Transcript UNIVERSITY OF IBADAN RESEARCH FOUNDATION _garlic

UNIVERSITY OF IBADAN RESEARCH
FOUNDATION
PROPOSAL FOR COLLABORATIVE RESEARCH PROJECT
INFLUENCE OF GARLIC (Allium sativum)
FEED INCLUSION ON PRODUCTION
PERFORMANCE AND HEALTH OF
COMMERCIAL CHICKENS
19 November, 2013
Research Team
1.
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.
3.
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Dr. Olufunke O. Ezekiel (Co-Principal Investigator)
Department of Food Technology,
University of Ibadan.
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4.
Dr. O.O. Esan
Department of Veterinary Medicine
University of Ibadan.
5.
Dr. Olutola Oyedele
Agricultural Extensionist,
National Horticultural Research Institute (NIHORT)
Idi-Ishin, Ibadan.
6.
MrsYemisi Adewale
Agronomist,
National Horticultural Research (NIHORT)
Idi-Ishin, Ibadan.
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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
optimal performance.
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 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).
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 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).
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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).
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 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.
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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.
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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).
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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.
ii.
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.
i.
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Expected outcome:
This project is expected to:
provide scientific explanations for some of the claims on
garlic.
II. provide information to poultry producers on the
potentials of garlic as a growth promoter with consequent
improvement in health and productivity of poultry.
III. reduce inadvertent use of antibiotics in poultry rearing as
well as production of safer meat and egg for human
consumption.
IV. 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.
I.
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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.
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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
ofOladele,
previously
prepared garlic meal (Patent No. 7/16/2015
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NG/P/2012/285) viz: 0.125% - group A, 0.25% -
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.
1.
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





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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
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a) Construction and renovation of poultry
experimental pens.
b) Quantitative assessment of the active components
of garlic varieties available in Nigeria.
c) Rearing of commercial layers.
d) Assessment of performance characteristics of
commercial
layers on garlic feed inclusion.
e) Assessment of immunity, haematology and serum
biochemistry of commercial layers on garlic feed
inclusion.
f)
Cost-benefit
analysis of garlic feed inclusion in
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commercial layer production.
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Year 2
a) Year 1 d and f to be continued.
b) Determination of disease susceptibility/tolerance
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
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inclusion.
Year 3
a) Year 2 b and c to be continued.
b) 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) @N89,100
7. Chick & grower mash (117 bags x 25kg) @ N2,500
8. Layer mash (713 bags x 25kg) @ N2,400
9. Broiler mash (100 bags x 25kg) @ N2,500
10. Garlic bulbs (4 bags x 30kg)
@ N25,000
11. Diesel 200ltrs (80; 60;60 ltrs)
@ N165
12. Biochemistry kits
TOTAL SUPPLIES
Animals (Chickens)
1. 500 day-old pullets
2. 590 day-old broilers
@ N200
@ N230
TOTAL ANIMALS
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100,000
50,000
16,000
200,000
1,200,000
160,000
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
4,071,150
409,900
59,900
100,000
0
0
135,700
178,200
292,500
1,711,200
0
50,000
13,250
100,000
100,000
135,700
0
0
0
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Equipment
1. 4 units Battery cage (120 birds/unit)
2. 2 units Poultry postmortem kits (red vinyl)
3. 30 units chick drinkers
4. 30 units chick feeders
5. 30 units grower drinkers
6. 30 units grower feeders
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
@ N90,000
@ N16,000
@ N200
@ N200
@ N400
@N400
TOTAL EQUIPMENT
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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
3,380,000
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
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Patient costs
Not Applicable
-
-
-
TOTAL PATIENT COSTS
Travel (do not include attendance at scientific meetings)
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
0
1,000,000
300,000
100,000
0
200,000
0
50,000
0
0
0
100,000
100,000
0
0
0
0
0
TOTAL OTHER
1,400,000
200,000
GRAND TOTAL
8,551,150
745,600
109,900
= N9,366,650.00
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Budget Justification
a)
b)
c)
d)
e)
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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.
Construction of a small poultry house is necessary
for the Layers project.
Conversion of an old kennel at Vet. Medicine to
poultry experimental house for infectivity studies.
An allowance is necessary for visits to farms and
meeting with PAN members for advocacy.
Allowance was made in budget for publications
since about five publications is expected from this
project.
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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. 52:159-161.
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.
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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
benefits and bioavalability. International Journal of food properties. Vol. 10:
245-268.
Sivam GP 2001. Protection against Helicobacter pylori and other bacterial infections
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, 23:327340.
2003.
Waibel, P.E., Abbott, OJ., Baurmann, CA. and Bird, HR. 1954.
Disappearance of the growth response of chicks to
antibiotics in
an “old” environment. Poultry Science. 33: 1141.
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Abstract Pictures
Sample of Product (garlic meal)
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Garlic meal (200g) to be mixed with
160kg poultry feed.
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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.
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Abstract Pictures
Closer picture of the
Nera Black layers on
garlic feed inclusion
immediately after
beak trimming.
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Abstract Pictures
Isa Brown layer
chicken on garlic
feed inclusion
showing lustrous
plumage and brightly
coloured combs and
wattles.
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