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Research Article
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Toxicity of Cadmium and Lead in Gallus gallus domesticus Assessment
of Body Weight and Metal Content in Tissues after Metal Dietary Supplements |
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Salwa A. Abduljaleel
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M. Shuhaimi-Othman
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ABSTRACT
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The influence of dietary cadmium on the accumulation and effects
of dietary lead, examined in chicken. This experiment was conducted to investigate
the toxic effects of dietary Cd and Pb on chicks body weight and organ,
content of the tissues of these two metals was also detected. One day age chicks
of Gallus gallus domesticus fed diet supplemented with 25, 50, 100 ppm
of Cd, second group exposure to 300, 500, 1000 ppm of Pb in feed daily during
4 weeks. The control groups were fed without supplementation of metals. The
concentrations of Cd and Pb resulted in increased of Cd and Pb content in liver,
gizzard and muscle. While Cd 100 ppm and Pb 1000 ppm were increased metals content
in feather. Body weight of chicks was not influenced by Cd treatment. In contrary
Pb treatment was significantly (p<0.05) decreased body weight of chicks after
dietary treatment. On the other hand, Liver weigh in chicks was significantly
(p<0.05) decreased after Cd and Pb treatments.
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Received: January 07, 2013;
Accepted: February 06, 2013;
Published: May 08, 2013
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INTRODUCTION
Lead is one of the heaviest metals toxic to the environment. It has a toxic
effects in the bird practically reduced the growth rate, growth retardation
and increased mortality. Lead causes reproductive effects such as reduced egg
production in Japanese quail and reproductive failure in cattle egret (Bubulcus
ibis). It can cause oxidative damage (DNA, lipids, proteins (Hoffman
et al., 1985; Grue et al., 1986;
Burger et al.,1992; Mateo
et al., 2003). According to Burger and Gochfeld
(1995) humans and other animals exposure to lead affects the development
anatomical, physiological, behavioral and intellectual. Up till now it is largely
unknown whether the effects occur gradually or are more pronounced if exposure
occurs at particular stages. There is ample evidence that lead have negative
effects on the resistance of the body of the disease.
Chronic Cd reduced reproductive success in bird by reduce the intake and egg
production in mallard, chicken, egret and increased susceptibility to stress,
disease and oxidative and histopathological damage (Leach
et al., 1979; Nicholson et al., 1983;
Scheuhammer, 1987; Burger et
al., 1992). It is hardly capable of generating free radicals directly,
but may increase the oxidation of lipids in tissues after exposure. Exposure
to Cd is known to cause harmful effects of different levels of the trophic chain,
because of bioaccumulation (Stoeppler, 1991). Toxic effects
of Cd on birds causes reduced egg production, kidney spoil, testicular damage
and modified behavioral reaction (Furness, 1996). Bioaccumulation
is defined as the accumulation of a metal in a tissue of interest or a whole
organism that results from exposure. In other term, bioaccumulation represents
the amount of metal adsorbed or absorbed by an organism from the bioaccessible
metal encountered by the organism. Bioaccumulation of metals comes from all
environmental sources, including air, water, solid phases and feed (McGeer
et al., 2004). While, metal bioavailability includes metal species
that are biologically available and are absorbed or adsorbed by an organism
with the potential for the distribution, metabolism, elimination and bioaccumulation
(McGeer et al., 2004). In this regard, the present
study was an investigation of the exposure to Cd and Pb during the developmental
period which includes chicks body weight and tissues content of metals. Few
studies aiming at enhanced understanding of the impact of heavy metals in birds
have been demeanor.
MATERIAL AND METHODS
Animals and experimental design: Gallus domesticus fertile eggs
were obtained from a commercial (poultry farm, Malaysia). The eggs were cleaned,
labeled and weighed 58.437±8.717 g, rang 52-64 g, then put in incubator.
After 21 days, eggs hatchling, one day age chicks were randomly assigned into
seven groups (n = 10 each) according to dietary cadmium and lead. (1) control,
(2) Cd 25 ppm, (3) Cd 50 ppm, (4) Cd 100 ppm, (5) Pb 300 ppm, (6) 500 ppm and
(7) Pb 1000 ppm. The animals were housed for 4 weeks in groups in stainless
steel cages home (90x60x50 cm) with warmth provided by OSRAM lamps (100 W).
Diet and water was available daily. Drinking water and commercial diets were
offered ad libitum.
The chicks were fed starter diet to 3 week of age and a grower diet until the
end of experiment (Table 1), The toxic concentration of metals
(Cd and Pb) was chosen depend on Nutrient Requirement of Poultry by National
Research Council (NRC, 1994). The first treatment groups
were fed the experimental diets which consisted of the basal diet supplemented
with 25, 50 and 100 ppm of Cd as (CdCl2. H2O) since the
toxic concentration of cadmium was 25 and 40 ppm for immature chicken (NRC,
1994). While the second groups were fed the food supplemented with 300,
500 and 1000 ppm Pb as Pb (NO3)2. Lead was added to the
diet at this level, since the toxic dietary concentration of lead was reported
at between 200-1000 ppm for chickens (NRC, 1994). Dietary
supplements to control group without addition of metals. Body-weight and data
were obtained bi-weekly. At the end of the 4 week exposure period, the chicks
were weighed and slaughtered and the liver, gizzard, heart and feather were
removed and samples of liver, gizzard breast muscle and feather were collected
and stored prior to analysis.
Determination of metal concentration in chick tissues: Sample of liver,
gizzard and breast muscle were thawed. The samples of feather were washed in
de-ionized water which was alternated with acetone to remove loosely adherent
external contaminations (Burger and Eichhorst, 2007).
All the tissue samples were dried in an oven at 70°C for 24 h or until a
constant dry mass was achieved (Van Eeden, 2004). Then
the dryer samples ground using a mortar to powder.
The feathers were homogenized using electric blender (Philips-HR-1741) (Rattner
et al., 2008). The samples weighted about 0.5 g of each tissue except
feather and blood were weighted 0.1 g (Burger and Gochfeld,
2001).
Table 1: |
Composition of poultry feed (starter and grower) |
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The individual tissue samples (including blood) were digested with 70% nitric
acid and 30% hydrogen peroxide (2:1), according to standard method of analytic
(AOAC, 1984) and left in room temperature overnight. The
samples were completely digested in block thermostat (150°C) for 4 h until
the solutions became clear. After cooling, the solution was made up to 50 and
25 mL with de-ionized water. Cd and Pb concentration determined using inductively
coupled plasma-mass spectrometry (ICP-MS, model Perkin-Elmer Elan 9000 A). Quality
assurance procedures included the analysis of reagent blanks and appropriate
standard reference material for lobster hepatopancreas (TORT-2, National Research
Council Canada). The recovery of Cd was 73.6% and that of Pb was 107.3%. The
analytical detection limit for Cd was 0.006 μg g-1 and that
for Pb was 0.05 μg g-1.
Statistic analysis: Data were expressed as Means±SD. The values were analyzed by Independent sample t-test, was performance to found significant differences between metals concentration in treatment group and control. Differences at p<0.05 were considered statistically significant. RESULTS AND DISCUSSION Body weight of the chicks in the control group and the groups treated with metals increased in pace from the age of one day to a ripe old age of 28 days. Whereas chicks have a lower average body weight weekly throughout the experimental period in four weeks compared with the control group. According to Table 2, the chicks weight after 2 week of dietary supplement Pb was decreased significantly (p<0.05) in group Pb 1000 ppm compare with control. While dietary Cd at concentration 5, 25 and 100 ppm did not affect significantly the final body weight. After 4 week dietary Pb supplementation in the diet at 300, 500, 1000 ppm were significantly affect the chicks weight gain compared with control. On the other hand, supplementation of either diet with 25, 50 and 100 ppm cadmium had slight effect on chicks weight and growth of the three groups of treated chicks.
Table 2: |
Mean body weight gains (g), after 2 and 4 week of feed supplement
metals concentration |
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*p<0.05 ANOVA between metal treatment and control |
Metallic lead is one of the main pollutants and it is known to be toxic for
birds especially during their early development (Pain, 1995).
Present result showed that body weight of chick was significantly decreased
than control after diet treated with Pb concentration. Same observation was
previously recorded by Abd EL-Galil et al. (2006)
who demonstrated that chicken fed diets supplemented with 1000 and 1500 ppm
of lead as lead chloride was decreasing body weight. In addition, similar results
have been reported by Dicheva and Ctanchev (1988) they
found that birds fed diets with 1000 mg kg-1 of lead the body weight
gain was 6% lower than the control. El-Sharrawi et al.
(1988) found that chickens of lead exposure at doses of 750 and 1500 ppm
diet caused a major reduces in body weight gain. In addition, Wittmann
et al. (1994) found that Pb 2400 and 3600 mg kg-1 resulted
diminution in body weight compared with controls in chicken. Further, Rahman
and Joshi (2009) concluded that poultry chicks treated with lead 250 and
400 ppm were gain lower average weekly body weight when compared with the control
group and there are undesirable effects of lead acetate on performance of broilers
in these concentrations. In this regard, Erdogan et
al. (2005) investigated the dietary lead exposure causes body weight
gain decrease and inhibitory effect on the growth of broilers.
In the same time, present result in contrary with Jeng
et al. (1997) who found body weights of laying ducks were not influenced
by Pb 10 or 20 mg kg-1 b.wt. treatment, he attributed that domestic
birds may be able to tolerate higher levels of Pb as well, the difference between
Pb tolerance domestic birds and wild birds can often be attributed to environment,
species and sex. Similar observation was recorded by Custer
et al. (1984) he conclude that weights of American kestrel did not
differ among treatment groups after feed treated with Pb (120, 212 and 448 ppm)
at 60 day. Body weight of Japanese quail showed significantly decreased rates
of weight gain at the level of 500 ppm of lead in the diet (Morgan
et al., 1975). Lowered body weights in treatment groups birds could
be due to reduce in the feed utilization or due to metabolic disarray related
with lead, such as inhibition of enzyme involved in the haem synthesis and the
oxidase system resulting in loss of cellular functions and tissue damage (Erdogan
et al., 2005).
Cadmium has less effect on chick weight gain after four week of treatment by
25, 50, 100 ppm compare to control as we detected in current study. Opposite
result was achieved by Sant'Ana et al. (2005)
who found that the exposure to CdCl2 100 ppm for 28 days significantly
reduced the body weight in Japanese quail. While present result is consistent
with (Sant'Ana et al., 2005) who revealed no
significant correlation between cadmium concentration and body mass of willow
tits (Poecile montanus). Sant'Ana et al.
(2005) explained the body weight of birds decreased after Cd exposure might
be associated with the action of Metallothioneins (MT). The major function of
MT in cadmium exposure is associated with increased preservation of Cd in tissues,
resulting in a protective mechanism. Sant'Ana et al.
(2005) found more toxic affects in bird after cadmium exposure such as induced
hepatic toxicity while kidney function and cellular immune response were not
affected by the Cd exposure.
Table 3, showed that the liver, gizzard and heart weights in chicks treated with Cd concentration and in the control treatment. It noted that the chicks treated Cd 25 ppm, Cd 50 ppm and 100 ppm of Cd was significantly (p<0.01) in liver weight decreased compared with control treatment. In contrast the weight of gizzard and heart did not affect significantly compare with control treatment. In addition chicks were treated Pb concentration seems that the gizzard and heart wear does not affect the concentration of Pb while the liver weight was significantly (p<0.05) decreased by Pb 300 ppm, 500 and 1000 ppm.
Bioaccumulation of trace metal in chicks tissues after metals feed supplement:
The effect of dietary cadmium on the cadmium content of selected tissues in
4 week old broiler chickens was studied in the experiment result summarized
in Table 4. All levels of cadmium resulted in significant
increases in the cadmium content of liver compare with control Cd content and
other tissues of Cd content.
Table 3: |
Organ body weight of chicks affected by nutritional supplement
Cd and Pb |
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*p<0.05 ANOVA between metal treatment and control |
Table 4: |
Effect of dietary cadmium on the tissue cadmium concentration
of liver, gizzard, muscle and feather of young chicks |
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*p<0.05 ANOVA between metal treatment and control |
Table 5: |
Effect of dietary lead on the tissue lead concentration of
liver, gizzard, muscle and feather of young chicks |
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*p<0.05 ANOVA between metal treatment and control |
Muscle showed a seriously reduced ability to accumulate cadmium although the
two levels of Cd in Cd 50 and Cd 100 ppm did result in significant increase
compare with control treatment. Gizzard occupy the second rank after liver in
Cd accumulation and it content from Cd in three Cd groups were significantly
(p<0.05) higher than its level at control. Followed by feather which accumulates
high level of Cd at Cd 100 ppm compare to control. Compare with other study,
these data are in agreement with Leach et al. (1979)
who revealed that liver and kidney in treated chicken with cadmium in diet were
accumulated higher concentration of cadmium while muscle appeared reduced ability
to accumulate cadmium. In contrary, Mamabolo et al.(2009)
found no significant different in metal (Pb) accumulation in tissues of chicken
(liver, muscle) after metal ingestion. While, Sharma et
al. (1979) revealed that no accumulation of cadmium occurred in egg
and bone of chicken after treated by Cd (0.3, 1.9 and 13.1 ppm) in their diet
for up to 6 months. A slight increase of Cd level in muscle. While liver and
kidney had the highest of Cd levels.
Table 5 showed the effect of dietary lead on the tissue lead
concentration of liver, gizzard, muscle and feather of young chicks. High level
of Pb content accumulated in feather than other tissues. pb content in feather
at group Pb 1000 was higher relatively (p<0.05) compare with control. In
contrary, less Pb accumulated in pectoral muscle, though, the pb levels in Pb
500 and 1000 ppm group were significantly high than control. Different levels
of supplemental Pb (300, 500, 1000 ppm) were effect in Pb content in liver and
gizzard of chicks and Pb accumulated in this tissue in higher level compare
to control. Present study demonstrated that tissues Pb concentration was significantly
increased after egg injection (Pb 100, 200 ppm as Pb (NO3)2
and food treatment (pb 500 and Pb 1000 ppm). Pb levels were higher in feather
followed by gizzard and liver while muscle was accumulated less concentration
of lead in Pb same observation was record in present study from chicken collected
from field which accumulated higher levels of lead in feather followed by gizzard
and liver and less levels found in muscle. In this regard, Zraly
et al. (2008) found significantly increased lead concentrations in
liver, kidneys, bones of chicken after food treated with lead acetate. Same
observation was detected by Jeng et al. (1997)
revealed that tissues of laying Tsaiya ducks was accumulate high levels lead
after food supplied with 20 mg kg-1 Pb. Similar finding recorded
by Custer et al. (1984) that American kestrels
treated by Pb (120, 212, 448 ppm) with diet did not significantly affect concentration
of lead in the muscle but there were significant differences among treatment
for kidney, liver, femur, brain and blood. Another study was showed the liver
of quail was accumulated high level of lead after 21 days of treatment while
less concentration in gonads (Mehrotra et al., 2008).
In addition, Scheuhammer (1987) found that tissues
level of Cd and Pb increased in direct proportion to dietary levels over a particular
dose range.
Briefly, the data indicate that dietary Cd did not affect body weight and organ (gizzard and heart) of chick. At the same time Cd was accumulated in high concentrations in young chicken tissues at the end of the experiment. While dietary Pb was decreased chick body weight and liver and accumulated in the tissues of chicken significantly. CONCLUSION It was observed from the current study that supplementation of cadmium in chicken diets at 25, 50 and 100 ppm and led at 300, 500 and 1000 ppm, was effect body and liver weight of chicks after 4 week of treatment. Bioavailability of Cd and Pb measured in terms of tissue concentration (liver, muscle, gizzard and feather) was directly related to their levels of supplementation in feed. ACKNOWLEDGMENT We take this opportunity to thank the Department of School of Environmental Science and Natural Resources, UKM, Malaysia for kindly providing all of the samples collected. The author wish to acknowledge the financial support provided by the Iraqi Ministry of Higher Education, Iraq.
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