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of 20 December 2007
concerning a financial contribution from the Community towards a survey on the prevalence of
Salmonella spp. and Methicillin-resistant Staphylococcus aureus in herds of breeding pigs to be
carried out in the Member States
(notified under document number C(2007) 6579)
THE COMMISSION OF THE EUROPEAN COMMUNITIES,
and to consider the best approach to evaluate in thefuture the achievement of such target, comparable dataon the percentage of Salmonella
infected holdings ofbreeding pigs in the Member States needs to be
Having regard to the Treaty establishing the European
available. Such information is not available and a
special survey should therefore be carried out tomonitor the prevalence of Salmonella
in breeding pigsover a suitable period in order to take account of
Having regard to Council Decision 90/424/EEC of 26 June
possible seasonal variations. The survey should be
1990 on expenditure in the veterinary field (1), and in particular
based on the Salmonella
report also recommends additional
sampling for the estimation of within-holding prevalence.
Such sampling should be carried out by a number ofMember States geographically representing the differentsituations in the Community.
Decision 90/424/EEC lays down procedures governing afinancial contribution by the Community towardsspecific veterinary measures, including technical andscientific measures. It provides for the Community to
undertake or assist Member States in undertaking the
infections have been recognised as an important risk in
technical and scientific measures necessary for the devel-
hospitals for several decades. MRSA is resistant to the
opment of Community veterinary legislation and for the
most commonly used antibiotics and it is particularly
development of veterinary education or training.
dangerous for patients with a decreased immunity. Thenumber of deaths attributed to MRSA in the UnitedKingdom has been estimated at around 3 000 per year.
Under Article 4 and Annex I of Regulation (EC) No
The costs for treatment per patient are estimated at EUR
2160/2003 of the European Parliament and of the
12 000 to 15 000. Additional expenses are incurred due
Council of 17 November 2003 on the control of
to hygiene and control programmes to prevent or limit
and other specified food-borne zoonotic
agents (2), a Community target is to be established forreducing the prevalence of Salmonella
in populations ofherds of breeding pigs.
A new strain of MRSA (ST398) has recently beendetected in production animals in several MemberStates. In particular, pigs have been recognised as an
The Task Force on Zoonoses Data Collection of the
important source of infection for pig farmers or their
European Food Safety Authority (EFSA) adopted on 30
relatives by direct contact with pigs. Infection with the
April 2007 a Report on a proposal for technical specifi-
new strain may also enter hospitals as previously MRSA
cations for a baseline study on the prevalence of
in breeding pigs (3) (the Salmonella
In order to increase awareness and to assess whether it is
In order to set the Community target for the reduction of
necessary to take measures to detect and control MRSA
the prevalence of zoonoses and zoonotic agents as
in order to reduce their prevalence and the risk they pose
foreseen in Article 4 of Regulation (EC) No 2160/2003
to public health, comparable data on the percentage ofMRSA (ST398) infected holdings of breeding pigs in the
(1) OJ L 224, 18.8.1990, p. 19. Decision as last amended by Regulation
Member States are needed. Such information is not
(EC) No 1791/2006 (OJ L 363, 20.12.2006, p. 1).
available and a special survey should therefore be
(2) OJ L 325, 12.12.2003, p. 1. Regulation as last amended
carried out to monitor the prevalence of MRSA in
by Commission Regulation (EC) No 1237/2007 (OJ L 280,24.10.2007, p. 5).
breeding pigs over a suitable period in order to take
(3) The EFSA Journal
(2007) 99, 1-28.
account of possible seasonal variations.
The Task Force on Zoonoses Data Collection of the EFSA
2005 on the financing of the common agricultural
adopted on 19 November 2007 a Report including a
policy (3), the conversion rate for expenditure in a
proposal for technical specifications for a baseline
currency other than euro should be the rate most
survey on the prevalence of Methicillin-Resistant Staphy-
recently set by the European Central Bank prior to the
(MRSA) in breeding pigs (the MRSA
first day of the month in which the application is
report) (1). The MRSA report makes recommendations
submitted by the Member State concerned. For reasons
with regard to the sampling frame, sample collection
of clarity and transparency, Decision 2007/636/EC
protocol, laboratory analytical methods and reporting.
should be repealed and a financial contribution from
The technical specifications for the survey provided for
the Community toward the surveys for the prevalence
in this Decision should be based on that report.
and MRSA should be laid down in thissingle Decision.
In accordance with Commission Decision 2007/636/EC
In order to ensure the coherence in carrying out the
of 28 September 2007 concerning a financial contri-
surveys, this Decision should apply from 1 January
bution from the Community towards a survey on the
2008, date of application of Decision 2007/636/EC.
prevalence of Salmonella
spp. in herds of breeding pigsto be carried out in the Member States (2), Member Statesare to carry out a survey in herds of breeding pigs from1 January 2008 until 31 December 2008 in order to
The measures provided for in this Decision are in
assess the prevalence of Salmonella
spp. Taking into
account the public health significance of MRSA, the
Committee on the Food Chain and Animal Health,
emerging risk of pigs as source of infection forhumans and the lack of comparable information onthe prevalence of MRSA in herds of breeding pigs indifferent Member States, additional sampling during the
survey provided for in Decision 2007/636/EC is the mostrapid and cost-effective way to evaluate the prevalence ofMRSA in herds of breeding pigs in the Community.
Subject matter and scope
The survey is to provide technical information necessary
This Decision lays down rules on the financial contribution
for the development of Community veterinary legislation
from the Community towards baseline surveys to be carried
as appropriate. Given the importance of collecting
out in the Member States to assess the prevalence of Salmonella
comparable data on the prevalence of MRSA in
spp. (the Salmonella
survey) and Methicilline-resistant Staphylo-
breeding pigs in the Member States, the Member States
(MRSA) (the MRSA survey) across the Community
should be granted a Community financial contribution
in breeding pigs sampled at farm level.
for implementing the specific requirements of thesurvey. It is appropriate to reimburse 100 % of thecosts incurred on the purchase of swabs and thelaboratory testing, subject to a ceiling. All other costs
incurred, such as costs for sampling, travelling and
For the purposes of this Decision, ‘competent authority’ shall bethe authority or authorities of a Member State as designatedpursuant to Article 3 of Regulation (EC) No 2160/2003.
The financial contribution from the Community shouldbe granted provided that the survey is carried out in
accordance with the relevant provisions of Communitylaw and that certain other conditions are complied with,
Scope of the surveys
including transmission of results within prescribeddeadlines.
The Member States shall carry out the Salmonella
accordance with Parts A and B of Annex I until 31 December2008.
For reasons of administrative efficiency all expenditurepresented
The Member States shall carry out the MRSA survey in
Community should be expressed in EUR. In accordance
accordance with Parts A and C of Annex I until 31 December
with Council Regulation (EC) No 1290/2005 of 21 June
(1) The EFSA Journal
(2007) 129, 1-14.
(3) OJ L 209, 11.8.2005, p. 1. Regulation as last amended by Regu-
lation (EC) No 1437/2007 (OJ L 322, 7.12.2007, p. 1).
the financial contribution to be paid by the Community shallbe reduced by 25 %.
Performance of sampling and analyses
Sampling and analysis shall be performed by the competentauthority or under its supervision in accordance with the
If the final report is submitted after 30 April 2009 but before
technical specifications set out in Annex I.
31 May 2009, the financial contribution shall be reducedby 50 %.
No financial contribution shall be paid if the final report is
Conditions for granting a Community financial
The financial contribution from the Community towards
the costs of analyses pursuant to this Decision shall be grantedto the Member States up to the maximum total amount for co-
Maximum amounts to be reimbursed
financing set out in Annex II to this Decision for the durationof the surveys provided for in this Decision.
The maximum amounts of the financial contribution from
the Community towards the costs to be reimbursed to theMember States for analyses covered by the Salmonella
surveyshall not exceed the following:
The financial contribution from the Community provided
for in paragraph 1 shall be granted to the Member Statesprovided that the Salmonella
and MRSA surveys are im-
(a) EUR 20 per test for bacteriological detection of Salmonella
plemented in accordance with the relevant provisions of
Community law, including the rules on competition and onthe award of public contracts, and subject to compliance withthe following conditions:
(b) EUR 30 for serotyping of the relevant isolates.
The maximum amounts of the financial contribution from
(a) the national laws, regulations and administrative provisions
the Community towards the costs to be reimbursed to the
required to implement the survey must enter into force on
Member States for analyses covered by the MRSA survey shall
the day of application of this Decision at the latest;
(b) a progress report containing the information listed in Part D
(a) EUR 30 per test for bacteriological detection of MRSA;
of Annex I and covering the first three months of thesurveys must be submitted to the Commission by 31 May2008 at the latest.
(b) EUR 8 per identification of the presence of MRSA by PCR;
(c) EUR 25 per Staphylococcus
type A typing (Spa typing);
(c) a final report on the implementation of the surveys,
together with supporting evidence for the costs incurredby the Member States for the analyses and the results
(d) EUR 150 per multi locus sequence typing (MLST) of
attained during the period from 1 January 2008 to 31
December 2008 must be submitted to the Commissionby 31 March 2009 at the latest;
(d) the surveys must be implemented effectively.
Collection of data, assessment and reporting
The supporting evidence for the costs incurred as referred to in
The competent authority responsible for preparing the
point (c) of the second paragraph shall comprise at least the
yearly national report pursuant to Article 9(1) of Directive
Council (1) shall collect and assess the results of the surveysand forward them to the Commission.
If the final report referred to in paragraph 2(c) is
(1) OJ L 325, 12.12.2003, p. 31. Directive as amended by Council
submitted after 31 March 2009 but before 30 April 2009,
Directive 2006/104/EC (OJ L 363, 20.12.2006, p. 352).
The Commission shall forward the national data and the
assessment referred to in paragraph 1 to the European FoodSafety Authority, which shall examine them.
This Decision shall apply from 1 January 2008.
National data and results shall be made publicly available
in a form that ensures confidentiality.
Conversion rate for expenditure
This Decision is addressed to the Member States.
Where a Member State’s expenditure is in a currency other thaneuro, the Member State concerned shall convert its expenditureinto euro by applying the most recent exchange rate set by theEuropean Central Bank prior to the first day of the month inwhich the application for the financial contribution from the
Community is submitted by the Member State.
Repeal of Decision 2007/636/EC
Decision 2007/636/EC is hereby repealed.
TECHNICAL SPECIFICATIONS REFERRED TO IN ARTICLE 3, ARTICLE 4 AND ARTICLE 5(2)(b)
Part A: Overview and sampling frame
Overview of the survey
The survey shall be performed according to the overview in Figure 1.
Overview of the survey
The survey shall be carried out on holdings harbouring at least 80 % of the breeding pig population in a MemberState. Preferentially holdings having 50 breeding pigs or more shall be sampled. However, if those holdingshaving 50 breeding pigs or more do not contain 80 % of the national herd of breeding pigs, then smallerholdings with less than 50 breeding pigs shall also be sampled.
Holdings with breeding pigs shall be classified either as ‘breeding holdings’ or as ‘production holdings’. Breedingholdings sell gilts and/or boars for breeding purposes. Typically, they sell 40 % or more of the gilts that they rearfor breeding whilst the remainder are sold for slaughter. In contrast, production holdings mainly sell pigs forfattening or slaughter.
and MRSA prevalence must be measured separately in breeding holdings (Part 1 of the Salmonella
and MRSA surveys) and in production holdings (Part 2 of the Salmonella
and MRSA surveys), representing theherds as indicated in Figure 2, but excluding weaner to finisher herds.
Overview of holdings
The sample and the sampling strategy
Both parts of the Salmonella
and MRSA surveys shall have a similar two-stage sampling design. In the first stage,a random sample of holdings shall be selected in every Member State from the breeding holdings and a secondrandom sample shall be selected from the production holdings group. The number of holdings required isdiscussed in section 2.3. In the second stage, a number of pens shall be selected for sampling within everyselected holding (see Section 2.2.2).
F i r s t s t a g e : s e l e c t i o n o f h o l d i n g s
Each Member State must create two sampling frames. The first shall list all eligible breeding holdings (usually,those holdings with at least 50 breeding pigs — see Section 2.1) and the second shall list all eligible productionholdings. The required number of holdings for each part of the Salmonella
and MRSA surveys will then beselected at random from each list. A random sample is intended to ensure that the surveys include holdings witha range of herd sizes and from all regions of a Member State where pig-keeping is practised. It is recognised thatin some Member States, there may be a few holdings (e.g. fewer than 10 % of all eligible holdings) with a verylarge herd size. Thus, random selection may by chance result in none of these very large herds being selected. AMember State may use a stratification criterion prior to holding selection — for example, to define a stratumcontaining the 10 % of largest herds and to allocate 10 % of the required sample size to this stratum. Similarly, aMember State may stratify the sample across administrative regions according to the proportion of eligible herdswithin each region. Any stratification that is considered should be described in the report that a Member Statesubmits to the Commission in accordance with Part D (1).
If a selected holding cannot be sampled (for example, if it no longer exists when sampling is carried out) a newholding shall be selected at random from the same sampling frame. If any stratification (e.g. on herd size orregion) was in operation, then the new holding should be selected from the same stratum.
The primary sample size (number of holdings to be sampled) shall be approximately equally distributed over theyear to cover the different seasons, as far as possible. Samples shall be taken from approximately one twelfth ofthe number of holdings each month.
Outdoor holdings must be included in the survey but there shall be no mandatory stratification on thisproduction type.
S e c o n d s t a g e : s a m p l i n g o n t h e h o l d i n g
In each selected breeding herd and production herd the pens, yards or groups of breeding pigs over six monthsof age to be sampled will be randomly selected.
The number of pens, yards, or groups to be sampled must be proportionally allocated according to the numbersof breeding pigs in the different stages of production (pregnant, non-pregnant, and other categories of breedingpigs). The exact age categories to be sampled are not prescribed, but this information shall be collected duringthe sampling.
Breeding pigs that have arrived recently to the herd and are held in quarantine shall not be included in theSalmonella
and MRSA surveys.
P r i m a r y s a m p l e s i z e ( f i r s t - s t a g e s a m p l e s i z e )
A regular primary sample size calculation shall be conducted for the breeding holdings and a second regularprimary sample size calculation shall be conducted for the production holdings. The primary sample size shall bethe number of breeding holdings to be sampled and the number of production holdings to be sampled in eachMember State and it shall be determined taking into account the following criteria, assuming simple randomsampling:
(a) the total number of breeding holdings (breeding holdings, Part 1 of the Salmonella
and MRSA surveys);
(b) the total number of production holdings (production holdings, Part 2 of the Salmonella
and MRSA surveys);
(c) annual expected prevalence (p): 50 %;
(d) desired confidence level (Z): 95 %, corresponding to a Zα value of 1,96;
The calculation shall be conducted firstly, for the breeding holdings and secondly, for the production holdings. Ineach case, the assumptions in steps c-e above are the same.
For practical purposes, if there are 100 000 or more holdings in either the breeding herds sampling frame or theproduction herds sampling frame then that population can be considered to be infinite and the number ofholdings to be randomly selected from that sampling frame is 171 (see Table 1). Where the number of breedingherds or production herds is less than 100 000 a finite population correction factor is applied and fewerholdings need to be sampled as shown in Table 1.
As an example, if there are in a Member State 1 000 holdings belonging to the production holding group and250 holdings belonging to the breeding holding group, 147 holdings must be sampled in the productionholding group and 102 holdings must be sampled in the breeding holding group.
Number of holdings with breeding pigs to sample in either part of the Salmonella and MRSA surveys as
a function of the finite population size (total number of holdings with breeding pigs in the Member
Number of holdings with breeding pigs (N)
Sample size (n) for finite population size 7,5 % accuracy
Non-response shall be anticipated e.g. by increasing the sample size in each group by 10 %. Any unsuitableholding shall be replaced in the process of the Salmonella
and MRSA surveys (see Section 2.2.1).
In case an estimation of the number of ‘breeding holdings’ is not possible prior to the start of the survey, anumber of holdings shall be selected for sampling as in Table 1 based on the total number of holdings withsows (X holdings). The number of holdings to be sampled shall be increased by at least 30 % ((X + 30 %)holdings). Prior to the survey, the competent authority shall identify a number of breeding holdings, equal toat least this additional 30 %. While visiting the farms, the holding will be classified as breeding or productionholding according to the definitions above.
S e c o n d a r y s a m p l e s i z e ( s e c o n d - s t a g e s a m p l e s i z e )
In each selected holding samples shall be collected from 10 randomly chosen pens, yards, or groups of breedingpigs. If necessary (for example, in farrowing accommodation or where sows are kept in small groups of less than10 individuals), a group can consist of more than one pen. At least 10 individual breeding pigs should contributeto each routine Salmonella
However, where on smallholdings or holdings with large numbers of breeding pigs kept outdoors in paddocks,the number of pens, yards or groups is less than 10, sampling of the same pen, yard, or group shall be requiredso that a total of 10 routine Salmonella
samples are submitted.
Part B: Sample collection and analysis for the Salmonella survey
Sample collection in the herds
Type and detail of the routine sample
The material collected for bacteriological analysis shall be freshly voided faeces representing the whole holding,which is the unit of interest. Since every holding is unique, it shall be decided, before starting the sampling,which pens, yards, or groups within the holding are sampled. The sample collected shall be placed in a separatecontainer avoiding cross-contamination and sent to the laboratory.
Each pooled sample shall total at least 25 g and two approaches may be employed to collect these pooled faecessamples:
1. where there is an accumulation of mixed faeces within an area of a pen or yard, a large swab (e.g.
20 cm × 20 cm) can be used to pass through the faecal mass, ensuring that at least 25 g of mixedmaterial is collected. This may be achieved by for example, moving the swab along a 2-metre zig-zagpath such that it is well coated with faecal material. If necessary, for example due to hot weather or onslatted flooring, then the swab may be moistened with an appropriate liquid such as drinking water.
2. where there is no such accumulation, for example in a field, large yard, in a farrowing house, or pens or other
accommodation with low numbers of pigs per group, then individual pinches shall be selected from indi-vidual fresh faecal masses or places so that a minimum of 10 individuals, contribute to a total sample volumeof at least 25 g. The sites from which these pinches are collected should be distributed in a representativemanner across the area concerned.
Approach 1 shall be preferred where practical. In this approach at least 10 individual pigs must contribute toeach sample taken, otherwise approach 2 shall be applied.
Additional sampling for the within-holding prevalence study
Together, 10 holdings, selected at random from the overall sample of breeding holdings and of productionholdings are subjected to more intensive sampling. On these holdings 10 routine samples shall be collected inthe same manner as described previously (Section 2.1 of Part A). In addition, 10 individual samples of at least30 g shall be collected in each selected pen and shall be identified in such a manner that these 10 individualsamples can be associated with the routine sample from that pen. Thus in total, 10 routine samples and 100(10 × 10) individual samples shall be collected from each of these 10 holdings. The processing of these samplesis described in Section 2.3.1.
This sampling should be applied in Czech Republic, Denmark, Romania, Slovenia, Sweden and the UnitedKingdom.
All relevant information available from the sample shall be recorded on a sampling form produced by thecompetent authority to enable the data requirements in Part D to be fulfilled.
Each sample and its sample form shall be labelled with a unique number which shall be used from sampling totesting, and with the code of the pen. The competent authority must arrange for the issue and use of a uniquenumbering system.
Samples shall be preferably kept between + 2 and + 8 °C and free of external contamination during transpor-tation. The samples shall be sent to the laboratory as quick as possible within 36 hours by fast mail or courierand shall reach the laboratory no later than 72 hours after sampling.
Laboratory analytical methods
Analysis and serotyping shall take place at the National Reference Laboratory (NRL). Where the NRL does nothave the capacity to perform all analyses or if it is not the laboratory that performs detection routinely, thecompetent authorities may decide to designate a limited number of other laboratories involved in official controlof Salmonella
to perform the analyses. These laboratories shall have proven experience of using the requireddetection method and have a quality assurance system complying with ISO 17025 and be submitted to thesupervision of the NRL.
At the laboratory, samples shall be kept refrigerated until bacteriological examination, which shall preferably becarried out within 24 hours after receipt but in any case no later than 96 hours after the sample was collected.
Member States shall guarantee that all involved parties have been sufficiently trained to carry out the analyses.
In the laboratory, routine samples shall be mixed carefully and thoroughly, before 25 g is collected for analysis.
For evaluation of the within-holding prevalence in accordance with paragraph 1.2, each of the individualcollected samples (30 g) needs to be divided into two parts. One part, weighing at least 25 g shall be mixedcarefully and thoroughly and subsequently cultured individually. The remaining second part is to be used toprepare an artificially pooled sample from the 10 individual samples in the selected pen, group or yard. Thislatter part shall be prepared by adding 10 times 2,5 g of the individual samples to create an artificially pooledsample of 25 g. The artificially pooled samples are mixed carefully and thoroughly before analysis. In total, 10routine samples, 10 artificially pooled sample and 100 individual samples shall be analysed from each of the 10holdings selected for the estimation of the within-holding prevalence.
D e t e c t i o n a n d i d e n t i f i c a t i o n m e t h o d s
The method recommended by the Community Reference Laboratory (CRL) for Salmonella
in Bilthoven, theNetherlands, shall be used. This method is described in the Annex D of ISO 6579: ‘Detection of Salmonella
spp. in animal faeces and in samples of the primary production stage’. The latest version of Annex D shall beused.
All strains isolated and confirmed as Salmonella
spp. shall be serotyped according to the Kaufmann-Whitescheme, by the NRL for Salmonella
For quality assurance, 16 typable strains and 16 non-typable isolates shall be sent to the CRL for Salmonella
. Aproportion of these isolates shall be sent to the CRL on a quarterly basis. If fewer strains have been isolated, allshall be sent.
220.127.116.11. Phage typing of Salmonella
In case isolates of Salmonella Enteritidis
and Salmonella Typhimurium
are phage typed (optional), the methodsdescribed by the WHO reference centre for phage typing of Salmonella
of the Health Protection Agency (HPA)Colindale, London, shall be used.
Part C: Sample collection and analysis for the MRSA survey
Type and detail of sample
Five dust samples shall be gathered using five dry sterile swabs of about 500 cm2 each from five of the 10 pensselected for sampling under part A. These five pens shall be chosen in a way that breeding pigs in differentproduction stages are included. For each pen dorsal surfaces of pen partition walls shall be swabbed. In case thereis not enough dust present, then ventilator ducts etc. shall be sampled in addition. After use, the soiled swabshall be placed in a sterile plastic bag.
The creation of aerosol in the building shall be avoided during sampling.
Each sample and its sample form shall be labelled with a unique number which shall be used from sampling totesting. The competent authority shall arrange for the issue and use of a unique numbering system.
Samples shall be kept at constant temperature between + 2 °C and 25 °C (room temperature) and free of externalcontamination during storage and transportation. The samples shall be sent to the laboratory as quickly aspossible and reach the laboratory no later than 10 days after sampling.
Laboratory analytical methods
Analysis and subtyping of MRSA shall take place in experienced laboratories. This shall preferably be theNational Reference Laboratories (NRL’s) for Staphylococcus aureus
and/or antimicrobial resistance in the MemberStates. In case the NRL does not have the capacity or experience to perform the analyses or if it is not thelaboratory that performs detection routinely, the competent authority shall decide to designate other experiencedlaboratories or a NRL in another Member State to perform the analyses. These laboratories shall have provenexperience of using the required methods and have an accreditation system in place according to ISO 17025. Anup-to-date list of authorised laboratories can be consulted on the website of the Community ReferenceLaboratory for antimicrobial resistance (CRL-AR) in Copenhagen, Denmark.
Samples arriving 10 days after sampling shall be discarded unless bacteriological examination can be startedwithin 13 days. At the laboratory, samples shall be kept at a constant temperature between + 2 °C and 25 °Cuntil bacteriological examination, which shall be carried out within 13 days after sampling.
In the laboratory the five dust swabs shall be pooled in a 100 ml of Mueller-Hinton broth supplemented with6,5 % NaCl and incubated at 37 °C for 16-20 h. One millilitre of this shall then be inoculated into 9 ml TryptoneSoy Broth + 3,5 mg/l cefoxitin and 75 mg aztreonam and shall be incubated for a further 16-20 h at 37 °C. Oneloop-full of this shall then be spread onto a chromogenic agar selective for MRSA and incubated for 24-48 h at37 °C. The specific agar recommended by the CRL-AR shall be used. This agar is described in the CRL-ARwebsite.
Based on colony morphology and colour, up to five colonies indicative for being MRSA shall be subcultivated onblood agar. Presumptive Staphylococcus aureus (S. aureus)
shall at this stage either be stored under appropriateconditions (– 80 °C) for later identification and characterisation or processed immediately.
Presumptive S. aureus
shall be identified as S. aureus
and MRSA by PCR. The identification shall be performedusing a multiplex PCR with simultaneous identification of the mecA-gene or two different PCR shall beperformed. To limit the amount of work only one of the five presumptive S. aureus
isolates shall initially beidentified. If this isolate is identified as MRSA, it shall be stored. No further testing of the remaining four isolatesis required if the first isolate is identified as MRSA and they can be discarded. If the first isolate is not identifiedas MRSA, the next of the initial five isolates shall be tested. This process shall continue until one MRSA has beenidentified or all five isolates have been tested. Alternatively, identification by PCR as a first step can be done on apool of the five presumptive colonies from a sample. In case of a positive PCR, the analysis shall be repeated onindividual colonies to identify a positive colony.
For quality assurance, 16 presumptive S. aureus
isolates which were not identified as MRSA, as well as 16 MRSAstrains, sampled over the whole year 2008 shall be sent to the CRL-AR. A proportion of these isolates shall besent to the CRL-AR on a quarterly basis. In case less than 16 isolates were verified as MRSA all these isolatesshall be sent.
S u b t y p i n g f o r p o s s i b l e l i n k t o h u m a n i s o l a t e s
Positive MRSA’s shall be tested for Staphylococcus
type A (Spa-typing). The typing shall be performed at the NRLor under its supervision, or isolates shall be forwarded to the CRL-AR, which shall then perform the typing.
On a subset of representative isolates (about 2 % of the number of pooled samples) MLST-typing shall beperformed by the NRL or the CRL-AR.
A n t i m i c r o b i a l s u s c e p t i b i l i t y t e s t i n g
Antimicrobial susceptibility testing is optional. If carried out, MRSA isolates shall be tested for antimicrobialsusceptibility using micro-dilution at least to the following antimicrobial agents: ciprofloxacin, erythromycin,fusidic acid, gentamicin, linezolide, mupirocin, sulphametoxazole, trimethoprim, tetracycline, chloramphenicol,vancomycin and quinupristin/dalfopristin. Reporting on antimicrobial susceptibility shall be carried out inaccordance with Article 9(1) of Directive 2003/99/EC.
The isolates shall be stored at the National Reference Laboratories (NRL’s) using the method for NRL culturecollection ensuring viability and no changes in properties of the strains for a minimum of five years. This is inorder to allow, for instance, later testing for antimicrobial susceptibility or other types of characterisation. Alsoisolates sent to the CRL-AR shall be stored for a minimum of five years. Isolates shall be stored under conditionsnot allowing changes in properties (– 80 °C). If the laboratory in charge do not have the available storagecapability, isolates shall be forwarded to the CRL-AR, which shall store these isolates.
Reporting from the laboratories
All analytical results shall be sent on a confidential basis only from the laboratory to the competent authority ofthe Member States where the dust samples were collected.
Part D: Reporting from the Member States
Overall description on the implementation of the Salmonella and MRSA surveys
A report in text format shall include at least:
(b) a description of the population of holdings with breeder pigs:
(iii) total number of multiplier holdings;
(iv) number of breeder holdings planned to be sampled, and number of breeder holdings actually
sampled; number of holdings planned for sampling but not sampled and the reason therefore;
(v) comments on the overall representativeness of the breeding holdings sampling programme;
(ii) total number of farrow to weaner/grower holdings;
(iii) total number of farrow to finish holdings;
(iv) number of production holdings planned to be sampled, and number of production holdings actually
sampled; number of holdings planned for sampling but not sampled and the reason therefore;
(v) possible comment on overall representativeness of the production holdings sampling programme;
(c) number of samples from the Salmonella
survey obtained and analysed:
(iii) from holdings sampled for within-holding prevalence study;
(d) overall results from the Salmonella
(i) prevalence of breeding holdings and of production holdings infected with Salmonella
, and serovars of
(ii) outcome of within-holding prevalence study;
(e) list of laboratories responsible in the Salmonella
(iii) phage typing (if carried out).
(f) number of samples from the MRSA survey obtained and analysed:
(g) overall results from the MRSA survey: prevalence of breeding holdings and of production holdings infected
with MRSA, based on detection and confirmation by PCR;
(h) list of laboratories responsible in the MRSA survey for:
Complete data on each holding sampled and corresponding tests results:
The Member States shall submit the results of the Salmonella
and MRSA surveys electronically to the Commissionin the form of raw data using a data dictionary and data collection requirements established and provided by theCommission.
The following information shall be collected in Member States and transmitted to the Commission for eachholding selected for sampling:
(i) indoor versus ‘any stage of the production kept outdoors’;
(ii) nucleus, multiplier, farrow to weaner, farrow to finish and farrow to grower;
(c) holding size: the number of breeding pigs present at the time of sampling (adult inventory);
(d) replacement policy: all replacement breeding pigs purchased; some replacement breeding pigs homebred or
(e) (voluntarily) clinical symptoms of diarrhoea: were there symptoms of diarrhoea within the three months
Information on all samples collected within the Salmonella survey
The following information shall be collected in Member States for each sample sent to the laboratory within theframe of the Salmonella
(b) code of the laboratory involved in initial analysis;
(e) detection of Salmonella
: qualitative result (positive/negative);
serotyping of Salmonella
: serovar(s) detected (may be more than one);
(g) age of the pigs: all gilts versus mixed age breeding pigs;
(h) sex: only sows; sows and boars or only boars;
production stage: maternity; mating, gestation (other?);
housing: slatted floor (fully/partly); solid floor; deep straw or other;
(k) diet: are pigs in this pen, yard or group fed compound feed exclusively;
feed supplement: is there any Salmonella
reducing substance added to the feed (like organic acid, a probiotic);
(m) systematic use of antibiotics: are antibiotics used in all animals of this group by any route of administration;
(n) the last date of administration of antimicrobials to the animals (within the last four weeks).
Additional information on the samples collected within the Salmonella survey for the within holding prevalence
The following addition information shall be collected in Member States for each individual sample sent to thelaboratory within the frame of the sampling for the within-holding prevalence:
(b) detection of Salmonella
in each individual sample: qualitative result (positive/negative);
(c) serotyping of Salmonella
in each individual sample: serovar(s) detected (may be more than one).
Information on the samples collected within the MRSA survey
The following information shall be collected in Member States and for each sample sent to the laboratory:
(b) the code/name of the laboratory involved in the detection;
(e) the result of detection of MRSA (positive/negative);
(f) the code/name of the laboratory involved in the PCR;
(h) the code/name of the laboratory involved in the Spa typing;
(j) code/name of the laboratory involved in the MLST-typing,
MAXIMUM COMMUNITY FINANCIAL CONTRIBUTION TO THE MEMBER STATES REFERRED TO IN
Maximum total amount for co-financing of analyses
CERTIFIED FINANCIAL REPORT ON THE IMPLEMENTATION OF THE BASELINE SURVEY ON THE
PREVALENCE OF SALMONELLA SPP. AND MRSA IN HERDS OF BREEDING PIGS
— . to . For the Salmonella
Statement on costs incurred in the survey and eligible for Community financial contribution:
Reference number of Commission Decision providing Community financial contribution:
Total costs of testing and swabs incurred
Costs incurred related to functions at/by
Bacteriology for Salmonella
Declaration by the beneficiary
— the costs listed above are genuine and have been incurred in carrying out the tasks laid down in Decision 2008/55/EC
and were essential for the proper performance of those tasks,
— all supporting documents for those costs are available for audit purposes,
— no other Community financial contribution has been requested for these surveys.
Hacking and Extending ACL2 ∗ ABSTRACT smaller, ACL2 authors were more able to adapt and ex-The ACL2 theorem prover provides the user a wide rangetend the system itself to suit user needs. A new reality hasof mechanisms for customization and extension while pre-set in, in which a growing number of sophisticated users areserving soundness. ACL2 researchers and power users, how-seeking t
Prevalence of Antimicrobial Resistances in Streptococcus pneumoniae Isolates in Australia, 2005: Report from the Australian Group on Antimicrobial Resistance (AGAR) South Metropolitan Area Thomas Gottlieb1, Peter Collignon2, Jennifer Robson3, Julie Pearson4, Jan Bell5, and the Australian Group on Antimicrobial Resistance 1Department of Microbiology and Infectious Diseases, Conc