2007/516/EC: Commission Decision of 19 July 2007 concerning a financial contribution from the Community towards a survey on the prevalence and antimicrobial resistance of Campylobacter spp. in broiler flocks and on the prevalence of Campylobacter spp. and Salmonella spp. in broiler carcasses to be carried out in the Member States (notified under document number C(2007) 3440)
Date: 2007-07-19
Institutions: European Commission
Subjects: EU financing, data collection, food safety, poultry, poultrymeat, public health, veterinary inspection, zoonosis

21.7.2007

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Official Journal of the European Union

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COMMISSION DECISION

of 19 July 2007

concerning a financial contribution from the Community towards a survey on the prevalence and
in broiler flocks and on the prevalence of
antimicrobial resistance of Campylobacter spp.
Campylobacter spp. and Salmonella spp. in broiler carcasses to be carried out in the Member States

(notified under document number C(2007) 3440)

(2007/516/EC)

THE COMMISSION OF THE EUROPEAN COMMUNITIES,

(4)

Having regard to the Treaty
Community,

establishing the European

Having regard to Council Decision 90/424/EEC of 26 June
1990 on expenditure in the veterinary field (1), and in particular
Article 20 thereof,

Whereas:

(1)

(2)

(3)

contribution by

the Community

Decision 90/424/EEC lays down procedures governing a
financial
towards
specific veterinary measures,
including technical and
scientific measures.
It provides for the Community to
undertake, or assist Member States in undertaking the
technical and scientific measures necessary for the devel-
opment of Community veterinary legislation and for the
development of veterinary education or training.

According to the Report of the European Food Safety
Authority (EFSA) on Trends and Sources of zoonoses,
resistance in the
zoonotic agents and antimicrobial
Community in 2005 (2), a total of 194 695 cases of
in humans were reported in 22
campylobacteriosis
Member States. Broiler meat
is considered the most
infection. Up to 66,4 % positive
common source of
samples in broiler meat were reported. In broiler flocks,
0,2 to 86 % of the reported samples were positive.

In addition, according to the EFSA report, a total of
168 929 cases of human salmonellosis were reported
in 22 Member States in 2005. Typical contamination
rates of fresh poultry meat vary from 4 to 10 %, being
the highest rates of all foodstuffs analysed.

(5)

(6)

(7)

resistant

and food were

The EFSA also indicates in its report that a relatively high
proportion of Campylobacter and Salmonella isolates from
animals
to antimicrobials
commonly used in treatment of human diseases. This
specially applies to the case of resistance to fluoroqui-
nolones in Campylobacter isolates from poultry, where up
to 94 % of
to cipro-
floxacin. Food-borne infections caused by these resistant
bacteria pose a particular risk to humans due to possible
treatment failure.

isolates were reported resistant

to be carried out

In accordance with Commission Decision 2005/636/EC
of 1 September 2005 concerning a financial contribution
by the Community towards a baseline survey on the
prevalence of Salmonella spp. in broiler flocks of Gallus
in the Member States (3),
gallus
comparable information was collected with regard to
the prevalence of Salmonella in such flocks.
is,
however, very difficult to compare prevalence of Campy-
flocks and broiler meat, and of
lobacter
Salmonella in broiler meat from different Member States
as there is no harmonised monitoring.

in broiler

It

Under Article 5 of Directive 2003/99/EC of the European
Parliament and of the Council of 17 November 2003 on
the monitoring of zoonoses
agents,
amending Council Decision 90/424/EEC and repealing
Council Directive 92/117/EEC (4), coordinated moni-
toring programmes may be established, especially when
to assess risks and to
specific needs are identified,
establish baseline
and
values
zoonotic agents at the level of Member States.

related to zoonoses

and zoonotic

a baseline

specifications

Scientific experts in collaboration with the EFSA prepared
study on a
for
technical
harmonised monitoring of Campylobacter
in broiler
flocks. Training was organised in 2006 for laboratory
staff in all Member States on the detection methods for
Campylobacter in such flocks and is scheduled in 2007
with regard to the enumeration method for Campylobacter
on carcasses.

(1) OJ L 224, 18.8.1990, p. 19. Decision as last amended by Decision

2006/965/EC (OJ L 397, 30.12.2006, p. 22).

(2) The EFSA Journal (2006) 94.

(3) OJ L 228, 3.9.2005, p. 14.
(4) OJ L 325, 12.12.2003, p. 31. Directive as amended by Council

Directive 2006/104/EC (OJ L 363, 20.12.2006, p. 352).

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21.7.2007

(8)

(9)

(10)

(11)

(12)

The Task Force on Monitoring of Zoonoses Data
Collection of EFSA adopted during its meeting on 16
and 17 October 2006 the report on proposed technical
specifications for a coordinated monitoring programme
for Salmonella and Campylobacter in broiler meat in the
EU (1).

The Task Force also adopted a Report
including a
proposal for a harmonised monitoring scheme of anti-
microbial resistance in Salmonella in fowl (Gallus gallus),
turkeys and pigs and Campylobacter jejuni and C. coli in
broilers (2) on 20 February 2007. The report makes
recommendations on a harmonised monitoring scheme
and harmonised methodology for susceptibility testing.

Pursuant to Article 7(3) and Annex II(B) of Directive
2003/99/EC, detailed rules should be laid down on the
antimicrobial resistance monitoring of Campylobacter jejuni
in poultry. Data needs to be
and Campylobacter
collected in order
to be laid down
such rules
Therefore, testing of the antimicrobial resistance should
be included in the survey in order to gather the necessary
data.

coli
for

on

antimicrobial

Taking into account the high number of Salmonella and
Campylobacter cases in humans, the importance of broilers
and broiler meat as source of infection and the increasing
concern
development,
comparable data on the prevalence of Campylobacter in
broilers and broiler meat, Salmonella in broiler meat in
the Member States should be collected to consider the
feasibility, cost and benefit of Community-wide
need,
control measures.

resistance

The survey is to provide technical information necessary
for the development of Community veterinary legislation
including on the use of antimicrobials in zoonoses
control programmes in poultry. Given the importance
of collecting comparable data on the prevalence of
in broilers and broiler
Salmonella and Campylobacter
meat and antimicrobial resistance of Campylobacter
in
broiler flocks in the Member States,
they should be
im-
granted a Community financial contribution for
plementing the specific requirements of the survey. It is
appropriate to reimburse 100 % of the costs incurred on
the laboratory testing, subject to a ceiling. All other costs
incurred, such as costs for sampling, travel and adminis-
tration should not be eligible for any Community
financial contribution.

(13) A financial contribution from the Community should be
in
to

granted provided that
accordance with Community
compliance with certain other conditions.

the survey is carried out
law and subject

(14) A financial contribution from the Community should be
granted insofar as the actions provided for are effectively
carried out and provided that the competent authorities
the necessary information within the time
furnish all
limits provided for in this Decision.

(15)

(16)

a

by

for

financial

contribution

For reasons of administrative efficiency all expenditure
presented
the
Community should be expressed in euro. In accordance
with Council Regulation (EC) No 1290/2005 of 21 June
2005 on the financing of
the common agricultural
policy (3),
the conversion rate for expenditure in a
currency other
than euro should be the rate most
recently set by the European Central Bank prior to the
first day of
the month in which the application is
submitted by the Member State concerned.

The measures provided for
accordance with
Committee on the Food Chain and Animal Health,

in this Decision are in
Standing
the

opinion

the

of

HAS ADOPTED THIS DECISION:

Article 1

Subject matter and scope

This Decision lays down rules on a financial contribution from
the Community towards a survey to be carried out
in the
Member States on the prevalence of:

(a) Campylobacter spp. in broiler flocks and their antimicrobial

resistance; and

(b) Campylobacter spp. and Salmonella spp. in broiler carcasses.

Article 2

Definitions

For the purposes of this Decision, the following definitions shall
apply:

(a)

‘flock’ means all poultry (such as broiler) of the same health
status kept on the same premises or in the same enclosure
and constituting a single epidemiological unit; in the case of
housed poultry,
includes all birds sharing the same
airspace;

it

(1) The EFSA Journal (2007) 96, 1-46.
(2) The EFSA Journal (2006) 403, 1-62.

(3) OJ L 209, 11.8.2005, p. 1. Regulation as last amended by Regu-

lation (EC) No 378/2007 (OJ L 95, 5.4.2007, p. 1).

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(b) ‘slaughter batch’ means a delivery of broilers which have
been raised in the same flock to a slaughterhouse on one
single day;

In such cases, the NRLs shall provide support and training to
the designated laboratories and ensure that they comply with
rules on quality controls by arranging regular ring tests.

(c)

‘competent authority’ means the authority or authorities of a
Member State as designated under Article 3 of Regulation
(EC) No 2160/2003 of the European Parliament and of the
Council (1).

Article 3

Zoonoses and zoonotic agents covered by the survey

The Member States shall carry out a survey to assess the
prevalence of the following zoonoses and zoonotic agents in
random in
samples
accordance with Annex I:

taken in slaughterhouses

selected at

The laboratories designated in accordance with the third
paragraph of this Article which perform testing must comply
which the following conditions:

(a)

they must have proven experience of using the methods
required for the testing;

(b) they must have a quality assurance system complying with

EN/ISO standard 17025;

(c)

they must be subjected to the supervision of the relevant
NRLs.

(a) Campylobacter spp. in broiler flocks and their antimicrobial

resistance;

Article 5

(b) Campylobacter spp. in broiler carcasses;

(c) Salmonella spp. in broiler carcasses;

the Community sampled in slaughterhouses. Only
across
broilers produced from day one within the Member State
shall be included in the survey.

Article 4

Performance of sampling and analyses

Sampling shall be performed by the competent authority
1.
or under its supervision in accordance with the technical speci-
fications set out in Annex I.

National reference laboratories (NRLs) for Salmonella spp.,
resistance testing shall
the analyses of samples and

2.
Campylobacter spp. and antimicrobial
perform the relevant parts of
isolates.

the

However,

competent

3.
to
designate other laboratories involved in official controls of
Salmonella spp., Campylobacter spp. and antimicrobial resistance
testing, to perform the analyses of samples and isolates.

authority may decide

Conditions for paying a Community financial contribution

1.
The Community financial contribution towards the costs
for sampling and analyses shall be paid to the Member States up
to the maximum total amount for the co-financing set out in
Annex II.

2.
The Community financial contribution provided for in
paragraph 1 shall be paid to the Member States provided that
the survey is implemented in accordance with the relevant
provisions of Community law, including rules on competition
and on the award of public contracts, and subject to compliance
with the following conditions:

(a)

the laws, regulations and administrative provisions required
to implement
the survey must enter into force by 31
December 2007 at the latest;

(b) a progress report containing the information listed in Part
E(1) to Annex I and covering the first three months of the
survey shall be submitted to the Commission by 31 May
2008 at the latest;

(c) a final

report on the implementation of

the survey
containing all information in points 1 and 2 of Part E to
Annex I, together with supporting evidence for the costs
incurred by the Member States
the sampling and
analyses and the results attained during the period from 1
January 2008 to 31 December 2008 shall be submitted to
the Commission by 28 February 2009 at the latest; and the
evidence as to costs incurred must comprise at least the
information set out in Annex III;

for

(1) OJ L 325, 12.12.2003, p. 1. Regulation as last amended by Council
Regulation (EC) No 1791/2006 (OJ L 363, 20.12.2006, p. 1).

(d) the survey must be implemented effectively.

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Failure to submit the final report referred to in paragraph
3.
2(c) by 28 February 2009 at the latest shall entail a progressive
reduction of the financial contribution to be paid, amounting to
25 % of the total amount by 30 March 2009, 50 % by 30 April
2009 and 100 % by 30 May 2009.

The Commission shall

forward those results obtained
2.
during the implementation of
the survey together with the
national aggregated data and the assessments thereof by the
Member States to the European Food Safety Authority, which
shall examine them.

Article 6

Maximum amounts to be reimbursed

Any use of the data submitted by the Member States for the
purpose other
to prior
agreement of the Member States.

than the survey shall be subject

The maximum amounts of the Community financial contri-
bution towards the costs to be reimbursed to the Member
States for sampling and analyses covered by the survey shall
not exceed the following:

(a) EUR 20 for each detection testing of Campylobacter and

Salmonella spp;

(b) EUR 30 for each confirmation, speciation and enumeration
serotyping of
isolates

and the

spp.
of Campylobacter
Salmonella spp. isolates;

National aggregated data and results

3.
available publicly in a form that ensures confidentiality.

shall be made

Article 8

Conversion rate for expenditure
Where a Member State’s expenditure is in a currency other than
the euro, the Member State concerned shall convert it into euro
by applying the most recent exchange rate set by the European
Central Bank prior to the first day of the month in which the
application is submitted by the Member State.

(c) EUR 30 for antimicrobial testing of Campylobacter isolates

from broiler flocks.

Article 9

Application

Article 7

Collection of data, assessment and reporting

the

collect

results of

and assess

The competent authority responsible for preparing the
1.
to Article 9(1) of Directive
yearly national report pursuant
2003/99/EC shall
the
sampling and analyses with regard to the prevalence of
Salmonella and Campylobacter carried out pursuant to Article 4
of this Decision and shall report all necessary data and its
assessment thereof by the Member States to the Commission
by 28 February 2009 at
the latest. The results from anti-
microbial resistance testing will be reported before the end of
May 2009 in the frame of the annual reporting in accordance
with Article 9(1) of Directive 2003/99/EC.

This Decision shall apply from 1 January 2008.

Article 10

Addressees

This Decision is addressed to the Member States.

Done at Brussels, 19 July 2007.

For the Commission

Markos KYPRIANOU
Member of the Commission

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ANNEX I

TECHNICAL SPECIFICATIONS REFERRED TO IN ARTICLE 4

PART A

Sampling frame

To avoid age-related effects the monitoring shall be carried out on slaughter batches at the slaughterhouse.

As prevalence of Campylobacter spp. has been shown to vary significantly depending on the season, this merits stratifi-
cation. For that purpose a 12-month period must be divided in 12 periods of one month. In each of those periods 1/12th
of the total sample size must be taken.

The sampling must otherwise be based on a random selection, both regarding slaughterhouses, sampling days each month
and which batches are to be sampled on a selected sampling day. In particular, the randomisation scheme shall guarantee
a selection of slaughter batches proportionate to the number of flocks fattened according to the different production types
(conventional, free-range, organic). In addition, the Salmonella spp. or Campylobacter spp. status, if known at slaughter,
must not bias the randomisation. The competent authority shall take responsibility for generating the randomization
scheme and ensuring it is implemented correctly. An example of a randomisation procedure is provided in the Report of
the EFSA Task Force on Monitoring of Zoonosis Data Collection on proposed technical specifications for a coordinated
monitoring programme for Salmonella and Campylobacter in broiler meat in the EU. Details of the randomisation scheme
shall be reported to the Commission.

PART B

Sample size

1. Primary sample size

(a) The primary sample size gives the number of slaughter batches to be tested.

(b) At least 384 slaughter batches shall be sampled. Non-response shall be anticipated by sampling about 10 % more

than the indicated numbers.

(c) By way of derogation from point (b), the following numbers of slaughter batches (1) shall be sampled in Estonia,

Latvia and Luxembourg:

(i) in Estonia, at least 96 slaughter batches;

(ii) in Latvia, at least 120 slaughter batches;

(iii) in Luxembourg, at least 12 slaughter batches.

2. Secondary sample size

The secondary sample size gives the number of individual broiler chickens per slaughter batch to be sampled. That
number shall be 10 birds for the detection of Campylobacter in caeca and one bird for the detection of Campylobacter
and Salmonella on carcasses. These caeca samples and carcass sample must be from the same slaughter batch.

PART C

Specimen collection, handling and analysis for the detection and antimicrobial testing of Campylobacter spp. in
broiler flocks

1. Collection and transport

Campylobacters are relatively fragile organisms, which die quickly outside the host gut. Therefore, care shall be taken to
ensure that samples are taken appropriately and analysed quickly. Extreme temperatures have to be avoided and
transport shall be as fast as possible.

Samples to be collected shall be intact caeca. Caecal samples shall be taken at the time of evisceration.

(1) Estimation: number of holdings (four in Estonia, five in Latvia) × two flocks per holding × two slaughter batches per flock × six rounds
per year. In Luxembourg, only broilers from three small flocks are slaughtered. A slaughter batch from each of them will be sampled
each trimester.

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Only staff trained in standard sampling procedures shall collect samples. The main objective shall be to minimise
external contamination from caecal content while sampling. That is best achieved by careful manual traction at the
junction with the intestine. One intact caecum shall be taken per bird, and samplers shall verify that the caecum is full
or it shall be disregarded. Birds shall preferably be sampled at random throughout the batch (avoiding the first part of
the batch to be slaughtered), collecting samples from non-consecutive birds. The 10 caeca collected may be placed in a
single sterile bag/pack for transport.

All relevant information available from the sample must be recorded on a sampling form produced by the competent
authority to enable the reporting requirements in Part E to be fulfilled. Each sample and its sample form must be
labelled with a unique number which must be used from sampling to testing. The competent authority must arrange
for the issue and use of a unique numbering system. The same identifier of the slaughter batch shall be used as the one
for the carcass sample.

Caecal samples shall be transported as intact caeca to the laboratory within 24 hours (i.e. overnight postage or courier)
and analysed there immediately. In cases where that cannot be managed, the samples shall be kept refrigerated at least
until transported to the laboratory and they shall be analysed no later than 72-80 hours after sampling. At the
laboratory, samples that cannot be tested on the day of arrival shall be kept refrigerated until analysis.

At the laboratory, the caecal contents shall be aseptically removed and pooled to one composite sample.

2. Diagnostic method

2.1. Culture

Direct culture on a selective medium provides a good estimate of the prevalence of Campylobacters. Direct culture
of the sample shall be carried out on a selective medium suitable for Campylobacter, (i.e. modified Campylobacter
blood free selective medium (CCDA); Karmali; or Preston Agar).

The plates shall be incubated at 41,5 ± 1 °C, in a micro-aerobic atmosphere, for at least 48 +/– 2 hours. Growth
may be detected after 24 hours.

The micro-aerobic atmosphere may be obtained in commercially available micro-aerobic incubators (gas mixture
10 % CO2/6 % O2). In the absence of such incubators, micro-aerobic culture systems can be used i.e. gas jars.
Commercial gas pack systems providing the appropriate micro-aerobic atmosphere are available.

Suitable positive and negative controls shall be included for each batch of samples cultured.

2.2. Confirmation and speciation of the genus Campylobacter

least one Campylobacter

Isolation and confirmation of Campylobacter organisms should be undertaken as described in ISO 10272-
1:2006(E). At
isolate per batch must be speciated using phenotypic methods as
described in ISO 10272-1:2006(E) or published molecular methods such as Polymerase Chain Reaction (PCR)
techniques. The method used shall be indicated. The isolate speciated should be used for subsequent antimicrobial
testing.

If a laboratory is less experienced in speciation, it shall store the isolate as set out in 2.4 pending additional
training or send it to a more experience laboratory in consultation of the Community reference laboratory for
Campylobacter.

2.3. Quality control

For quality assurance, a proportion of Campylobacter spp. isolates with a maximum of eight isolates shall be sent
to the Community reference laboratory for Campylobacter for confirmation and speciation.

A proportion of those isolates shall be sent to that laboratory either in one batch or on a quarterly basis. If
isolates are to be transported between laboratories, appropriate conditions (for example charcoal swabs) shall be
used.

2.4. Storage

At least one isolate per positive sample shall be stored at the NRLs using the normal method for NRL culture
collection, as long as it ensures viability of the strains for a minimum of two years.

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2.5. Antimicrobial resistance testing

The number of Campylobacter isolates to be included in the antimicrobial resistance monitoring per Member State
shall be 170. Not more than one isolate per Campylobacter species from the same slaughter batch shall be included
in the monitoring.

In those Member States where, in any given year, a lower number of isolates than the target sample size is
available, all these isolates shall be included in the antimicrobial resistance monitoring.

In those Member States where a higher number of isolates is available all isolates, or a representative random
selection equal or larger than the target sample size, shall be included.

Member States shall test at least the antimicrobials that are specified in Table 1, using the cut-off values given and
an appropriate concentration range to determine the susceptibility of Campylobacter.

Campylobacter
Jejuni

Campylobacter
Coli

Table 1

Antimicrobial

Erythromycin

Ciprofloxacin

Tetracycline

Streptomycin

Gentamicin

Erythromycin

Ciprofloxacin

Tetracycline

Streptomycin

Gentamicin

Cut-off value (mg/L) R >

4

1

2

2

1

16

1

2

4

2

Dilution methods shall be performed according to the methods described in CLSI guidelines M31-A3 — Third
Edition, Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from
Animals and M100-S16, Performance Standards for Antimicrobial Susceptibility testing; Sixteenth International
Supplement.

PART D

Specimen collection, handling and analysis for the detection of Campylobacter spp. and Salmonella Spp. in broiler
carcasses

1. Collection and transport

One whole carcass per slaughter batch shall be collected immediately after chilling but before further processing such
as freezing, cutting or packaging. In some slaughterhouses this may mean that samples are taken after pre-chilling
when this is the last step before further processing.

The sample collected shall be placed in a separate sterile plastic bag, avoiding cross-contamination and be sent to the
laboratory where skin sampling shall be undertaken.

Cross-contamination by other carcasses or caeca samples must be avoided during the collection of the carcasses.
Precautions must therefore be taken at all stages to ensure that the equipment used during sampling, transport and
storage are not contaminated with the pathogens investigated in the survey.

All relevant information available from the sample shall be recorded on a sampling form produced by the competent
authority to enable the recording requirements in Part E to be fulfilled.

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Each sample and its sample form shall be labelled with a unique number which shall be used from sampling to testing.
The competent authority must arrange for the issue and use of a unique numbering system. The same identifier of the
slaughter batch shall be used as the one for the caeca samples.

The samples shall be kept at between + 2 to 8 °C and free of external contamination during transportation.

All samples shall ideally reach the laboratory within 24 hours of sampling. In exceptional situations (for example, long
journeys, weekends and public holidays) that period may be extended to 80 hours.

In the case that different laboratories are used for Campylobacter and Salmonella testing then the laboratory testing
Campylobacter should take preference in receipt of the sample.

2. Sampling in the laboratory and the analytical methods

2.1. Receipt of samples

On receipt of the samples, laboratories shall check the information recorded by the sampler and complete the
relevant sections of the sample form.

Samples shall be held at + 2-8 °C in the laboratory and the laboratory sampling procedure shall begin as soon as
possible after the arrival of the samples at the laboratory and in any case within 72-80 hours from the time of
sampling.

2.2. Sample preparation

All samples received shall be examined to ensure that the transport packaging is intact before testing.

Handlers must avoid cross-contamination between samples and from the surrounding environment at all stages.

Wearing disposable gloves, the chicken shall be removed from its sample bag, taking care not to contaminate the
outer surface of the chicken.

Using a sterile instrument and aseptic technique, the neck skin shall be removed, if present, together with the skin
from one side of the carcass avoiding any fat to make a 27 g test portion and placed into a stomacher bag (or
pulsifier).

2.3. Initial suspension

The 27 g test portion shall be transferred to NINE volumes (243 ml) buffered peptone water (BPW), brought to
room temperature before adding. The mixture shall be treated in a stomacher or pulsifier for approximately ONE
minute (27 g are needed to perform analyses for Salmonella spp. and Campylobacter spp. from one sample in
parallel). Foaming shall be avoided by removing the air from the stomacher bag as much as possible.

This initial suspension shall be used as follows:

(a) 10 ml (~1g) shall be transferred to 90 ml enrichment medium for Campylobacter spp. detection;

(b) 10 ml (~1g) shall be transferred to an empty sterile tube; 1 ml is used for the enumeration of Campylobacter

spp. on selective plates.

The rest of the initial suspension (250 ml ~ 25g) shall be used for the detection of Salmonella spp.

2.4. Detection, identification methods for Salmonella spp.

2.4.1. D e t e c t i o n o f S a l m o n e l l a s p p .

The detection of Salmonella spp. shall be done according to ISO 6579-2002 (E).
animal feeding stuffs — Horizontal method for the detection of Salmonella spp.’

‘Microbiology of food and

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2.4.2. S e r o t y p i n g o f S a l m o n e l l a s p p .

At least one isolate from each positive sample shall be typed by the National Reference Laboratory for
Salmonella, using the Kaufmann-White scheme.

For quality assurance, a proportion of the non-typeable isolates shall be sent to the Community Reference
Laboratory for Salmonella, with a maximum of 16 non-typeable isolates. A proportion of those isolates shall be
sent to that laboratory on a quarterly basis.

2.4.3. P h a g e t y p i n g o f S a l m o n e l l a s p p .

For S. Enteritidis and S. Typhimurium it is recommended that at least one isolate from each positive sample shall
be phage typed, using the protocol defined by the Health Protection Agency (HPA), Colindale, London.

2.5. Detection, identification and quantification methods for Campylobacter spp.

2.5.1. D e t e c t i o n o f C a m p y l o b a c t e r s p p .

Isolation and confirmation of Campylobacter organisms should be undertaken as described in ISO 10272-
1:2006(E). At least one Campylobacter isolate per batch must be speciated using phenotypic methods as
described in ISO 10272-1:2006(E) or published molecular methods such as Polymerase Chain Reaction
(PCR) techniques. The method used shall be indicated.

For quality assurance, a proportion of Campylobacter spp. isolates with a maximum of eight isolates shall be
sent to the Community reference laboratory for Campylobacter for confirmation and speciation.

A proportion of those isolates shall be sent to that laboratory on a quarterly basis. If isolates are to be
transported between laboratories, appropriate conditions (for example charcoal swabs) shall be used.

2.5.2. Q u a n t i f i c a t i o n o f C a m p y l o b a c t e r s p p .

The quantitative detection of Campylobacter spp. shall be carried out according to ISO/TS 10272-2:2006
‘Microbiology of food and animal feeding stuffs — Horizontal method for detection and enumeration of
Campylobacter spp. Part 2: Colony-count technique’. Starting with 10 ml initial suspension 0,1 ml of this initial
suspension and of further dilutions thereof shall be examined to allow the enumeration of up to 106 cfu/g. In
addition, 1 ml of the undiluted initial suspension shall be examined to obtain a limit of enumeration of 10
cfu/g. All plate determinations shall be done in duplicate.

To enable correct comparison and judgement of data (for future risk assessment) the measurement uncertainty
(MU) of the quantitative determination method shall be estimated for each laboratory.

To estimate the MU the technical specification ISO/TS 19036:2006 shall be used with the exception that the
parallel dilutions from the initial suspension is applied for estimation of the MU.

The MU is derived from the intra-laboratory standard deviation of reproducibility. Data on MU estimation shall
be collected from May to September in order to ensure positive samples. A total of 12 positive samples shall
be examined in duplicate and parallel dilutions prepared from the 10 ml initial suspension. Raw data on MU
estimation shall be reported separately as part of the overall description on the implementation of the survey
as set out in Part E.

3. Storage of isolates

In order to allow for e.g. later testing for antimicrobial susceptibility storage of a representative subset of isolates is
recommended. One isolate per positive sample shall be stored. The Campylobacter isolate obtained from the quantitative
analysis must be preferred. The isolates must be stored at the NRLs using the normal method for NRL culture
collection, as long as it ensures viability of the strains for a minimum of two years.

PART E

Reporting

Reports shall be made including at least the following information:

1. Overall description on the implementation of the survey:

— slaughterhouses: total number per country and the number that were sampled,

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— primary sample size realised,

— description of the stratification and randomisation procedures,

— description of the quality control activities,

inclusive a report on the 12 MU estimations per laboratory for

Campylobacter quantification,

— overall results.

2. Specific information with regard to prevalence data

Member States shall submit the results of the investigation in the form of raw data using a data dictionary and data
collection forms provided by the Commission.

That data shall include at least the following information:

— name/code of the slaughterhouse,

— identifier of the slaughter batch,

— name/code of the holding (farm) of origin of the slaughter batch,

— holding size if known,

— Salmonella vaccination status of the flock if known,

— age of the broilers at sampling (slaughter),

— information regarding if this was the first or a subsequent batch to be slaughtered from the flock (preceding

thinning or not),

— production type (i.e. conventional, free-range, organic),

— results of previous Salmonella and Campylobacter testing in the same flock,

— date of sampling,

— number of birds slaughtered per year in that slaughterhouse,

— type of chilling method used (air, immersion, spray),

— details of transport protocol (as specified: Y/N),

— date received at laboratory,

— date of testing,

— identification of the laboratory,

— type of sample,

— description of the culture methods used, in particular the selective medium/media,

— Campylobacter isolate: Method used for speciation,

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— Campylobacter: result of the bacteriological testing, including speciation from the caecal sample,

— Campylobacter: result of the bacteriological testing, including speciation and quantification from the carcass sample,

— Salmonella: result of bacteriological testing and serotyping,

— time between sampling and analysis (per 12 hour period).

3. Specific information with regard to antimicrobial resistance testing of Campylobacter isolates of caecal samples.

The results of the antimicrobial resistance monitoring shall be assessed and reported, in accordance with Article 9 of
Directive 2003/99/EC, in the yearly report on trends and sources of zoonoses, zoonotic agents and antimicrobial
resistance.

Without prejudice to the provisions of Annex IV of Directive 2003/99/EC the following information shall be reported:

— origin of isolates i.e. baseline study, control programme, passive surveillance,

— number of isolates susceptibility tested per Campylobacter species,

— number of isolates found to be resistant per antimicrobial per Campylobacter species, and

— number of fully-susceptible isolates and number of isolates resistant to 1, 2, 3, 4 and > 4 antimicrobials listed in

Table 1 per Campylobacter species.

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ANNEX II

Maximum Community financial contribution to the Member States

Member State

Maximum total amount for co-financing
of sampling and analyses

(EUR)

Belgium — BE

Bulgaria — BG

Czech Republic — CZ

Denmark — DK

Germany — DE

Estonia — EE

Ireland — IE

Greece — EL

Spain — ES

France — FR

Italy — IT

Cyprus — CY

Latvia — LV

Lithuania — LT

Luxembourg — LU

Hungary — HU

Malta — MT

Netherlands — NL

Austria — AT

Poland — PL

Portugal — PT

Romania — RO

Slovenia — SI

Slovakia — SK

Finland — FI

Sweden — SE

United Kingdom — UK

58 092

58 092

58 092

58 092

58 092

14 688

58 092

58 092

58 092

58 092

58 092

58 092

18 360

58 092

1 836

58 092

58 092

58 092

58 092

58 092

58 092

58 092

58 092

58 092

58 092

58 092

58 092

Total

1 429 092

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ANNEX III

Certified financial report on the implementation of a survey on the prevalence of Campylobacter spp. in broiler
flocks and their antimicrobial resistance and on the prevalence of Campylobacter spp. and Salmonella spp. in
broiler carcasses

Reporting period:

............................................................................................. to .............................................................................................

Statement on costs incurred on the survey and eligible for Community financial contribution:

Reference number of Commission Decision providing Community financial contribution:

......................................................................................................................................................................................................................................

......................................................................................................................................................................................................................................

Costs incurred related to functions at/by

Number of tests

Total costs of testing incurred during
reporting period
(national currency)

Bacteriological detection of Campylobacter spp.

Bacteriological detection of Salmonella spp.

Confirmation of Campylobacter spp.

Speciation of Campylobacter isolates

Enumeration of Campylobacter isolates

Serotyping of Salmonella isolates

Antimicrobial
isolates

resistance testing of Campylobacter

We certify that:

Declaration by the beneficiary

— the above costs are genuine and have been incurred in carrying out the tasks laid down in this Decision and were

essential for the proper performance of those tasks;

— all supporting documents supporting for the costs are available for audit purposes;

— no other Community contribution was requested for this programme.

Date:

.........................................................................................................................................................................................................................

Person financially responsible:

........................................................................................................................................................................

Signature:

................................................................................................................................................................................................................