BENHA VETERINARY MEDICAL JOURNAL, VOL. 25, NO.

1:29‐45, SEPTEMBER 2013

The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

Adel Mohamed Bakeer1, Ayman Samir Farid2 and Mohamed Farouk GadElKarim1*
1
Department of Pathology, Faculty of Veterinary Medicine, Cairo University, Cairo Egypt.2 Department
of Clinical Pathology, Faculty of Veterinary Medicine, Benha University, Moshtohor, Toukh 13736,
Qalyubia, Egypt. mohamedfarouk2013@yahoo.com

ABSTRACT

This study has been carried out to highlight the effects of some mycotoxins (aflatoxin, ochratoxin and
their combination) on pathological and clinicopathological picture in broiler chickens. Sixty Arber Acers
broiler chicks (one day old) were classified into 4 groups each 15 chickens. The 1st group was kept as a
control, the 2nd group was fed 2 ppm aflatoxin contaminated ration, the 3rd group was fed 4 ppm ochratoxin
contaminated ration, and the 4th group was fed 2 ppm aflatoxin + 4 ppm ochratoxin (AF+OTA) contaminated
ration from the first day of age till 6 weeks. At 2nd, 4th and 6th week of experiment, 5 birds of each group
were sacrificed and samples of blood, serum and tissue specimens were collected for histopathological,
hematological and biochemical investigations. Histopathological examination of liver from aflatoxin-
treated group showed Kupffer cells activation, focal areas of sinusoidal dilatation and periacinar hepatic
necrosis as well as hepatocellular vacuolations with mononuclear cells infiltration. As ochratoxins were
mainly nephrotoxic, the most conspicuous changes in kidneys were tubular nephrosis, subcapsular
hemorrhage, distension of some renal tubules with cellular casts with marked connective tissue proliferation
and mononuclear cells infiltration, and atrophied renal tubules were noticed. Histopathological changes
were more pronounced in group 4 (AF+OTA). The pathological changes increase in correlation with
prolonged administration of toxins. Serum biochemical results confirmed hepatotoxic effects of these
mycotoxins in chicken, which manifested by significant increases in ALT and AST values that associated
with significant elevations in total leukocytes counting. Therefore, our results indicate that mycotoxins
(aflatoxin, ochratoxin and their combination) have hepatoxic and nephrotoxic effects on growing broilers.

KEY WORDS: Mycotoxin, Broiler, Hepatotoxicity, Nephrotoxicity.

(BVMJ‐25(1):29‐45, 2013)

1- INTRODUCTION

M
ycotoxins are secondary food is a global problem because more than
metabolites present worldwide in 25% of world grain production is
agricultural commodities and contaminated by mycotoxins (3). Mycotoxins
produced by filamentous fungi that cause a according to their chemical structure exert a
toxic response (mycotoxicosis) when broad variety of biological effects (4).
ingested by animals (1). These mycotoxins Mycotoxins can be acutely or chronically
are pharmacologically active substances that toxic, or both, depending on the kind of toxin
produced in a strain-specific way that elicit and the dose. Membrane-active properties of
some complicated toxicological activities (2). various mycotoxins determine their toxicity.
Mycotoxin contamination of the feed and Incorporation of mycotoxins into cell

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 Bakeer et al., 2013

membrane structures lead to alterations in water ad libitum according to breed manual.

membrane functions. In general, mycotoxins All protocols were approved by the
effects on DNA, RNA, protein synthesis and institutional review board for animal
the pro-apoptotic action causing changes in experiments of Cairo University.
physiological functions including growth,
2.2. Experimental design:
development and reproduction (5). Almost
certainly, the main human and veterinary Chicks were randomly allocated into four
health burden of mycotoxin exposure is groups (25 animals per group). The groups
related to chronic exposure (e.g., cancer were as follows: (1) group 1; chicks were fed
induction, kidney toxicity, immune on a regular ration, mycotoxins free, and
suppression). However, the best-known served as a control group, (2) group 2; chicks
mycotoxin episodes are manifestations of were fed 2 part per million (ppm) aflatoxin
acute effects (e.g., turkey X syndrome, contaminated ration for 6 weeks, (3) group 3;
human ergotism, stachybotryotoxicosis) (2). chicks were fed 4 ppm ochratoxin
Mycotoxicosis, which can occur in both contaminated ration for 6 weeks, and (4)
industrialized and developing countries, arise group 4; chicks fed 2 ppm aflatoxin plus 4
when environmental, social and economic ppm ochratoxin contaminated ration for 6
conditions combine with meteorological weeks.
conditions (humidity, temperature), which
favour the growth of moulds (6). Aflatoxins 2.3. Samples collections:
and ochratoxins (OTA) are natural At the 2nd, 4th and 6th weeks of age, five
contaminants of feed stuff that produced by chicks from each group were randomly
Aspergillus and Penicillium species that can collected and blood samples were collected
be present as contaminant in a variety of from wing vein on EDTA as anticoagulant for
agricultural products. OTA was shown to hematological examinations and on plane
cause a diversity of toxic effects in different tube for sera separation that used for
animal species; it acts as a potent nephrotoxic biochemical investigations, then the birds
and renal carcinogen and it was also reported were killed with cervical dislocation and
to cause hepatotoxicity, immunosuppression examined for the gross lesions. Specimens of
and teratogenicity (7). Mycotoxin the liver, kidneys, spleen, thymus and bursa
contamination can reduce the birds' ability to of Fabricius, were collected on formalin
withstand stress by inhibiting the immune buffered saline (pH 7.2) for histopathological
system (8,9). Therefore, this study was aimed examination.
to investigate pathological and
clinicopathological effects of mycotoxins on 2.4. Preparation of mycotoxin contaminated
liver, kidney and immune tissues of broiler ration:
chickens.
Aflatoxin and/or ochratoxin contaminated
2- MATERIALS AND METHODS diet was prepared by artificial contamination
of the crushed yellow corn by Aspergillus
2.1. Experimental animals: parasiticusm and/or A. ochraceus,
One hundred one day-old Arbor Acher respectively and left to grow on for 14 days at
broiler chicks were used in this experiment. 28°c.
All chicks were maintained under continuous 1. For induction of aflatoxicosis, group 2
lighting for the experimental period (i.e. 6 chicks were fed on the toxicated diet, which
weeks) and given a standard diet and clean prepared by mixing of broiler ration with

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 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

artificially aflatoxicated corn to provide a 3. RESULTS

final toxin concentration of 2 mg AF/kg feed.
3.1. Histopathological examination:
2. For induction of ochratoxicosis, group 3
chicks were fed on toxicated ration prepared 1- Aflatoxin administration:
by mixing of broiler ration with artificially
ochratoxicated corn to provide a final a. Liver: Two weeks after feeding aflatoxin,
concentration of 4 mg OA/kg feed. the liver of chicks showed focal areas of
necrosis with lymphocytic cellular
3. For induction of combined aflatoxicosis aggregation. There were vacuolar and
and ochratoxicosis, contaminated ration was hydropic degeneration of the hepatocytes
prepared by mixing of broiler ration with with swollen, pale, vacuolated cytoplasm
artificially aflatoxicated and ochratoxicated and inflammatory cellular infiltration in
corn to a final toxin concentration of 2 mg AF between. The portal areas revealed
+ 4 mg OA/kg feed. mononuclear cellular infiltration and
hyperplasia of the biliary epithelium.
2.5. Hematological and biochemical analysis Then changes progressed by the 4th week
into multifocal areas of mononuclear
Red blood cell and total leukocytic count cellular aggregation in between the
was carried out according to the method hepatocytes. By the end of experiment
described previously (10). Differential (6th week), liver showed massive numbers
leukocytic count, Hb concentration and PCV of inflammatory cells infiltration
value were measured following the previous surrounding the bile duct in the portal
method (11). Serum biochemical analysis for area. Focal coagulative necrosis of the
measuring alanine aminotransferase (ALT), hepatocytes accompanied by
aspartate aminotransferase (AST) and uric inflammatory cellular infiltration mainly
acid was carried out using diagnostic lymphocytes, and macrophages were
commercial kits containing all chemicals noticed (Figure 1.a).
required as recommended by manufacturer. b. Kidney: Two weeks after feeding
2.6. Histopathological analysis: aflatoxin, there was congestion of the
cortical blood vessels associated with
To evaluate the histopathological hypercellularity of the glomerular tufts
changes, sections from each specimen (liver, due to proliferation of the endothelial and
kidney, spleen, bursa and thymus) were mesangial cells of glomerular capillaries.
stained with hematoxylin and eosin following The tubules showed coagulative necrosis
the standard method (12). of lining of epithelial cells with
inflammatory cells infiltration in
2.7. Statistical analysis: between. By the 4th week, focal
aggregates of lymphocytes were observed
Statistical analysis was performed with
in between the degenerated tubules. After
the statistical software package SPSS for
6 weeks of treatment, focal inflammatory
Windows (version 18.0; SPSS Inc., Chicago,
cells infiltration was observed in between
Ill.). The significance of differences between
the glomeruli and tubules in association
treated groups and control were evaluated by
with focal areas of hemorrhage and
student t test. A P value of less than 0.05 was
congestion in blood vessels (Figure 1.b).
considered significant. Data were grouped
and reported as means ± standard errors of the
means (SE).

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 Bakeer et al., 2013

Figure (1). Histopathological changes in different organs 6 weeks after 2 ppm aflatoxin feeding. a.
Liver of chicken showing inflammatory cells infiltration surrounding the bile duct in bile duct in
portal area. (H&E x64). b. Kidneys of chicken (group2) at 6weeks showing focal inflammatory
cells infiltration in between the tubules with congestion and hemorrhage. (H&E x40). c. Spleen of
chicken showing swelling and oedema in the media of follicular blood vessels. (H&E x64). d.
Bursa of Fabricius of chicken showing severe lymphoid depletion with fibrosis in between. (H&E
x64). e. Thymus of chicken showing slight lymphoid depletion with congested blood vessels in the
medullary portion. (H&E x40)

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 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

Figure (2). Histopathological changes in different organs 4 and 6 weeks after 4 ppm ochratoxin
feeding. a. Liver of chicken at 6weeks showing focal circumscribed round aggregation of
mononuclear leucocytes inflammatory cells infiltration in portal area with sever dilatation of portal
vein with fibrous connective tissue proliferation and degeneration in hepatocytes . (H&E x40).
b. Kidney of chicken at 6weeks showing focal area of fibrosis with inflammatory cells infiltration
in between the necrosed renal tubules. (H&E x40). c. Spleen of chicken at 6weeks showing oedema
with hypertrophy in the vascular wall of blood vessels with multiple focal microscopic nodules
formation. (H&E x64). d. Bursa of Fabricius of chicken at 6weeks showing depletion in the central
portion of the lymphoid follicles. (H&E x64). e. Thymus of chicken at 4weeks showing sever
hemorrhages and congestion in the medullary portion. (H&E x40)

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 Bakeer et al., 2013

Figure (3). Histopathological changes in different organs 6 weeks after 2 ppm aflatoxin + 4 ppm
ochratoxin combined feeding. a. Liver of chicken showing focal round aggregation of mononuclear
leucocytic inflammatory cells in the portal area with severs congestion in the portal vein. (H&E
x40). b. Kidney of chicken showing focal mononuclear leucocytic inflammatory cells infiltration
in between the glomeruli and degenerated and necrosed tubules. (H&E x40). c. Spleen of chicken
showing focal circumscribed round aggregations of mononuclear leucocytes replacing the splenic
tissue. (H&E x40). d. Bursa of Fabricius of chicken showing mucosal epithelium hyperplasia with
polyps formation and inflammatory cells infiltration in the medullary lamina propria with lymphoid
depletion of the follicles with mild fibrosis. (H&E x64). e. Thymus of chicken showing congestion
and hemorrhage in the medullary portion. (H&E x40)

34
 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

Table (1) Erytherogram parameters from control and mycotoxin-fed chickens. Data are presented as mean values ± standard error of mean.

RBCs (×106/µl) Hb (gm/dL) PCV (%) MCV (fL) MCHC (%)

Group/Toxin & Dose
2 wks 4 wks 6 wks 2 wks 4 wks 6 wks 2 wks 4 wks 6 wks 2 wks 4 wks 6 wks 2 wks 4 wks 6 wks
2.93± 1.60± 1.40 7.86± 5.23± 8.56 19.33 17.67 15.00 65.83± 116.66 ± 108.48 40.88 ± 30.27 ± 59.88 ±
Group 1 (Control)
0.03 0.30 ±0.32 0.12 0.26 ±2.35 ±0.33 ±1.72 ±2.64 0.43 17.85 ± 4.67 0.13 3.95 18.10

3.13± 1.93 2.26 7.66 4.70 8.36 20.33 18.66 20.66 65.51± 104.04 ± 92.60± 37.63 ± 23.64 ± 40.49 ±
Group 2 (AF; 2 ppm)
0.31 ±0.33 ±0.20 ±0.58 ±0.17 ±0.34 ±1.20 ±0.66 ±0.88 2.86 1.16 6.40 0.78* 1.55 2.62

3.00± 1.83 2.23 8.06 4.00± 6.9 16.66 18.66 18.00 68.96± 102.71 ± 80.42± 38.88 ± 20.97 ± 39.45 ±
Group 3 (OT; 4 ppm)
0.25 ±0.03 ±0.23 ±0.90 1.74 ±0.55 ±0.88 ±0.88 ±2.08 0.87 1.38 2.48 1.24 5.24 6.50

Group 4 ( AF; 2 ppm 2.92± 1.76± 2.21± 6.26± 3.63± 7.46± 15.96 18.06 17.89 67.98± 101.81 ± 79.82± 37.98 ± 20.08 ± 38.85 ±

+OT; 4 ppm 0.18 0.28 0.21 0.08* 0.48 0.44 ±0.86 ±0.08 ±1.88 0.77 1.08 2.28 1.14 4.84 6.20

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 Bakeer et al., 2013

Table 2. Leukogram parameters from control and mycotoxin-fed chickens. Data are presented as mean values ± standard error of mean.

WBCs (×103/µl) Heterophils (×103/µl) Lymphocytes (×103/µl)

Group/Toxin & Dose
2 wks 4 wks 6 wks 2 wks 4 wks 6 wks 2 wks 4 wks 6 wks
29.00± 97.66± 80.00± 29.00± 16.66± 13.33± 29.00± 28.66± 32.66±
Group 1 (Control)
5.00 1.45 6.08 19.00 1.76 2.40 1.00 4.37 6.35
55.00± 256.00± 67.00± 17.33±6. 11.33± 16.66± 42.00±6. 32.66± 24.00±
Group 2 (AF; 2 ppm)
2.51* 3.05* 12.42 56 2.90 1.76 00* 4.05 5.29
44.33± 199.33± 78.66± 11.33± 13.33± 26.00± 23.33±7. 37.33± 8.00±
Group 3 (OT; 4 ppm)
4.70 1.20* 10.71 4.66 0.66 1.15* 33 3.71* 1.15*
Group 4 (AF; 2 34.66± 163.33± 58.00± 14.00± 18.66± 7.33± 26.00± 23.33± 28.66±
ppm+OT; 4 ppm) 3.17 1.76* 13.11 4.61 5.69 1.76 2.30 4.80 1.76

Table 3. Serum biochemical parameters from control and mycotoxin-fed chickens. Data are presented as mean values ± standard error
of mean.

Group/Toxin & Dose ALT (U/µl) AST (U/µl) Uric acid (mg/dl)
2 wks 4 wks 6 wks 2 wks 4 wks 6 wks 2 wks 4 wks 6 wks
10.1 10.2 10.3 15.4 16.1 13.9 6.24 ± 6.13 ± 6.03 ±
Group 1 (Control)
±0.35 ±0.65 ±0.83 ±0.53 ±1.02 ±1.12 0.63 1.51 1.49
20.3 19.8 24.8 30.5 61.6 65 11.07 ± 9.89 ± 5.79 ±
Group 2 (AF; 2 ppm)
±2.11* ±0.91* ±1.15* ±3.17* ±2.82* ±3.00* 1.10 1.62 2.07
25.9 28.9 32.8 40.1 70.8 85 8.03± 5.87 ± 13.39 ±
Group 3 (OT; 4 ppm)
±1.20* ±1.33* ±1.33* ±1.85* ±3.27* ±3.44* 0.66 1.30 5.03
Group 4 (AF; 2 30.1 37.8 55.9 38.9 99.8 121.5 9.63 ± 6.77 ± 12.27 ±
ppm+OT; 4 ppm) ±1.91* ±1.75* ±2.90* ±2.47* ±4.61* ±4.31* 4.10 2.78 4.76

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 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

c. Spleen: There were thickening and hepatocytes, while marked congestion of

hypertrophy in the tunica media, and the central, portal veins and hepatic
swelling of lining endothelium of the sinusoids associated with circumscribed
intima of the follicular blood vessels. areas of round mononuclear
Circumscribed round areas of inflammatory cells aggregation were seen
mononuclear cellular aggregations adjacent to the portal area in the
forming microscopic nodules and degenerated hepatic parenchyma by the
surrounded by fine connective tissue 6th week of feeding (Figure 2.a).
capsule with lymphoid depletion were g. Kidney: The examined kidneys revealed
noticed after 2 weeks of feeding. While congestion of the cortical blood vessels
after 4th week, there were oedema and and intertubular capillaries. The
hypertrophy in the tunica media of the interstitium connective tissue revealed
follicular blood vessels. By the end of focal aggregates of lymphocytes and
experiment, marked hypertrophy in the fewer plasma cells. The renal glomeruli
tunica media of some blood vessels and were occasionally enlarged and exhibited
swelling of the lining endothelial cells hypercellularity of the glomerular tufts
were recorded (Figure 1.c) due to proliferation of the endothelial
d. Bursa of Fabricius: Lymphoid depletion cells of glomerular capillaries by the end
and necrobiosis in the lymphoid follicles of 2nd week of feeding ochratoxin. While
with hypertrophy and hyperplasia in the by the end of 4th week, the interstitium
lining mucosal epithelium were recorded connective tissue revealed massive
after 2 weeks of aflatoxin feeding. aggregates of inflammatory cells mainly
Thickening of the interstitial tissue due to lymphocytes. The renal glomeruli were
oedema and fibrous tissue proliferation enlarged and exhibited hypercellularity of
were detected after 4 weeks of aflatoxin the glomerular tufts and swelling as well
feeding. While by the 6th week, there was as proliferation of the endothelial cells of
marked lymphoid depletion in the glomeruli. Vacuolar and hydropic
follicles associated with interfollicular degeneration of the lining epithelial cells
fine fibrous tissue proliferation (Figure of some proximal and distal convoluted
1.d). tubules were observed.
Circumscribed areas of mononuclear
e. Thymus: Two weeks after feeding cellular aggregations, focal areas of
aflatoxin, there were congestion in the fibrous tissue proliferation were observed
blood vessels and focal hemorrhages in in between the necrotic tubules infiltrated
the medulla. Lymphoid depletion of the by moderate numbers of inflammatory
medulla with congestion in blood vessels cells mainly lymphocytes and
was noticed at 4 and 6 weeks post- macrophages were seen by the end of
aflatoxin administration (Figure 1.e). experiment (Figure 2.b).
h. Spleen: The spleen showed congestion of
2. Ochratoxin administration: splenic blood vessels with thickening and
hypertrophy in the tunica media after 2
f. Liver: by the 2nd and 4th weeks of feeding
weeks of experiment. While, after 4
ochratoxin, the liver showed congestion
weeks, there was marked hypertrophy in
of the central and portal veins with
the tunica media of follicular blood
multifocal areas of mononuclear cellular
vessels with swelling in the lining
aggregation as well as vascular and
endothelial cells. By the end of
hydropic degeneration in the surrounding

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 Bakeer et al., 2013

ochratoxin feeing (6th week), there were wall of bile ducts due to fibrous tissue
hypertrophy in the tunica media of some proliferation and massive inflammatory
blood vessels and swelling of the lining cellular infiltration mainly lymphocytes,
endothelial cells associated with plasma cells and fewer macrophages. At
multifocal circumscribed areas of 6 weeks, congestion was observed in the
mononuclear cellular aggregations portal vein with massive number of
forming microscopic nodules and leukocytic inflammatory cells infiltration
surrounded by fine connective tissue in the portal area. Multifocal areas of
capsule, swelling and oedema were mononuclear cellular aggregation in
observed in splenic capsule (Figure 2.c). between the hepatocytes were prominent
i. Bursa of Fabricius: After 2 weeks, there (Fig.3.a).
was lymphoid depletion of the lymphoid b. Kidney: After 2 weeks, there were
follicles with hyperplasia in the lining congestion of the blood vessels with focal
mucosal epithelium. Moreover, aggregates of lymphocytes and fewer
subepithelial and interstitial tissue was plasma cells. Circumscribed areas of
infiltrated by inflammatory cells with lymphoid cells aggregation were seen in
oedema. By 4th week, lymphoid depletion between degenerated and necrotic
of the follicles with thickening of the tubules. While, after 4 weeks, the
interstitial tissue by fibrous tissue interstitium tissue revealed focal
proliferation was noticed. While at the inflammatory cellular infiltration and
end of experiment, there was lymphoid extravasated erythrocytes. The renal
depletion of the follicles with mucosal glomeruli were enlarged and exhibited
hyperplasia of the lining epithelium and hypercellularity of the glomerular tufts
thickening of the interstitial tissue by due to swelling and proliferation of the
oedema (Fig.2,d). endothelial cells of glomerular
j. Thymus: After 2 weeks, congested blood capillaries. At the end of experiment, the
vessels with focal hemorrhages were interstitial and glomerular changes were
observed in the medulla. At the end of markedly observed. Multifocally, the
experiment, the thymus showed lining epithelial cells of proximal and
congested blood vessels and focal distal convoluted tubules were
hemorrhages were observed in both occasionally swollen with vacuolated
cortex and medulla (Figure 2.e). cytoplasm (degeneration); or they were
hyper eosinophilic with shrunken,
3. Combined administration of aflatoxin and pyknotic nuclei, loss of cellular detail and
ochratoxin: rarely karyorrhexis (necrosis).
Inflammatory cells mainly lymphocytes,
a. Liver: Two weeks after feeding,
and macrophages were observed in
multifocal areas of mononuclear cellular
between the degenerated and necrosed
aggregation were observed in between the
tubules (Figure 3.b).
hepatocytes. While after 4 weeks,
c. Spleen: 2 weeks after feeding, the red
Activation of Von Kupffer cells with the
pulp was expanded by large numbers of
presence of few numbers of macrophages
erythrocytes while the white pulp showed
and lymphocytes were noticed in between
lymphoid depletion. While after 4 weeks
the hepatic acini. The portal areas
from start of combined aflatoxin and
revealed hyperplasia of the biliary
epithelium with formation of newly ochratoxin fedding, marked hypertrophy
in the tunica media of some blood vessels
formed bile ductules, thickening of the

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 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

was characteristic. Multifocal areas of Blood hemoglobin (Table 1) showed a

hemorrhages in the red pulp and significant lower value in chicken fed 2 ppm
lymphoid depletion of the white pulp, AF+4ppm OA (group 4) at 2 weeks after
characterized by low numbers of mature treatment compared with control, while
small lymphocytes was observed. The packed cell values (Table 1) of chicken fed
splenic architecture was distorted by focal AF or OA or combination groups did not
circumscribed areas of inflammatory cells showed significant changes than control
aggregation mainly lymphocytes and group at 2, 4 and 6 weeks. Mean corpuscular
macrophages. At the end of experiment, volume in all treated groups did not show
multifocal circumscribed areas of significant changes in comparison with those
mononuclear cellular aggregations of control (group 1) at 2, 4 and 6 weeks (Table
forming microscopic nodules and 1). Mean corpuscular hemoglobin
surrounded by fine connective tissue concentration in blood of chicken fed 2 ppm
capsule were observed (Figure 3.c). AF+4ppm OA (group 4) (Table 1) were
d. Bursa of Fabricius: After 2 weeks, there significantly lower than control at the 4th
were lymphoid depletion and necrosis of week. Other groups did not show significant
the follicles with hypertrophy and changes in MCHC. Total leucocytes count in
hyperplasia in the lining mucosal blood of chicken fed aflatoxin (Table 2) was
epithelium. After 4 weeks, the changes significantly higher than the control at 2 and
were accompanied by proliferations of 4 weeks. The OA and combination groups
fibrous tissues in between necrotic areas. resulted in a significant increase in TLC than
At the end of experiment, there were the control at 4 weeks. Values of heterophils
lymphoid depletion of the follicles with count in blood of chicken fed OA groups
mucosal hyperplasia of the lining (Table 2) were significantly higher than the
epithelium and inflammatory cellular control group at the 6th week. Lymphocytes
infiltration of the lamina propria (Figure count in blood of chicken fed AF were
3.d). significantly higher than the control group
e. Thymus: After 2 weeks, the thymus (Table 1) at 2 weeks, while lymphocytes of
revealed mild lymphoid depletion at the chicken fed OA showed significant increase
medulla associated with focal at 4 weeks which changed to a significant
hemorrhage and congestion in blood decrease at 6 weeks . Values of ALT and AST
vessels, while after 4 weeks, hyalinization (Table 3) in sera of chicken fed Aflatoxin or
of Hussels corpuscles in the medullary Ochratoxin were significantly higher than the
portion was seen. At the end of control group at 2 to 6 weeks. Combination of
experiment, lymphoid depletion and focal AF+OA group resulted in higher values
hemorrhages were detected in medullary followed by OA and AF groups. Values of
portion (Figure 3. e). Uric acid (Table 3) shows non-significant
changes than the control in all groups.
3.2. Clinicopathological findings:
4. DISCUSSION
Red blood cell count of chicken fed 2 ppm
Aflatoxin (group 2), 4 ppm Ochratoxin Mycotoxins are one of the major
(group 3) and 2 ppm AF+4 ppm OA (group factors suppressing poultry productivity (13).
4) did not show significant changes in Therefore, the present study was carried out
comparison with those of control (group 1) at to investigate the pathological and
2, 4 and 6 weeks (Table 1). hematological changes in the different
organs. Mycotoxicosis was induced in broiler

39
 Bakeer et al., 2013

chicken by feeding aflatoxins (2ppm) and agreement with the results of previous studies
ochratoxins (4ppm) singly and in (14,16,17). Meanwhile, these results were in
combination. In regard to aflatoxin effect, partial agreement with other studies (19),
histopathologically, at two weeks post where thickening of glomerular basement
aflatoxin administration, liver revealed focal membrane and degenerative changes in
areas of necrosis, vacuolar and hydropic severe degree characterized by desquamation
degeneration of hepatocytes, mononuclear of epithelial tubular cells. Our results
cellular aggregation in between hepatocytes revealed also thickening in the wall of splenic
with leukocytic cellular infiltration in the blood vessels and nodulesof mononuclear
portal area biliary hyperplasia. At four weeks, cellular aggregations in spleen. Lymphoid
multifocal areas of moderate mononuclear depletion was observed in Bursa of Fabricius
cellular aggregation in between the and thymus at all periods of the experiment.
hepatocytes were common. While at six Moreover, focal areas of necrosis were
weeks; periductal massive numbers of detected in these organs at four weeks; while
inflammatory cells infiltration and focal fibrous tissue proliferation was seen in Bursa
coagulative necrosis of the hepatocytes of Fabricius, focal medullary hemorrhage in
accompanied by inflammatory cellular thymus, at six weeks. These results agreed
infiltration were detected. These findings with the results previous works (17,20). Our
agreed with earlier findings (14,15), results revealed that there were non-
meanwhile, our results were in partial significant changes in red blood cells (RBCs)
agreement with some other authors (16,17) counts at week 6, while total leukocytic count
where fibrosis, trabecular derangement of (TLC) counts significantly increased
varying degree were also observed. The especially heterophilis at week 4. These
degenerative and necrotic changes observed findings agreed with the earlier findings (16).
in aflatoxicosis could be attributed to the A drastic increase in the total leukocytic
damage of critical cellular macromolecules count, heterophils and lymphocytes were
(lipids, DNA and proteins) through the observed during cases of aflatoxicosis. This
oxidative stress of aflatoxins, which may increase suggests that the toxin elicited an
result in the peroxidation of lipids and inflammatory response (21). Our results
oxidative damage of DNA. Moreover, the revealed that the level of ALT significantly
accumulation of intracellular calcium in cases increased in aflatoxicated birds, while uric
of aflatoxicosis causes mitochondrial acid level remains within normal level. These
dysfunction and reduces adenosine results agreed with the previous results
triphosphate (ATP) generation, hyperplasia (15,22,23), and in partial agreement with
of bile ducts, which is a characteristic or other results (24,25). Increased AST activity
pathognomic lesion may be due to direct during aflatoxicosis, increased serum activity
effect of aflatoxins on biliary cells or the of enzymes, mycotoxicosis is believed to be
production of prostaglandins during the sequel of hepatocyte degeneration and a
peroxidation of lipids (18). The Kidneys subsequent leakage of enzymes into the
revealed congested blood vessels, glomerular circulation (26).
hypercellularity and coagulative necrosis of
renal tubules at two weeks postaflatoxin Concerning the effects of ochratoxin;
administration; while focal areas of grossly, at two weeks post ochratoxin
lymphocytic aggregation and hemorrhage in administration, the kidneys and livers were
between renal tubules were noticed at four swollen and pale which changed to tan color,
and six weeks. These results were in occasionally with yellowish coloration, and
have rounded borders. At four and six weeks,
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 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

the kidneys and livers were more enlarged with the earlier findings (31,32), meanwhile,
and pale in color with presence of minute our results were in partial agreement with
grayish white foci on the surfaces and cut other findings (34) where fibrosis,
sections. The ureters were markedly swollen hemorrhage, allergic reaction and defense of
and distended with urates. Decrease in the the organ against ochratoxin effect.
size of bursa, thymus lobules and spleen were Microscopical findings in immune system
also observed. These lesions were in (spleen, thymus, and bursa of Fabricius) at
complete agreement with previous results two weeks post ochratoxin administration
(27), meanwhile, were in partial agreement revealed that congested blood vessels, with
with other works (28,29). Where there were hypertrophy of their tunica media in spleen;
secondary air sacculitis, enlargement of lymphoid depletion of lymphoid follicles,
gizzard and proventriculus, pancreas and interstitial inflammatory oedema in bursa of
heart, enlarged gall bladder, hemorrhage on Fabricius; focal medullary hemorrhage in
the thigh muscles, hyperemic meninges with thymus. At four weeks, similar lesions were
oedematous changes in brain, Moreover, our detected in spleen and thymus; interstitial
results disagree with previous work (30), fibrous tissue proliferation was found in bursa
where both bursa of Fabricius and spleen of Fabricius. At six weeks, mononuclear
were not affected. Histopathologically, the cellular aggregations forming microscopic
current study revealed that microscopical nodules and surrounded by fine connective
examination of renal lesions in kidneys, at tissue capsule were seen in spleen; lymphoid
two weeks post ochratoxin administration, depletion of lymphoid follicles, focal
were characterized by congested blood hemorrhages and interstitial inflammatory
vessels, and capillaries, focal aggregation of oedema were observed in bursa of Fabricius
lymphocytes, hypercellularity of glomerular and thymus. Our microscopical findings in
tufts, vacuolar and hydropic degeneration of immune system (Spleen and Bursa of
convoluted tubules. At four weeks, similar Fabricius) agreed with the earlier results
lesions were noticed, while at six weeks, sub- (30,35).
acute to chronic interstitial nephritis
characterized by circumscribed areas of Our results revealed that PCV, Hb,
mononuclear cellular aggregations and focal MCV and MCHC showed non-significant
areas of fibrous tissue proliferation in changes, while TLC counts showed a
between the necrotic tubules. These results significant increase after 4 weeks of feeding
were in complete agreement with results of ochratoxin. This increment in TLC mainly is
previous studies (31) and in partial agreement due to increased lymphocyte. These findings
with others (32), where hemorrhage and agreed with the earlier findings (23,28,35).
fibrosis were detected, while no agreement Also, our results revealed that there is a
with the results (33) found normal kidneys. significant increase in ALT level, and non-
Microscopical examination of examined significant increase in AST level in
livers in the present study, revealed similar ochratoxicated birds. These results agreed
lesions at all periods of the experiment (2, 4 with the earlier results (23,30). Uric acid a
and 6 weeks post ochratoxin administration). non-significant increase in ochratoxicated
Congested blood vessels, focal areas of birds. These results agreed with the results
mononuclear aggregation, degenerated mentioned previously (36). In regard to
hepatocytes and inflammatory cells combined effects of aflatoxin and ochratoxin,
aggregation of portal area were the postmortem examination of broilers
predominant lesions. These findings agreed administrated with 2 ppm aflatoxins+4 ppm
ochratoxin revealed enlarged pale swollen
41
 Bakeer et al., 2013

kidneys which changed to a tan color, depletion, focal medullary hemorrhage at 2

demarcated ureters and filled with urates, pale and 6 weeks and hyalinization of Hussles
enlarged livers occasionally with yellowish corpuscle at 4 weeks. Cecal tonsils showed
coloration with focal hemorrhage, and marked lymphoid depletion of lymphoid
rounded borders, at two weeks post follicles at 2, 4 and 6 weeks. These
administration. Similar lesions in addition to microscopical findings in immune system
grayish white foci on the surfaces of liver and (spleen, bursa of Fabricius and thymus)
kidneys were seen at four weeks. Meanwhile, agreed with the previous results (38). Our
at six weeks, the livers became grayish results revealed that the level of ALT
yellow in color and firm in consistency. The significantly increased in
ureters were markedly distended with urate aflatoxin+ochratoxin administered birds from
while bursa, thymus and spleen were 2weeksof toxins administration then increase
decreased in size. These lesions were in during the 4 and 6 weeks administration.
complete agreement with the results These results agreed with the previous results
mentioned previously (37) in addition to (37). In addition to, there is no change in the
hematomas along the borders of liver (38). level of uric acid after 6 weeks of
Histopathologically, at 2 weeks post administration.
administration liver revealed multifocal areas
of mononuclear cells aggregation in between In conclusion, the previous results
hepatocytes; at 4 and 6 weeks, portal area indicate that mycotoxins (aflatoxin,
revealed hyperplasia of biliary epithelium ochratoxin or its combination) have
with fibrous tissue proliferation and hepatotoxic and nephrotoxic effects on grown
inflammatory cells infiltration, these findings chickens and every endeavor should be
agreed with the earlier findings (38). Our adopted to reduce feed contamination by
results indicated that kidney lesions were these toxins.
characterized microscopically by congested
blood vessels, focal areas of lymphoid cells 4. ACKNOWLEDGEMENT
aggregation in between degenerated and The authors thank all members of the
necrotic tubules at 2 weeks post department of Pathology, Faculty of
administration. Similar lesions as well as Veterinary Medicine, Cairo University,
hypercellularity of glomerular tuft, Egypt for their helps during this work.
degeneration and necrosis of renal epithelium
were seen at 4 and 6 weeks. These results 5. References:
were in agreement with previous results (39).
Microscopical examination of lymphoid 1. Jard, G., Liboz, T., Mathieu, F.,
organs in the present study revealed Guyonvarc'h, A., and Lebrihi, A. 2011.
hemorrhage of red pulb, and lymphoid Review of mycotoxin reduction in food and
depletion in white pulb with nodules of feed: from prevention in the field to
mononuclear cellular aggregation surrounded detoxification by adsorption or
by fine connective tissue capsule in spleen at transformation. Food Addit Contam Part A
2, 4, and 6 weeks post administration, bursa Chem Anal Control Expo Risk Assess
of Fabricius revealed lymphoid depletion and 28:1590.
2. Bennett, J.W. and Klich, M. 2003.
necrosis of follicles at 2 weeks, with
Mycotoxins. Clin Microbiol Rev 16:497.
hyperplasia of mucosal epithelium, and 3. 1996. Residues of veterinary drugs and
interstitial fibrous tissue proliferation at 4 and mycotoxins in animal products : new
6 week. Thymus revealed also lymphoid

42
 The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens

methods for risk assessment and quality aflatoxicosis in broiler chicks. Poult Sci
control. Wageningen Pers. 89:2147.
4. Fink-Gremmels, J. 1999. Mycotoxins: their 16. Del Bianchi, M., Oliveira, C.A.,
implications for human and animal health. Albuquerque, R., Guerra, J.L., and Correa,
Vet Q 21:115. B. 2005. Effects of prolonged oral
5. Jha, A., Shah, K., and Verma, R.J. 2012. administration of aflatoxin B1 and fumonisin
Aflatoxin-induced biochemical changes in B1 in broiler chickens. Poult Sci 84:1835.
liver of mice and its mitigation by black tea 17. Karaman, M., Ozen, H., Tuzcu, M.,
extract. Acta Pol Pharm 69:851. Cigremis, Y., Onder, F., and Ozcan, K. 2010.
6. Peraica, M., Radic, B., Lucic, A., and Pathological, biochemical and
Pavlovic, M. 1999. Toxic effects of haematological investigations on the
mycotoxins in humans. Bull World Health protective effect of alpha-lipoic acid in
Organ 77:754. experimental aflatoxin toxicosis in chicks.
7. Pfohl-Leszkowicz, A. and Manderville, R.A. Br Poult Sci 51:132.
2007. Ochratoxin A: An overview on 18. Hashem, M.A. and Mohamed, M.H. 2009.
toxicity and carcinogenicity in animals and Haemato-biochemical and pathological
humans. Mol Nutr Food Res 51:61. studies on aflatoxicosis and treatment of
8. Bray, G.A.E. and Ryan, D.H.E. 1991. broiler chicks in Egypt. Vet Ital 45:323.
Mycotoxins, cancer and health. Lousiana 19. Valdivia, A.G., Martinez, A., Damian, F.J.,
State University Press. Quezada, T., Ortiz, R., Martinez, C., Llamas,
9. Thompson, C. and Henke, S.E. 2000. Effect J., Rodriguez, M.L., Yamamoto, L.,
of climate and type of storage container on Jaramillo, F., Loarca-Pina, M.G., and Reyes,
aflatoxin production in corn and its J.L. 2001. Efficacy of N-acetylcysteine to
associated risks to wildlife species. J Wildl reduce the effects of aflatoxin B1
Dis 36:172. intoxication in broiler chickens. Poult Sci
10. Natt, M.P. and Herrick, C.A. 1952. A New 80:727.
Blood Diluent for Counting the Erythrocytes 20. Shivachandra, S.B., Sah, R.L., Singh, S.D.,
and Leucocytes of the Chicken. Poultry Kataria, J.M., and Manimaran, K. 2003.
Science 31:735. Immunosuppression in broiler chicks fed
11. Schalm, O.W. 1962. Practical Veterinary aflatoxin and inoculated with fowl
Hematology. Can Vet J 3:116. adenovirus serotype-4 (FAV-4) associated
12. Bancroft, J.D. and Stevens, A. 1990. Theory with hydropericardium syndrome. Vet Res
and practice of histological techniques, 3rd Commun 27:39.
edn. Churchill Livingstone, Edinburgh ; 21. Donmez, N., Donmez, H.H., Keskin, E., and
New York. Kisadere, I. 2012. Effects of aflatoxin on
13. Yunus, A.W., Razzazi-Fazeli, E., and Bohm, some haematological parameters and
J. 2011. Aflatoxin B(1) in affecting broiler's protective effectiveness of esterified
performance, immunity, and gastrointestinal glucomannan in Merino rams.
tract: a review of history and contemporary ScientificWorldJournal 2012:342468.
issues. Toxins (Basel) 3:566. 22. Raju, M.V. and Devegowda, G. 2000.
14. Tessari, E.N., Oliveira, C.A., Cardoso, A.L., Influence of esterified-glucomannan on
Ledoux, D.R., and Rottinghaus, G.E. 2006. performance and organ morphology, serum
Effects of aflatoxin B1 and fumonisin B1 on biochemistry and haematology in broilers
body weight, antibody titres and histology of exposed to individual and combined
broiler chicks. Br Poult Sci 47:357. mycotoxicosis (aflatoxin, ochratoxin and T-
15. Zhao, J., Shirley, R.B., Dibner, J.D., Uraizee, 2 toxin). Br Poult Sci 41:640.
F., Officer, M., Kitchell, M., Vazquez-Anon, 23. Aravind, K.L., Patil, V.S., Devegowda, G.,
M., and Knight, C.D. 2010. Comparison of Umakantha, B., and Ganpule, S.P. 2003.
hydrated sodium calcium aluminosilicate Efficacy of esterified glucomannan to
and yeast cell wall on counteracting counteract mycotoxicosis in naturally
contaminated feed on performance and

43
 Bakeer et al., 2013

serum biochemical and hematological ochratoxin A on egg production of laying

parameters in broilers. Poult Sci 82:571. hens. Res Vet Sci 88:486.
24. Oguz, H., Kececi, T., Birdane, Y.O., Onder, 32. Stoev, S.D., Gundasheva, D., Zarkov, I.,
F., and Kurtoglu, V. 2000. Effect of Mircheva, T., Zapryanova, D., Denev, S.,
clinoptilolite on serum biochemical and Mitev, Y., Daskalov, H., Dutton, M.,
haematological characters of broiler Mwanza, M., and Schneider, Y.J. 2012.
chickens during aflatoxicosis. Res Vet Sci Experimental mycotoxic nephropathy in
69:89. pigs provoked by a mouldy diet containing
25. Oguz, H., Kurtoglu, F., Kurtoglu, V., and ochratoxin A and fumonisin B1. Exp Toxicol
Birdane, Y.O. 2002. Evaluation of Pathol 64:733.
biochemical characters of broiler chickens 33. Schlosberg, A., Elkin, N., Malkinson, M.,
during dietary aflatoxin (50 and 100 ppb) Orgad, U., Hanji, V., Bogin, E., Weisman,
and clinoptilolite exposure. Res Vet Sci Y., Meroz, M., and Bock, R. 1997. Severe
73:101. hepatopathy in geese and broilers associated
26. Gibson, R.M., Bailey, C.A., Kubena, L.F., with ochratoxin in their feed.
Huff, W.E., and Harvey, R.B. 1990. Impact Mycopathologia 138:71.
of L-phenylalanine supplementation on the 34. Sandhu, B.S., Singh, H., and Singh, B. 1995.
performance of three-week-old broilers fed Pathological studies in broiler chicks fed
diets containing ochratoxin A. 1. Effects on aflatoxin or ochratoxin and inoculated with
body weight, feed conversion, relative organ inclusion body hepatitis virus singly and in
weight, and mortality. Poult Sci 69:414. concurrence. Vet Res Commun 19:27.
27. Stoev, S.D., Anguelov, G., Ivanov, I., and 35. Elaroussi, M.A., Mohamed, F.R., El
Pavlov, D. 2000. Influence of ochratoxin A Barkouky, E.M., Atta, A.M., Abdou, A.M.,
and an extract of artichoke on the vaccinal and Hatab, M.H. 2006. Experimental
immunity and health in broiler chicks. Exp ochratoxicosis in broiler chickens. Avian
Toxicol Pathol 52:43. Pathol 35:263.
28. Stoev, S.D., Daskalov, H., Radic, B., 36. Denli, M., Blandon, J.C., Guynot, M.E.,
Domijan, A.M., and Peraica, M. 2002. Salado, S., and Perez, J.F. 2009. Effects of
Spontaneous mycotoxic nephropathy in dietary AflaDetox on performance, serum
Bulgarian chickens with unclarified biochemistry, histopathological changes,
mycotoxin aetiology. Vet Res 33:83. and aflatoxin residues in broilers exposed to
29. Stoev, S.D., Stefanov, M., Denev, S., Radic, aflatoxin B(1). Poult Sci 88:1444.
B., Domijan, A.M., and Peraica, M. 2004. 37. Campbell, M.L., Jr., May, J.D., Huff, W.E.,
Experimental mycotoxicosis in chickens and Doerr, J.A. 1983. Evaluation of
induced by ochratoxin A and penicillic acid immunity of young broiler chickens during
and intervention with natural plant extracts. simultaneous aflatoxicosis and
Vet Res Commun 28:727. ochratoxicosis. Poult Sci 62:2138.
30. Hanif, N.Q., Muhammad, G., Siddique, M., 38. Trenk, H.L., Butz, M.E., and Chu, F.S. 1971.
Khanum, A., Ahmed, T., Gadahai, J.A., and Production of ochratoxins in different cereal
Kaukab, G. 2008. Clinico- products by Aspergillus ochraceus. Appl
pathomorphological, serum biochemical and Microbiol 21:1032.
histological studies in broilers fed ochratoxin 39. Verma, J., Johri, T.S., Swain, B.K., and
A and a toxin deactivator (Mycofix Plus). Br Ameena, S. 2004. Effect of graded levels of
Poult Sci 49:632. aflatoxin, ochratoxin and their combinations
31. Stoev, S.D. 2010. Studies on some feed on the performance and immune response of
additives and materials giving partial broilers. Br Poult Sci 45:512.
protection against the suppressive effect of

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 ‫‪The Hepatotoxic and Nephrotoxic Effects of Mycotoxin in Broiler Chickens‬‬
‫ﻋﺪد ‪ 45-29 :(1) 25‬ﺳ ﺘﻤ ﺮ ‪2013‬‬ ‫ﻣﺠﻠﺔ ﺑ ﺎ ﻟﻠﻌﻠﻮم اﻟﻄﺒﻴﺔ اﻟﺒﻴﻄﺮ ﺔ‬

‫اﻟﺗﺄﺛﯾر اﻟﻛﺑدي واﻟﻛﻠوي اﻟﺿﺎر ﻟﻠﺳﻣوم اﻟﻔطرﯾﺔ ﻋﻠﻰ دﺟﺎج اﻟﺗﺳﻣﯾن‬

‫‪1‬‬
‫ﻋﺎدل ﻣﺣﻣد ﺑﻛﯾر‪1‬؛ أﯾﻣن ﺳﻣﯾر ﻓرﯾد‪2‬؛ ﻣﺣﻣد ﻓﺎروق ﺟﺎد اﻟﻛرﯾم‬
‫‪1‬ﻗﺳم اﻟﺑﺎﺛوﻟوﺟﯾﺎ‪-‬ﻛﻠﯾﺔ اﻟطب اﻟﺑﯾطري‪-‬ﺟﺎﻣﻌﺔ اﻟﻘﺎﻫرة‪2 ،‬ﻗﺳم اﻟﺑﺎﺛوﻟوﺟﯾﺎ اﻹﻛﻠﯾﻧﯾﻛﯾﺔ‪-‬ﻛﻠﯾﺔ اﻟطب اﻟﺑﯾطري‪-‬ﺟﺎﻣﻌﺔ ﺑﻧﻬﺎ‬

‫اﻟﻣﻠﺧص اﻟﻌرﺑﻲ‬

‫ﺍﺳﺘﻬﺪﻓﺖ ﻫﺬﻩ ﺍﻟﺪﺭﺍﺳﺔ ﻣﻌﺮﻓﺔ ﺍﻷﺛﺎﺭ ﺍﻟﺴﻤﻴﺔ ﺍﻟﻐﺮﺽ ﻟﻜﻞ ﻣﻦ ﺳﻢ ﺃﻓﻼﺗﻮﻛﺴﻴﻦ ‪ 2‬ﺟﺰء‪ /‬ﺍﻟﻤﻠﻴﻮﻥ ﻭﺳﻢ ﺍﻷﻛﻮﺭﺍﺗﻮﻛﺴﻦ ‪ 4‬ﺟﺰء‬
‫‪/‬ﺍﻟﻤﻠﻴﻮﻥ ﺃﻭ ﻛﻼﻫﻤﺎ ﻋﻠﻰ ﺑﺪﺍﺭﻯ ﺍﻟﺘﺴﻤﻴﻦ‪ .‬ﺍﺟﺮﻳﺖ ﺍﻟﺘﺠﺮﺑﺔ ﻋﻠﻲ ‪ 100‬ﻛﺘﻜﻮﺕ ﻋﻤﺮ ﻳﻮﻡ ﺗﻢ ﺗﻘﺴﻴﻤﻬﺎ ﺇﻟﻲ ﺍﺭﺑﻊ ﻣﺠﻤﻮﻋﺎﺕ )‪ 25‬ﻛﺘﻜﻮﺕ‬
‫‪ /‬ﻣﺠﻤﻮﻋﺔ( ﻭﻛﺎﻧﺖ ﺍﻟﻤﺠﻤﻮﻋﺎﺕ ﻛﺎﻵﺗﻲ‪ :‬ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻷﻭﻟﻲ )ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻟﻀﺎﺑﻄﺔ( ﻟﻢ ﺗﻌﺎﻟﺞ ﺑﺄﻱ ﻣﻌﺎﻟﺠﺎﺕ ﻭﺗﻤﺖ ﺗﻐﺬﻳﺘﻬﺎ ﻋﻠﻲ ﻋﻠﻴﻘﺔ‬
‫ﺴﻤﻮﻡ ﺍﻟﻔﻄﺮﻳﺔ – ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻟﺜﺎﻧﻴﺔ ﺗﻢ ﺗﻐﺬﻳﺘﻬﺎ ﻋﻠﻲ ﻋﻠﻴﻘﺔ ﺗﺤﺘﻮﻱ ﻋﻠﻲ ‪ 2‬ﺟﺰء‪ /‬ﺍﻟﻤﻠﻴﻮﻥ ﺃﻓﻼﺗﻮﻛﺴﻴﻦ – ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻟﺜﺎﻟﺜﺔ‬ ‫ﺧﺎﻟﻴﺔ ﻣﻦ ﺍﻟ ُ‬
‫ﺗﻢ ﺗﻐﺬﻳﺘﻬﺎ ﻋﻠﻲ ﻋﻠﻴﻘﺔ ﺗﺤﺘﻮﻱ ﻋﻠﻲ ‪ 4‬ﺟﺰء ‪ /‬ﺍﻟﻤﻠﻴﻮﻥ ﺃﻭﻛﺮﺍﺗﻮﻛﺴﻴﻦ – ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻟﺮﺍﺑﻌﺔ ﺗﻢ ﺗﻐﺬﻳﺘﻬﺎ ﻋﻠﻲ ﻋﻠﻴﻘﺔ ﺗﺤﺘﻮﻱ ﻋﻠﻲ ‪ 2‬ﺟﺰء‬
‫‪ /‬ﺍﻟﻤﻠﻴﻮﻥ ﺃﻓﻼﺗﻮﻛﺴﻴﻦ ‪ 4 +‬ﺟﺰء ‪ /‬ﺍﻟﻤﻠﻴﻮﻥ ﺍﻭﻛﺮﺍﺗﻮﻛﺴﻴﻦ ﻟﻤﺪﺓ ‪ 6‬ﺃﺳﺎﺑﻴﻊ ﻭﺗﻢ ﺫﺑﺢ ‪ 5‬ﻁﺎﺋﺮ ﻣﻦ ﻛﻞ ﻣﺠﻤﻮﻋﺔ ﻓﻲ ﺍﻷﺳﺒﻮﻉ ﺍﻟﺜﺎﻧﻲ‬
‫ﻭﺍﻟﺮﺍﺑﻊ ﻭﺍﻟﺴﺎﺩﺱ ‪ .‬ﻭﻟﻘﺪ ﺗﻢ ﺗﺠﻤﻴﻊ ﻋﻴﻨﺎﺕ ﺍﻟﺪﻡ ﻣﻊ ﺍﺿﺎﻓﺔ ﻣﺎﻧﻊ ﺗﺠﻠﻂ – ﻋﻴﻨﺎﺕ ﺳﻴﺮﻡ ﻭﻛﺬﻟﻚ ﻋﻴﻨﺎﺕ ﻣﻦ ﻣﺨﺘﻠﻒ ﺍﻷﻧﺴﺠﺔ ﻣﺜﺒﺘﺔ ﻓﻲ‬
‫ﻣﺤﻠﻮﻝ ﻓﻮﺭﻣﺎﻟﻴﻦ ﻭﺫﻟﻚ ﻹﺟﺮﺍء ﺩﺭﺍﺳﺎﺕ ﺗﺤﻠﻴﻞ ﺍﻟﺪﻡ ﻭﺍﻟﺘﺤﺎﻟﻴﻞ ﺍﻟﺴﻴﺮﻭﻟﻮﺟﻴﺔ ﻭﺍﻟﺒﺎﺛﻮﻟﻮﺟﻴﺔ ﻭﺫﻟﻚ ﻓﻲ ﺍﻷﺳﺎﺑﻴﻊ ﺍﻟﺜﺎﻧﻲ ﻭﺍﻟﺮﺍﺑﻊ ﻭﺍﻟﺴﺎﺩﺱ‬
‫ﺣﺘﻲ ﻧﻬﺎﻳﺔ ﺍﻟﺘﺠﺮﺑﺔ‪ .‬ﻭﻟﻘﺪ ﺃﻅﻬﺮ ﺍﻟﻔﺤﺺ ﺍﻟﻨﺴﻴﺠﻲ ﻟﻠﻤﺠﻤﻮﻋﺔ ﺍﻟﺜﺎﻧﻴﺔ ﺣﺪﻭﺙ ﺗﺤﻠﻞ ﻓﻲ ﺍﻟﺨﻼﻳﺎ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﻭﻧﻘﺺ ﺷﺪﻳﺪ ﻓﻲ ﻋﺪﺩﻫﺎ ﻓﻲ‬
‫ﺍﻟﻄﺤﺎﻝ ﻭﻏﺪﺓ ﻓﺎﺑﺮﻳﺸﺲ ﻭﺍﻟﻐﺪﺓ ﺍﻟﺰﻋﺘﺮﻳﺔ )ﺍﻟﺜﺎﻳﻤﺲ( ﻭﺍﻟﺘﺠﻤﻌﺎﺕ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﺑﺎﻷﻣﻌﺎء‪ ،‬ﻭﺟﻮﺩ ﺍﺭﺗﺸﺎﺡ ﺑﺎﻟﺨﻼﻳﺎ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﻓﻲ ﺍﻟﻤﻨﻄﻘﺔ‬
‫ﺍﻟﺒﺎﺑﻴﺔ ﻟﻠﻜﺒﺪ ﻣﻊ ﻭﺟﻮﺩ ﻧﺨﺮ ﻟﺒﻌﺾ ﺍﻟﺨﻼﻳﺎ ﻭﻛﺬﻟﻚ ﻭﺟﻮﺩ ﻧﻤﻮﺍﺕ ﺟﺪﻳﺪﺓ ﻟﻠﻘﻨﻮﺍﺕ ﺍﻟﺼﻔﺮﺍﻭﻳﺔ‪ ،‬ﺍﻣﺎ ﺍﻟﻜﻠﻲ ﻓﻘﺪ ﻟﻮﺣﻆ ﻭﺟﻮﺩ ﻧﺨﺮ ﻟﺒﻌﺾ‬
‫ﺧﻼﻳﺎ ﺍﻷﻧﺎﺑﻴﺐ ﺍﻟﻜﻠﻮﻳﺔ‪ .‬ﻋﻨﺪ ﺍﻟﻔﺤﺺ ﺍﻟﻨﺴﻴﺠﻲ ﻟﻠﻄﻴﻮﺭ ﻓﻲ ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻟﺜﺎﻟﺜﺔ ﻟﻮﺣﻆ ﺣﺪﻭﺙ ﻧﺨﺮ ﻓﻲ ﺍﻟﺨﻼﻳﺎ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﻭﻧﻘﺺ ﻓﻲ‬
‫ﻋﺪﺩﻫﺎ ﻓﻲ ﺍﻟﻄﺤﺎﻝ ﻭﻏﺪﺓ ﻓﺎﺑﺮﻳﺸﺲ ﻭﺍﻟﻐﺪﺓ ﺍﻟﺰﻋﺘﺮﻳﺔ )ﺍﻟﺜﺎﻳﻤﺲ( ﻭﺍﻟﺘﺠﻤﻌﺎﺕ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﺑﺎﻷﻣﻌﺎء‪ ،‬ﻭﺟﻮﺩ ﻧﺨﺮ ﺷﺪﻳﺪ ﺑﺨﻼﻳﺎ ﺍﻟﻜﺒﺪ‪ ،‬ﺃﻣﺎ‬
‫ﺍﻟﻜﻠﻲ ﻓﻘﺪ ﻟﻮﺣﻆ ﺣﺪﻭﺙ ﻓﺮﺍﻏﺎﺕ ﺑﺎﻟﺴﻴﺘﻮﺑﻼﺯﻡ ﻟﺒﻌﺾ ﺍﻟﺨﻼﻳﺎ ﺍﻟﻤﺒﻄﻨﺔ ﻟﻸﻧﺎﺑﻴﺐ ﺍﻟﻜﻠﻮﻳﺔ ﺍﻟﻌﻠﻮﻳﺔ ﻣﻊ ﻭﺟﻮﺩ ﻧﺨﺮ ﻓﻲ ﺑﻌﺾ ﺍﻟﺨﻼﻳﺎ‬
‫ﺍﻷﺧﺮﻯ ﺑﺠﻤﻴﻊ ﻣﺮﺍﺣﻠﻪ‪ .‬ﻋﻨﺪ ﺍﻟﻔﺤﺺ ﺍﻟﻨﺴﻴﺠﻲ ﻟﻠﻄﻴﻮﺭ ﻓﻲ ﺍﻟﻤﺠﻤﻮﻋﺔ ﺍﻟﺮﺍﺑﻌﺔ ﻟﻮﺣﻆ ﺣﺪﻭﺙ ﻧﺨﺮ ﻓﻲ ﺍﻟﺨﻼﻳﺎ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﻭﻧﻘﺺ ﻓﻲ‬
‫ﻋﺪﺩ ﻫﺬﻩ ﺍﻟﺨﻼﻳﺎ ﻓﻲ ﺍﻟﻄﺤﺎﻝ ﻭﻏﺪﺓ ﻓﺎﺑﺮﻳﺸﺲ ﻭﺍﻟﻐﺪﺓ ﺍﻟﺰﻋﺘﺮﻳﺔ )ﺍﻟﺜﺎﻳﻤﺲ( ﻭﺍﻟﺘﺠﻤﻌﺎﺕ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﺑﺎﻷﻣﻌﺎء‪ ،‬ﻭﻛﺬﻟﻚ ﻭﺟﻮﺩ ﺍﺭﺗﺸﺎﺡ‬
‫ﺑﺎﻟﺨﻼﻳﺎ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﻓﻲ ﺍﻟﻤﻨﻄﻘﺔ ﺍﻟﺒﺎﺑﻴﺔ ﺑﺎﻟﻜﺒﺪ ﻣﻊ ﻭﺟﻮﺩ ﻧﺤﺮ ﻟﺒﻌﺾ ﺍﻟﺨﻼﻳﺎ ﻭﻛﺬﻟﻚ ﻭﺟﻮﺩ ﻧﻤﻮﺍﺕ ﺟﺪﻳﺪﺓ ﻟﻠﻘﻨﻮﺍﺕ ﺍﻟﺼﻔﺮﺍﻭﻳﺔ‪ ،‬ﺃﻣﺎ ﺍﻟﻜﻠﻲ‬
‫ﻓﻘﺪ ﻟﻮﺣﻆ ﺣﺪﻭﺙ ﻧﺨﺮ ﻟﺒﻌﺾ ﺧﻼﻳﺎ ﺍﻷﻧﺎﺑﻴﺐ ﺍﻟﻜﻠﻮﻳﺔ‪ .‬ﻭﺟﺪ ﺃﻳﻀﺎ ً ﺃﻧﻪ ﻓﻲ ﺍﻟﻤﺠﻤﻮﻋﺎﺕ ‪ 4-3-2‬ﻧﻘﺺ ﻏﻴﺮ ﻣﻌﻨﻮﻱ ﻓﻲ ﺍﻟﻌﺪﺩ ﺍﻟﻜﻠﻲ ﻓﻲ‬
‫ﻛﺮﺍﺕ ﺍﻟﺪﻡ ﺍﻟﺤﻤﺮﺍء ﺑﺎﻹﺿﺎﻓﺔ ﺍﻟﻲ ﻧﻘﺺ ﻣﻠﺤﻮﻅ ﻓﻲ ﻧﺴﺒﺔ ﺍﻟﻬﻴﻤﻮﺟﻠﻮﺑﻴﻦ ﻣﻊ ﻭﺟﻮﺩ ﺯﻳﺎﺩﺓ ﻓﻲ ﺍﻟﻌﺪﺩ ﺍﻟﻜﻠﻲ ﻓﻲ ﻛﺮﺍﺕ ﺍﻟﺪﻡ ﺍﻟﺒﻴﻀﺎء ﺑﺎﻻ‬
‫ﺿﺎﻓﺔ ﺍﻟﻲ ﺯﻳﺎﺩﺓ ﻓﻲ ﺍﻟﺨﻼﻳﺎ ﺍﻟﻠﻴﻤﻔﺎﻭﻳﺔ ﻭﺧﻼﻳﺎ ﺍﻟﻬﻴﺘﻴﺮﻭﻓﻴﻞ‪ .‬ﻛﻤﺎ ﻭﺟﺪﺕ ﺯﻳﺎﺩﺓ ﻏﻴﺮ ﻣﻌﻨﻮﻳﺔ ﻓﻲ ﻣﺴﺘﻮﻱ ﺍﻟﺴﻴﺮﻡ ﻣﻦ ﺣﻤﺾ ﺍﻟﻴﻮﺭﻳﻚ‪،‬‬
‫ﺴﻤﻮﻡ ﺍﻟﻔﻄﺮﻳﺔ‬ ‫ﻣﻊ ﺯﻳﺎﺩﺓ ﻭﺍﺿﺤﺔ ﻓﻲ ﻧﺸﺎﻁ ﺍﻧﺰﻳﻢ )‪ (ALT‬ﻣﻊ ﺯﻳﺎﺩﺓ ﻏﻴﺮ ﻣﻌﻨﻮﻳﺔ ﻓﻲ ﻧﺸﺎﻁ ﺍﻧﺰﻳﻢ ) ‪ .(AST‬ﻣﻤﺎ ﺳﺒﻖ ﻧﺴﺘﻨﺘﺞ ﺃﻥ ﺍﻟ ُ‬
‫ﺴﻢ ﻭﻳﺘﻤﺮﻛﺰ ﺗﺄﺛﻴﺮ ﺍﻻﻓﻼﺗﻮﻛﺴﻴﻦ ﻋﻠﻲ ﺍﻟﻜﺒﺪ ﻛﻤﺎ ﻳﺘﻤﺮﻛﺰ ﺗﺄﺛﻴﺮ ﺍﻷﻭﻛﺮﺍﺗﻮﻛﺴﻴﻦ ﻋﻠﻲ ﺍﻟﻜﻠﻰ‬ ‫ﻟﻬﺎ ﺗﺄﺛﻴﺮ ﺿﺎﺭ ﻏﻠﻲ ﺍﻷﻧﺴﺠﺔ ﺍﻟﻤﺨﺘﻠﻔﺔ ﺑﺎﻟﺠ ُ‬
‫ﻣﻊ ﺯﻳﺎﺩﺓ ﻭﺍﺿﺤﺔ ﻓﻲ ﻧﺸﺎﻁ ﺍﻧﺰﻳﻢ )‪ (ALT‬ﻭﺯﻳﺎﺩﺓ ﻏﻴﺮ ﻣﻌﻨﻮﻳﺔ ﻓﻲ ﻧﺸﺎﻁ ﺍﻧﺰﻳﻢ )‪. (AST‬‬

‫)ﻣﺟﻠﺔ ﺑﻧﻬﺎ ﻟﻠﻌﻠوم اﻟطﺑﯾﺔ اﻟﺑﯾطرﯾﺔ‪ :‬ﻋدد ‪ ,45-29:(1)25‬ﺳﺑﺗﻣﺑر ‪(2013‬‬

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