Effect of Different Levels of Perlite on Mucosal Lipase Enzymes Activity in Small Intestine of Broiler Chicks
Jamshid Ghiasi Ghalehkandi,
Mohammad Reza Valilu,
Lipase is an enzyme responsible for digestion and absorption of lipids in the small intestine and effect of perlite in the diet of broilers on lipase enzyme had not been investigated. This study had been designed and performed to evaluate effect of different levels of perlite on lipase enzyme activity in small intestine of male broiler chicks. The experiment design was arranged as randomized complete bocks in 4 x 2 factorial arrangement of treatment. The 180 male broilers of Ross 308 commercial hybrid was designated into 3 groups (0, 2 and 4%). Three replicates of 20 birds were assigned to each treatment. Control treatments were fed base diet and treatment groups with the same base diet plus 2 or 4% perlite. Animals were slaughtered at 21, 28, 36 and 42 days and different segments of small intestine (at 1,10,30,50,70 and 90% of total length of the small intestine) were taken from each replicates (n=2). Lipase enzyme activity was measured and recorded. Data were analyzed by SAS (p<0.05). Data showed that intake of perlite, significantly increased Lipase enzyme activity at different weeks and sites of the small intestine of the broiler chicks (p<0.05). These data suggested that 2% perlite administration had significantly affected lipase activity as compared with control treatment.
to cite this article:
Jamshid Ghiasi Ghalehkandi, Mohammad Reza Valilu, Jalal Shayegh, Behrad Eshratkhah, Eilyad Issabeagloo and Saeed Ghaemmaghami, 2011. Effect of Different Levels of Perlite on Mucosal Lipase Enzymes Activity in Small Intestine of Broiler Chicks. Asian Journal of Animal and Veterinary Advances, 6: 838-843.
Received: January 03, 2011;
Accepted: April 24, 2011;
Published: June 29, 2011
Perlite is one of the volcanic, aluminum- silicate minerals which are hydrated
and clear in color and there can be found tiny holes inside. Raw perlite is
transparent and light grey or gloss black and if it is put in the temperature
of 871 degrees centigrade will increase 4 to 20 times in volume and its color
will change to snow white or grey white. Perlite has neutral pH and it was confirmed
by the official congress of controlling animals as a feed additive in U.S. Its
usage as an additive is also confirmed in Europe. Concerning the chemical constituent,
it contains aluminum and silicate components (Talebali and
Farzinpour, 2006). Dietaru addition of inorganic compounds such as natural
zeolite and minerals caused considerable morphological and enzymatic changes
in intestine of broiler chickens (Incharoen et al.,
2009; Ruttanavut and Yamauchi, 2010). There are
limited number of studies on the use of perlite as an adsorbent for removal
of dyes such as methylene blue, methyl violet victoria blue and also removal
of metal icons such as copper (II) and cadmium (Demirbas
et al., 2002; Dogan and Alkan, 2003a, b;
Dyer et al., 2004). Perlite is essentially a
metastable amorphous aluminum silicate, and has recently been used as an aflatoxin
binder and adsorbent or controlling of wet litter and also decrease level of
chloride in blood serum (Talebali and Farzinpour, 2006).
Tangkawanit et al. (2005) have studied analcime
synthesized perlite for its potential use as an ion exchanger for removal of
the toxic metals such as Cu 2+, Ni2+, Pb2+
and Zn2+. In other study the use of perlite in swine feed were experimented
(Talebali and Farzinpour, 2006) A comparison was made
between pigs fattened with traditional feeds and those fattened with the same
feeds combined with perlite. He concluded that perlite fed pigs achieved a daily
weight gain higher (197 g) and duration of breeding period lower by 23 days
with the same feed utilization as the ration-fed control animals. Sakai
and Nagao (1985) used three levels of perlite (1, 10, 20%) for 8 weeks for
feeding 21 male and 21 female mice and concluded that the mices behavior,
mortality and food consumption were not affected by the experimental food and
there was no significant change in the biochemical parameters of blood and urine,
the weight of the limbs, autopsy findings and pathology of tissue, However,
the male mice fed by 10 and 20 % of perlite did not grow well but one % of perlite
was reported to be the appropriate dosage for the growth of mice. Alkan
and Dogan (2001) and Sheila (1990) reported that
perlite is responsible for breakdown of feces and absorbent of moisture and
it acts like a damper between the earth and birds and increases growth along
with decreasing the respiratory diseases, thigh bruise and bump in the breast.
Lipase is one of the most importance enzymes that play a role in fat digestion.
This enzyme facilitates hydrolysis of bonds between 1-monoglysirid 3-monoglysirid
But its action on 2-monoglysirid is very slowly, therefore, major products of
this enzyme is free fatty acids and monoglysirid. This enzyme acts on fats which
is previously emulsified (Duke, 1996).
Lipolysis for first time was established by Shif. Amylase, saccharidase, peptidase
and lipase are secreted by intestinal mucosa. Studies on Depancreatized dogs
reported that intestinal mucosa is able to hydrolyze fat, its activity increased
by biliary salts (Duke, 1996; Kihara
and Sakata, 1997). The activity of intestinal mucosal lipase is approximately
same with pancreatic lipase activity. Thus, from quantity aspect, this enzyme,
intestinal lipase, has importance role on hydrolyzing and fat absorption. In
fact about 50% of absorbed fat from gastrointestinal is mediated by intestinal
lipase. Because after obstruction of pancreatic duct, about 50% of fat hydrolyzed
and absorbed (Kihara and Sakata, 1997). Optimum pH for
activity of intestinal lipase is 9. Duodenal lipase activity was moderated for
24 h at 37°C in pH 7 and in pH more than 10 and less than 5 activity of
lipase decreased less than 10 min. EDTA caused the decreasing of lipase activity
(Osman, 1982). This study had been designed and performed
to evaluate effect of different levels of perlite on lipase enzyme activity
in small intestine of male broiler chicks.
MATERIALS AND METHODS
Birds and diet: A total number of 180 male broilers of commercial hybrid
(Ross 308) were divided into 2 experimental groups (0, 2 and 4% perlite). Each
treatment group was divided into 3 replicates of 20 birds. Birds in each replicate
were kept in cages separately next to each other and on litter. All conditions
were same for all replicates. Chicks diets were formulated according to
NRC (1994). The control treatment group was fed basal
diet (with 0.0% level of perlite) throughout the experimental period.
The other two treatment groups were fed diets supplemented with 2 and 4% of
perlite respectively. Food and water were provided ad-libitum (Table
Sample collection: In rearing period, all conditions such as temperature,
humidity, light, ventilation and management were appropriate and similar for
all broilers at the age of 21, 28, 3 and 42, 2 broilers from every group were
slaughtered after 5 h of starvation, (18 chickens for each sampling). Abdominal
cavity was opened and the entire gastrointestinal tract was removed. Small intestine
was isolated and its length was measured. The positions at 1, 10, 30, 50, 70
and 90% of the length of small intestine for analyzing the lipase enzyme activity
were separated with specific scissors (8 cm sample was taken). The samples for
lipase determination were cut open lengthwise, rinsed carefully with phosphate
buffer saline (pH = 7), blotted dry, enveloped in vacuum pack and stored at
-80°C until enzyme analysis (Teshfam, 1984).
Enzyme assay: After thawing, all of vacuum packed samples were opened
and then using a sensitive scale, 0.05 g of the mucosal small intestine was
weighed and along with 10 mL phosphate buffer saline (pH = 7) was formed into
a homogenized solution using sonic Vibracell Sonics (VCX 130 TE USA) device.
Enzyme activity of Lipase was measured according to the procedure (calorimetric
method) (AOAC, 1995). For detection of enzyme activity
it was needed to measure total protein by Pirogallol (calorimetric) method (Teshfam,
1984; Watanaba et al., 1986). Enzyme activity
level of each sample was divided into the amount of its total protein so that
activity level of the enzyme is calculated according to the IU in liter/gram
Statistical analysis: The results of the study have been statistically
analyzed using the linear model of SAS software (SAS Institute,
Analysis of variance according to the model:
||All dependent variable
||The fixed effect of RRO levels (i = 1, 2, 3)
||The effect of experimental error
Values of different parameters were expressed as the Mean±Standard deviation
(X±SD). When significant difference among means was found, means were
separated using Duncans multiple range tests.
According to Table 2, adding different levels of perlite to broilers diet at different ages caused different effects on activity of Lipase enzyme on different parts of the small intestine. Adding 2% perlite to the diet at the ages of 21, 28, 35, 42 days demonstrated a significant increase in 1, 10, 30, 50, 70 and 90% of small intestine in comparison with other treatment (4%) and control groups (p<0.05). Results showed that perlite had significant effect on Lipase activity in other ages and segments of small intestine. In part of 1% of small intestine in days 21, 35 and 42, there is a significant increase in 2% treatment than control and 4% treatments. But in part of 10% of small intestine in days 21, 35 and 42, there is a significant increase in 2% treatment than control group and in control than 4% treatment. In part of 30% of small intestine in day 21, there is a significant increase in 2% treatment than control and in control than 4% treatment whereas, in days 28 and 35, there isn't significant increase among control and 4% treatment. In part of 50% of small intestine in day 21, there is a significant increase in 2 and 4% treatments than control group whereas, in days 35 and 42, there is a significant increase in control and 2% treatment than 4% treatment. In part of 70% of small intestine in days 21 and 28, there is a significant increase in 2% treatment than control and 4% treatments. Finally, in part of 90% of small intestine in days 21 and 35, there is a significant increase in 2% treatment than control and 4% treatment.
||Comparison of average lipase activity between treatments in
different periods and 1, 10, 30, 50, 70 and 90 percent segments of small
intestine in broiler chicks (IU g-1 protein)
|Means in the same column with different superscripts differ
significantly X±SD (p<0.05)
Researchers showed role of perlite on different animal species in breakdown
of feces through transmission of moisture and it acts like a damper between
the earth and the birds and increased the growth and decreased respiratory diseases,
thigh bruises and bumps in the breast (Alkan and Dogan,
2001; Sheila, 1990). The swine which were fed daily
by perlite, were heavier (197 g) comparing with the control treatment and it
resulted in the reduction of the fattening period (Talebali
and Farzinpour, 2006). Three levels of perlite (1, 10 and 20%) were used
for 8 weeks for feeding 21 male and 21 female mice and results showed that the
mices behavior, mortality and food consumption were not affected by the
experimental food and there was no significant change in the biochemical parameters
of blood and urine, the weight of the limbs, autopsy findings and pathology
finding of tissue. However the male mice fed by 10 and 20 levels of perlite,
did not grow well and level 1 of perlite was reported to be the appropriate
dosage for the growth of mice (Sakai and Nagao, 1985).
The usage of perlite in the diet of broilers can decrease toxicity of Aflatoxin
in the body and the amount of chloride in blood serum (Talebali
and Farzinpour, 2006). In a study on chicks performance, it was concluded
that the appropriate perlite level for broiler diet was 1 to 3 percent and for
laying hens was 3 percent (Talebali and Farzinpour, 2006).
In this study, the level of mucosal enzyme activity per IU g-1 protein
was calculated in different regions of small intestine broiler chicken. Lipase
is of the most important enzymes in fat digestion which decomposes it to fatty
acids and glycerol. The activity of intestinal mucosal lipase is approximately
same with pancreatic lipase. Researches shown that secretion of lipase in hen
to six days of age is low and increased from 7 to 21 days of age. Optimum pH
for intestinal lipase is 9 and EDTA causes decreasing of lipase activity (Kihara
and Sakata, 1997; Osman, 1982). In other research
were done by Hulan and Bird (1972) revealed that enhancement
of carbohydrates and fat receiving through diet, increases the activity of amylase
and lipase enzymes.
With Adding 2% perlite to broiler diet has significance increased on lipase activity in several parts of small intestine. Perlite absorbs hydrogen ions instance other buffers and causes increasing of gastrointestinal pH. Increasing of intestinal pH is supplies sufficient environment for most enzymes activity especially intestinal amylase and lipase.
In conclusion based on result of present study and literatures data, it was suggested that adding of 2% of perlite can increase broiler performance and had stimulatory effects on intestinal lipase activity in broilers.
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