INFLUENCE%20OF%20GARLIC%20(Allium%20sativum)%20FEED%20INCLUSION%20ON%20PRODUCTION%20PERFORMANCE%20AND%20HEALTH%20OF%20COMMERCIAL%20CHICKENS

1
UNIVERSITY OF IBADAN RESEARCH FOUNDATIONPROPOSAL
FOR COLLABORATIVE RESEARCH PROJECT

• INFLUENCE OF GARLIC (Allium sativum) FEED
  INCLUSION ON PRODUCTION PERFORMANCE AND HEALTH OF
  COMMERCIAL CHICKENS
• 19 November, 2013

2
Research Team

• Dr. Omolade A. Oladele (Principal investigator)
• Avian Diseases Unit,
• Department of Veterinary Medicine,
• University of Ibadan.
• 2. Dr. O. A. Abu (Co-Principal Investigator)
• Department of Animal Science,
• University of Ibadan.
• Dr. Olufunke O. Ezekiel (Co-Principal
  Investigator)
• Department of Food Technology,
• University of Ibadan.

3

• Dr. O.O. Esan
• Department of Veterinary Medicine
• University of Ibadan.
• Dr. Olutola Oyedele
• Agricultural Extensionist,
• National Horticultural Research Institute
  (NIHORT)
• Idi-Ishin, Ibadan.
• 6. Mrs Yemisi Adewale
• Agronomist,
• National Horticultural Research (NIHORT)
• Idi-Ishin, Ibadan.
•  

4
INTRODUCTION

• Inclusion of growth promoters in poultry feed is
  old common worldwide (Waibei et al., 1954
  Libby Schaible, 1955).
• Usually antibiotics due to abundance of
  pathogenic microorganisms in the environment.
• Microorganisms subclinical infections
  sub-optimal performance.

5

• The use of antibiotics as growth promoters in
  poultry has faced intense criticism for public
  health reasons
• i. Antibiotic resistant strains of pathogens
  have evolved
• thereby challenging effective therapy in
  both poultry
• and humans (Campylobacter, Enterococcus,
  Salmonella
• spps and E. coli Luo et al., 2003
  Graham et al., 2007).
• European Union,1999 banned, most
  antibiotics (Casewell et al., 2003).
• U.S., 2004 banned, Enrofloxacin in food
  animals (Graham et al., 2007).
• ii. Antibiotic residues poultry meat eggs
  made available for human consumption in Nigeria
  (Dipeolu et al., 2000 2002 2004).

6

• Poultry producers have therefore been searching
  for alternatives (Javandel et al., 2008).
• Non-conventional growth promoters
• - improve nutrient digestibility,
• - control pathogenic micro-organisms,
• - facilitate favourable intestinal microbial
  balance and
• - enhance nutrient absorption through
  increased absorptive capacity (Al-Harthi, 2002
  El-Deek et al., 2003).

7
Introduction Garlic (Allium sativum)

• A well known spice and herbal medicine for the
  prevention and treatment of a variety of diseases
  (Adimoradi et al., 2006).
• A member of the onion family Alliceae having
  antimicrobial, antioxidant and antihypertensive
  properties (Konjufca et al., 1997 Sivam, 2001).
• Allicin (diallyl-thiosulfinate) is the major
  organosulfur cpd biologically active (Raham,
  2007).

8

• In poultry
• - improve broiler growth
• - decreased mortality rate.
• The use of GP in poultry production is of
  particular importance in the tropics
• - high environmental temperature,
• - high relative humidity and
• - poor biosecurity
• are optimal conditions for disease agents to
  thrive.

9
Justification

• Increasing pressure to reduce/eliminate the use
  of antibiotics as growth promoters in poultry
  feed.
• Prevalence of antibiotic-resistant bacteria
  pathogens of poultry has made control a challenge
• sub-optimal production.
• Thus, further depletion of the already inadequate
  protein supply in Nigeria (FAO, 2010)., thereby
  hampering food security (MDG).
• Search for alternative growth promoters
  preferably from organic sources for
  sustainability.

10
Preliminary Studies

• Preliminary studies have shown
• Increased body weight and higher Newcastle
  disease vaccinal immune response.
• Increased villi length width as well as cryptal
  depth
• In commercial broilers fed garlic inclusion
  (Oladele Bakare, 2011 Oladele et al., 2012).
• (The garlic meal used has been patented (Patent
  No. NG/P/2012/285 of 19 July, 2012).

11

• Aim To evaluate the potentials of garlic as a
  growth promoter in chickens via the assessment of
  production performance and markers of immune
  status and disease tolerance.

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Specific Objectives

• To determine quantitatively, the active
  components of garlic varieties available in
  Nigeria.
• To determine the effect of garlic feed inclusion
  on
• a. performance characteristics of
  commercial layers.
• b. immunity (humoral and cellular),
  haematology and
• serum biochemistry of commercial
  chickens.
• c. disease susceptibility/tolerance of
  commercial
• chickens.
• d. the organoleptic properties of poultry
  meat.
• iii) To conduct a cost-benefit analysis of garlic
  feed inclusion in commercial chicken production.
• iv) To partner with poultry producers and assess
  the acceptability of this product.

13
Expected outcome

• This project is expected to
• provide scientific explanations for some of the
  claims on garlic.
• provide information to poultry producers on the
  potentials of garlic as a growth promoter with
  consequent improvement in health and productivity
  of poultry.
• reduce inadvertent use of antibiotics in poultry
  rearing as well as production of safer meat and
  egg for human consumption.
• stimulate the adoption of the use of garlic as an
  organic growth promoter by producers with
  consequent reduction in the emergence of
  antibiotic-resistant bacteria.
•  

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• Experimental Design and Methods
• Phase 1
•  
• 1. Quantitative assessment of the active
  components of garlic varieties available in
  Nigeria
• Available garlic varieties in Nigeria will be
  sourced and their active components i.e. allicin
  and alliin will be quantitatively determined
  using the high-performance liquid chromatography
  and/or the method of Miron et al., 2002.
•  
•  

15

• 2.Rearing of Commercial Layers
• 500 day-old pullet chicks will be purchased and
  reared in a designated poultry house at the
  Teaching Research Farm.
• These chicks will be randomly separated into four
  groups (A, B, C and D) each consisting of two
  replicates
• designated as 1 and 2 from day-old and
  administered all necessary prophylaxis i.e.
  vaccines and multivitamins.
• Appropriate mash will be served with the
  inclusion of previously prepared garlic meal
  (Patent No. NG/P/2012/285) viz 0.125 - group A,
  0.25 - group C, 0.5 - group C and 0 - group D
  (control).

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• 3. Assessment of performance characteristics of
  commercial layers on garlic feed inclusion
• Performance characteristics to be assessed
  include morbidity, mortality, onset of lay,
  production, egg quality (size, weight, shell
  colour and thickness, yolk colour and size,
  albumen and yolk viscosity) and spent hen value.
• Shelf-life of eggs laid will also be evaluated.

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• 4. Assessment of immunity, haematology and serum
  biochemistry of commercial layers on garlic feed
  inclusion
• Cell mediated immunity will be assayed using
  lymphocyte proliferation ELISA kit (BrdU
  colorimetric, Roche Applied Science, UK).
• Humoral immunity (Newcastle disease and
  infectious bursal disease vaccinal response) will
  be assayed using the ELISA technique.
• Haematology (PCV, RBC counts, total and
  differential WBC counts) and serum biochemistry
  (protein, enzymes and lipid levels) will be
  determined using standard procedures (Campbell,
  1997).

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• 5. Determination of organoleptic properties of
  poultry meat from commercial layers on garlic
  feed inclusion
• Sensory evaluation of poultry meat will be
  conducted in a sensory testing room equipped with
  positive air pressure, individual stations and
  proper light (ISO, 1988).
• Coded samples of freshly prepared poultry meat
  will be served in separate, well-lit cubicles to
  40 panelists at room temperature.
• A descriptive 9-point hedonic scale rating will
  be used to assess the meat for appearance/colour,
  aroma, chewiness, juiciness, toughness and
  overall acceptability.
• 1 representing like extremely and 9 dislike
  extremely (Osman and Aldosari, 2006).

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• 6. Cost-benefit analysis of garlic feed inclusion
  in commercial layer production
• The cost of rearing these chickens will be
  determined and compared across the different
  groups considering inputs, disease control,
  mortalities, etc.
• This will be compared with gains from egg
  production and spent layers across the groups to
  determine profitability.

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Phase 2

• Determination of disease susceptibility/
  tolerance of commercial broilers on garlic feed
  inclusion
• Three experiments to determine disease
  susceptibility/tolerance to three classes
• of pathogens (virus, bacteria and protozoan) will
  be conducted separately.
• a) Experimental infection with Infectious
  bursal disease (IBD) virus
• 200 day-old broilers will be purchased and reared
  in an isolated facility.
• Random separation into 6 groups (A, B, C, D, E
  and F) from day-old and administered all
  necessary prophylaxis i.e. vaccines (except IBD)
  and multivitamins.
• Garlic inclusion, IBD vaccine and IBD virus will
  be administered as
• follows

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• Group A Garlic IBD vaccine
• Group B Garlic IBD virus
• Group C Garlic IBD vaccine IBD virus
• Group D IBD vaccine IBD virus
• Group E IBD virus only
• Group F IBD vaccine only
• Garlic inclusion will be served from day-old at a
  level pre-determined from Phase 1 as most
  desirable.
• IBD vaccination will be at the recommended age
  and route.
• Experimental infection with IBD virus will be at
  5 weeks of age via intraocular route.
• LD50 would have been pre-determined using 30 four
  week-old broiler chicks.
• Clinical signs, mortality and pathology will be
  monitored and recorded until 10 days
  post-infection.
•  

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b) Experimental infection with Salmonella
gallinarum

• 180 commercial broilers will be purchased. Thirty
  of them will be
• separated for the determination of LD50 of
  the bacterial inoculum.
• 150 chicks will be reared in five different
  groups of 30 each and administered garlic
  inclusion and S. gallinarum as follows
• Group A 0.125 garlic S. gallinarum
• Group B 0.25 garlic S. gallinarum
• Group C 0.5 garlic S. gallinarum
• Group D 0 garlic S. gallinarum
• Group E Garlic inclusion only (at the rate most
  desirable from Phase 1).
• Experimental infection will be orally at 5-6
  weeks of age.
• Clinical signs, mortality and pathology will be
  monitored and recorded until 14 days
  post-infection. 

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• c) Experimental infection with oocysts of Eimeria
  spp
• As in 1c above.
• 2. Determination of organoleptic properties of
  meat from commercial broilers on garlic feed
  inclusion
• Uninfected broilers from control groups will be
  used for this study.
• Same procedure as Phase 1.5. will be adopted.

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Possible limitations

• a) incessant power outage.
• b) delays in supply of reagents and equipment.

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Summary of Plan of Work

• Year 1
•  
• a) Construction and renovation of poultry
• experimental pens.
• Quantitative assessment of the active components
  of garlic varieties available in Nigeria.
• Rearing of commercial layers.
• Assessment of performance characteristics of
  commercial
• layers on garlic feed inclusion.
• Assessment of immunity, haematology and serum
• biochemistry of commercial layers on
  garlic feed inclusion.
• f) Cost-benefit analysis of garlic feed
  inclusion in commercial layer production.

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• Year 2
• a) Year 1 d and f to be continued.
• b) Determination of disease susceptibility/toler
  ance of
• commercial broilers on garlic feed
  inclusion.
• Experimental infection with infectious bursal
  disease virus.
• Experimental infection with Salmonella
  galllinarum.
• Experimental infection with oocysts of Eimeria
  spp.
• c) Determination of organoleptic properties of
  meat from commercial layers and broilers on
  garlic feed inclusion.

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• Year 3 
• Year 2 b and c to be continued.
• Consultations with poultry producers through the
  Poultry Association of Nigeria to encourage the
  use of garlic as growth promoter.
• b) Project round-off and final report will be
  written.

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Supplies 1. Brooding materials 2. Vaccines and medicaments 3. Disinfectant 4. Sampling materials (tubes, bottles, Needles Syringes, cotton swabs, methanol, etc) 5. Reagents (Buffers, L-glutamine, Ficoll, Trypan blue, Neutral formalin, Brilliant cresyl blue, standards, etc) 6. Lymphocyte proliferation ELISA kit (Roche) 7. Newcastle and infectious bursal diseases antibody ELISA kits (1 each) _at_N89,100 7. Chick grower mash (117 bags x 25kg) _at_ N2,500 8. Layer mash (713 bags x 25kg) _at_ N2,400 9. Broiler mash (100 bags x 25kg) _at_ N2,500 10. Garlic bulbs (4 bags x 30kg) _at_ N25,000 11. Diesel 200ltrs (80 6060 ltrs) _at_ N165 12. Biochemistry kits 100,000 50,000 16,000 200,000 1,200,000 160,000 178,200 292,500 1,711,200 0 50,000 13,250 100,000 50,000 50,000 0 0 0 0 0 0 0 250,000 50,000 9,900 - 50,000 0 0 0 0 0 0 0 0 0 9,900
TOTAL SUPPLIES 4,071,150 409,900 59,900
Animals (Chickens) 1. 500 day-old pullets _at_ N200 2. 590 day-old broilers _at_ N230 100,000 0 0 135,700 0 0
TOTAL ANIMALS 100,000 135,700 0
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Equipment 1. 4 units Battery cage (120 birds/unit) _at_ N90,000 2. 2 units Poultry postmortem kits (red vinyl) _at_ N16,000 3. 30 units chick drinkers _at_ N200 4. 30 units chick feeders _at_ N200 5. 30 units grower drinkers _at_ N400 6. 30 units grower feeders _at_N400 7. 1 unit ELISA reader 8. 1 unit Deep freezer 10. 1 unit Venier caliper 11. 1 unit Micrometer screw guage 12. 1 unit Digital weighing scale 13. 1 unit Inverter (7.5KVA 48V) 14. 1 unit Inverter Battery 15. 1 unit Multichannel pipette (0.5 -10µl) 16. 1 unit Multi-channel pipette (10 -100µl) 17. 1unit Single-channel pipette (1 10µl) 18. Viscometer 19. pH meter 20. Binocular microscope with camera 360,000 32,000 6,000 6,000 12,000 12,000 800,000 85,000 16,000 17,000 150,000 350,000 60,000 160,000 160,000 60,000 70,000 64,000 320,000 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
TOTAL EQUIPMENT 3,380,000 0 0
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Patient costs Not Applicable - - -
TOTAL PATIENT COSTS
Travel (do not include attendance at scientific meetings) 0 0 50,000
Other (specify) 1. Construction of Poultry pen - 10 x18 m. (Teaching Research Farm) 2. Renovation of experimental house (Dept. of Veterinary Medicine) 3. Specialized laboratory analysis ( Histopathology, HPLC, etc) 4. Publication of research findings 5. Miscellaneous 1,000,000 300,000 100,000 0 200,000 0 0 0 100,000 100,000 0 0 0 0
TOTAL OTHER 1,400,000 200,000 0
GRAND TOTAL 8,551,150 745,600 109,900


N9,366,650.00
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Budget Justification

1. There is sparsity of equipment and reagents in
   the Poultry Diseases Laboratory at the Dept. of
   Veterinary Medicine at present such that so much
   is needed for this project.
2. Construction of a small poultry house is
   necessary for the Layers project.
3. Conversion of an old kennel at Vet. Medicine to
   poultry experimental house for infectivity
   studies.
4. An allowance is necessary for visits to farms and
   meeting with PAN members for advocacy.
5. Allowance was made in budget for publications
   since about five publications is expected from
   this project.

32
References

• Adimoradi, M., Navidshad, B., Saif, J., Royan, M.
  2006. Effect of dietary garlic meal on
• histological structure of small intestine
  of Broiler chickens. Poult. Sci., 43 378
  383.
• Al-Harthi MA. Efficacy of vegetable diets with
  antibiotics and different types of spices or
  their mixtures on performance, economic
  efficiency and carcass traits of broilers. J.
  Agr. Sci. Mansoura University 2002 27 3531
  3545.
• Campbell, TW Avian Hematology and Cytology.
  Ames, IA, Iowa State University Press, 1997.
• Casewell, M., Friis, C., Marco, E., McMullin,
  P., Phillips, I. 2003. The European-ban on
• growth promoting antibiotics and
  emerging consequences for human and animal
• health. J. Antimicrob. Chemother.
  52159-161.
• Dipeolu, MA., Akpan, NJ. and Olutayo, A. 2000.
  Residues of Tetracycline antibiotic in
• turkey and egg of chickens sold for
  human consumption. Poultry Science
• Journal. 1(1) 4-11.
• Dipeolu, MA., Eruvbetine, D. and Sowunmi, KS.
  2002. Tetracycline residue deposition in egg of
  layers fed antibiotics and enzyme supplemented
  feed. Proceedings of the
• 27th Annual Conference of the Nigerian
  Society for Animal Production. 29 292- 294.
• Dipeolu, MA., Adebayo, AJ. and Oke, MO. 2004.
  Residues of Streptomycin antibiotic in
• commercial layers in Abeokuta and
  Ibadan metropolis. Nigerian Journal of
• Animal production, 31(1) 130-134.

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• Graham, J.P., Boland, J.J. and Silbergeld, E.
  2007. Growth promoting antibiotics in food
  animal production An economic analysis. Public
  Health Reports, 122 79-87.
• ISO (International Standard Organisation). 1988.
  Sensory Analysis-General Guidance for the Design
  of Test Rooms. ISO 8589, 1-9 .
• Javandel F, Navidshad B, Seifdavati J, Pourrahimi
  GH, Baniyaghoubi S. The favourite dosage of
  garlic meal as a feed additive in broiler chicken
  ratios. Pak J Biol Sci 2008 11(13) 1746-1749.
• Konjufca VH, Pest GM and Bakalli RI 1997.
  Modulation of cholesterol levels in Broiler meat
  by dietary garlic and copper. Poultry Science 76,
  1264 1271.
• Libby, DA, Schaible, PJ. 1955. Observations on
  growth responses to antibiotics and arsenic acids
  in poultry feeds. Science 121 733.
• Luo, N., Sahin, O., Lin, J., Michel, L.O., Zang,
  Q. 2003. In vivo selection of Campylobacter
  isolates with high levels of fluoroquinolones
  resistance associated with gyrA
• mutations and the function of the cmeABC
  efflux pump. Antimicrob. Agents Chemother. 47
  390-394.

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• Miron, T., Shin, I., Feigenblat, G., Weiner, L.,
  Mirelman, D., Wilchek, M., Rabinkov, A. (2002). A
  spectrophotometric assay for allicin, alliin, and
  alliinase (alliin lyase) with a chromogenic
  thiol reaction of 4-mercaptopyridine with
  thiosulfinates. Anal
• Biochem. 307(1)76-83.
• Oladele Omolade and Bakare Hauwa (2011). Effects
  of garlic (Allium sativum) on
• growth performance and vaccinal immune
  response in commercial broilers.
• Conference Proceedings 5th Pan
  Commonwealth Veterinary Association. Journal
• of Commonwealth Veterinary
  Association.Special issue. Vol. 27(2), 243-247.
• www.commonwealthvetassoc.org/
• Oladele OA, Emikpe BO, Hauwa B. Effects of
  dietary garlic (Allium sativum)
• supplementation on body weight and gut
  morphometry of commercial broilers.
• Int J Morph 2012, 30(1), 238-240.
  http//www.scielo.cl/revistas/ijmorphol
• Osman, BM, Aldosari, MN. 2006. Sensory evaluation
  of Awassi and Nadji lambs meat.
• Scientific Journal of faisal
  University (Basic and Applied Sciences). Vol.
  7(2) 1427H.
• Raham, M.S. 2007. Allicin and other functional
  active components of garlic Health
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  International Journal of food properties. Vol.
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• 245-268.
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• by garlic. Journal of Nutrition 131,
  1106S 1108S.
• .
•  

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• Tollba, AAH and Hasan, MSH. 2003.Using some
  natural additives to
• improve physiological and productive
  performance of broiler
• chicks under high temperature
  conditions. Black cumin (Nigella
• sativa) or garlic (Allium sativum).
  Poultry Science, 23327-340.
• 2003.
• Waibel, P.E., Abbott, OJ., Baurmann, CA. and
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  Science. 33 1141.

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Abstract Pictures
Sample of Product (garlic meal)
Garlic meal (200g) to be mixed with 160kg poultry
feed.
37
Abstract Pictures
Two chicken layers (Nera Black) on garlic feed
inclusion showing lustrous plumage and brightly
coloured combs and wattles in spite of being
reared on old litter.
38
Abstract Pictures
Closer picture of the Nera Black layers on garlic
feed inclusion immediately after beak trimming.
39
Abstract Pictures
Isa Brown layer chicken on garlic feed inclusion
showing lustrous plumage and brightly coloured
combs and wattles.