EN    EN  COMMISSION OF THE E UROPEAN COMMUNITIES 
Brussels, 
C(2008)  
Draft 
COMMISSION REGULATION 
of 
amending Regulation (EC) No 2160/2003 of the European Parliament and of the Council 
and Regulation (EC) No 1003/2005 as re gards the control and testing of Salmonella  in  
breeding flocks of Gallus gallus and turkeys  
  
(Memorandum from Ms A.Vassilou)  
EN 2   EN Draft 
COMMISSION REGULATION 
of 
amending Regulation (EC) No 2160/2003 of the European Parliament and of the Council 
and Regulation (EC) No 1003/2005 as re gards the control and testing of Salmonella  in  
breeding flocks of Gallus gallus and turkeys  
 
 
 (Text with EEA relevance) 
THE COMMISSION OF THE EUROPEAN COMMUNITIES, 
Having regard to the Treaty esta blishing the European Community, 
Having regard to Regulation (EC) No 2160/2003 of the European Parliament and of the 
Council of 17 November 2003 on the control of Salmonella  and other specified food-borne 
zoonotic agents1 and, in particular Article 5(6) and Article 13 thereof; 
Whereas: (1) The purpose of Regulation (EC) No 2160/2003 is  to ensure that proper and effective 
measures are taken to detect and control Salmonella  and other zoonotic agents at all 
relevant stages of producti on, processing and distribution, a nd particularly at the level 
of primary production, in order to reduce th eir prevalence and the risk they pose to 
public health. 
(2) Pursuant to Regulation (EC) No 2160/ 2003, specific require ments concerning 
breeding flocks of Gallus gallus  apply whenever certain anal ysis of samples indicates 
the presence of Salmonella  Enteritidis or Salmonella  Typhimurium  in such flocks . The 
purpose of these requirements is to prevent the spread of infection in the egg and 
broiler meat production chain, namely fr om breeders to their progeny. Similar 
requirements should be applied to turk ey production in order to prevent the 
transmission of infection in the turkey meat productio n chain. Regulation (EC) No 
2160/2003 should therefore be amended accordingly.  
(3) Commission Regulation (EC) No 1003/ 2005 of 30 June 2005 implementing 
Regulation (EC) No 2160/2003 as  regards a Community target for the reduction of the 
                                                
 
1 OJ L 325, 12.12.2003, p. 1.   
EN 3   EN prevalence of certain Salmonella  serotypes in breeding flocks of Gallus gallus2 lays 
down a Community target for the reduction of the prevalence of certain Salmonella 
spp. in breeding flocks of Gallus gallus. In addition, the Annex to that Regulation sets 
out the testing scheme necessary to verify  the achievement of the Community target. 
(4) Pursuant to Article 2 of Regulation (EC) No 1003/2005, the Commission should 
review the Community target in the light  of the results of the first year of 
implementation of the national control programmes approved in accordance with 
Regulation (EC) No 2160/2003. The year 2007 was the first year of implementation. 
(5) Pursuant to Directive 2003/99/ EC of the European Parliame nt and of the Council of 17 
November 2003 on the monitoring of zoonoses and zoonotic agents3, Member States 
has transmitted the results of their monito ring for the year 2007 to the Commission. In 
the light of those results, it does not appear necessary to amend the Community target.  
(6) In view of the efficient allocation of reso urces, Member States that have achieved the 
Community target should be permitted to reduce the number of official controls. Regulation (EC) No 1003/2005 should th erefore be amended accordingly.  
(7) A review of the testing scheme set out in  the Annex to Regulation (EC) No 1003/2005 
revealed difficulties in the implementation of sampling instructions and new information on the sensitivity of testing schemes has become available. The testing scheme should thus be amended.  
(8) Regulations (EC) Nos 2160/2003 and 1003/ 2005 should therefore be amended 
accordingly. 
(9) The measures provided for in this Regulati on are in accordance with the opinion of the 
Standing Committee on the Food Chain and Animal Health,  
HAS ADOPTED THIS REGULATION: 
Article 1 
 In Part C of Annex II to Regulation (EC) No  2160/2003, the title and point 1 is replaced by 
the following: 
 " C.  Specific requirements co ncerning breeding flocks of Gallus gallus and breeding 
turkeys  
1.  The measures laid down in points 3 to 5 must be taken whenever the analysis of 
samples taken in accordance with part B, or in accordance with the testing schemes 
set out in the A nnexes to Commission Regulations (EC) No 1003/2005* and 
584/2008**, indicates the presence of Salmonella  Enteritidis  or Salmonella 
Typhimurium in a breeding flock of Gallus gallus  or breeding turkeys in the 
circumstances set out in point 2.  
                                                
 
2  OJ L 170, 1.7.2005, p. 12.  
3  OJ L 325, 12.12.2003, p. 31.  
EN 4   EN *OJ L 170, 1.7.2005, p. 12. 
** OJ L 162, 21.6.2008, p. 3." 
Article 2 
The Annex to Regulation (EC) No 1003/2005 is re placed by the text in the Annex to this 
Regulation.  
   
Article 3 
Entry into force 
This Regulation shall enter into force on the
 twentieth day following that of its publication in 
the Official Journal of  the European Union . 
However, Article 2 shall apply from1 Ap ril 2009 and Article 1 from 1 January 2010   
This Regulation shall be binding in  its entirety and directly applicable in all Member States. 
Done at Brussels, 
 For the Commission 
 Androulla VASSILOU  Member of the Commission  
EN 5   EN ANNEX   
'ANNEX 
 
'Testing scheme necessary to verify the ac hievement of the Community target for the 
reduction of Salmonella Enteritidis, Salmonella  Hadar, Salmonella  Infantis, Salmonella  
Typhimurium and Salmonella  Virchow  in adult breeding flocks of Gallus gallus  
1. S AMPLING FRAME  
The sampling frame shall cover all adu lt breeding flocks of domestic fowl ( Gallus 
gallus)  comprising at least 250 bi rds (‘breeding flocks’). 
2. M ONITORING IN BREEDING FLOCKS  
2.1. Location, frequency and status of sampling 
 Breeding flocks shall be sampled at the initiative of the operator and as part of 
official controls. 
2.1.1. Sampling at the initia tive of the operator 
 Sampling shall take place every two weeks at the place designated by the 
competent authority from the following two possible options: 
(a) at the hatchery; or (b) at the holding. 
 The competent authority shall implement one of the options referred to in 
points (a) or (b) to the whole testing scheme for all broiler breeder flocks 
and one of those options for layer breeding flocks. It shall set up a procedure so that the detection of Salmonella  serotypes referred to in 
Article 1(1) (‘relevant Salmonella ’) during the sampling at the initiative 
of the operator is notified without de lay to the competent authority by the 
laboratory performing the analyses. Ti mely notification of the detection 
of Salmonella , including the serotype, shall remain the responsibility of 
the operator and the laboratory performing the analyses. 
By way of derogation, if the Commun ity target has b een reached during 
the last two calendar years, sampli ng at the holding may be extended to 
take place every three weeks, at the discretion of the competent authority. 
The competent authority may decide to revert to a two week testing 
interval in the case of detection of  a positive flock on the holding pr in 
any case deemed suitable by the competent authority.  
2.1.2. Official control sampling 
 Without prejudice to Annex II, Part C.2 of Regulation (EC) No 
2160/2003, official sampling shall consist of:  
EN 6   EN 2.1.2.1. If sampling at the initiative of  the operator takes place at the 
hatchery: 
(a) routine sampling every 16 week s at the hatchery, which shall 
on that occasion replace the corresponding sampling at the initiative of the operator;  
(b) routine sampling at the holding on two occasions during the 
production cycle, the first one  being within four weeks 
following moving to laying pha se or laying unit and the 
second one being towards the end of the laying phase, not earlier than eight weeks befo re the end of the production 
cycle if the Community target  has not been reached during 
the last two calendar years. 
If the Community target has been achieved during the last 
two calendar years, the compet ent authority may sample at 
the holding only at one occasion anytime during the production cycle;  
(c) confirmatory sampling at the holding, following the detection 
of relevant Salmonella  from sampling at the hatchery. 
2.1.2.2. If sampling at the initiative of  the operator takes place at the 
holding, routine sampling shall be carried out on three occasions 
during the production cycle: 
(a) within four weeks follow ing moving to laying phase or 
laying unit; 
(b) towards the end of the laying  phase, not earli er than eight 
weeks before the end of the production cycle; 
(c) during the production, at any ti me sufficiently distant from 
the samples referred to in points (a) and (b). 
By way of derogation from the this paragraph, the competent 
authority may decide to carry out only one official sampling 
anytime during the production cycle if the Community target has been achieved for at least two calendaryears.  
2.2. Sampling protocol 
2.2.1. Sampling at the hatchery For each breeding flock, the sample shall consist of at least:  
(a) one composite sample of visibly so iled hatcher basket liners taken at 
random from five separate hatcher baskets or locations in the hatcher, to reach 
a total sampling surface of at least 1m²; however,  if the hatching eggs from a 
breeding flock occupy more than one hatcher, then one such composite sample per 50 000 eggs shall be taken containi ng material from each hatcher; or  
EN 7   EN   (b) one sample taken with a moistened fabric swab of at least 900 cm2 surface 
area, taken immediately after the rem oval of the chickens from the whole 
surface area of the bottom of at least a total of five hatcher baskets, or from 
fluff from five places in all hatchers w ith eggs from the flock ensuring that at 
least one sample per flock from whic h eggs are derived, is taken; or 
(c) 10 g broken eggshells taken from a total of 25 separate hatcher baskets 
in all hatchers with eggs from the fl ock, crushed, mixed and taken from a 25 g 
subsample. 
The procedure set out in points (a), (b ) and (c) shall be followed for sampling 
at the initiative of the operator as well as for official sampling. 
2.2.2. Sampling at the holding: 
2.2.2.1. Routine sampling at the initiative of the operator 
 Sampling shall primarily consist of  faecal samples and shall aim to 
detect a 1% within flock prevalen ce, with a 95% confidence limit. 
To that effect, the samples shall comprise one of the following: 
(a) Pooled faeces made up of separate samples of fresh faeces 
each weighing not less than 1 g taken at random from a number of sites in the building in which the flock is kept, or where the flock has free access to more than one building on a particular holding, from each group of buildings on the holding in which the flock is kept. Faeces may be pooled for 
analysis up to a minimum of two pools. 
 The number of sites from which separate faeces samples are 
to be taken in order to make a pooled sample shall be as follows: 
Number of birds kept in the flock Number  of faeces samples to be taken in the 
flock 
250-349 200 
350-449 220 
450-799 250 
800-999 260 
1000 or more 300 
 
(b) Boot swabs and/or dust samples:  
EN 8   EN  Boot swabs used shall be sufficiently absorptive to soak up 
moisture. Tubegauze ‘socks’ shall also be acceptable for that purpose. 
 The surface of the boot swab shall be moistened using 
appropriate diluents (such as  0,8% sodium chloride, 0,1% 
peptone in sterile deionised wate r, sterile water or any other 
diluent approved by the competent authority).  
  
 The samples shall be taken while walking through the 
premises using a route that will produce representative samples for all parts of the premise or the respective sector. This shall include littered and sl atted areas provided that slats 
are safe to walk on. All separate pens within a house shall be included in the sampling. On co mpletion of sampling in the 
chosen sector, boot swabs must be removed carefully so as not to dislodge adherent material. 
The samples shall consist of: (i)  five pairs of boot swabs, representing each about 20% of 
the area of the house; the swabs may be pooled for analysis into a mini mum of two pools; or, 
(ii)  one pair of boot swabs representing the whole area of 
the house and an additional dus t sample collected from 
multiple places throughout the house from surfaces with visible presence of dust. A mo istened fabric swab of at 
least 900 cm
2 surface area shall be used to collect this 
dust sample. 
 (c) In cage breeding flocks, sampling may consist of naturally 
mixed faeces from dropping belts, scrapers or deep pits, depending on the type of house.  Two samples of at least 
150 g shall be collected to  be tested individually: 
(i) droppings belts beneath each tier of cages which are 
run regularly and discharged into an auger or conveyor system; 
(ii) droppings pit system in which deflectors beneath the 
cages are scraped into a deep pit beneath the house;  
(iii) droppings pit system in a step cage house when cages 
are offset and faeces fall directly into the pit. 
 There are normally several st acks of cages within a house. 
Pooled faeces from each stack shall be represented in the overall pooled sample. Two pooled samples shall be taken  
EN 9   EN from each flock as described in the following third to sixth 
subparagraphs: 
 In systems where there are belts or scrapers, these shall be 
run on the day of the sampling before sampling is carried out. 
 In systems where there are deflectors beneath cages and 
scrapers, pooled faeces which has lodged on the scraper after 
it has been run, shall be collected. 
 In step-cage systems where there is no belt or scraper system 
it is necessary to collect pooled  faeces from the deep pit.  
 Droppings belt systems: pooled  faecal material from the 
discharge ends of the belts shall be collected. 
2.2.2.2. Official sampling 
(a) Routine sampling shall be performed as described in point 
2.2.2.1. 
(b) Confirmatory sampling followi ng the detection of relevant 
Salmonella from sampling at the hatchery shall be performed 
as described in point 2.2.2.1. Ad ditional samples shall be 
collected for the possible testing of antimicrobials or bacterial 
growth inhibitors as follows: birds shall be  taken at random 
from within each house of birds on the farm, normally up to five birds per house, unless th e authority deem s it necessary 
to sample a higher number of bird s. The test shall at least be 
carried out if infection is not confirmed. If anti-microbials or 
bacterial growth inhibitors are detected, the Salmonella 
infection shall be cons idered as confirmed.  
 In case the presence of the relevant Salmonella  is not 
detected but anti-microbials or  bacterial growth inhibitory 
effect are, sampling of the breeding flock for relevant 
Salmonella  and bacterial growth inhi bitory effect shall be 
repeated until no bacterial growth inhibitory effect is detected, or the breeding flock is  destroyed. In the latter case, 
the breeding flock shall be counted as an infected breeding flock for the purpose of the Community target.  
 (c) Suspect cases 
 In exceptional cases where the competent authority has 
reasons to call the result into question (false pos itive or false 
negative results, it may decide to repeat the testing in accordance with point (b).  
3. EXAMINATION OF THE SAMPLES  
3.1. Preparation of the samples  
EN 10   EN 3.1.1. Hatcher basket liners: 
(a) place in 1 litre Buffered Pept one Water (BPW) which has been 
prewarmed at room temperature and mix gently;  
(b) continue the culture of the samp le by using the detection method 
described in 3.2. 
3.1.2. Boot swabs and dust samples: 
(a) carefully unpack the pair(s) of  boot swabs (or “socks”) or dust 
sample to avoid dislodging adhere nt faecal material or loose dust 
material and place in 225 ml of BPW which has been prewarmed to room temperature; 
 (b) where five pairs of boot swabs are pooled into two samples, place 
five individual samples into a mi nimum of 225 ml BPW and ensure 
that all samples are totally emerged in the BPW. 
(c) swirl to fully saturate the sample and continue the culture by using 
the detection method described in 3.2. 
3.1.3. Other faecal material samples: 
(a) The faeces samples shall be pooled and thoroughly mixed and a 25 
gram sub-sample shall be collected for culture. 
(b) The 25 gram sub-sample shall be added to 225 ml of BPW which   
has been pre-warmed to room temperature; 
(c) The culture of the sample shall be continued by using the detection 
method described in 3.2.  
If ISO standards on the preparati on of faeces for the detection of 
Salmonella are agreed on they shall be applied and replace the above 
provisions on sampling preparation   
3.2. Detection method 
 The detection of Salmonella  spp. shall be carried out  according to Amendment 
1 of EN/ISO 6579-2002/Amd1:2007. “Mic robiology of food and animal 
feeding stuffs - Horizontal method for the detection of Salmonella  spp. – 
Amendment 1: Annex D: Detection of Salmonella  spp. in animal faeces and in 
environmental samples from the primary production stage”.   
 As regards the boot swabs samples and other faecal material samples referred 
to in paragraph 3.1, it is possible to pool incubated BPW enrichment broth for 
future culture. To do that, incubate both samples in BPW as normal. Take 1 ml 
of incubated broth from each sample and mix thoroughly then take 0,1 ml of 
the mixture and inoculate the MS RV plates in the usual way. 
3.3. Serotyping  
EN 11   EN  At least one isolate from each sample  showing a positive reaction shall be 
typed, following the Kaufmann-White scheme. 
4. R ESULTS AND REPORTING  
A breeding flock shall be considered in fected for the purpose of verifying the 
achievement of the Community target , when the presence of relevant Salmonella  
(other than vaccine strains) wa s detected in one or more fa ecal samples (or if there is 
a secondary official confirmation in the Memb er State, in the relevant faecal samples 
or birds organ samples), taken at the hol ding. This shall not apply in exceptional 
cases of suspect breeding flocks where sa lmonella detection at the holding at the 
initiative of the operator was not  confirmed by official sampling. 
For statistical purpose, an infected flock shall be c ounted once only regardless of 
how often Salmonella have been detected in this fl ock during the production period.   
Reporting shall include: (a) detailed description of the options implemented for the sampling scheme and 
the type of samples taken, as appropriate; 
(b) number of existing breedi ng flocks and those tested; 
(c) results of the testing; (d) explanations on the results, in particular concerning exceptional cases.'