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Effect of a Specific Combination of
Mannan-Oligosaccharides and β-
Glucans Extracted from Yeast Cell
Wall on the Health Status and
Growth Performance of
Ochratoxicated Broiler Chickens
Journal of American Science, 2011;7(3)
Introduction
Gastrointestinal tract (GIT) of poultry harbors microflora,which is
formed immediately after the bird is hatched and is an important
barrier against colonization of potentially pathogenic
microorganisms. The bird’s microflora is potentially depleted for
a period of time at hatching and following any medication with
an anti-microbial product
(OTA) is the major component of a group of secondary
metabolites produced by several fungi such as
Aspergillus ochraceus or Penicillium verrucosum.
• One of the major deleterious effects of ochratoxinOne of the major deleterious effects of ochratoxin
The immunosuppressive effect of ochratoxinsThe immunosuppressive effect of ochratoxins
in chicken immune system will lead to immunein chicken immune system will lead to immune
dysfunction that can lead to exacerbationdysfunction that can lead to exacerbation
of diseasesof diseases
Eventually, it has already known that many
seases/disorders,that have immunomodulated
mponents, can be modified by administration
biological compounds that activate key pathways
the immune system. They strengthen the defense
nd immune mechanisms of the body.
Aim of workAim of work
An attempt to investigate the possible effect
of a specific combination of Mannan-
oligosaccharides (MOS) and β-glucans
(AGRIMOS®) extracted from the yeast cell
wall of Saccharomyces cerevisiae on
productive performance, ochratoxicosis
and immune dysfunction caused by
ochratoxin in broiler chickens.
Materials and MethodsMaterials and Methods
Experimental design
Three hundred and sixty, one day-old Arbor Acres
plus broiler chickens were used in this study. The
birds were allotted into 4 equal groups (I-IV) of 90
birds assigned to 3 replicates of 30 each. Those of
groups I and III were fed on ration containing
ochratoxin for the first 3 weeks of age (OTA and
OTA+AGRIMOS®
groups, respectively), while those
of groups II and IV were fed on plain ration ad
libitum (control and AGRIMOS®
treated groups,
respectively).
•At 35 days of age, 10 chickens from each group
were challenged with velogenic viscerotropic
Newcastle disease virus (VVNDv) at a dose of
106.8 EID50 / ml / bird by intramuscular
injection and kept under close observation for
clinical signs and mortality for 2 weeks. At the
end of the observation period, dead as well as
sacrificed birds (at 49 days) were subjected to
post-mortem examination for lesion scoring of
Newcastle disease virus (NDv)
Measured parameters:
Chicken zootechnical performances were determined
according to North and bell (1990) for Body Weight (g),
Body Weight Gain (g), Feed Consumption (g/day/bird),
Feed Conversion (g feed/g live body weight), and
Performances Indexes defined as follows:
- Point Spread= (Live body weight in pounds) - (Feed
conversion) X 100.
- Performance Index = Live body weight (Kg) / Feed
conversion X 100.
- European Performance Efficiency Factor (EPEP): A /
B x 10000
Where: A= Average live weight (kg) X Livability.
B= Marketing age (days) X Feed conversion.
Birds were individually weighed.
Immunoassays:
• The possible effect of AGRIMOS®
on the cell
mediated immunity was investigated using
phagocytic activity of macrophages, lysozyme
and Nitric oxide activities on blood samples
taken at 3 and 5 weeks of age on 5 birds
randomly chosen per group.
•The possible effect of AGRIMOS®
on the
humoral immunity was assessed through
haemaglutination inhibition (HI) test for
determining antibody titers against ND
Histopathology assay:
Liver, kidneys, Bursa of Fabricius, spleen
and thymus glands were collected from the
sacrificed 5 chickens per replicate at 3 and
5 weeks of age and fixed in 10% buffered
formalin. Paraffin-embedded sections were
routinely prepared and stained with
Hematoxylin and Eosin (Bancroft et al.
1996), and scored for histopathological
lesions according to the method described
by Rosales et al. (1989).
Results  I
OTA
II
Control
III
OTA+AGRIM
O
IV
AGRIMOS
SEM P value
Body weight (g)
on d 1
wk 1
wk 2
wk 3
wk 4
wk 5
 
40.4
149.7b
360.0b
746.2b
1454.2b
2022.7c
 
39.8
153.2a
404.1a
813.0a
1507.3ab
2061.2bc
 
40.7
152.1a
405.4a
838.4a
1542.3a
2157.2a
 
39.6
156.9a
414.7a
823.6a
1549.4a
2120.4ab
   
Body gain (g)
wk 0-1
wk 1-2
wk 2-3
wk 3-4
wk 4-5
wk 1-5
 
109.3
210.7b
381.0b
691.9
578.4
1982.3c
 
113.2
250.9a
408.9ab
660.8
554.0
2021.5bc
 
111.4
253.4a
427.1a
688.5
586.2
2116.9a
 
117.3
258.3a
409.3ab
710.8
571.0
2080.7ab
   
Daily feed intake (g/head)
d 1-35
 
157.1
 
145.5
 
148.3
 
157.1
   
FCR
d 1-35
 
1.577
 
1.477
 
1.480
 
1.537
   
Mortality (%)
wk 1
wk 2
wk 3
wk 4
wk 5
wk 1-5
 
0.0
0.0
0.0
6.07b
3.03
9.1
 
0.0
3.03
6.07
0.0a
0.0
9.1
 
3.03
3.03
0.0
0.0a
0.0
6.07
 
0.0
0.0
6.07
0.0a
3.03
9.1
   
Point spread (%) 288.2b
306.5ab
329.2a
314.2ab
   
Performance Index 283.5b
308.3ab
322.8a
305.0ab
   
EPEF 317.0 342.9 359.9 351.7    
   
Age
I
OTA
II
Control
III
OTA+AGR
IMOS
IV
AGRIMO
S
 
SE
M
 
P 
value
Phagocytic % 3 wk 58.33b
61.25b
61.00b
65.50a
5wk 59.00b
60.50b
63.75b
71.00a
Phagocytic
index
3 wk 0.080b
0.123b
0.133b
0.253a
5wk 0.100b
0.140b
0.160b
0.258a
Lysozyme
(µg/ml)
3 wk 9.85ab
2.73b
17.00a
9.85ab
5wk 9.85a
3 6.28a
9.85a
7.53a
Nitric oxide
(µg/ml)
3 wk 10.75c
13.25bc
17.75ab
19.50a
5wk 17.50a
21.25a
24.50a
17.50a
Macrophage activity, serum lysozyme activity and
Nitric oxide content at 3 and 5 weeks of age.
Figure 1. Haemaglutination inhibition (HI) against Newcastle disease
virus (NDv) during the first 35 days of chickens’ life.
Figure 2. Bursa weight and Bursa/Body weight indexes of
ochratoxicated and non-ochratoxicated, AGRIMOS®
treated
and untreated chickens versus blank control chicken
groups.
Figure 3. Results of macroscopic lesion scores of velogenic
viscerotropic Newcastle disease virus (VVNDv) challange of
ochratoxicated and non-ochratoxicated AGRIMOS® treated
and untreated chickens versus blank chicken group.
Histopathological results
Photo 1: Liver (gr.I) showing chronic
cholangitis. Notice the fibrous
connective tissue proliferation and
massive inflammatory cells
infiltration in the wall of bile duct
(arrow) (H&E x200)
Photo 2: Liver (gr.I) showing focal
hepatic necrosis replaced by
mononuclear leucocytes (arrow)
(H&E x200)
Photo 3: Liver (gr.IV) showing
vacuolar degeneration of centrolobular
hepatocytes (arrow) (H&E x200)
Photo 4: Liver (gr.III) showing
vacuolar degeneration of
hepatocytes, slight thickening in the
wall of bile ducts associated with
leucocytic cells infiltration (arrow)
(H&E x200)
Photo 5: Kidney (gr.I) showing
massive interstitial haemorrhage
(arrow) (H&E x100)
Photo 6: Kidney (gr.I) showing
multiple focal areas of necrosis
completely replaced by massive
leucocytes (arrow) (H&E x100)
Photo 7: Kidney (gr. III) showing
peritubular leucocytic cells infiltration
(arrow) (H & E x200)
Photo 8: Bursa of Fabricius (gr. I)
showing vaculations of lymphoid
follicles (arrow) (H & E x200)
Photo 9: Bursa of Fabricius (gr. III &
IV) showing no histopathological
changes (H & E x100)
Photo 10: Spleen (gr. I) showing
atrophy of lymphoid follicles
(arrow) (H & E x200)
Photo 11: Spleen (gr. III & IV) showing
no histopathological changes (H & E
x200)
Photo 12: Thymus gland (gr. I) showing
focal thymic haemorrhage (arrow) (H &
E x100)
Photo 13: Thymus gland (gr. III & IV)
showing no histopathological alterations
(H & E x100)
Conclusion
Administration of a specific combination of Mannan
oligosaccharides and β-glucans extracted form yeast cell wal
(AGRIMOS®
) to chickens improved zootechnical parameters and
had a potent immunomodulatory effect in the form of evoking
immune response and enhancing vaccination effectiveness.
It helps not only in controlling chicken
ochratoxicosis but also can play a positive role
in treating chicken immune dysfunction.
‫استماعكم‬ ‫لحسن‬ ‫شكرا‬‫استماعكم‬ ‫لحسن‬ ‫شكرا‬
Thank you for your attentionThank you for your attention

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Effect of a specific combination of mannan oligosaccharides and β-glucans extracted from yeast cell wall on the health status and growth performance of ochratoxicated broiler chickens

  • 1. Effect of a Specific Combination of Mannan-Oligosaccharides and β- Glucans Extracted from Yeast Cell Wall on the Health Status and Growth Performance of Ochratoxicated Broiler Chickens Journal of American Science, 2011;7(3)
  • 3. Gastrointestinal tract (GIT) of poultry harbors microflora,which is formed immediately after the bird is hatched and is an important barrier against colonization of potentially pathogenic microorganisms. The bird’s microflora is potentially depleted for a period of time at hatching and following any medication with an anti-microbial product (OTA) is the major component of a group of secondary metabolites produced by several fungi such as Aspergillus ochraceus or Penicillium verrucosum.
  • 4. • One of the major deleterious effects of ochratoxinOne of the major deleterious effects of ochratoxin The immunosuppressive effect of ochratoxinsThe immunosuppressive effect of ochratoxins in chicken immune system will lead to immunein chicken immune system will lead to immune dysfunction that can lead to exacerbationdysfunction that can lead to exacerbation of diseasesof diseases Eventually, it has already known that many seases/disorders,that have immunomodulated mponents, can be modified by administration biological compounds that activate key pathways the immune system. They strengthen the defense nd immune mechanisms of the body.
  • 5. Aim of workAim of work An attempt to investigate the possible effect of a specific combination of Mannan- oligosaccharides (MOS) and β-glucans (AGRIMOS®) extracted from the yeast cell wall of Saccharomyces cerevisiae on productive performance, ochratoxicosis and immune dysfunction caused by ochratoxin in broiler chickens.
  • 6. Materials and MethodsMaterials and Methods Experimental design Three hundred and sixty, one day-old Arbor Acres plus broiler chickens were used in this study. The birds were allotted into 4 equal groups (I-IV) of 90 birds assigned to 3 replicates of 30 each. Those of groups I and III were fed on ration containing ochratoxin for the first 3 weeks of age (OTA and OTA+AGRIMOS® groups, respectively), while those of groups II and IV were fed on plain ration ad libitum (control and AGRIMOS® treated groups, respectively).
  • 7. •At 35 days of age, 10 chickens from each group were challenged with velogenic viscerotropic Newcastle disease virus (VVNDv) at a dose of 106.8 EID50 / ml / bird by intramuscular injection and kept under close observation for clinical signs and mortality for 2 weeks. At the end of the observation period, dead as well as sacrificed birds (at 49 days) were subjected to post-mortem examination for lesion scoring of Newcastle disease virus (NDv)
  • 8. Measured parameters: Chicken zootechnical performances were determined according to North and bell (1990) for Body Weight (g), Body Weight Gain (g), Feed Consumption (g/day/bird), Feed Conversion (g feed/g live body weight), and Performances Indexes defined as follows: - Point Spread= (Live body weight in pounds) - (Feed conversion) X 100. - Performance Index = Live body weight (Kg) / Feed conversion X 100. - European Performance Efficiency Factor (EPEP): A / B x 10000 Where: A= Average live weight (kg) X Livability. B= Marketing age (days) X Feed conversion. Birds were individually weighed.
  • 9. Immunoassays: • The possible effect of AGRIMOS® on the cell mediated immunity was investigated using phagocytic activity of macrophages, lysozyme and Nitric oxide activities on blood samples taken at 3 and 5 weeks of age on 5 birds randomly chosen per group. •The possible effect of AGRIMOS® on the humoral immunity was assessed through haemaglutination inhibition (HI) test for determining antibody titers against ND
  • 10.
  • 11. Histopathology assay: Liver, kidneys, Bursa of Fabricius, spleen and thymus glands were collected from the sacrificed 5 chickens per replicate at 3 and 5 weeks of age and fixed in 10% buffered formalin. Paraffin-embedded sections were routinely prepared and stained with Hematoxylin and Eosin (Bancroft et al. 1996), and scored for histopathological lesions according to the method described by Rosales et al. (1989).
  • 12. Results  I OTA II Control III OTA+AGRIM O IV AGRIMOS SEM P value Body weight (g) on d 1 wk 1 wk 2 wk 3 wk 4 wk 5   40.4 149.7b 360.0b 746.2b 1454.2b 2022.7c   39.8 153.2a 404.1a 813.0a 1507.3ab 2061.2bc   40.7 152.1a 405.4a 838.4a 1542.3a 2157.2a   39.6 156.9a 414.7a 823.6a 1549.4a 2120.4ab     Body gain (g) wk 0-1 wk 1-2 wk 2-3 wk 3-4 wk 4-5 wk 1-5   109.3 210.7b 381.0b 691.9 578.4 1982.3c   113.2 250.9a 408.9ab 660.8 554.0 2021.5bc   111.4 253.4a 427.1a 688.5 586.2 2116.9a   117.3 258.3a 409.3ab 710.8 571.0 2080.7ab     Daily feed intake (g/head) d 1-35   157.1   145.5   148.3   157.1     FCR d 1-35   1.577   1.477   1.480   1.537     Mortality (%) wk 1 wk 2 wk 3 wk 4 wk 5 wk 1-5   0.0 0.0 0.0 6.07b 3.03 9.1   0.0 3.03 6.07 0.0a 0.0 9.1   3.03 3.03 0.0 0.0a 0.0 6.07   0.0 0.0 6.07 0.0a 3.03 9.1     Point spread (%) 288.2b 306.5ab 329.2a 314.2ab     Performance Index 283.5b 308.3ab 322.8a 305.0ab     EPEF 317.0 342.9 359.9 351.7    
  • 13.     Age I OTA II Control III OTA+AGR IMOS IV AGRIMO S   SE M   P  value Phagocytic % 3 wk 58.33b 61.25b 61.00b 65.50a 5wk 59.00b 60.50b 63.75b 71.00a Phagocytic index 3 wk 0.080b 0.123b 0.133b 0.253a 5wk 0.100b 0.140b 0.160b 0.258a Lysozyme (µg/ml) 3 wk 9.85ab 2.73b 17.00a 9.85ab 5wk 9.85a 3 6.28a 9.85a 7.53a Nitric oxide (µg/ml) 3 wk 10.75c 13.25bc 17.75ab 19.50a 5wk 17.50a 21.25a 24.50a 17.50a Macrophage activity, serum lysozyme activity and Nitric oxide content at 3 and 5 weeks of age.
  • 14. Figure 1. Haemaglutination inhibition (HI) against Newcastle disease virus (NDv) during the first 35 days of chickens’ life.
  • 15. Figure 2. Bursa weight and Bursa/Body weight indexes of ochratoxicated and non-ochratoxicated, AGRIMOS® treated and untreated chickens versus blank control chicken groups.
  • 16. Figure 3. Results of macroscopic lesion scores of velogenic viscerotropic Newcastle disease virus (VVNDv) challange of ochratoxicated and non-ochratoxicated AGRIMOS® treated and untreated chickens versus blank chicken group.
  • 17. Histopathological results Photo 1: Liver (gr.I) showing chronic cholangitis. Notice the fibrous connective tissue proliferation and massive inflammatory cells infiltration in the wall of bile duct (arrow) (H&E x200) Photo 2: Liver (gr.I) showing focal hepatic necrosis replaced by mononuclear leucocytes (arrow) (H&E x200)
  • 18. Photo 3: Liver (gr.IV) showing vacuolar degeneration of centrolobular hepatocytes (arrow) (H&E x200) Photo 4: Liver (gr.III) showing vacuolar degeneration of hepatocytes, slight thickening in the wall of bile ducts associated with leucocytic cells infiltration (arrow) (H&E x200)
  • 19. Photo 5: Kidney (gr.I) showing massive interstitial haemorrhage (arrow) (H&E x100) Photo 6: Kidney (gr.I) showing multiple focal areas of necrosis completely replaced by massive leucocytes (arrow) (H&E x100)
  • 20. Photo 7: Kidney (gr. III) showing peritubular leucocytic cells infiltration (arrow) (H & E x200) Photo 8: Bursa of Fabricius (gr. I) showing vaculations of lymphoid follicles (arrow) (H & E x200)
  • 21. Photo 9: Bursa of Fabricius (gr. III & IV) showing no histopathological changes (H & E x100) Photo 10: Spleen (gr. I) showing atrophy of lymphoid follicles (arrow) (H & E x200)
  • 22. Photo 11: Spleen (gr. III & IV) showing no histopathological changes (H & E x200) Photo 12: Thymus gland (gr. I) showing focal thymic haemorrhage (arrow) (H & E x100) Photo 13: Thymus gland (gr. III & IV) showing no histopathological alterations (H & E x100)
  • 23. Conclusion Administration of a specific combination of Mannan oligosaccharides and β-glucans extracted form yeast cell wal (AGRIMOS® ) to chickens improved zootechnical parameters and had a potent immunomodulatory effect in the form of evoking immune response and enhancing vaccination effectiveness. It helps not only in controlling chicken ochratoxicosis but also can play a positive role in treating chicken immune dysfunction.
  • 24. ‫استماعكم‬ ‫لحسن‬ ‫شكرا‬‫استماعكم‬ ‫لحسن‬ ‫شكرا‬ Thank you for your attentionThank you for your attention