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Dr. Vicent Yusà. [email protected] / [email protected]
CONTENTS
Introduction
European Framework
Organization of the Monitoring System in Spain
Sampling PlanSampling strategyRepresentativeness of samplingSampling procedures
Analytical Methods
Results (2014)
Conclusions
USE OFANTIMICROBIALS
IN FARMS
Presence of Antimicrobials
Residues in food at levels > MRL
AMRChemical hazard
BiologicalHazard
USE OF ANTIMICROBIALS: A FOOD SAFETY ISSUE
Very few non-compliant samples Low riskHigh budget
Raising threat for public health.
Very low budget
~ 25, 000 isolates (2014)
~123, 000 samples (2014)
Control of
Veterinary Residues in
Food
HORMONS
(88.600 samples)
β-AGONISTS
(41,300 samples)
ANTIBACTERIALS
(122,959 samples)
Prohibited substances
(80,570 samples)
Others
(100.392)
Monitoring of veterinary medicinal product residues in food (2014)
6600
18221593
2038
629367 342
208 179 89
0
1000
2000
3000
4000
5000
6000
7000
Pigs Bovines Poultry Sheep andgoats
Milk Rabbit meat Eggs Honey Aquaculture Horse
Number of total samples for antibacterial residues in food (Spain 2014). Total: 13,947
MONITORING OF ANTIBACTERIAL RESIDUES (B1) (Directive 96/23/EC)
EUROPE: 122,959 samples analysed for antibacterials; only 0.18 % non-compliant
Nº isolates for AMR: ~ 1000 (2014)
0,15
0,27
0,19
0,93
0,16
0,27
0
0,96
0
0
0 0,2 0,4 0,6 0,8 1 1,2
Pigs
Bovines
Poultry
Sheep and goats
Milk
Rabbit meat
Eggs
Honey
Aquaculture
Horse
Europe Spain
Non-compliant samples (%) for antibacterials (B1). 2014
MONITORING OF ANTIBACTERIAL RESIDUES (Directive 96/23/EC)
ANIMAL Substance Number samples
Non-compliant
BOVINES Amoxycillin (P) 628 2 (0.3 %)
Benzylpenicillin(Penicillin G)
6 1 (16.7%)
Doxycycline (T) 556 1 (0.2 %)
Enrofloxacin (FQ) 596 1 (0.2%)
Oxytetracycline (T) 642 2 (0.3%)
PIGS Amoxicillin (P) 1910 1 (0.1%)
Doxycycline (T) 1947 7 (0.4 %)
Lincomycin (L) 1314 1 (0.1%)
Oxytetracycline (T) 1948 1 (0.1%)
POULTRY Doxycycline (T) 290 2 (0.7 %)
Enrefloxacin (FQ) 250 1 (0.4 %)
MILK Spiramycin (M) 319 1 (0.3%)
Antibacterials (B1) detected in non-compliant samples . Spain, 2014
THERE IS A LOW EXPOSURE TO ANTIMICROBIALS RESIDUES FROM THE DIET LOW RISK FOR THE GENERAL POPULATION
CONTROL OF VETERINARY RESIDUES IN FOOD
(Directive 96/23/EC)
AMR in food(Decision 2013/652/EU)
ANTIMICROBIALS IN FOOD SAFETY
FARM ABATTOIR RETAIL
Modules of the food (meat) chain
ANTIBIOTICS7982 tonnes
/year (EU)
Antibiotic Resistant Bacteria in animals and food
Foodborne pathogens: Salmonella
Campylobacter
Indicator bacteria:
E.Coli; Enterococci
How muchFOOD CHAIN
contributesto
resistance transfer
from animals to
human?
Reservoir of resistance genes with potential for transferring resistance to pathogenic bacteria.
Permit a more accurate study of the trends and the effects of patterns of antimicrobials use (all animal carry them)
Infection agents transferred from animals to human
One Health
ANTIBIOTICS in humans: 3399 tonnes per year (EU)
RESISTANT BACTERIA IN HUMANS (≥25 % o more)25.00 people die each year in Europe
The link is well documented
European Surveillance of Veterinary Antimicrobial Consumption (ESVAC). October 2016population correction unit (PCU)
SALES
AMR monitoring should cover both zoonotic agents (Salmonella and Campylobacter, and indicator organisms of the commensal flora
.
Commensal bacteria that contaminate food may be also considered a potential AMR hazard, as they can harbour transferable resistance genes
LEGAL AND TECHNICAL REFERENCES
Legal Reference Title
Reg. 882/2004 Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 on official controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules
Dec. 2013/652/EU 2013/652/EU: Commission Implementing Decision of 12 November 2013 on the monitoring and reporting of antimicrobial resistance in zoonotic and commensal bacteria
Dir. 2003/99/EC Directive 2003/99/EC of the European Parliament and of the Council of 17 November 2003 on the monitoring of zoonoses and zoonotic agents, amending Council Decision 90/424/EEC and repealing Council Directive 92/117/EEC
Reg. 2073/2005 Commission Regulation (EC) No 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs
EU framework
EFSA Guidance Documents
EFSA. 2012 - Technical specifications on the harmonised monitoring and reporting of antimicrobial resistance in Salmonella, Campylobacter and indicator Escherichia coli and Enterococcus spp. bacteria transmitted through food.In EFSA Journal. http://www.efsa.europa.eu/en/efsajournal/pub/274.htm
EFSA. 2012 - Technical specifications for the analysis and reporting of data on antimicrobial resistance (AMR) in the European Union Summary Report.In EFSA Journal. http://www.efsa.europa.eu/en/efsajournal/pub/2587.htm
EFSA. 2014 - Technical specifications on randomised sampling for harmonised monitoring of antimicrobial resistance in zoonotic and commensal bacteria.In EFSA Journal. http://www.efsa.europa.eu/en/efsajournal/pub/3686.htm
EFSA. 2015 - Data dictionaries-guidelines for reporting data on zoonoses, antimicrobial resistance and food-borne outbreaks using the EFSA data models for the Data Collection Framework (DCF) to be used in 2015, for 2014 data.In EFSA. http://www.efsa.europa.eu/en/supporting/doc/776e.pdf
EFSA. 2015 - Manual for reporting on antimicrobial resistance within the framework of Directive2003/99/EC and Decision 2013/652/EU for information deriving from the year 2014.In EFSA. http://www.efsa.europa.eu/en/supporting/pub/771e.htm
PREVALENCE OF FODDBORNE AMR
DETERMINATION OF TRENDS AND SOURCES OF
AMR
DATA FOR CONDUCTING RISK ANALYSIS
(humans and animals)
INFORMATION FOR EVALUATING AM
PRESCRIPTIONS AND PRACTICES
MONITORING OF AMR IN
FOOD
EU framework
HARMONIZED MONITORING
of AMR
COMBINATIONS
OF
BACTERIAS
ANIMALS/FOODS
SUSCEPTIBILITY TESTING METHOD
(phenotypic methods; panel
of antimicrobials)
SAMPLING PLANS
REPORTING OF THE DATA
Decision 2013/652/EU
EU framework
Randomised sampling strategy
(reduction of sampling bias)
Representativeness
(data from a sample reflect
the population under study)
Stratified
sampling approach
(allocation of the sample numbers per strata: slaughterhouses, NUTS-3,
laboratories)
Sampling time
Even distribution of collected samples (quarters)
Sampling strategy(Prospective sampling/Retrospective
sampling)
QUALITY OF THE SAMPLING
Ministry of
Agriculture, Food
and Environment
(MAGRAMA)
Ministry of Health
(MSSSI)
SGSE (PIFs)
AGRICULTURAL DEPARTMENT OF
AACC
HEALTH DEPARTMENT OF
AACC
AUTONOMOUS
COMMUNITIES
(AACC)
Official control of primary production and food quality
Official control of food safety after primary production
Sallmonella andESBL-E.Coli
sampling at slaughterhouses
and retail
Monitoring of AMR in primary production including caecalsamples at slaughterhouses
ORGANIZATION OF THE MONITORING SYSTEM
CENTRAL AUTHORITIES
COMPULSORY SAMPLING PLAN 2014-2020(domestic production)
Year Animal populations/Type of meat
Where to collect Salmonella(number of ISOLATES)
ESBL-or AmpC- producing E.Coli(number of samples)
2014, 2016, 2018, 2020
Broilers Slaughterhouse Carcases (170)
Retail Fresh meat (300)
Fattening turkeys
Slaughterhouse Carcases (170)
2015,2017, 2019
Fattening pigs Slaughterhouse Carcases (170)
Retail Meat (300)
Calves under 1 year
Slaughterhouse Carcases (170)
Retail Meat (300)
SALMONELLA isolates from carcasses
at slaughterhouses(≥60% of the specific
processed population)
ESBL-AmpC producing E. COLI
from meat at retail
Retrospectivesampling strategy
Isolates from official samples
collected to verify FBO compliance
withProcess Hygiene
Criteria(R 2073/2005)
AntimicrobialSusceptibility Testing (AST)
Sampledby FBO
(R. 2073/2005)
FBO: food business operators
Collected by Competent Authorities
If not sufficient
Prospective sampling plan
Fresh meat (chilled meat) including wrapped,
vacuum-wrapped/controlled
atmosphere
SAMPLING STRATEGY
Food category Micro-organisms
Sampling plan (1)
Limits Analytical referencemethod
Stage where thecriterion applies
2.1.3 Carcasses of cattle
Salmonella n=50 (5)c=2 (6)
Absence in the area tested per carcase
EN/ISO 6579 Carcases after dressingBut before chilling
2.1.4. Carcases of pig
Salmonella n=50 (5)c=5 (6) Absence in the
area tested per carcase
EN/ISO 6579 Carcases after dressingbut before chilling
2.1.5 Poultry carcases of broilers and turkeys
Salmonella n=50 (5)c=7 (6) Absence in 25
g of a pooled sample of neck skin
EN/ISO 6579 Carcases after chilling
(1) n = number of units comprising the sample; c = number of sample units giving values between m and M.(5) The 50 samples are derived from 10 consecutive sampling sessions in accordance with the sampling rules and frequencies laid down in this Regulation ; (6) The number of samples where the presence of salmonella is detected. The c value is subject to review in order to take into account the progress made in reducing the salmonella prevalence. Member States or regions having low.
SAMPLING at SLAUGHTERHOUSES (R. 2073/2005)
Isolation of Salmonella
EN/ISO 6579
SALMONELLA
Send to
AntimicrobialSusceptibility Testing (AST)(ISO 20776)
Public Health Laboratories of the AACC ISO 17025
National Reference Laboratory (CNA)(ISO 17.025)
SEROTYPES (Typhimurium, Enteritidis,…)
Broilers, turkeys.
Pig and cattle
SAMPLING IN ABATTOIR(ISO 17604)
ISOLATION (EN/ISO 6579)
GEOGRAPHICAL DISTRIBUTION OF SAMPLES OF PIG AND CATTLE CARCASES (300 + 300)
13896
100
116
32
24
24
24
22
24
The 600 samples from slaughterhouses distributed between 13 provinces (NUTS-3) that represent more than 60 % of total animal production(should cover at least 60% of the animal production)
National Reference Laboratory (CNA)(ISO 17.025)
Samples collected at retail byAACC inspectors
Send to NRL
< 24 hTransport : +2 ºC/ +8ºCBefore Date of ExpireEach sample from different lot
ESBL- E. Coli
SAMPLING at retail
Isolation (McConkey + cefotaxime) and specific identification
ESBL: extended spectrum β-lactamase
Antimicrobial Susceptibility Testing (AST)
GEOGRAPHICAL DISTRIBUTION OF ESBL-E.Coli FROM RETAIL (300 samples)
69
48
50
58
16
12
12
12
1212
The 300 samples from retail distributed between 13 provinces (NUTS-3) that represent more than 60 % of total population(should cover at least 80% of the population)
SAMPLING PROCEDURE FOR POULTRY CARCASES
EXAMPLES OF SAMPLING SESSIONS
CARCASES AFTER CHILLING
< 3.5 ºC
Carcasses shall be sampled at random during each sampling session and after chilling
1 sample: 25 g of a pooled sample of neck skin
Each sample contains neck skin from 3 carcases from the same flock (Epidemiological unit)
A minimum of 15 carcases shall be sampled at random during each sampling session
A piece of approx. 10 g from each carcase
Each sampling session includes 5 samples (n=5) from different flocks
Transport the samples in an insulated box with frozen freezer blocks
Transport: 1-8 ºC
Put the container with the sample in an plastic bag and close it with the official seal
NON-DESTRUCTIVE SAMPLING METHOD (ISO 17604): Abrasive sponge sampling
SAMPLING KIT
Peptone water (25 mL)
Before sampling
Moist the sponge in 10 mL of peptone water
After sampling
Pour the rest of the peptone water
SAMPLING PROCEDUE FOR PIGS
spongeSquare template
SAMPLING SITES(ISO 17604)
NON-DESTRUCTIVE SAMPLING METHOD (ISO 17604): Abrasive sponge sampling
Five carcases shall be sampled at random during each sampling session.Sampling sites (4) should be selected taking into account the slaughter technology used in each plant and the recommendations of ISO 17604
Template: 100 cm2
Press the template hard on to the surface.
Rub the sponge over the whole area 10 times in each direction (horizontally and vertically)
More than one site may be sampled using the same swab
NON-DESTRUCTIVE SAMPLING METHOD (ISO 17604): Abrasive sponge sampling
Five carcases shall be sampled at random during each sampling session.Sampling sites (4) should be selected taking into account the slaughter technology used in each plant and the recommendations of ISO 17604
ANALAYTICAL METHODS IN ROUTINE MONITORINGCLSI method (ISO 20776-1:2006)
Antimicrobial susceptibility testing (AST)
Antimicrobial susceptibility testing is performed with phenotypic methods.
The basis of phenotypic methods is the Minimum Inhibitory Concentration (MIC, mg/l).
DILUTION procedures are the reference methods for AST
DILUTION METHOD shall be performed according with ISO 20776-1:2006
Panel 1 of antimicrobial substances to be included in AMR monitoring
Epidemiological cut-off (mg/L)
ANTIMICROBIAL Salmonella E.Coli
Ampicillin >8 >8
Cefotaxamine >0,5 >0,25
Ceftazidime >2 >0.5
Meropenem >0.125 >0.125
Nalidixic acid >16 >16
Ciprofloxacin >0.064 >0.064
Tetracicline >8 >8
Colistin >2 >2
Gentamicin >2 >2
Trimethoprim >2 >2
Sulfamethoxazole NA >64
Chloramphenicol >16 >16
Azithromycin NA NA
Tigecycline >1 >1
Stepwise strategy for testing for specific monitoring of ESBL-producing Salmonella and E.Coli
14 antimicrobials
8antimicrobials
REPORTING
Data Collection Framework(EFSA)
Submission of data in the form of raw isolate-data in the electronic collection form provided by EFSA.The submitted reports include the compulsory information for each individual isolate
RESULTS (2014)
132 isolates from broilers 101 res ≥1 31 sus to all
Salmonella spp.- S. Kentucky-S. Enteritidis-S. Anatum
resistance rate (%)
RESULTS (2015)
128 isolates from pigs 104 res ≥1 24 sus to all
Salmonella spp.- S. Typhimurium-S. Derby-S. Rissen
Resistance rate (%)
RESULTS (2015)
E.Coli
ESBL: 27 (all res ≥1 )ESBL: 36 (all res ≥1)
301 isolates from pig meat299 isolates from bovine meat
resistance rate (%)resistance rate (%)
TAKE HOME MESSAGES
The sales of antibiotics in Spain for food producing animals are the highest in Europe (29) and need to be reduced
To ensure representativeness, the sampling program should be distributed evenly over the year
The samples taken at retail should include the necessary information to exclude isolates from the same repeated epidemiological unit (batch of product)
Increase the number of official labs accredited for AST
Increase the participation of food business operators with isolates from their own control
Include more zoonotic and commensal bacterial in the food monitoring program
Reduce ANTIMICROBIAL residues control, and INCREASE AMR monitoring