (AST) and alanine amino transferase (ALT) were measured. Different graphs and multivariate
analysis revealed that two types of feeds produce significant difference in weight, lipid profile levels and liver
function measurements.
Key words: Broiler, Lipid profile, Liver function, Multivariate Analysis of Variance,......
1 Introduction
One of the major livestock sub-sectors is poultry industry in Bangladesh which can ensure to supply cheap
sources of good quality nutritious animal protein to the nation. Poultry meat contributes approximately 37 percent of
the total animal protein in Bangladesh (Islam, 2005). Poultry meat industry focuses mainly on optimizing the profit
through improving body weight and feed efficiency, but the consumer’s health issues are not taken into
consideration seriously. A high level of toxic elements of blood and also lipid profile (i.e., total cholesterol) level in
the chickens’ body can eventually lead to serious consequences on consumers’ health. The concentration levels of
these blood parameters are changeable due to different types of poultry feeds. Lipid profile is the risk indicators of
coronary heart disease (Eqbal, et al., 2011).
Issa et al., (2012) inspected the effect of feeding garlic powder (GP) on the performance, digestibility, digestive
organs and lipid profile of broilers and found that GP could provide positive advantages in broilers performance as it
significantly decreased total cholesterol (COL), triglycerides (TG), low density lipoprotein (LDL) and increased
high density lipoprotein (HDL) levels compared to control birds and these result supported the findings of Prasad
et.al.,(2009). The effects of alpha lipoic acid on the performance and serum lipid profile in broiler chicken was
found to have no effect on serum cholesterol, triglycerides and VLDL-cholesterol levels, but reduced the LDL-
cholesterol level and increased the HDL-cholesterol level of broilers compared to control group (Murali, et al.,
2014). Tohala (2010) studied the effect of high diet feed on lipid profile levels of chickens.
The food which is be free from all types of health hazards and have good taste and necessary nutritional diet for
human growth and development is healthy food. Now-a-days food security and food safety is the top most public
health concern worldwide. Important aspects of food quality on health safety are destroyed by entering the
hazardous type of pollution in the food chain. Increasing the LDL and total cholesterol level the risk of serious
problem of human like as stroke, heart attack and atherosclerosis also increases. Since the poultry fattening at the
lower age for better profit, it is important to know the concentration level of these above parameters at different
types of feed of chickens as well as identify the available better feed in the market.
In view of lack of available information of poultry feed effects on bio-chemical parameters, this study was
undertaken to investigate the effect of feed on lipid profile, liver function and others parameters in chicken. For the
purpose, seventy poultry chickens were split into two groups and were given two different feeds of this study. One
of the feed was supplied by the hatcher firm and other was widely used feed available in the local market. Blood
sample was collected from the chickens at the age of 35 days and lab experiment was performed. Lipid profile and

liver function’s readings were considered as group of variables while other measurements were regarded as
univariate. As the data did not violate classical assumptions, simple t-tests were performed for univariate tests,
whereas multivariate analysis of variance (MANOVA) was resorted for grouped-variables. Chickens’ weight and
some biochemical parameters were found to differ significantly for two types of feeds.
2 Experimental design and method
2.1 Experimental units and treatments
Randomly selected seventy broiler chicks from 600 (Cobb-500) were randomly distributed into two groups each
belongs of 35 birds. Chicks were collected for the study belonged to the same batch and the same breeding stock,
reared under similar and strict hygienic conditions. Rooms, brooder battery and cages were thoroughly cleaned with
2.5% phenol and subsequently fumigated with formaldehyde gas. Sunlight and electric bulbs were used as source of
heat and light, electric fan were used for controlling the room temperature. Birds of the both groups were vaccinated
against disease according to the direction of the expert of the poultry science department of Sylhet Agricultural
University (SAU), Sylhet. Two different feed i.e., Feed-A, Feed-B (starter, grower and finisher) were supplied to the
two groups from the first day to 35th day of the experiment. Water was supplied ad libitum during the entire
experimental period. Composition of the diets and its nutrient composition for the above-mentioned phases are
presented below.
Table 1: Ingredients and nutrient compositions of experimental diets
Nutrient compositions Feed-A (Standard feed available in Market) Feed-B (Supplied by Hatcher)
Starter period (day 01 to day 14)
Crude Protein Min 21.50% 22.00-23.00%
Fat Min 3.50% 5.00-6.00%
Crude Fiber Max 5.00% 3.00-4.00%
Moisture Max 12.00% 10.00-11.00%
Fiber (Ash) 7.00-8.00%
Calcium Min 1.00%
Phosporus Min 0.45%
Lycin Min 1.40%
Mithieonin Min 0.58%
Bipakio shakti 3050 killoCal/kg
Grower period (day 15 to day 25)
Crude Protein Min 20.00% Min 21.00-22.00%
Fat Min 3.00% 5.00-6.00%
Crude Fiber Max 5.00% 3.00-4.00%
Moisture Max 12.00% 10.00-11.00%
Fiber (Ash) Max 9.00%
Calcium Min 0.95%
Phosporus Min 0.45%
Lycin Min 1.30%
Mithieonin Min 0.56%
Bipakio shakti 3150 killoCal/kg
Finisher period (day 26 to day 35)
Crude Protein Min 19.00% Min 21.00-22.00%
Fat Min 3.00% 5.00-6.00%
Crude Fiber Max 5.50% 3.00-4.00%
Moisture Max 12.00% 10.00-11.00%
Fiber (Ash) Max 9.00%
Calcium Min 0.95%
Phosporus Min 0.45%
Lycin Min 1.30%
Mithieonin Min 0.56%
Bipakio shakti 3150 killoCal/kg
2.2 Collection of blood sample for the bio-chemical parameters
At 35th day 3ml blood sample were collected from every birds of both group from the wing veins for lipid profile,
liver function and other bio-chemical parameter test. We collected the blood sample at very early morning for the
fasting stage of the bird with the help of expert personnel and also measured the weight of each sampled bird. Blood
samples were transferred to the Physiology and Toxicology department laboratory of SAU, Sylhet within two hours

of collection and serum were separated by centrifugation (3000g, for 10 minutes at room temperature) and stored in
eppendorf tubes for analysis.
2.3 Studies of bio-chemical parameters of blood
The concentrations of the biochemical parameters estimation was carried out by the chemistry semi-auto analyzer
AUTOPAK by routine methods. Commercially available kits (RANDOX, County Antrim, United Kingdom) were
used to analyse the serum for total cholesterol (TC) in enzymatic endpoint method, total triglycerides (TG) in GPO-
PAP method, low density lipoprotein (LDL) in enzymatic (colorimetric) method, high density lipoprotein (HDL),
calcium in colorimetric method, creatinine in colorimetric method and values were expressed as mg/dL.
Furthermore, aspartate amino transferase (AST) and alanine amino transferase (ALT) for liver functioning were
measured and expressed as U/L.
2.4 Methodology
Explanatory data analysis was performed as a descriptive analysis using the data obtained from the laboratory
experiment. In inferential data analysis t-test, ANOVA and MANOVA were applied for the difference of two groups
according to two feeds due to individual blood parameter and also clustered parameters. All of these analyses were
done using statistical software R and SPSS.
3 Results and Discussion
3.1 Exploratory data analysis
We resorted to boxplots of all the variables for exploratory data analysis. The boxplot in Figure 1(a) shows that
different median levels of weights were produced by two types of feeds. Figure 1(b) shows that the calcium levels
are almost similar for two feeds, although there are couple of outlying observations for feed A.

(a) (b)
Figure 1. Box-plot representation for the feed associated changes in (a) weight and (b) calcium concentration of broilers serum
Figure 2. Box-plot representation for the feed associated changes in blood lipid profile concentration of broilers serum
The boxplot in Figure 2 compares four indicators of lipid profile. The levels are not quite same for both feed
types, especially the boxplot for triglycerides demonstrated that feed A has apparently lower level. The box plot of
liver function parameters (Figure 3) shows that feeds produce different median levels of ALT and AST. Moreover,
there is much less variation in reading for ALT than for AST.
Figure 3. Box-plot representation for the feed associated changes in liver function concentration of broilers serum

3.2 Results of t-test and MANOVA
In order to search the significance difference of biochemical parameters and their clusters due to two feed of
poultry chicken we performed t-test and MANOVA.
Table 2. Effects of dietary supplementation on weight and biochemical composition (calcium and creatinine) of broiler
chickens at finisher period
Blood Parameters Feed Mean±SE t p-
value
Mean
difference
95% Confidence Interval for
mean difference
Lower Bound Upper Bound
Weight (gm) A 2159.77±44.05 -2.631 0.011 -150.650 -264.910 -36.410
B 2310.43±36.58
Calcium (mg/dl) A 10.31±0.18 -0.095 0.924 -0.020 -0.500 0.460
B 10.34±0.15
Creatinine A 0.43±0.01 1.900 0.062 0.030 -0.002 0.064
B 0.39±0.01
Independent samples t-test in Table 2 shows that the mean weight is significantly different for two feeds. The mean
weight of experimental birds fed two dietary treatments A and B were 2159.77gm and 2310.43gm respectively.
However, we did not find significant difference for the mean levels of calcium and creatinine for two different feeds.
Table 3. Effects of dietary supplementation on blood Lipid profile and Liver function of broiler chickens at finisher period
(Multivariate Tests using Pillai's Trace)
Effect Response F p-value Observed Power
Feed Lipid profile (TC,TG,HDL & LDL) 3.50 0.012 0.840
Liver function (ALT & AST) 15.18 0.000 0.990
Using the Pillai’s Trace test of the multivariate analysis of variance (MANOVA), the lipid profile levels and the
liver function level of poultry chickens were significantly different for two types of feeds. We were also interested in
finding out the specific components of lipid profile and liver function that varied significantly.
Table 4: The influence of dietary supplementation on serum lipid profile and liver function of broilers
Blood Parameters Types of
Feed Mean±SE F p-value Mean
difference
95% Confidence Interval for
mean difference
Lower Bound Upper Bound
Cholesterol (mg/dl) A 209.46±7.63 0.00 0.995 -0.09 -24.84 24.67
B 209.54±9.78
Triglycerides (mg/dl) A 151.14±9.21 7.87 0.007 -34.74 -59.46 -10.03
B 185.89±8.28
HDL(mg/dl) A 126.14±8.47 0.66 0.419 -10.34 -35.73 15.04
B 136.49±9.49
LDL(mg/dl) A 77.51±6.38 0.94 0.337 -8.86 -27.13 9.41
B 86.37±6.57
ALT(U/L) A 9.89±0.83 25.21 0.000 4.66 2.81 6.51
B 5.23±0.42
AST(U/L) A 198.06±11.29 12.99 0.001 55.51 24.77 86.26
B 142.54±10.48
The results of the analysis of variance (Table 4) revealed that only mean triglycerides was significantly different
for two feeds. This also confirms the result Zhan et al., (2006) serum triglyceride level is important indicator of fat
metabolism. On the other hand, both parameters observed in serum liver function i.e., mean ALT and mean AST
level were significantly different for two feeds.
4 Conclusion
A study with necessary lab experiment on seventy randomly selected broiler chickens was conducted to explore the
feed effect on biochemical parameter of the chickens’ body and some functional relationship among the components.

Analyses and graphs show that the feed provided by the hatcher produces significantly higher weight than that by
other balanced feed available in the market which supported the findings of Tohala, S. H. (2010) that birds fed high
(ME) diet showed heavier body weight (BW). However, similar test revealed no significant difference in the calcium
and creatinine levels produced by two feeds. As the variables in lipid profile levels and liver functioning are in
multivariate format, MANOVA results show that although higher weight was gained through the feed from hatcher,
this feed significantly increases the triglyceride level. Nevertheless, similar test uncovers that ALT and AST levels
were significantly lower due to the feed provided by hatcher.
Acknowledgments: Authors are thankful to Shahjalal University of Science & Technology, Research Centre for
funding and to the department of Physiology and Toxicology, department of Poultry Science; Sylhet Agricultural
University for technical help and laboratory facilities rendered to this research work.

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