ANTIBIOTIC RESIDUE ANALYSIS IN EDIBLE MEAT USING HIGH PERFORMANCE LIQUID

CHROMATOGRAPHYBy

Betty KeruboI56/76366/2014

Supervisors: Dr. Gabriel AbogeDr. George Obiero

Introduction • Administration of antibiotics has facilitated successful

management of bacterial infections.

• Today with the advancement in technology their use has been vast in growth promotion.

• Antibiotic efficacy is dependent on judicious and appropriate administration otherwise residual in food producing animals.

• There is evidence of the existence of residual antibiotics in food producing animals.

Introduction…• Low antibiotic residue levels in high income regions such as

Europe recording ˂ 1% of reported cases (Food & Authority, 2013).

• The food safety inspection service monitoring plan 2013 reported about 0.13% residues in meat of healthy animals (Are et al., 2013).

• Samples analyzed for antibiotic residues in meat in Nairobi slaughter houses in Kenya reported 20% tetracycline (Muriuki et al., 2001).

• Despite this trace antibiotic levels recorded by regulatory agencies, the rate of consumption will lead to accumulation over time and adverse effects (Boeckel et al., 2014).

Introduction…• Allergic reactions in sensitive individuals are few

consequences of residual antibiotics.

• This problem is recurrent in countries with limited regulation agencies and lack of awareness by regulatory bodies (Bedada et al., 2012).

• Resistance arising from residual antibiotics will burden the human population to discard existent antibiotics for newer ones which tend to be expensive (Boeckel et al., 2014).

• Regulatory agencies such as WHO and FDA have made efforts to manage these residues setting MRLs for various antibiotics (Nisha, 2008).

Problem statement• Antibiotic residues are as a result of inappropriate

antibiotic use and lack of awareness by regulatory bodies.

• Research has been done in Kenya to analyse the levels of antibiotic residues but still this data is inadequate.

• Trends in antibiotic use have seen increments 36% between the years 2000 to 2010(Fao, Workshop, & Drugs, n.d.).

• Low and middle income countries account for about 76% of this, with last resort antibiotic accounting for 58%

• Research to establish the quantities of residual antibiotics needs to be addressed to provide supplement the already existing data.

Hypothesis

There are higher antibiotic residue levels in edible meat from Kenyan slaughter houses compared to the maximum residue limits (MRLs) set by WHO/FDA.

ObjectivesGeneral objective

To determine the presence of oxytetracycline streptomycin and diminazene in kidney, liver and other edible meat.

Specific objectives

•To monitor levels of streptomycin, oxytetracycline and diminazene in kidney, liver and other edible meat;•To quantify the amount of streptomycin, diminazene and oxytetracycline;•To analyze contraindications associated with residual antibiotics to enhance prospect development of consensus and judicious antibiotic residue tolerant levels

Justification of the studyFindings from this study will

•Provide the current quantities of residual antibiotics in edible meat;

•Assess the extent of hazard presented to the consumers of food producing animals.

Expected outcomes

Finding from this study will

• Help to possibly to suggest and recommend new MRLs to regulatory agencies;

• Help to prioritize programs that will ensure judicious use of antimicrobials through concerted efforts by the international community.

Materials and Methods

Statistical analysis For H0 : ρ = Ɵ VS H1 : ρ < /> Ɵ

zc = Ƶ - µƵ δƵ Where Ƶ = ½ ln (1+r) (1- r)  µƵ= 1.1513 log (1+ ρ0) (1 –ρ0) δƵ= 1 1 √(n-3) if zc ˂ Ƶα accept H0 at 0.05% L.O.S 

Timeline Activity Septemb

er 2015 October 2015

November 2015

December 2015

January 2016

February 2016

March 2016

April 2016

Proposal developmentSample collection and Meat extractionMeat HPLC clean up

Data analysis

Thesis write up and submission

Budget Item Quantity Cost

Acetonitrile hplc grade 5 - 2.5 litres @ 6500/= 32500/=Methanol hplc grade 5- 2.5 litres @ 3000/= 15000/=Nylon membrane filters 0.45mm by 47mm 10000/=Nylon syringe filters acrodiscs 1000 pieces @20/= 20000/=5ml plastic pipette (disposable) 400 pieces @ 30/= 12000/=1.5 mls autosampler vials 300pieces @ 1000/= 30000/=Hplc grade water 6500/= 6500/=Whatman filter papers (no. 1) 3 packets @4500 13500/=2 mls Syringes hypodermic 500 pieces @ 5/= 2500/=10 mls Syringes 500 pieces @ 10/= 5000/=Nitrogen gas 120M3 7500/=Blue micropipete tips 2 packets @ 1000/= 2000/=yellow micropipete tips 2 packets @ 1000/= 2000/=White /clear micropipete tips 2 packets @ 1500/= 3000/=15 mls Centrifuge tubes PPE 300 pieces @ 25/= 7500/=Paper towels 8 Pairs @ 300/= 2400/=Sub total 171400/=Contigency (10% of sub total) 17140/=Grand total (contigency +sub total)

188540/=

Acknowledgement

Dr. Gabriel Aboge

Dr. George Obiero

University of Nairobi ILRI