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July 2013

Technical information note


Antibiotic test methods
Antimicrobial residues, or antibiotic residues as they are more commonly referred to within the Australian dairy industry, are able to be detected in milk by using a range of test methods. Antibiotic residue testing of milk and dairy products can be split into two main categories: screening test methods (e.g. qualitative and semi-quantitative test methods) and confirmatory test methods (quantitative test methods).

What factors to consider when choosing a screening test method


Test matrix Most of the commercial antibiotic residue screening test kits are designed for use with cows milk. Some matrices, such as goats, ewes, and buffalos milk; highly viscous products such as cream; or high fat products such as butter, may not be suitable to test with some kits or may require variations to the standard kit instructions in order to achieve a valid result. Prior to using a test kit on a matrix other than cows milk, it is advisable to check with the test kit manufacturer or test kit distributor to make sure that it is suitable to use on that matrix, and whether or not variations in the standard test kit instructions are necessary.

Screening test methods


Screening tests are further divided into either broad or narrow spectrum test methods. A broad spectrum test detects a range of antibiotic classes, whereas a narrow spectrum test detects a limited number of antibiotic classes. For example, a narrow spectrum test may only cover beta-lactams, or it may cover both beta-lactams and aminoglycosides. A wide range of commercial screening test kits are available for use. This technical information note provides general details on the test methods used by the Australian dairy industry for the screening of antibiotic residues in milk and dairy products. The main antibiotic residue screening test methods currently used are listed in Table 1. This note is intended to provide guidance only and may not include all of the kits that are currently available in Australia. The availability of the kits listed may also change. Screening test methods are generally qualitative tests, and are designed to give a positive or negative result to indicate whether particular antibiotic residues may be present or absent in the milk or dairy product. Screening test methods do not identify the specific antibiotic residue present, nor are they specifically designed to indicate whether or not a particular antibiotic residue is present at a level above the regulated maximum residue limit (MRL). A broad spectrum test detects a range of antibiotic classes. Some of these antibiotic classes may include beta-lactams (penicillin falls within this group), cephalosporins, aminoglycosides, macrolides, tetracyclines and sulphonamides.

Approvals
Many of the commercial antibiotic residue screening test kits have been formally approved or certified by regulatory bodies or internationally recognised organisations that undertake evaluations, validations and/or performance testing of test kits. Note that DFSV does not perform approvals or certifications of test kits. When choosing a commercial antibiotic residue screening test kit, it is important to check that it has appropriate approval or certification. For each of the antibiotic residue screening test kits shown in Table 1, their relevant approval or certification is listed.

Sensitivities
Sensitivities (also referred to as detection limits) vary between each of the commercial screening test kits for the different antibiotic residues. These different sensitivities should be considered carefully when deciding which test kit to use. For further details on the sensitivities of each of the kits listed in the table, refer to the technical information note Detection limits of antibiotic residue screening test kits.

Benefits of screening test methods


Screening tests are a useful tool to indicate whether or not antibiotic residues are present. Compared to confirmatory methods they are more rapid, easy to use and relatively inexpensive.

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Other factors
Other important factors to consider when choosing which commercial test kit to use are: The time taken to yield a result ease of use the cost of set-up equipment and consumables. It is suggested that the Australian test kit distributor be consulted to provide this information.

Further information
Other technical information notes on antibiotics and dairy food safety topics are available at www.dairysafe.vic.gov.au Or contact Dairy Food Safety Victoria on (03) 9810 5900 or info@dairysafe.vic.gov.au
Copyright Dairy Food Safety Victoria 2013

Key points to consider


Antibiotic residues can be detected in milk and milk products using a range of test methods. Screening test methods are generally qualitative tests, designed to give a positive or negative result to the presence of antibiotic residues. However, they do not identify the specific antibiotic residue present, nor are they designed to indicate whether or not a particular antibiotic residue is present at a level above the MRL. Most of the commercial antibiotic residue screening test kits are designed for use in cows milk. Prior to using a test kit on a matrix other than cows milk, check its suitability with the test kit manufacturer or distributor. Consider the different sensitivities (detection limits) of the test kits when deciding which to use.

This document is intended to be used as a general guide only and is not a comprehensive statement of all the relevant considerations with respect to your particular circumstances, nor does it comprise, or substitute for, legal or professional advice. DFSV does not guarantee the accuracy, reliability, currency or completeness of the information. DFSV accepts no legal liability arising from, or connected to, any reliance on this document.

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Table 1: Antibiotic residue screening test methods available for use in Australia
Test method BetaStar Charm II 1 Australian distributor CHR Hansen Pty Ltd Merck Pty Ltd Antibiotic classes detected Beta-lactams and cephalosporins Beta-lactams, cephalosporins, sulfa drugs, tetracyclines, macrolides, Aminoglycosides, others (novobiocin, chloramphenicol, thiamphenicol and dapsone) Sulfonamide, Sulfadimethoxine, Sulfamethazine Current certifications/approvals AFNOR, USFDA and AOAC Argentina, AOAC, NSW Food Authority and USFDA4

Suitable matrices* Raw cow/sheep/goat milk Raw cows milk and liquid milk. Goat/ sheep milk, cream, condensed milk, whey and dairy powders Raw cows milk, liquid milk and permeate. Goat/sheep milk, colostrum, cream and condensed milk

Merck Pty Ltd Charm ROSA MRL Sulfonamide, Sulfamethoxine (SDSM), Sulfamethazine (SMZ)2 Cowside II Merck Pty Ltd

AOAC, USFDA

Beta-lactams, cephalosporins, sulfa drugs, tetracyclines, macrolides, aminoglycosides, others (chloramphenicol, thiampheniicol, trimethoprim and dapsone) Beta-lactams, Cephalosporins, sulfa drugs, tetracyclines, macrolides, Aminoglycosides, others (chloramphenicol, thiamphenicol, trimethoprim and dapsone) Beta-lactams, cephalosporins, sulfa drugs, tetracyclines, macrolides, Aminoglycosides, others (chloramphenicol, trimethoprim and dapsone) Beta-lactams and cephalosporins Beta-lactams and cephalosporins

AOAC, USFDA

Raw cows milk, liquid milk and permeate. Goat/sheep milk, colostrum, cream and condensed milk Raw cows milk and liquid milk. Goat/ sheep milk and milk powders

Copan

CHR Hansen Pty Ltd

NSW Food Authority and MPI NZ

Delvotest SPNT

Rejuva Care International Pty Ltd

AOAC, MPI NZ, NSW Food Authority, Germany, UK, Argentina, Denmark and Switzerland

Raw cows milk and liquid milk

SNAP MRL Betalactam Ballya Q-Beta

ThermoFisher Scientific Pacific Laboratory Products Pacific Laboratory Products

NSW Food Authority None

Raw cows milk Raw cows, sheep and goat milk Raw cows, sheep and goat milk Raw cows milk. Goat/sheep milk and other dairy products

Ballya SensorBT

Beta-lactams, cephalosporins and tetracyclines

None

Disc Assay3

Not applicable Penicillin and non-specific for other beta-lactams and non-beta-lactams

Not applicable

All sensitivities are based on those declared by kit manufacturers/distributors Australian information brochures and are current as of the date of this publication. These sensitivities correspond to when the kits are used according to the kit manufacturer/distributor instructions to give maximum sensitivities (e.g. for Delvotest SP-NT- running the kit at control time). KEY * Suitable matrices listed are obtained from manufacturers written materials only. Manufacturers are advised to contact distributors for suitability of other matrices of interest that are not listed. Matrices other than raw cows milk and liquid milks generally require pre-treatment of samples prior to testing. Refer to the test kit distributor for further details. Charm II systems also allow for semi-quantitative analysis. The SMZ test detects sulfamethazine only. Can be used as qualitative and quantitative methods for penicillin G. Approval applicable for particular Charm II test kits only.

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Abbreviations: AOAC AOAC International AFNOR Association Francaise de Normalisation, MPI NZ Ministry for Primary Industries New Zealand, NSW New South Wales, USFDA The United States Food & Drug Administration. Note: All of the information provided in this table for antibiotic classes detected, current certifications/approvals and suitable matrices is based on the information provided by kit manufacturer/distributors Australian information brochures.

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