osama kornah abou emera master presentation

بسم الله الرحمن الرحيم سُبْحَانَكَ لاَ عِلْمَ لَنَا إِلاَّ مَا عَلَّمْتَنَا إِنَّكَ أَنتَ الْعَلِيمُ الْحَكِيمُ صدق الله العظيم: 

11/14/2011 1 بسم الله الرحمن الرحيم سُبْحَانَكَ لاَ عِلْمَ لَنَا إِلاَّ مَا عَلَّمْتَنَا إِنَّكَ أَنتَ الْعَلِيمُ الْحَكِيمُ صدق الله العظيم

USE OF SCANNING ELECTRON MICROSCOPY TECHNIQUES FOR PREDICTING VARIATIONS in eggshell quality of chickens: 

11/14/2011 2 USE OF SCANNING ELECTRON MICROSCOPY TECHNIQUES FOR PREDICTING VARIATIONS in eggshell quality of chickens إستخدام تقنيات الفحص بالميكروسكوب الإلكترونى للتنبؤ بالإختلافات فى جودة قشرة بيض الدجاج

Prof. Dr. Moataz Mohamed Fathi Prof. of Poultry Breeding, Faculty of Agriculture, Ain Shams University (Principal Supervisor) Prof. Dr. Usama Mohamed Ali Prof. of Poultry Breeding, Faculty of Agriculture, Ain Shams University Dr. Ahmed Galal El-Sayed Associate Prof. of Poultry Breeding, Faculty of Agriculture, Ain Shams University: 

11/14/2011 3 Prof. Dr. Moataz Mohamed Fathi Prof. of Poultry Breeding, Faculty of Agriculture, Ain Shams University (Principal Supervisor) Prof. Dr. Usama Mohamed Ali Prof. of Poultry Breeding, Faculty of Agriculture, Ain Shams University Dr. Ahmed Galal El-Sayed Associate Prof. of Poultry Breeding, Faculty of Agriculture, Ain Shams University Under the supervision of

Slide 4: 

Scenario 1. Aim of the study 2. Brief memorandum about experimental chickens 3. Genetic flock husbandry 4. Measurements and observations 5. Results 6. Conclusion

Aim of the study: 

11/14/2011 5 Aim of the study The main purpose that thesis aimed at is assessment the genetically variations related to eggshell quality in two native breed (Dandarawi and Fayoumi) vs. local synthesized strain (Norfa) and commercial strain (Hy-line) chickens. This study was an attempting to recognize finite elements of shell structure, factors that affecting the overall quality of the eggshell and relationship among eggshell microstructure elements, phenotypic traits and mechanical properties that’s can be use as an indicator of eggshell ability to confrontation environmental and mechanical insults.

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Scenario 1. Aim of the study 2. Brief memorandum about experimental chickens 3. Genetic flock husbandry 4. Measurements and observations 5. Results 6. Conclusion

2. Brief memorandum about experimental chickens : 

11/14/2011 7 1- Fayoumi breed Originally an ancient native breed from the Egyptian city of Fayoum bred for egg production. Several names have were been given to this breed. However, the most popular one is the "Fayoumi". The "Bigawi" and "Ramadi" are other names. 2. Brief memorandum about experimental chickens

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11/14/2011 8 2- Dandarawi breed These native breed chickens are found in Upper Egypt of Dandrah/Qena. The males are black with white hackle and saddle and some white on the body and wings. Females are Wheaten-looking, reddish-brown or gray. 2. Brief memorandum about experimental chickens

Slide 9: 

11/14/2011 9 3- Norfa strain These chickens are Norwegian-Egyptian strain invented for improving local breeds of laying hens in Egypt via crossing Fayoumi with two high performance lines of the White Leghorn breed imported from Norway. to develop new strain with acceptable adaptability and reasonable good production performance. 2. Brief memorandum about experimental chickens

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11/14/2011 10 4- Hy-Line strain Brown Hy-Line strain is the world’s most balanced brown egger. Hen's produces over 320 rich brown eggs to 74 weeks, peaks in the mid-90’s and begins lay early with optimum egg size. 2. Brief memorandum about experimental chickens

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Scenario 1. Aim of the study 2. Brief memorandum about experimental chickens 3. Genetic flock husbandry 4. Measurements and observations 5. Results 6. Conclusion

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11/14/2011 12 3- Genetic flocks husbandry Total 392 female chicks (75 Dandarawi, 97 Fayoumi, 150 Hy-Line and 70 Norfa) were examined in the current study. At hatching time, chicks were weighed to the nearest gram, wing tagged for their identification and brooded in electrical brooding batteries up to 4 weeks of age. Then, they were housed in floor pens. At 16 weeks of age, females were placed in individual cages to the end of the experiment. All chicks were brooded and reared under identical environmental, biosecurity and nutritional conditions. Feed and water for all birds were provided ad libitum .

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Scenario 1. Aim of the study 2. Brief memorandum about experimental chickens 3. Genetic flock husbandry 4. Measurements and observations 5. Results 6. Conclusion

4. Measurements and observations 4. 1. Growth related body weight : 

11/14/2011 14 4. Measurements and observations 4. 1. Growth related body weight Body weight (in grams) was recorded individually using top loaded scale balance for Dandarawi, Fayoumi and Brown Hy-line chicks at 0, 2, 4, 6, 8, 10, 12, 14 and 16 weeks of age.

4. Measurements and observations 4. 2. Maturation measurements : 

11/14/2011 15 4. 2. 1. Body weight 4. 2. 2. Body measurements 4. 2. 3. Shank length 4. 2. 4. Body depth 4. 2. 5. Keel length 4. 2. 6. Head appendages (Comb and wattle) lengths 4. 2. 7. Age at sexual maturity 4. 2. 8. First egg weight 4. Measurements and observations 4. 2. Maturation measurements

4. Measurements and observations 4. 3. Egg production measurements : 

11/14/2011 16 4. 3. 1. Egg number 4. 3. 2. Egg weight 4. 3. 3. Egg mass 4. 3. 4. Cracked egg 4. 3. 5. Abnormal eggs 4. 3. 6. Double yolk egg 4. Measurements and observations 4. 3. Egg production measurements

4. Measurements and observations Egg quality measurements 4. 4. Internal egg quality : 

11/14/2011 17 4. 4. 1. Yolk quality measurements 4. 4. 1. A. Yolk weight 4. 4. 1. B. Yolk percentage 4. 4. 1. C. Yolk index 4. 4. 2. Albumin quality measurements 4. 4. 2. A. Albumin weight 4. 4. 2. B. Albumin percentage 4. 4. 2. C. Haugh unit 4. Measurements and observations Egg quality measurements 4. 4. Internal egg quality

4. Measurements and observations Egg quality measurements 4. 5. External egg quality : 

11/14/2011 18 4. Measurements and observations Egg quality measurements 4. 5. External egg quality 4. 5. 1. Specific gravity 4. 5. 2. Egg volume 4. 5. 3. Breaking strength with Quasi Static Compression (QSC) 4. 5. 3. A. Instron apparatus [breaking strength (N) & deformation (mm)] 4. 5. 3. B. Fathi and El-Sahar apparatus (kg/cm 2 ) Side view

4. Measurements and observations Egg quality measurements 4. 5. External egg quality (Continued) : 

11/14/2011 19 4. 5. 4. Shell weight (wet and dry) 4. 5. 5. Shell percentage 4. 5. 6. Shell thickness (with and without membranes) 4. 5. 7. Shell surface area 4. 5. 8. Shell index 4. Measurements and observations Egg quality measurements 4. 5. External egg quality (Continued)

4. Measurements and observations 4. 6. Scanning Electron Microscopy (SEM) : 

11/14/2011 20 4. Measurements and observations 4. 6. Scanning Electron Microscopy (SEM) The ultrastructural diversity intra-mammillary layer was evaluated according to the technical terms and methodology developed via Poultry Research Unit, Univ. of Glasgow (Bain, 1990, 1992 and Solomon, 1991).

4. Measurements and observations 4. 6. Scanning Electron Microscopy (SEM) : 

11/14/2011 21 4. Measurements and observations 4. 6. Scanning Electron Microscopy (SEM) The ultrastructure evaluation of eggshell quality by accurate inspection within mammillary layer elucidate presence of 12 variance (each one estimated according to its impact positive or negative increase or decrease shell resistance to breakage) and total score (the whole measurements values) can influence on shell stiffness and its ability to tolerate the impacts.

4. Measurements and observations 4. 6. Scanning Electron Microscopy (SEM) : 

11/14/2011 22 4. Measurements and observations 4. 6. Scanning Electron Microscopy (SEM) 1-Early fusion. 2-Good cap formation (e.g., close binding between aragonite and inorganic fractions of shell). 3-cuffing. 4-confluence mammillae. 5-a high mammillary density. 1-Late fusion. 2-Type Bs. 3-Type As. 4-Aragonite. 5-Pitting: depressions, erosions, pin holes. 6-Alignment. 7-Cubics. 8-Changed membrane. 9-A low mammillary density. Increase resistance Decrease resistance

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Scenario 1. Aim of the study 2. Brief memorandum about experimental chickens 3. Genetic flock husbandry 4. Measurements and observations 5. Results 6. Conclusion

5. Results 5. 1. Growth related body weight: 

11/14/2011 24 5. Results 5. 1. Growth related body weight body weight development for Dandarawi, Fayoumi and Hy-line chicks at 2, 4, 6, 8, 10, 12, 14, 16 wks from hatching. 0 200 400 600 800 1000 1200 1400 0 2 4 6 8 10 12 14 16 Age (wk) Body weight, g Dandarawi Fayoumi Hy-line

5. Results 5. 3. Egg production measurements 5. 3. 1. Egg Number : 

11/14/2011 25 5. Results 5. 3. Egg production measurements 5. 3. 1. Egg Number Egg production rate (%) for native breed (Dandarawi and Fayoumi) and strain (Hy-line and Norfa) chickens. 60 65 70 75 80 85 90 95 100 P 1 P 2 P 3 Age (day) Egg production rate (%) Dandarawi Fayoumi Norfa Hy-line

5. Results 5. 3. Egg production measurements 5. 3. 2. Egg weight : 

11/14/2011 26 5. Results 5. 3. Egg production measurements 5. 3. 2. Egg weight Means ± SE of egg weight, Ewt (g) for native breed (Dandarawi and Fayoumi) and strain (Norfa and Hy-line) chickens. 46.72 45.65 50.45 60.51 30 35 40 45 50 55 60 65 70 Dandarawi Fayoumi Norfa Hy-line Breed or strain Ewt, g

5. Results 5. 3. Egg production measurements 5. 3. 4. Cracked egg: 

11/14/2011 27 5. Results 5. 3. Egg production measurements 5. 3. 4. Cracked egg Means ± SE of cracked egg, CE (no.) for native breed (Dandarawi and Fayoumi) and strain (Norfa and Hy-line) chickens. 2.38 0.86 0.51 0.71 0 0.5 1 1.5 2 2.5 3 Dandarawi Fayoumi Norfa Hy-line Breed or strain CE Average

5. Results 5. 4. Egg quality measurements 5. 4. 1. Internal egg quality – Yolk index : 

11/14/2011 28 5. Results 5. 4. Egg quality measurements 5. 4. 1. Internal egg quality – Yolk index Means of yolk index, YI (%) for native breed (Dandarawi and Fayoumi) and strain (Norfa and Hy-line) chickens. 44.82 44.25 46.91 48.86 43.00 44.00 45.00 46.00 47.00 48.00 49.00 50.00 Dandarawi Fayoumi Norfa Hy-line Breed or Strain Yolk index

5. Results 5. 4. Egg quality measurements 5. 4. 1. Internal egg quality – Haugh unit: 

11/14/2011 29 5. Results 5. 4. Egg quality measurements 5. 4. 1. Internal egg quality – Haugh unit Means of Haugh unit, HU for native breeds (Dandarawi and Fayoumi) and strain (Norfa and Hy-line) chickens. 81.30 82.65 77.94 87.84 76.00 78.00 80.00 82.00 84.00 86.00 88.00 90.00 Dandarawi Fayoumi Norfa Hy-line Breed or strain Haugh unit

5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – Specific gravity: 

11/14/2011 30 5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – Specific gravity Means of specific gravity, (sp gr) for native breed (Dandarawi and Fayoumi) and strain (Norfa and Hy-line) chickens. 1.090 1.089 1.092 1.088 1.087 1.088 1.088 1.089 1.089 1.090 1.090 1.091 1.091 1.092 1.092 Dandarawi Fayoumi Norfa Hy-line Breed or Strain Specific gravity

5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – Breaking strength: 

11/14/2011 31 5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – Breaking strength Means of breaking strength, QSC (Fathi and El-Sahar apparatus) kg/cm 2 Vs. Instron load (N) for native breed (Dandarawi and Fayoumi). 4 4.2 4.4 4.6 4.8 5 5.2 38 46 54 62 Age (wk) Breaking strength ( kg / cm 2 ) 41 42 43 44 45 46 47 48 49 50 51 Breaking strength (N) Fathi's load, kg / cm 2 Instron load, N Danadrawi 5.2 5.4 5.6 5.8 6.0 6.2 38 46 54 62 Age (wk) Breaking strength ( kg / cm 2 ) 49 50 51 52 53 54 55 56 57 58 59 Breaking strength (N) Fathi's load, kg / cm 2 Instron load, N Fayoumi

5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – Breaking strength: 

11/14/2011 32 5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – Breaking strength Means of breaking strength, QSC (Fathi and El-Sahar apparatus) kg/cm 2 Vs. Instron load (N) for native breed (Norfa and Hy-line). 3.6 3.8 4 4.2 4.4 4.6 4.8 5 38 46 54 62 Age (wk) Breaking strength ( kg / cm 2 ) 35 36 37 38 39 40 41 42 43 44 45 Breaking strength (N) Fathi's load, kg / cm 2 Instron load, N Norfa 3.2 3.3 3.4 3.5 3.6 3.7 3.8 3.9 4 38 46 54 62 Age (wk) Breaking strength ( kg / cm 2 ) 32 33 34 35 36 37 38 39 40 Breaking strength (N) Fathi's load, kg / cm 2 Instron load, N Hy-line

5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – STWM: 

11/14/2011 33 5. Results 5. 4. Egg quality measurements 5. 4. 2. External egg quality – STWM Means of shell thickness with membranes, STWM (mm) for native breed (Dandarawi and Fayoumi) and strain (Norfa and Hy-line) chickens. 0.38 0.40 0.37 0.35 0.33 0.34 0.35 0.36 0.37 0.38 0.39 0.40 0.41 Dandarawi Fayoumi Norfa Hy-line Breed or strain STWM, mm

5. Results 5. 5. Scanning Electron Microscopy (SEM): 

11/14/2011 34 5. Results 5. 5. Scanning Electron Microscopy (SEM) Means± SE of calcified columns lengths (vertical section), µm and percentages (%) for Dandarawi and Fayoumi eggs at 62 weeks of age. 0.01 18.20 b 19.09 a Cap % 0.01 81.80 a 80.91 b Palisade % NS 66.01±6.64 65.14±3.25 Cap thickness (CT), µm 0.0001 296.64 a ±22.92 276.02 b ±6.90 Palisade thickness (PT), µm 0.0001 362.65 a ±25.99 341.16 b ±9.51 Total thickness (TT), µm F D Prob. Breed Trait Palisade % = (palisade length / total length x 100), Cap % = (cap length / total length x 100). a and b Means within the same row with different letters are significantly differed, NS= Non significant.

5. Results 5. 5. Scanning Electron Microscopy (SEM): 

11/14/2011 35 5. Results 5. 5. Scanning Electron Microscopy (SEM) Effect of breeds on ultrastructural variants of eggshell mammillae for Dandarawi and Fayoumi chicken eggs. 0.02 1.56 23.33 b 29.17 a Total score NS 0.00 1.00 1.00 Changed membranes NS 0.44 1.67 2.00 Type A's 0.05 0.32 3.00 b 4.00 a Caps NS 0.00 1.00 1.00 Aragonite NS 0.00 1.00 1.00 Cubics NS 0.45 2.00 2.00 Erosion NS 0.28 2.33 1.50 Depression NS 0.47 3.00 3.67 Type B's 0.02 0.44 2.00 b 3.67 a Alignment NS 0.39 2.00 2.67 Cuffing 0.03 0.68 1.67 b 4.00 a Fusion NS 0.33 2.67 2.67 Confluence F D Prob. Pooled SEM Breed Variant a and b Means within the same row with different letters are significantly differed. NS= Non significant

5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's: 

11/14/2011 36 5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's Photo. 1. High density of pointed mammillary caps in Dandarawi breed. Photo. 2. Low density of rounded mammillary caps in Fayoumi breed.

5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's: 

11/14/2011 37 5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's Photo. 3. Type A's of basal mammillary caps and late fusion in Dandarawi breed. Photo. 4. Early fusion and good confluence of mammillae in Fayoumi breed.

5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's: 

11/14/2011 38 5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's Photo. 5. Disorganized mammillary bodies and extensive type B's in Dandarawi breed. Photo. 6. Good mammillary caps in Fayoumi breed.

5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's: 

11/14/2011 39 5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's Photo. 7. Isolated caps with confluent mammillary area in Dandarawi breed. Photo. 8. Extensive confluence covering mammillary caps in Fayoumi breed.

5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's: 

11/14/2011 40 5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's Photo. 9. Erosion in mammillae and poorly structured area of eggshell (Dandarawi breed). Photo. 10. Cuffing (extra crystallization material between caps) in Fayoumi breed.

5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's: 

11/14/2011 41 5. Results 5. 5. Scanning Electron Microscopy (SEM) Photo's Photo. 11. Extensive alignment in Dandarawi eggshell. Photo. 12. Early fusion and low mammillary alignment in Fayoumi eggshell

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Scenario 1. Aim of the study 2. Brief memorandum about experimental chickens 3. Genetic flock husbandry 4. Measurements and observations 5. Results 6. Conclusion

6. Conclusion: 

11/14/2011 43 6. Conclusion Finally, from these findings it could be deduces that Fayoumi breed had stiffer eggshells than Dandarawi one, owned best ultrastructure figures which contains superior well structures that magnifies eggshell elements cohesion and lower harmful structures which dismantle elements attachments of which weaken eggshell compared to Dandarawi breed chickens.

6. Conclusion: 

11/14/2011 44 And can be with a little of genetically improvement in egg production traits and raising interior egg quality characteristics, utilizing it in selection and crossbreeding programs or identifying and segregating genes responsible to good eggshell configurations. 6. Conclusion

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11/14/2011 45 Thank you