MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU

Miljø- og Fødevareudvalget 2015-16
MOF Alm.del Bilag 148
Offentligt

Annual Report on Zoonoses

in Denmark 2014

Annual Report on Zoonoses in Denmark 2014

Edited by:

Anne Wingstrand, Anna Irene Vedel Sørensen and

Birgitte Helwigh

Danish Zoonosis Centre

National Food Institute

Technical University of Denmark

Luise Müller

Statens Serum Institut

This is an official publication from the

National Food Institute, Technical

University of Denmark, the Danish

Veterinary and Food Administration, and

Statens Serum Institut.

Text and tables may be cited and reprinted only with

reference to this report.

Suggested citation:

Anonymous, 2015. Annual Report on Zoonoses

in Denmark 2014, National Food Institute,

Technical University of Denmark.

Reprints can be ordered from:

Danish Zoonosis Centre

National Food Institute

Technical University of Denmark

Mørkhøj Bygade 19

DK - 2860 Søborg

Denmark

Phone: +45 35 88 74 95

Fax:

+45 35 88 70 28

E-mail: [email protected]

Layout: Birgitte Helwigh og Susanne Carlson

Photos: Colourbox

Printing: GraphicCo.

2. edition with minor corrections

ISSN: 1600-3837

The report is also available at:

www.food.dtu.dk

MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU

Contents

Introduction

1. Trends and sources in human salmonellosis

2. Food- and waterborne outbreaks

3. Listeria

4. Verotoxin-producing

E. coli

(VTEC)

5. New insights in the European epidemiology of

Salmonella

and 20

Campylobacter

6. Vectorborne zoonoses

Status on National and EU targets

7.1 National targets

7.2 EU targets

8. International topics

8.1 Trichinella

8.2 Ebola and imported foods

9. Surveillance and control programmes

9.1 Surveillance of human disease

9.2 Outbreaks of zoonotic gastrointestinal infections

9.3 Surveillance and control of animals and animal products

9.4 Official testing of zoonotic pathogens in foodstuffs

Appendix

Trends and sources in human salmonellosis

Human disease and outbreak data

Monitoring and surveillance data

Monitoring and surveillance programmes

Population and slaughter data

MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU

Introduction

Overall, the number of human infections with

Campy-

lobacter

(3,782 cases) and

Salmonella

(1,122 cases) remains

at the same level in 2014 as in the last two years. Compared

to 2013, fewer foodborne outbreaks were registered in 2014

(60 outbreaks compared with 73), with a total of 2,209 cases

registered of which 295 were confirmed in the laboratory.

Although the number of human

Salmonella

cases was

similar to 2013, some changes occurred. The number

of infections with

Enteritidis was reduced by 22.5%

compared to 2013, where a large travel-related outbreak

with this serotype occurred, despite three outbreaks with

Enteritidis in 2014, including the first outbreak from

domestic eggs since 2009. Overall, human

Typhimurium

cases (incl. monophasic

1,4,[5],12:i:-) increased by 26.7%

in 2014 compared to the low level in 2013, primarily due to

an 81.1% increase in monophasic

1,4,[5],12:i:- infections

mainly related to five outbreaks. In four of these outbreaks,

the source was domestic pork or beef. Thus, in 2014 mo-

nophasic

1,4,[5],12:i:- became the second most common

human

Salmonella

serotype and for the first time exceeded

the number of genuine

Typhimurium infections.

In animals

1,4,[5],12:i:-has been more common than

genuine

Typhimurium in domestic pigs and pork since

2012, and in 2014, this serotype was also the most common

Salmonella

serotype in the domestic broiler production

and in imported pork.

An outbreak of

Agona running in 2013 and 2014

had an uncommon age and gender distribution. Extensive

investigations concluded that whey powder was a possible

outbreak source, but the final identification of the source

was not possible.

In the 2014

Salmonella

source account, Multi-Locus

Variable number tandem repeat Analysis (MLVA) was in-

troduced for subtyping of

Dublin similar to the existing

MLVA-typing of

Typhimurium and

Enteritidis. The-

refore, in this years’ report, top-10 MLVA types for these

serotypes are reported. The

Salmonella

source account 2014

attributed more cases to domestic pork, eggs and imported

broilers and fewer cases to imported pork and beef than in

2013. For the first time since 2011,

Salmonella

was detected

in the surveillance of domestic broiler meat, and human

cases were attributed to this source.

Several isolations of

Typhimurium DT40 and DT41

in the broiler and egg production from the end of 2013

and the beginning of 2014 were investigated. A common

source of introduction was not identified, and the impact

on human disease appeared to be limited.

Another characteristic of the year 2014 was a 84.0%

increase in Listeria

monocytogenes

cases including four

outbreaks, which are described in Chapter 3. In particular

one serious outbreak of listeriosis was in focus in 2014

with a total of 41 cases reported and a mortality of 41.4%.

Overall, the number of cases with VTEC and

Yersinia

enterocolitica

increased by 33.3% and 25.2%, respectively.

Increased awareness and improved diagnostic methods

play an important role in the increasing number of VTEC

cases. For both pathogens, outbreaks contributed to the

increase, but only for one small VTEC outbreak the source

of infection was identified (beef of Danish origin).

The number of norovirus outbreaks in 2014 was similar

to the number in 2013, while the number of patients in

these outbreaks more than doubled. An increasing number

of cases was caused by transmission of norovirus from

healthy carriers, ill persons among kitchen staff or kitchen

staff attending ill persons at home prior to handling food.

New initiatives on

Listeria

In 2014, Whole Genome Sequencing (WGS) was intro-

duced as the routine typing of

Listeria

isolates in Denmark,

and parallel on-time testing of human and food isolates

with WGS and Multi-Locus sequence Typing (MLST) was

introduced. Aided by WGS, the outbreak source of the

large and serious outbreak of

Listeria monocytogenes

in the

summer 2014 was identified to be “rullepølse”, a Danish

cold cut ready-to-eat speciality, from a specific producer.

Following the outbreak, the Danish Ministry of Food,

Agriculture and Fisheries initiated a critical review in or-

der to evaluate and improve the existing Danish

Listeria

efforts and initiatives, and new initiatives will be instigated

from 2015-2018 targeted for groups at risk and health care

The Annual Report on Zoonoses presents a summary of the trends and sources of zoonotic infections in

humans and animals, as well as the occurrence of zoonotic agents in food and feeding stuffs in Denmark in

2014. Greenland and the Faroe Islands are not represented. The report is based on data collected according to

the Zoonoses Directive 2003/99/EC, supplemented by data obtained from national surveillance and control

programmes as well as data from relevant research projects. Corrections to the data may occur after publication

resulting in minor changes in the presentation of historical data in the following year’s report. The report is also

available at www.food.dtu.dk.

Annual Report on Zoonoses in Denmark 2014

MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU

professionals, kitchens, food businesses and the official

control. Other initiatives support

Listeria

typing, tracing

and sampling. Additionally, several research initiatives

were propossed, including development of predictive tools,

studies of the genetic diversity and use of indicators for

Listeria

and studies of factors related to the high incidence

Listeria

in Denmark.

Risk ranking of human Verotoxin-producing

E. coli

Investigation of associations between the virulence

factors of human Verotoxin-producing

E. coli

(VTEC)

and occurrence of haemolytic uraemic syndrome (HUS)

in a Danish cohort of 2001 VTEC cases from 1983-2012

indicates, that primarily the verotoxin subtype

vtx2a

associated with HUS. The well-documented association

between

vtx2

and the gene for either attaching and effacing

properties (the

eae

gene) or the ability to colonize and

persist in the gut (regulated by the

aggR

gene) has resulted

in a basic and primary definition of HUS associated

coli

from humans for first line public health action:

vtx2

in a background of an eae or aggR positive

E. coli.

Such

isolates are

vtx

subtyped and further characterized. Other

VTEC are generally considered low risk and are treated

as other enteropathogens, although careful evaluation of

each patient is important in order to consider other risk

factors for progression to severe disease. This approach

is generally accepted and practical and easy to use in an

operational environment.

Campylobacter

and

Salmonella

seroincidence

As an alternative to defining the occurrence of human

infections with

Salmonella

and

Campylobacter

from the

incidence of registered culture confirmed cases which is

biased from the passive laboratory surveillance, Statens

Serum Institut, has calculated the seroincidence in the ge-

neral population from the levels of antibody classes against

Salmonella

and

Campylobacter

based on the kinetics of

the antibody decay. Measuring the seroincidences for a

selection of European countries and comparing these to

e.g. the national incidence of these infections, and the oc-

currence of

Salmonella

and

Campylobacter

in food animals

has provided new information on the exposure from these

sources and their relative importance.

Surveillance of vectors and vector-borne zoono-

ses

The National Veterinary Institute, Tecnical University

of Denmark, monitors vectors and vector borne diseases

in Denmark. Surveillance of mosquitoes and biting midges

is running and constitutes an early warning system for

vector borne diseases, which are currently not present in

Denmark. These diseases have been detected elsewhere

in Europe, even as close as in Germany. The surveillance

systems have identified new vectors, and DNA screening

of vectors using a new DNA screening chip has revealed

new pathogens carried by the vectors, and e.g. a widespread

occurrence of

Neoerlichia Mikurensis,

which has been as-

sociated with severe human disease in Sweden. Weekly

surveillance data are available at the internet.

National and EU targets on zoonoses

In this years’ report, chapter 7 gives an overview of the

targets for the national and EU action plans and programs

Salmonella

and

Campylobacter

and assess the achieve-

ment of the targets in 2014.

Whereas most targets were met in 2014, the occurrence

of specific targeted

Salmonella

serotypes in breeding flocks

Gallus gallus

exceeded the EU-target of a maximum of

1% positive flocks in 2014. From 4 out of 5 positive flocks

Typhimurium or

1,4,12:i:- were detected.

Assessment of Ebola risk from imported bush

meat

The outbreak of Ebola virus in West Africa in 2014, led

the European Food Safety Agency (EFSA) to assess the risk

from handling and consuming bush meat. Ebola is prima-

rily transmitted between humans from blood or bodily

fluids, but has a reservoir in certain wild animals, and a

large illegal import of bush meat to Europe occurs. EFSA

concluded, that the risk for introduction and transmission

of Ebola virus via bush meat to Europe is currently low,

although the size of the risk could not be estimated due to

lack of data and knowledge. The Danish border control is

aware of the risk.

Annual Report on Zoonoses in Denmark 2014

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1. Trends and sources in human

salmonellosis

By Leonardo de Knegt ([email protected]) and Tine Hald

Salmonella enterica

is an important cause of foodborne

disease in humans throughout the world and is a significant

cause of morbidity, mortality, and economic loss [1]. In 2014,

a total of 1,122 human cases of salmonellosis were reported

in Denmark, and the incidence was 19.9 cases per 100,000

inhabitants, which is similar to 2013. The incidence of

Typhimurium was 7.6/100.000 (Appendix Table A2), and

for the first time with a predominance of the monophasic

Typhimurium strains 1,4,[5],12:i:- (4.1/100.000) [2]. The

observed incidence for

Enteritidis was 4.8/100,000, which

is a decrease when compared to 2013 (6.2/100,000), where a

large travel related

Enteritidis outbreak took place.

Since the mid-nineties,

Salmonella

control strategies have

been guided by the results of a

Salmonella

source attribution

model (SSA). The routine application of such model has

helped the identification of the main animal-food sources of

human salmonellosis, as well as the monitoring of shifts in

importance among those sources throughout time. This has

identified broilers, pigs or laying hens as the main reservoirs

for this pathogen at different time points in the last 30 years,

thus allowing Danish risk managers to define different control

strategies [3].

The method currently used for source attribution in

Denmark compares the number of human cases caused

by different

Salmonella

subtypes with the distribution of

the same subtypes isolated from the various animal-food

sources. Additionally to serotyping of all

Salmonella,

isolates

Typhimurium and

Enteritidis have, since 2013, been

subtyped using multiple-locus variable-number tandem

repeat analysis (MLVA). MLVA-typing of

Dublin was first

presented by Kjeldsen et al. in 2014 [4], and is being used

for the first time in the

Salmonella

source account in 2014.

MLVA profiles for

Salmonella

are defined by the number of

repetitions observed in five independent loci, namely SE1,

SE5, SE2, SE9 and SE3 for

Enteritidis, STTR9, STTR5,

STTR6, STTR10 and STTR3 for

Typhimurium, and SE1,

SE2, SE5 and SD1 for

Dublin. To better fit source attri-

bution purposes, the level of discrimination of the original

schemes was adjusted by simplifying the MLVA profile for

Typhimurium to STTR9|STTR10|STTR3, and for

Dublin

to SE5|SE2|SE1, according to observations on loci stability

and the epidemiological value of MLVA discrimination [5,6].

The complete scheme SE1|SE5|SE2|SE9|SE3 was used for

Enteritidis. Finally, antimicrobial resistance profiles of

Typhimurium and

Dublin isolates was included, to further

distinguish between similar MLVA types. In the source ac-

count model, strains of

1,4,[5],12:i:- were separated from

genuine

Typhimurium.

8.0

Incidence per 100,000

Figure 1.1. Total incidence of human

salmonellosis and estimated human incidence due

to domestic broilers, pork, table eggs and imported

meat products in Denmark, 1988 to 2014

4.0

100.0

80.0

Incidence per 100,000

60.0

40.0

20.0

0.0

Source: Danish Zoonoses Centre,

National Food Institute

04 05 06 07 08 09 10 11 12 13 14

88 89 90 91 92 93 94 95 96 97 98 99 00 01 02 03 04 05 06 07 08 09 10 11 12 13 14

Broilers

Pork

Table eggs

Total Import

Source: Danish Zoonosis Centre, National Food Institute.

Total cases

Annual Report on Zoonoses in Denmark 2014

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Salmonella

source account 2014

The overall trend in human salmonellosis cases attri-

butable to the major food-animal sources is presented in

Figure 1.1.

As in 2013, domestic pork was estimated to be the most

important food source of salmonellosis in Denmark in 2014

(Figure 1.2 and Appendix Table A1), with 15.4% of laboratory-

confirmed cases. Three outbreaks related to domestic pork

contributed 4.6%, while sporadic cases accounted for the

remaining 10.7%. More than 60% of the sporadic cases at-

tributed to pork belong to the three subtypes involved in the

outbreaks. The fraction of cases attributed to domestic beef

in 2014 (2.2%) was similar to 2013, but as opposed to 2013,

more than 75% of the cases from domestic beef in 2014 were

related to an outbreak. Domestic table eggs were estimated

to be responsible for 3.0% of human cases in 2014. Appro-

ximately half of the cases were part of an outbreak and 1.4%

were sporadic cases, which is similar to 2012 and 2013. For

the first time since 2010,

Salmonella-positive

Danish broiler

carcasses were detected by the surveillance system, and as a

result, 2.0% of cases was attributed to this reservoir. A total

of 2.8% of cases were estimated as attributable to imported

chicken, with 0.7% belonging to an

Infantis outbreak and

0.4% to an

Enteritidis outbreak (Appendix Table A4).

Imported duck was estimated as the reservoir of 2.0%

of cases, which cannot be compared to last year, as samples

from this source were not available in 2013. This number is,

however, in accordance with 2.4% estimated in 2011 and 1.9%

in 2012, suggesting that no major changes have occurred. Im-

ported pork was estimated as the source of 1.1% of cases. No

positive samples were observed from imported turkey meat

in 2014, therefore no cases were attributed to this source. No

samples were available from domestic ducks or turkey meat,

and consequently no attribution estimates are presented for

these sources. A total of 48% of patients were estimated to have

travelled abroad within seven days prior to onset of symptoms.

This number varied from 40.3% to 46.9% in the last seven

years, except from 2008 and 2009, where extraordinary large,

domestically acquired outbreaks took place.

A total of 19.2% of reported

Salmonella

cases were spo-

radic cases which could not be associated with any of the

included food sources. This fraction has been relatively stable

since 2008 and may be caused, for example, by imported or

domestically produced fruits and vegetables (Appendix Table

A22), food-animals not included in the national surveillance

or by non-food sources of infection, such as direct contact

with pet animals.

Of the 268 reported

Enteritidis cases, the SSA estimated

77.9% to be related to international travel and 10.1% were

part of three outbreaks. One of those outbreaks (outbreak

number: FUD1379), corresponding to 6.7% of

Enteritidis

cases, had Danish eggs identified as the source. This is the

first

Enteritidis outbreak related to Danish eggs since

2009. Of the remaining two

Enteritidis outbreaks, one was

associated with imported chicken, whereas the source of the

third outbreak could not be identified (see Chapter 2 for more

information).

A total of 427

Typhimurium cases were reported in

2014, including 197 cases of classical

Typhimurium and

230 cases of monophasic

1,4,[5],12:i:- strains. In total, 98

cases were part of six outbreaks with

Typhimurium, of

which 86 cases (87.8%) were monophasic. Sixty-five cases of

1,4,5,12:i:- were implicated in four outbreaks, of which two

were connected to pork, one to beef and one to an unknown

source. Monophasic

1,4,12:i:- was isolated in one pork-

related outbreak with 22 registered cases, and a classical

Typhimurium strain was responsible for five cases in a third

pork-related outbreak (see Chapter 2 and Appendix Table A4

for more information). Among cases infected with

Typhi-

murium, 97 cases (22.7 %) were estimated to have travelled

abroad prior to disease onset, with a predominance of classical

strains over monophasic (approximately 2:1).

Antibiotic resistance in

Typhimurium

Of the 134

Typhimurium cases (including monophasic

strains) attributed to domestic food products, with resi-

stance information available, 22.8% were caused by strains

susceptible to all tested antimicrobial agents, 74.4% by strains

resistant to one to three antimicrobial agents and 2.8% by

strains resistant to four or more antimicrobial agents (multi-

resistant) (Figure 1.3). No quinolone-resistant strains were

observed in cases attributed to domestic food animal sources

Figure 1.2. Estimated sources of 1,122 cases of human salmonellosis in Denmark, 2014. (See also Appendix Table A1)

Sporadic cases, source unknown (15.9-22.4%)

Domestic pork (6.6-14.4%)

Domestic pork, outbreak-related (4.6%)

Domestic beef (0.1-1.1%)

Domestic beef, outbreak-related (1.7%)

Domestic eggs (0.3 -2.6%)

Domestic eggs, outbreak-related (1.6%)

Domestic broilers (0.1 -6.2%)

Imported pork (0.0-3.9%)

Imported beef (0.0- 0.6%)

Imported broilers (0.2-3.7%)

Imported broilers, outbreak-related (1.1%)

Outbreak cases, source unknown (4.0%)

Travel (47.0-49.0%)

Source: Danish Zoonosis Centre, National Food Institute.

Imported duck (1.0-3.1%)

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Trends and sources in human salmonellosis

in 2014, returning to a situation observed between 2010 and

2012, and interrupted in 2013, when 1.2% of

Typhimurium

cases, attributed to domestic sources, were resistant to this

antibiotic group.

Large annual changes in the relative distribution of resi-

stance patterns are observed among

Typhimurium cases

attributed to imported food sources 2012-2014 (Figure 1.3).

However, these changes must be interpreted with caution, as

the number of

Typimurium cases attributed to imported

food sources these years were limited, and available data from

these sources changed, with no data from imported beef or

imported duck available in 2013 (Appendix Table A1).

From the 94

Typhimurium cases estimated to be

acquired abroad, for which resistance information was avai-

lable, 33.4% were caused by resistant types, 37.4% by types

susceptible to all tested antimicrobial agents, 15.7% by types

resistant to quinolones and 13.5% by multi-resistant types.

References

1. Hald T, Wegener HC (2013). “Salmonella – epidemio-

logy and public health impact” in Foodborne Infections and

Intoxications, 4th edition. Eds. Morris G & Potter M. Elsevier,

Academic Press. ISBN 97801244160415..

2. Panel on Biological Hazards (BIOHAZ) (2010). Scienti-

fic Opinion on monitoring and assessment of the public

health risk of “Salmonella Typhimurium-like” strains. EFSA

Journal 8(10):1826.

3. Wegener HC (2010). Danish initiatives to improve the

safety of meat products. Meat Science 84:276-283.

4. Kjeldsen MK, Torpdahl M, Campos J, Pedersen K,

Nielsen EM (2014). Multiple‐locus variable‐number tandem

repeat analysis of

Salmonella enterica subsp. enterica

serovar

Dublin. J Appl Microbiol 166:1044-1054.

5. Petersen RF, Litrup E, Larsson J, Torpdahl M, Sørensen

G, Müller L, Nielsen EM (2011). Molecular characterization of

Salmonella

Typhimurium highly successful outbreak strains.

Foodborne Path Dis 8:655-661.

6. Litrup E, Christensen H, Nordentoft S, Nielsen EM, Da-

vies RH, Helmuth R, Bisgaard M (2010). Use of multiple-locus

variable-number tandem-repeats analysis (MLVA) typing to

characterize

Salmonella

Typhimurium DT41 broiler breeder

infections. J Appl Microbiol 109:2032-2038.

Figure 1.3. Distribution of antimicrobial resistance

Typhimurium, including S. 1,4,[5],12:i:-, from human infec-

tions attributed to domestic or imported food sources, or travel in the

Salmonella

source account, 2011-2014

2014

2013

2012

2011

2014

2013

2012

2011

2014

2013

2012

2011

No. of cases

120

Resistant S. Tm

Import

Travel

160

Susceptible S. Tm

Multi-resistant S. Tm

Quinolone-resistant S. Tm.

a) Resistant: Resistant towards one to three antimicrobial agents; Multi-resistant: Resistant towards four or more antimicrobial agents.

Antimicrobials in the resistance profile for the

Salmonella

source account were: ampicillin, ceftiofur or cefotaxime/ceftazidine, chloramp-

henicol, ciprofloxacin, gentamicin, nalidixic acid, sulphamethoxazole, tetracycline and trimethoprim.

Source: Danish Zoonosis Centre, National Food Institute.

Where do we acquire

Salmonella

infections?

By Luise Müller ([email protected])

In 2014, as in the previous years, Statens Serum Institut attempted to interview all registered

Salmonella

cases

where no travel information was reported by the general practitioner. The patients were asked about the date of disease

onset and whether they had travelled abroad within a seven-day period prior to disease onset. This information was

complemented with information from general practitioners’ reports. Travel information was obtained from a total of

71.2 % of the

Salmonella

cases in 2014. Among the cases with known travel history, 46.4 % were infected abroad (Table

1.1). However, the proportion of travel-related cases varied greatly between the different serotypes, hence 77.2 % of

the

Enteritidis cases, 31.2 % of the

Typhimurium cases, 12.7 % of the monophasic

1,4,[5],12:i:- cases and 54.1%

of cases with other serotypes were infected abroad. In 2014, the majority of travel-related cases travelled to Thailand

(17.5%), Turkey (15.4%), and Spain (6.4%). In contrast to 2013, no travel-related outbreaks due to

Enteritidis were

registered (Figure 1.4).

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Trends and sources in in human salmonellosis

Trends and sources human salmonellosis

Table 1.1. Top 10

Salmonella

serotypes in humans and information about travel aboad, 2013-2014

2014

Enteritidis

1,4,[5],12:i:-

Typhimurium

Infantis

Dublin

Stanley

Newport

Virchow

Agona

Kentucky

Other serotypes

Total

Number of

patients (%)

268 (23.9)

230 (20.5)

197 (17.6)

38 (3.4)

21 (1.9)

19 (1.7)

18 (1.6)

16 (1.4)

278 (24.8)

1,122 (100)

% of patients

infected

Abroad

Domestically

77.2

12.7

31.2

26.1

81.3

50.0

77.8

28.6

58.3

59.0

46.4

22.8

87.3

68.8

73.9

100

18.8

50.0

22.2

71.4

41.7

41.0

53.6

2013

Enteritidis

Typhimurium

1,4,[5],12:i:-

Dublin

Newport

Stanley

Agona

Infantis

Virchow

Corvallis

Other serotypes

Total

Number of

patients (%)

346 (30.5)

210 (18.5)

127 (11.2)

27 (2.4)

23 (2.0)

22 (1.9)

17 (1.5)

13 (1.1)

302 (26.6)

1,136 (100)

% of patients

infected

Abroad

Domestically

79.0

17.5

32.5

62.5

84.6

25.0

42.9

75.0

87.5

51.1

52.1

21.0

82.5

67.5

100

37.5

15.4

75.0

57.1

25.0

12.5

48.9

47.9

a) Patients with unknown travel information (28.8 of all patients in 2014 and 32.2% of all patients in 2013) were excluded from the

percent calculations.

b) Infected abroad is defined as travel abroad in a seven-day period prior to disease onset.

Source: Statens Serum Institut.

Figure 1.4. Monthly distribution of

Enteritidis,

Typhimurium,

and monophasic S. 1,4,[5],12:i:-

cases, 2010-2014

Number of human cases

120

100

Enteritidis

1 3 5 7 9 11 1 3 5 7 9 11 1 3 5 7 9 11 1 3 5 7 9 11 1 3 5 7 9 11

2010

2011

2012

2013

2014

Number of human cases

120

100

Typhimurium and monophasic

1,4,[5],12:i:-

1 3 5 7 9 11 1 3 5 7 9 11 1 3 5 7 9 11 1 3 5 7 9 11 1 3 5 7 9 11

2010

2011

2012

2013

2014

Domestic

Domestic outbreak related cases

Travel status unknown

Travel

Travel related outbreak cases

Source: Statens Serum Institut.

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2. Food- and waterborne outbreaks

By the Central Outbreak Management Group

Food- and waterborne outbreaks in Denmark are re-

ported in the Food- and waterborne Outbreak Database

(FUD). Outbreaks that occurred in 2014 are presented in

Appendix Table A4. Figure 2.1 shows the relative distribu-

tion of these outbreaks by the different causative agents.

Household outbreaks and clusters that could not be verified

as common source outbreaks are not included. The out-

break investigation procedures in Denmark are described

in further details in Chapter 8.

In total, 60 foodborne outbreaks were reported to FUD

in 2014 (Appendix Table A4) which is a decrease from

2013, where 74 foodborne outbreaks were reported. There

were no notifications of waterborne outbreaks in 2014. The

outbreaks were mainly regional or local outbreaks (80%)

and only 12 outbreaks were considered national outbreaks.

The largest national outbreak was caused by

Listeria mono-

cytogenes,

MLST224 (outbreak number in FUD: FUD1373).

In total, the number of persons affected by foodborne

outbreaks was 2,209, with a median of 11 persons per

outbreak (range 2 – 430). The largest outbreak involving

430 persons was an outbreak caused by Norovirus (NoV)

in a canteen serving a buffet meal. The source of the illness

appeared to be one of the kitchen staff who had attended

a sick child at home with the same symptoms. Analyses of

stool samples from both the child and some of the guests

that became ill showed the same type of NoV.

As in previous years NoV was the most frequent cause

of foodborne outbreaks (24 outbreaks), and in total, 1,339

persons were affected by NoV outbreaks. The transmission

routes for NoV causing foodborne outbreaks were multiple.

In Table 2.1 a breakdown of the number of outbreaks and

the number of people affected per route of transmission

for 2013 and 2014 are shown. The most common ways of

infection with NoV in 2014 were contamination from ill

kitchen staff or kitchen staff attending ill family members

before working in the kitchen. Less often, the infection

originated from contaminated ready-to-eat products like

oysters and frozen berries.

In 2014,

Clostridium perfringens

was less frequently

associated with foodborne outbreaks than in 2013. In

total, seven outbreaks of

C. perfringens

affecting a total of

528 people were reported in 2014, compared to 16, 8 and

7 outbreaks caused by this agent in 2013, 2012 and 2011

respectively.

When dividing the outbreaks into reported settings, the

most frequent setting was restaurants (40%) with 24 outb-

reaks affecting 363 people (mean: 15 people per outbreak).

Outbreaks taking place in workplace canteens and catering

Figure 2.1. Aetiology of the 60 foodborne disease outbreaks reported with a causative agent in the Food- and water-

borne Outbreak Database (FUD), 2014 . Percentage of total outbreaks indicated in brackets

Other

Salmonella

serotypes (5.0%)

Typhimurium

(10.0%)

. Enteritidis (5.0%)

Shigella sonnei

(1.7%)

E. coli

(3.3%)

VTEC (5.0%)

Yersinia

(1.7%)

Unknown (1.7%)

Bacillus cereus

(1.7%)

Campylobacter

(3.3%)

Clostridium perfringens

(11.7%)

Histamine (1.7%)

Listeria

(6.7%)

Norovirus (40.0%)

Typhimurium includes the

Salmonella

strains 1,4,[5],12:i:-.

Source: Food- and waterborne Outbreak Database (FUD).

Lectines (1.7%)

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(9 outbreaks) also affected a high number of people (1,262

people) and affected on average 140 persons per outbreak.

Composite meals (20 outbreaks) and buffet meals (nine

outbreaks) were the most frequently reported sources of

outbreaks in 2014 and most often these outbreaks were as-

sociated with NoV or

C. perfringens

(Appendix Table A4).

2.1 Outbreaks with

Listeria

The most severe foodborne outbreak in 2014 was caused

Listeria monocytogenes

(FUD1373). with a total of 41 ca-

ses reported. Seventeen cases died within 30 days from the

laboratory sample date. The outbreak vehicle was identified

as ”rullepølse” – a Danish cold cut ready-to-eat speciality

made from pork. Apart from this outbreak, listeriosis has

been in focus in 2014, where the implementation of new

laboratory methods with whole-genome sequencing (WGS)

have improved identification and comparison of clusters

over long periods of time. Read more about

Listeria

Chapter 3.

2.2 Outbreaks with

Salmonella

In 2014, eight outbreaks of

Salmonella

with patients

in two or more regions were reported in FUD. The first

outbreak - caused by

Agona - started in August 2013

and lasted 14 months (FUD1317). In total, 21 cases were

registered from August 2013 to October 2014 with

Agona

PFGE0022. Isolates were analysed by WGS which supported

the identification of a cluster already identified by PFGE and

in addition, sub-clustered isolates from patients being less

than one year old. The age distribution was characterized

by more than half of the cases being under five years of age,

a group of men aged 20-24 years and a few elderly people

above 70 years old (Figure 2.2). Seven cases were infants 4-9

months old and infant formula was therefore suspected as

the source. Hypothesis generating interviews showed that

young male cases were working out to build muscles and

they consumed protein shakes made from protein powder.

Furthermore, two of the elderly cases had consumed protein

drinks during hospital admissions while recovering from

illness. These observations led to the hypothesis of whey

powder as the source of the outbreak, as whey powder is

used in infant formula, protein powder and in products for

special nutritional purposes e.g. protein drinks used by ill

and elderly people with a low calorie intake. This hypothe-

sis was tested in a case-control study, which showed that

the young adult cases were more likely than age matched

controls to consume protein products. In addition, infant

cases were more likely to consume infant powder and/or

pre-made baby food than the infant controls – however, the

study had its limitations due to a small number of cases in

the different age groups. Samples of protein powder from

two patients’ homes were tested, but

Salmonella

was not

The role of asymptomatic food handlers in calicivirus outbreaks

New research has shown that one of five calicivirus outbreaks is caused by contamination from asymptomatic food

handlers. Data from FUD on 191 calicivirus (189 norovirus and 2 sapovirus) outbreaks that occurred from 2005-2011

was reviewed and categorized in order to clarify the routes of contamination. For 51 (27%) outbreaks, the contamination

had occurred during production, and the most common food items implicated were frozen berries, lettuce and oysters.

Another 55 (29%) outbreaks had supposedly occurred after an ill guest had attended a self-serve buffet. Contamination

from food handlers took place during the preparation or serving of the food in 64 (34%) of the outbreaks of which 41

(64%) were caused by asymptomatic food handlers – either food handlers who had contact to ill household members

(but remained asymptomatic), or retrospectively were found to be in the incubation- or recovery period at the time of

handling the food. [1]

Table 2.1. Norovirus outbreaks in 2014 per route of transmission based on number of cases or number of outbreaks

2014

Transmission route/source

Ill kitchen staff or healthy carrier of virus among kitchen staff

Kitchen staff tending to ill persons at home before entering the kitchen

Ill person/guest attending a buffet

Seafood (oysters)

Frozen raspberries

Norovirus in total

Source: Food- and waterborne Outbreak Database (FUD).

2013

No. of

outbreaks persons ill

226

129

261

655

No. of

outbreaks persons ill

507

729

1,339

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Food- and waterborne outbreaks

detected. No tested animal, food or feed isolates matched

the PFGE type. Trace back investigation of whey powder

used for infant formula, baby food, protein powder and the

products for special nutritional purposes mentioned by the

patients was conducted. The result of the tracing was very

complex and showed that the whey powder used originated

from the major producers on the market with an even more

complex supplier network to these establishments. No

specific product or supplier of whey powder or raw mate-

rial for the production of whey powder could be pointed

out to be the outbreak source. The long lasting outbreak

indicates that the vehicle was a low-contaminated product

with a long shelf life or a contaminated factory with occa-

sionally contaminated products or a combination of both.

In conclusion, the investigation pointed at whey powder

as a possible source for several reasons: The specific age

distribution, the fact that

Agona is previously known to

have caused outbreaks in infant formula [2, 3], the long shelf

life of the product and the case-control study supporting

the hypothesis although the epidemiological evidence from

the case-control study was not strong.

Four outbreaks with patients in two or more regions

were caused by

Typhimurium or monophasic

Salmonella

1,4,[5],12:i:-. Three of the outbreaks were found to have

been caused by Danish meat. For the fourth outbreak, no

source could be identified.

From March to May 2014 an outbreak occurred with

22 cases of monophasic

Salmonella

1,4,12:i:- MLVA0007

fully sensitive to antibiotics (FUD1368). Patients were 1-81

years old, 13 were male and the majority of cases were from

Jutland. Hypotheses generating interviews were performed

with 20/22 cases. Results indicated that the source most

likely was fresh pork sold either from the retail part of a

specific cutting plant or in a small supermarket chain at the

same geographical locations as the cases. Trace back inve-

stigation showed that the supermarket chain was supplied

with fresh meat and minced meat from the cutting plant.

Furthermore, it was possible to support the hypothesis of

pork from the supermarket chain with information from

one patient’s shopping receipts showing that the patient had

bought pork from the supermarket chain. However, it was

not possible to point out a specific time period or a specific

batch of meat that could have caused the outbreak and thus

no meat could be withdrawn from the market.

Another outbreak caused by monophasic

Salmonella

1,4,5,12:i:- MLVA0201 with the resistance pattern: Am-

picillin, Streptomycin, Sulfamethoxazole, and Tetracy-

cline (ASSuT) occurred between May and September

2014 (FUD1372). This outbreak was discovered, when an

unusually high number of food isolates from one specific

slaughterhouse and one cutting plant in late May tested

positive for the same monophasic

Salmonella

type. The

slaughterhouse had received pigs from a farm, where the

herd was categorized as

Salmonella

positive. The pigs from

this farm were slaughtered as the last herd on May 22, 2014

and sampled as scheduled in the legislation. The carcasses

were all retained until the analysis results were known. The

results were positive and the carcasses consequently sent for

heat treatment. However, insufficient cleaning and disinfec-

tion of the plant and equipment led to contamination with

the same

Salmonella

type of carcasses from pigs slaughte-

red the following day. The carcasses were cut in a cutting

plant, and pork sampled as part of the normal procedures

prior to export was found to contain the same strain of

Salmonella.

Consequently, both establishments were closed

down for extra cleaning and disinfection. Environmental

Figure 2.2. Salmonella Agona PFGE0022 by age and sex, (FUD1317, n=21)

Number of cases

0-4

5-9

10-14

15-19

20-24

25-29

30-34

35-39

40-44

45-49

50-54

55-59

60-64

65-69

70-74

75-79

80-84

Age group (years)

Female

Source: Statens Serum Institut

Male

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Food- and waterborne outbreaks

Outbreaks of special interest

samples taken after cleaning showed satisfactory results. No

further positive samples were seen. The meat from these

pigs was not withdrawn from the market, as there with

few exceptions is no legal requirement demanding absence

Salmonella

in fresh meat. In the beginning of July, the

first eight patients infected with the same

Salmonella

type

were identified. In total, 25 cases were reported from May

to September scattered throughout Denmark. Interviews

with cases showed that 18/20 had consumed fresh pork in

the week prior to disease onset – eleven of these had eaten

minced meat. Two cases had tasted raw minced meat. Eight

patients had bought the minced pork in a local butcher shop

– four in the same butcher chain. No other food item was

identified as eaten by the majority of cases. In conclusion,

this outbreak was very likely caused by fresh meat from this

specific slaughterhouse.

Another outbreak, in which the agent had already been

detected in foodstuff at the time a patient cluster was identi-

fied, was an outbreak of monophasic

Salmonella

1,4,5,12:i:-

MLVA1277 not previously seen in Denmark (FUD1374).

A total of 19 cases were identified in the outbreak with

disease onset from June to August 2014. The foodstuff

isolate originated from minced beef from a specific meat

production establishment, which had been tested as part

of the company’s own check program. According to Reg.

(EC) No 2073/2005 concerning microbiological criteria for

certain food items the isolation of

Salmonella

in minced

meat should have resulted in an immediate recall, but un-

fortunately the contaminated batch was not recalled from

the market and consumers were not informed until much

later. The food consumption pattern found through inter-

views with 10 cases and the trace forward investigation of

the batch of minced meat supported, that cases could have

been exposed to meat from this specific establishment.

Three outbreaks of

Enteritidis were reported in 2014.

The first outbreak was caused by

Enteritidis MLVA0206

and a total of 12 cases were reported from November 2013

to January 2014 (FUD1327). Patient interviews with a focus

on egg consumption did not reveal the source, but it seemed

unlikely that eggs should be the source, since some cases

did not eat eggs in the week prior to disease onset, and

those who did, reported consumtion of eggs from different

producers and of different types.

The second outbreak with

Enteritidis outbreak

investigation started in August 2014, when a large egg

producing unit in Denmark experienced severe clinical

illness among their laying hens and the herd was found

positive for

Enteritidis phage type 21 (FUD1379). The

eggs were immediately recalled from the consumers. In

August and September, 13 patients without travel history

outside Denmark in the week prior to disease onset, were

identified with the same phage type, identical by MLVA ty-

ping (MLVA0019) to the poultry isolate. Interviews showed

that all patients - but one - had consumed or handled eggs

from the recalled batch - after the time of the recall. The

outbreak was considered to be over and successfully and

timely controlled and thereby presumably several cases of

salmonellosis had been prevented. However, a few weeks

later another five cases with the same type were registered.

This second peak of cases indicated a new introduction

of the same

Salmonella

type, but interviews showed that

cases had not eaten eggs from the same producer and 3/5

cases had eaten eggs from local farms in the area near the

large producer.

The third outbreak with

Enteritidis MLVA0017

(FUD1391) took place in September to October 2014. WGS

proved useful to specify the case-definition, as different

resistance profiles were observed for cases. In total, four

patients were related to this outbreak. Relation between

human isolates and an isolate from a batch of fresh chicken

breast filet, originating from Poland, and tested positive in

the case-by-case control program, was detected by SNP

analysis of WGS results (Table A17). When the batch of

chicken was found to be contaminated, it was recalled from

the consumers. Fresh chicken breast filet seemed to be a

plausible source of this outbreak since three interviewed

cases reported that they had consumed chicken prior to

disease onset. However, this could not be finally proved

since the places of purchase reported by the cases differed

from the places where the incriminated batch had been sold.

The

Salmonella

outbreaks in 2014 emphasize the im-

portance of close collaboration between the surveillance

of human

Salmonella

and the surveillance of

Salmonella

food, especially when the food item (e.g. pork products) is

consumed by many inhabitants on a daily or weekly basis

which limits the potential of epidemiological investigation.

Therefore, knowledge of a possible match between food

isolates and human isolates is essential in the outbreak

investigation in order to prevent new cases of illness.

2.3 References

1. Franck KT, Lisby M, Fonager J, Schultz AC, Böttiger

B, Villif A, Absalonsen H, Ethelberg S (2015). Sources of

Calicivirus Contamination in Foodborne Outbreaks in

Denmark, 2005-2011 – The role of the Asymptomatic Food

Handler. J Infect Dis. 2015 Feb 15;211(4):563-70.

2. Cahill SM, Wachsmuth IK, Costarrica Mde L, Ben

Embarek PK (2008). Powdered Infant Formula as a Source

Salmonella

Infection in Infants. Clin Infect Dis. 2008 Jan

15;46(2):268-73.

3. Brouard C, Espié E, Weill FX, Kérouanton A, Bri-

sabois A, Forgue AM, Vaillant V, de Valk H (2007). Two

consecutive large outbreaks of

Salmonella enterica

serotype

Agona infections in infants linked to the consumption

of powdered infant formula. Pediatr Infect Dis J. 2007

Feb;26(2):148-52.

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Listeria

In 2014, an increase in the number of human listerio-

sis cases was observed mainly due to several foodborne

outbreaks caused by

Listeria monocytogenes.

Thorough

investigations were carried out to find the sources to the

outbreaks, and a critical review was initiated to come up

with recommendations to improve the general control and

management of

Listeria

in the food production.

3.1 Whole-genome sequencing as part of the

Listeria

surveillance

By Eva Møller Nielsen ([email protected]) and Charlotta Löfström

3.2 Listeria outbreaks in 2014

By Luise Mûller ([email protected]) on behalf of the Central

Outbreak Management Group

3.2.1 Extensive

Listeria

outbreak caused by Danish

cold cut (”rullepølse”)

During 2014, several foodborne outbreaks of

Liste-

ria

were detected using the whole-genome sequencing

(WGS). WGS was introduced in routine typing for sur-

veillance of listeriosis in Denmark in September 2013 and

has increased the discrimination of isolates. During 2014,

procedures with parallel, on-time, analysis of clinical and

food isolates have been introduced. The new technique

and procedures have increased the ability to find the link

between human cases and food sources

During 2014, a 1-year pilot project was initiated in a

collaborative effort between Statens Serum Institut, Natio-

nal Food Institute at the Technical University of Denmark,

and the Danish Veterinary and Food Administration,

where

L. monocytogenes

food isolates obtained from the

official control programs were sequenced and analysed in

parallel with the human clinical isolates and results were

compared. WGS was performed on a weekly basis on

both human and food isolates. The WGS data analysis was

done by initial determination of the classical multi-locus

sequence type (MLST) according to the established inter-

national nomenclature [1]. Isolates with the same MLST

were then compared in a single nucleotide polymorphism

(SNP)-based analysis using a reference genome within the

group (i.e., same MLST). For more information see chapter

3 in Annual Report 2013.

During the course of the project, 28 isolates from the

routine food control were sequenced along with the 92

human isolates from 2014. Furthermore, as part of outb-

reak investigations, 68 isolates were obtained by sampling

in several food producing companies, e.g. as follow-up on

the large outbreak of MLST sequence type 224 (ST224)

(FUD 1373). These isolates were also subjected to WGS.

The use of WGS and the procedure with parallel, on-time

analysis of clinical and food isolates, allowed identification

of the source of several foodborne outbreaks of

Listeria

during 2014.

On the 26

of June 2014 a cluster of three recent

Liste-

ria monocytogenes

ST224 patients similar to four known

patients from 2013 was detected by WGS. An outbreak

investigation was initiated to reveal the source in order to

stop the outbreak (FUD1373). As part of a project initiated

in 2013, all listeriosis patients were routinely followed up

with extensive contact to the hospitals for further clinical

information and, if possible, patients or relatives were

interviewed with a trawling questionnaire on symptoms,

food intake in the 30 days prior to disease onset, place of

food purchase and food handling habits at home.

In total, 41 patients were reported (four from 2013 and

37 from 2014 (Figure 3.1)). The patients were 43-90 years

old with a median age of 72 years, and 23 (56%) were wo-

men. Seventeen patients (41%) died within 30 days from

the sample date. Clinical information was obtained for all

patients, and interviews were possible for 25 patients. The

place of exposure could be established for 35 patients and

showed, that five patients had been exposed in hospital/

elderly home, 14 in private homes and 16 in either one of

these. All patients had underlying diseases, in particular

cancers and haematological diseases, and many were in

treatment with immunosuppressive drugs rendering them

more susceptible to listeriosis.

The 16

of July, a match was found by WGS between

the human isolates and

Listeria

isolated from ”rullepølse

”– a Danish cold cut speciality - made from pork from

a Danish manufacturer (Company A). In the interviews

18/23 patients confirmed that they had consumed ”rulle-

pølse” in the 30 days prior to disease onset. A production

and distribution ban for Company A was induced and an

extensive withdrawal including downstream businesses

was initiated. In total, approximately 6000 companies in

–5

distribution layer were affected, and all these were

contacted by telephone. Furthermore, 600 companies were

inspected physically.

The outbreak investigation included extensive sampling

with more than 800 samples from Company A and down-

stream businesses. In total, 32/43

Listeria

positive samples

were detected positive with ST224, and these included

samples from Company A and from the production env-

ironment of a large cutting plant receiving products from

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Company A. These findings led to further product recall.

In conclusion, this outbreak was successfully con-

trolled and supposedly, further illnesses/deaths were

prevented, despite the fact that outbreaks of listeriosis

are often very difficult to investigate because of the long

incubation period and patients often being severely ill.

The outbreak demonstrates that surveillance of

Listeria

food as well as typing of isolates from food and patients

are crucial to identify the source.

As a consequence of this large outbreak, the Danish

Ministry of Food, Agriculture and Fisheries initiated a cri-

tical review of the Danish efforts against

L. monocytogenes

which resulted in several recommendations for initiatives

to improve the control of

Listeria

in Denmark. These inclu-

ded increased knowledge on

Listeria

risk in food handlers

preparing food to vulnerable groups, e.g. in hospitals, and

increased information on

Listeria

to risk groups, increased

knowledge on

Listeria

risk in food companies and in the

food control, and optimized procedures for sampling and

source tracing. See 3.3 [2].

3.2.2 Other

Listeria

outbreaks

In 2014, three additional outbreaks/clusters of

Listeria

monocytogenes

detected by WGS were investigated. In

April, a cluster of four cases with

L. monocytogenes

ST391

was reported with three cases from 2013, the first one in

June 2013 (FUD1376). No known food isolates matched

this type, and the next months a new case occurred every

second month. The latest ST391 case was detected in No-

vember 2014 and in total, eight cases were identified. It is

uncertain whether more cases will occur. One case was a

child aged 12 years and the other seven were aged 47-89

years. Four were women and three cases died within 30

days from the sample date. Interviews were possible for

only three cases, which was not enough to identify a com-

mon exposure.

Another

Listeria

cluster dating back to 2013 was

detected in September 2014. This was an outbreak of

L. monocytogenes

ST399 (FUD1384). The possibility of

hospital-acquired infection was discussed already in July

2013 and the occurrence of new cases supported this hy-

pothesis. Furthermore, the same type was found in an own

control sample of asparagus soup from the hospital kitchen

and, later on, in a sample of frozen meatballs taken before

being added in the soup. Improper heating of the soup

after addition of the frozen meatballs was identified as a

factor contributing to the possibility for

Listeria

to survive

in the soup. From March 2013 to September 2014, six cases

could be related to this outbreak – four of these could be

identified as possibly infected in the hospital. Because of

this outbreak, the hospital kitchen now changed the pro-

cedures for re-heating pre-cooked soup.

In September, a cluster of

L. monocytogenes

ST6 was

detected (FUD1385). Again, cases had occurred over a

long period with date of onset for the first case in May 2013

and the latest in November 2014. Six cases were identified;

Figure 3.1 Week of sampling for

Listeria

ST224 patients, April to October 2014

, n=37

Number of patients

Outbreak detected

Investigation started

Full recall and

withdrawal

Isolate from Company A

matches outbreak strain 100%

14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41

a) Four cases from 2013 are not included in the figure.

Source: Statens Serum Institut.

Week, April-October, 2014

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Listeria

five women and one man, aged 43-89 years old. Two cases

died within 30 days from the sample date. In September

2014, food isolates of

L. monocytogenes

from a Danish

producer were found to be closely related based on WGS.

The isolates were from smoked trout and halibut and these

products were withdrawn from the market. Furthermore,

the producer temporarily stopped the production of smo-

ked halibut. For the five patients, who became ill in 2014,

a link was established with eating smoked fish products

purchased before the withdrawal in supermarkets, which

were selling products from the producer.

3.3 New initiatives on

Listeria/Critical

review

By Stine Thielke ([email protected])

on the risk of

L. monocytogenes

must be communicated to

groups at risk and healthcare professionals in contact with

these groups. The intention is among others to get a better

distinction of the groups at risk and thereby to a greater

extent target the relevant information, recommendations

and precautions.

Theme 2 - Knowledge of

Listeria

in kitchens prepa-

ring food primarily to groups at risk

Following the large outbreak of

Listeria monocytogenes

during the summer 2014 the Danish Ministry of Food,

Agriculture and Fisheries initiated a critical review of the

Danish

L. monocytogenes

effort. The critical review was

initiated in order to evaluate and improve current efforts

and initiatives as well as to identify future focus areas

relevant to

L. monocytogenes.

National and international experts took part in the

evaluation and critical assessment of the Danish effort

including representatives from Danish research institutes

and universities, the food industry, the Danish consumer

council, the authorities and two international experts from

Belgium and Austria. The critical review was based on an

initial workshop followed by comprehensive assessment of

identified focus areas. The result was a report presenting

six overall themes and associated initiatives based on the

assessments (Table 3.1).

The initiatives within each theme are presented below.

The initiatives in theme 1-4 will be instigated during the

next four years as part of a political agreement. The indu-

stry has committed itself to instigate the initiatives within

theme 5, and an effort is put into commencing the initia-

tives within theme 6 through research funds. Overall, the

initiatives are intended to improve the general control and

management of

Listeria

in the food production as well as to

enhance the knowledge and skill base of the food control,

the food businesses and food handlers, and groups at risk

in relation to

Listeria

with the general aim to secure safe

food and reduce the number of cases of listeriosis.

Theme 1 – Knowledge of

Listeria

and information for

groups at risk

Kitchens and food handlers preparing food or delive-

ring food to groups at risk need to understand and manage

the risk of

Listeria

since errors during production can be

fatal. Hence one of the initiatives is to develop tools and

guidance material specifically for this food sector. Mo-

reover the Danish Veterinary and Food Administration

will conduct a control campaign directed at kitchens pre-

paring food to groups at risk, and the control frequency

in food businesses preparing food to groups at risk will

be evaluated.

Theme 3 – Knowledge of

Listeria

in the food control

and in the food businesses

It is necessary to continuously increase the knowledge,

both within to the food control and the food businesses, on

how to manage and control

Listeria

in the food production.

Consequently the existing guidance material on

Listeria

will be adjusted and extended, i. e. with practical tools,

to better match the needs of the food businesses and the

food control. The elaboration of online guidance material

will be based on the results of a communication project,

which will be conducted to obtain insight and information

on how to ensure efficient distribution, communication

and design of written material. The Danish Veterinary

and Food Administration will also convene specialization

courses for the food control inspectors and establish a

group of specialists within the food control.

Theme 4 – Source tracing and sampling

It is imperative to increase the knowledge and aware-

ness of

Listeria

for groups at risk to minimize the risk of

infections with

L. monocytogenes.

The initiatives will be a

revision of the current recommendations to groups at risk

and definitions of groups at risk. Concurrently, information

Typing of food isolates have proved to be valuable

for source tracing and outbreak investigation and can

also provide important information to food businesses

in relation to the management of

L. monocytogenes

the food production. Therefore, a collaboration between

the authorities and research institutes will be initiated to

determine, which isolates from official samples that should

be typed using whole genome sequencing. Parallel to this

initiative a collaboration between the authorities, the

research institutes and the food industry will take place

in order to investigate the possibilities to use own control

samples and whole genome sequencing in the official

control. Sampling is an important part of the control and

management of

L. monocytogenes

in the food production

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Listeria

if it is performed correctly and based on a risk assessment

and as an integrated part of a HACCP plan, also referred

to as intelligent sampling. Hence another initiative will be

the development of guidance material and description of

principles of intelligent sampling to the food businesses.

The Danish Veterinary and Food Administration will also

elaborate guidelines for official sampling in withdrawal and

recall cases to better ensure efficient corrective measures

by the food businesses and a close follow up.

Theme 5 – Initiatives by the food industry

Theme 6 – Areas of research

A number of initiatives will be conducted by the food

industry, including cross-disciplinary development of tools

and practical guides for the food businesses, evaluation

and if necessary adjustment of the labour market training

courses to ensure, that the education of the food handlers

sufficiently covers the subject of

L. monocytogenes,

focus on

L. monocytogenes

in the collaboration with the

production equipment industry, and more focus on

L. mo-

nocytogenes

at the annual seminars between the food indu-

stry and the Danish Veterinary and Food Administration.

L. monocytogenes

comprise a complex problem regar-

ding food production and food safety, and more research

is needed in several areas. Research in these areas can

contribute to increase the overall level of knowledge and to

continuously improve the effort towards

L. monocytogenes.

Among the identified research areas considered beneficial

are further development of existing predictive models, de-

termination of the genetic diversity of

Listeria

isolates from

food and humans, indicators for

Listeria

and identification

of factors influencing the high incidence of listeriosis in

Denmark matched to other comparable countries.

3.4 References

1. Ragon M, Wirth T, Hollandt F, Lavenir R, Lecuit M,

et al. (2008). A New Perspective on

Listeria monocytogenes

Evolution. PLoS Pathog 4(9): e1000146.

2. Ministry of Food, Agriculture and Fisheries (2015).

Rapport om kritisk eftersyn af

Listeria-indsats

20. februar

2015 (available on www.fvm.dk in Danish).

Table 3.1 Overview of the 6 Listeria monocytogenes themes and initiatives to be instigated from 2015-2018

1. Knowledge of

Listeria

and information for groups at

risk

2. Knowledge of

Listeria

for kitchens preparing food

primarily to groups at risk

3. Increased knowledge to the food businesses and the

official control

4. Source tracing and sampling

5. Initiatives by the food industry

6. Areas of research

Source: The Danish Veterinary and Food Administration.

•

A better definition of groups at risk

•

Targeted information and recommendations to groups

at risk and healthcare professionals

•

Improved guidance on risks related to

Listeria

•

Intensified control

•

Control campaign on riscs of food to groups at risk

•

Online

Listeria

guideline

•

Increase of

Listeria

specialists within the official control

•

Increased knowledge on how to efficiently communicate

guidance material

•

Typing by whole genome sequencing

•

Guideline to intelligent sampling

•

Official samples in relation to outbreaks and withdra-

wal/recall

•

Practical tools and guides to control

Listeria

•

Evaluation of labour market training courses

•

Focus on

Listeria

in relation to production equipment

•

Annual seminars between the food industry and the

Danish Veterinary and Food Administration

•

Predictive models

•

Genetic diversity of

Listeria

isolates

•

Indicators

•

Factors related to high incidence in Denmark

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4. Verotoxin-producing

E. coli

(VTEC)

By Flemming Scheutz ([email protected]) and Luise Müller

In 2014, a total of 280 episodes of verocytotoxin- pro-

ducing

Escherichia coli

(VTEC) were notified. VTEC iso-

lates were obtained from 229 episodes, of which 37 (16%)

were caused by O157 (Appendix Table A3). A total of 12

patients developed haemolytic uraemic syndrome (HUS).

Nineteen patients were tested positive for VTEC by PCR

techniques but isolates were not obtained or submitted for

further characterization. Laboratory confirmed VTEC in-

fection thus had an incidence of 4.4 per 100,000 (Appendix

Table A2 and A3). Further 32 VTEC cases (including 4

HUS cases) were notified but without information on the

method and validity of the diagnose. Overall, the annual

number of episodes has been increasing during the last

decade. Improved diagnostic methodologies and increased

awareness play an important role in this increase.

The incidence of VTEC infections in children less than

five years old is approximately 10 fold higher than the inci-

dence in adults (Table 4.1). Nine of the 12 HUS cases were

in children less than 10 years. This is the highest number

of HUS cases yearly since HUS became notifiable in 2000.

4.1 Outbreaks with Verocytoxin-producing

E.coli

In 2014, three outbreaks – an unusual high number,

were registered with verocytotoxin-producing

E. coli

(VTEC). On average, less than one outbreak per year has

been registered in Denmark the last five years, and the last

outbreak of VTEC was in October 2012. The first outbreak

in 2014 was detected in August, when PFGE typing revea-

led a cluster of VTEC O103 cases (FUD1377). Hypothesis

generating interviews showed, that two cases were from

the same household, but no further connection with the

third case could be established. Four weeks later, two new

cases with the same PFGE pattern were identified, and new

interviews were conducted. The five cases were 0-71 years

old and lived in different parts of Denmark. No common

events or environmental sources were evident, and the

outbreak was closed as presumably foodborne, where the

source could not be identified.

In October-November, an outbreak of VTEC O157:H-,

was identified in a day-care centre in Jutland (FUD1392).

Three children in the day-care were diagnosed with VTEC,

of which a two-year old child developed HUS. A fourth

case – a child living more than 100 km away from the

day-care centre had no connection to the other cases. The

medical officer and the Food Control Office in Northern

Jutland put intensive investigation and control measures

at the day care facilities in place. The outbreak source was

not identified, but either a common food, environmental

exposure or person-to-person transmission in the day-care

centre could have played a role.

In November-December an outbreak of VTEC

O157:H7 occurred (FUD1409). In total, seven cases were

identified, of which four were laboratory-confirmed. The

age distribution was unsual for this agent, as cases were

young people aged 14-27 years. The cases were from the

areas of Copenhagen and Aarhus and they had all been

eating kebab (beef) from kebab restaurants. A Swedish

kebab restaurant in Malmö had also had a small outbreak

of VTEC O157 and comparison of the human isolates by

whole genome sequencing (WGS) showed related types.

The Danish Veterinary and Food Administration under-

took trace-back investigation, but no common batch of

Table 4.1. Number of Verocytoxin-producing

E. coli

(VTEC) cases and HUS-cases (haemolytic uraemic syndrome) by age

group and incidence per 100,000 population, 2014

Age group

<1 year

1-4 years

5-9 years

10-19 years

20-29 years

30-39 years

40-49 years

50-59 years

60-69 years

>69 years

Total

VTEC cases

No.

280

VTEC cases

Incidence per 100,000

31.5

32.7

5.4

4.1

3.7

2.4

2.5

3.1

1.9

5.4

4.9

HUS cases

No.

HUS cases

Incidence per 100,000

2.9

0.6

0.1

0.2

Source: Statens Serum Institut.

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meat or producer could be found. As a control measure,

The Danish Veterinary and Food Administration visited

all restaurants serving kebab in Denmark in January and

February 2015, to ensure proper heating of kebab meat

and proper hygiene procedures in the kitchens. Appro-

ximately 1,200 establishments were visited during this

campaign. No further outbreak cases were reported after

this period. An overview of the outbreaks can be seen in

appendix Table A4.

4.2 Virulence factors and HUS

VTEC infection and HUS were made notifiable in

Denmark in 2000, but the dataset from the Danish Cohort

presented below includes patients from 1983 to 2012. The

virulence profile for each strain has been related to the

clinical data, in particular during the period 1997-2006,

where detailed information was obtained on all cases to

investigate the cause of disease and complications.

For the period 1983 to 2012, VTEC infection was

complicated with HUS in three per cent (70/2,001) of the

patients registered in the Danish Cohort (Table 4.2); 6%

(44/698) in children less than five years old, 5% (12/229)

in children aged 5-15 years and one per cent (14/1,074) in

adults. However, great variation was found among different

virulence types. The

vtx2

gene together with the

eae

gene

is particularly often associated with HUS in children less

than 14 years old. Furthermore, preliminary results (data

not presented) indicate, that it is primarily the VT subtype

vtx2a,

which is associated with HUS. Only 0.7% (4/552)

of the patients developed HUS following infection with a

strain of genotype

vtx1

and

eae,

and they all had several

predisposing or underlying risk factors, e.g. antibiotic treat-

ment during the acute phase, nephrotic syndrome. For each

case, it is therefore important that unusual associations

between HUS and specific virulence profiles are carefully

examined for underlying disease, epidemiologic relation

and antibiotic treatment during the acute phase of illness.

The observed and well documented association

between

vtx2

and either presence of either the

eae

gene

(attaching and effaing properties) or the ability to colo-

nize and persist in the gut, e.g. enteroaggregative

E. coli

(EAEC), has resulted in a basic and primary definition

of HUS associated

E. coli (HUSEC)

for first line public

health action:

vtx2

in a background of an

eae

aggR

(master regulator gene in EAEC). For such human strains

vtx

is subtyped and further characterized. VTEC with the

virulence profile

vtx1, vtx1

and

eae, vtx2, vtx1

and

vtx2

regardless of serotype are considered “low-risk” VTEC and

are treated as other enteropathogens like e.g.

Salmonella

Campylobacter.

This simplified approach has been generally accepted

by the primary diagnostic clinical microbiology labora-

tories and public health officers in Denmark. Though not

complete in the characterization of each VTEC isolate,

this approach is practical and easy to use in an operational

environment, which is expected to quickly:

•

evaluate the risk of progression of the disease in indi-

viduals,

•

minimize transmission of HUSEC,

•

rehabilitate individuals in order to prevent the worse-

ning of an individual’s health, and

•

reduce the socio-economic impact on affected families.

However, such a simple approach should be applied

with prudence because each individual case is unique and

each patient should be carefully evaluated with regard to

predisposing factors, general clinical condition, contact

with other VTEC infected individuals, and possible link

to an outbreak. The identification of “low-risk” VTEC does

not per se exclude the risk of progression to severe disease,

dehydration or HUS, and patients with bloody stools and/

or affected kidney function must be carefully monitored.

Table 4.2. The Danish Cohort: HUS-cases (haemolytic uraemic syndrome) in the period 1983-2012 among Danish cases

with Verocytoxin-producing

E. coli

(VTEC) stratified according to virulence types and age

< 5 years

Virulence type

eae + vtx2

eae + vtx1 + vtx2

eae + vtx1

vtx2

vtx1 + vtx2

vtx1

Total

VTEC cases

No.

180

332

698

HUS cases

No. (%)

34 (18.9)

6 (6.3)

3 (0.9)

1 (3.0)

44 (6.3)

5-14 years

VTEC cases

No.

229

HUS cases

No. (%)

7 (14.3)

3 (7.8)

1 (2.0)

1 (4.0)

12 (5.2)

> 14 years

VTEC cases

No.

120

110

169

233

187

255

1,074

HUS cases

No. (%)

4 (3.3)

2 (1.8)

8 (3.4)

14 (1.3)

a) These patients were treated with antibiotics during the acute phase of illness, had several predisposing or underlying risk

factors, or (one case) had a double VTEC infection (also infected with O157:H7 (eae +

vtx1

vtx2).

One patient was positive for

VTEC O13, O73:K1:H18 (vtx2d).

b) Eight of 25 (32%) patients with culture confirmed EAEC-VTEC O104:H4 (vtx2a) developed HUS and were part of the German

outbreak in 2011.

Source: Statens Serum Institut.

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5. New insights in the European

epidemiology of

Salmonella

and

Campylobacter

By Kåre Mølbak ([email protected]), Hanne-Dorthe Emborg and Steen Ethelberg

Measuring human incidence of foodborne infections

including

Salmonella

and

Campylobacter

infections is af-

fected by biases inherent in passive laboratory surveillance

as reflected in the surveillance pyramid. The pyramid

describes the chain of events that have to occur so that a

case of

Salmonella

Campylobacter

in the population will

become a reported case, including factors like health-se-

eking behavior, clinical and laboratory practices regarding

microbiological diagnostics and finally reporting rules and

compliance. The attributes of the surveillance pyramid are

different in different countries and therefore it is often mis-

leading to compare reported figures of culture confirmed

cases between countries. To overcome this problem, we

hypothesized that seroepidemiologic methods could pro-

vide a stringent approach to measure the force of infection,

i.e. the rate at which humans are exposed to

Salmonella

Campylobacter

to a degree that this exposure is recognized

by the immune system as an antibody response.

Based on initial work in the European Med-Vet-Net

network of excellence, we have developed methods and

models to estimate the force of infections in defined popu-

lations. The advantage is, that this measure is independent

of the sensitivity of public health surveillance. Because this

measure is different from the incidence of clinical infec-

tions, we have named it seroincidence. The seroincidence

is calculated from the analysis of the levels of antibody

classes against non-typhoid

Salmonella

(S. Typhimurium

and

Enteritidis) or

Campylobacter jejuni

in the general

population, and the results of these examinations are con-

verted into a single measure (the seroincidence) based on

the kinetics of the antibody decay [1-5].

5.1 Seroepidemiology of

Salmonella

infections

We applied the model to almost 10,000 serum samples

randomly selected from individuals representing the gene-

ral healthy population in 13 different European countries.

The year of collection ranged from 2000 to 2012. There

was a 10-fold difference in the seroincidence; lowest in

Sweden (0.06 infections per person-year), Finland (0.07),

and Denmark (0.08) and highest in Spain (0.61) followed

by Poland (0.55). The figures did not correlate with the

reported national incidence of

Salmonella

infections in

humans (Figure 5.1), but correlated with prevalence data of

Salmonella

in laying hens (p < 0.001) (Figure 5.2), broilers

(p < 0.001) (data not shown) and slaughter pigs (p=0.03)

(data not shown) as published by the European Food Safety

Authority [7-9]. The seroincidence also correlated with

Swedish figures of country-specific risk of travel-associated

Salmonella

infections [10] (p=0.001) (Figure 5.3).

5.2 Seroepidemiology of

Campylobacter

infec-

tions

We applied the model to approximately 8,000 se-

rum samples collected from populations in 11 different

countries. Again, the samples represented the general

population and were collected over the same time span

as for the

Salmonella-sera.

For

Campylobacter,

only a

two-fold difference was observed: Greece had the lowest

seroincidence (0.55) and Poland the highest (1.11). The

figures had an inverse correlation with the reported na-

tional incidence of

Campylobacter

infections in humans

(p=0.008), and did not correlate with prevalence data of

Campylobacter

in broilers (11). Seroincidence tended to

correlate with Swedish figures of country-specific risk of

travel-associated

Campylobacter

infections (12) (p=0.09).

Data on seroincidence of

Campylobacter

is not presented.

5.3 Conclusions

While

Campylobacter

and

Salmonella

remain as com-

mon bacterial foodborne zoonoses, these studies highlight

some important similarities and differences. For both

infections, the actual number of infections is much higher

than suggested in the official surveillance figures. The mul-

tipliers are very different across Europe. Indeed, the official

numbers are mainly an indicator of the overall function of

public health microbiology rather than the real incidence

of infections. Hence, national surveillance is useful for fol-

lowing trends within a country and to detect events such

as outbreaks or emergence of new subtypes. But the official

figures should never be used in a comparison of disease

burden between countries.

It is of importance to underline that the observed

differences between reported figures and seroincidence

estimates are not only due to underreporting and under

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Figure 5.1. Salmonella seroincidence and the incidence of culture-confirmed cases reported to the European Union

Note: Each point represents one country that took part in the study except that The Netherlands is represented by two serum col-

lections (1998-2002 and 2006-2007). Spearmans rho = -0.367, P=0.197. Reproduced from [6].

Source: Statens Serum Institut.

Figure 5.2. Figure 2. Salmonella seroincidence and the observed prevalence of salmonella-positive holdings of laying

hens in 12 European countries

Note: Each point represents one country. Spearmans rho = 0.893, p = 0.001. Reproduced from [6].

Source: Statens Serum Institut.

Figure 5.3. Salmonella seroincidence and risk of Salmonella infection in Swedish travellers

Note: Each point represents one of 12 European countries. Spearmans rho = 0.811, p = 0.001. Reproduced from [6].

Source: Statens Serum Institut.

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Campylobacter

and

Salmonella

serology in humans

ascertainment. Any activation of the immune system inclu-

ding asymptomatic infections and mild illness will also be

captured by the seroincidence measure, while only a frac-

tion of infections causing clinical illness will be included

in the reported laboratory surveillance incidence estimate.

For

Salmonella

it is clear that countries such as the

Nordic countries, that implemented control activities

several decades ago, have a much lower rate of infections

than countries in the Eastern and the Southern Europe,

where many countries only started the

Salmonella

control

programmes in poultry when they became mandatory in

the European Union from 2008. The predominant mode

Salmonella-transmission

is through the food chain, and

there are striking correlations between the prevalence of

Salmonella

in food animals and the seroincidence. This

correlation was present even though the samples were

collected over a considerable time span and the EFSA

baseline surveys were conducted in specific years. This

may indicate, that seroincidence is a robust indicator of

the level of exposure to

Salmonella.

In contrast to

Salmonella,

there was not much differen-

ce in the seroincidence of

Campylobacter

infections across

the participating countries. This shows that there across

Europe is less variation in the force of

Campylobacter-

infections than the in the force of

Salmonella-infections.

It was a surprise that there was no correlation between

Campylobacter

seroincidence and prevalence of conta-

minated broiler carcasses in the corresponding countries.

This may indicate that the prevalence of

Campylobacter

not an appropriate measure of the risk of infection but that

other measures, e.g., the degree of contamination may be

suitable. Furthermore, there are other routes of transmis-

sion than poultry. This suggests that seroepidemiology is

not suitable to address the impact of control activities of

Campylobacter

aimed at the poultry reservoir. These data

support the notion of

Campylobacter-transmission

from a

number of reservoirs, including environment and various

animal sources, may be a driving factor for seroconversion

and immunity whereas high-dose exposures from poultry

(and during travel abroad) may still be a leading cause of

clinical illness.

ECDC has developed a software tool that can be used

to convert measurements from cross sectional data to a

seroincidence figure. The tool has been developed in R

and can be downloaded from the www.ecdc.europa.eu.

The measurements should be aligned with the reference

curves (decay profiles) in order to make meaningful use

of the tool.

5.4 References

1. Strid MA, Engberg J, Larsen LB, Begtrup K, Mølbak

K, Krogfelt KA (2001). Antibody responses to

Campylobac-

ter

infections determined by an enzyme-linked immuno-

sorbent assay: 2-year follow-up study of 210 patients. Clin

Diagn Lab Immunol; 8:314–9.

2. Ang CW, Krogfelt K, Herbrink P, Keijser J, van Pelt

W, Dalby T, et al. (2007). Validation of an ELISA for the

diagnosis of recent

Campylobacter

infections in Guillain-

Barré and reactive arthritis patients. Clin Microbiol Infect;

13:915–22.

3. Simonsen J, Mølbak K, Falkenhorst G, Krogfelt KA,

Linneberg A, Teunis PFM (2009). Estimation of incidences

of infectious diseases based on antibody measurements.

Stat Med; 28:1882–95.

4. Simonsen J, Teunis P, Van Pelt W, Van Duynhoven Y,

Krogfelt KA, Sadkowska-Todys M, et al. (2011). Usefulness

of seroconversion rates for comparing infection pressures

between countries. Epidemiol Infect; 139:636–43.

5. Teunis PFM, Van Eijkeren JCH, Ang CW, Van

Duynhoven YTHP, Simonsen JB, Strid MA, et al. (2012).

Biomarker dynamics: estimating infection rates from se-

rological data. Stat Med; 31: 2240–8.

6. Mølbak K, Simonsen J, Jørgensen CS, Krogfelt KA,

Falkenhorst G, Ethelberg S, et al. (2014). Seroincidence of

human infections with non-typhoid

Salmonella

compared

with data from public health surveillance and food animals

in 13 European countries. Clin Infect Dis; 59:1599-606.

7. EFSA (2007). Report of the task force on zoonoses

data collection on the analysis of the baseline study on the

prevalence of

Salmonella

in holdings of laying flocks of

Gallus gallus.

The EFSA Journal; 97.

8. EFSA (2007). Report of the task force on zoonoses

data collection on the analysis of the baseline study on the

prevalence of

Salmonella

in broiler flocks of

Gallus gallus,

Part A. The EFSA Journal; 98: 1-85.

9. EFSA (2008). Report of the task force on zoonoses

data collection on the analysis of the baseline study on

the prevalence of

Salmonella

in slaughter pigs, Part A. The

EFSA Journal; 135: 1-111.

10. De Jong B, Ekdahl K (2006). The comparative

burden of salmonellosis in the European Union member

states, associated and candidate countries. BMC Public

Health; 6: 4.

11. EFSA (2010). Analysis of the baseline survey on the

prevalence of

Campylobacter

in broiler batches and of

Cam-

pylobacter

and

Salmonella

on broiler carcasses, in the EU,

2008, Part B:

Campylobacter.

The EFSA Journal; 8(8):1522.

12. Ekdahl K, Giesecke J (2004). Travellers returning

to Sweden as sentinels for comparative disease incidence

in other European countries, campylobacter and giardia

infection as examples. Euro Surveill; 9(9): 6-9.

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Salmonella

Dublin in cattle

6. Vectorborne Zoonoses

By René Bødker ([email protected]), Birgit Kristensen and Carsten Kirkeby

The National Veterinary Institute, Tecnical University

of Denmark (National Veterinary Institute), monitors

vectors and vector borne diseases in Denmark on behalf

of the Danish Veterinary and Food Administration.

Denmark is considered free of zoonotic diseases

transmitted by mosquitoes (Culicidae) and biting midges

(Culicoides). But tick borne zoonotic disease transmitted

Ixodes ricinus

are common and, due to increasing num-

bers of the tick vector, considered an increasing problem.

The increased tick density is most likely linked to growing

populations of roe deer that functions as the main host for

adult egg laying ticks.

In recent years, Denmark had an outbreak of Blueton-

gue virus serotype 8 (2008) and an outbreak with Schmal-

lenbergs virus (2012); both outbreaks were in ruminants

and transmitted via biting midges (Culicoides sp.). Further,

an ongoing spread of exotic invasive mosquito species has

been observed in Europe and since 2000, and outbreaks

of mosquito borne West Nile virus, Dengue fever, Chi-

kungunya fever virus, malaria and

Dirofilaria

parasites

in humans have been reported in Europe. As a response

to the two outbreaks in ruminants in Denmark and due

to the increasing risk of introduction of mosquito borne

diseases in Denmark, two vector surveillance programs

for the abundance of mosquitoes and biting midges were

implemented in 2011 and 2012, respectively.

During transmission season the vector abundance

of two species of biting midges (Obsoletus and

Pulicaris

groups) and five species of mosquitoes (Aedes,

Anophe-

les, Culex, Culiseta

and

Coquillettidia)

are monitored at

sentinel sites. Weekly surveillance data are available at

www.myggetal.dk with a three days delay throughout the

season. Most diseases transmitted by mosquitoes and

biting midges have epidemic potential, and monitoring

of temperatures and vector abundance followed by a

calculation of the disease transmission potential (R0 - the

number of new cases arising from each case) forms an

important component of the early warning system at the

Technical Univerity of Denmark. This potential for disease

transmission must be interpreted in parallel with the risk of

introduction. The two main zoonotic introduction threats

are presently considered to be the mosquito borne Usutu

virus and the mosquito borne

Dirofilaria

worms, with

outbreaks recorded in Germany in 2010 [1] and 2014 [2],

respectively.

The surveillance of mosquitoes showed a low to medi-

um abundance of

Aedes, Anopheles, Culex

and

Culiseta

for

the 2014 season caused by the dry summer, and a normal

Table 6.1 Overview of vector borne diseases presented in the text

Vector

Ticks

Pathogen

Bacteria

Rickettsia helvetica

Neoehrlichia mikurensis

Anaplasma phagycytophilum

Borrelia

sp.

Babesia

sp.

Chikungunya fever

Dengue fever

Usutu virus

West Nile virus

Malaria

Dirofilaria

(worms)

Bluetongue

Schmallenberg

Transmission

Zoonotic

Only between humans

Zoonotic

Only between humans

Zoonotic

Only between ruminants

Transmitted in Denmark

Yes

Yes (several species)

Yes (two species)

Yes (2007)

Yes (2012)

Mosquitoes

Virus

Parasite

Biting Midges

Virus

Source: National Veterinary Institute, Technical University of Denmark.

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Vectorborne zoonoses

Salmonella

Dublin in cattle

abundance of

Coquillettidia,

a species not affected by low

rainfall in Denmark. However, the high temperatures in

late summer gave rise to three generations of biting midges

compared to the usual two generations, and resulting in

midge abundances in mid-October comparable to peak

abundances normally measured during summer (Figure

6.1). This illustrates that small increase in temperatures

may have disproportional large impact on vector biology.

The mosquito surveillance program also identified a

new mosquito species in Denmark,

Culex modestus

(Cx.

modestus,

’nilfeber-myg’ in Danish). This is the most

northern recording of this species in Europe [3]. This

species has in recent years been reported moving north in

Europe and was in 2012 reported in large numbers south

of London in United Kingdom [4].

Cx. modestus

is a highly

competent vector for zoonotic West Nile virus with reser-

voir in wild birds. Denmark has large populations of both

Cx. pipiens

and

Cx. torrentium

mosquitoes that effectively

transmit the virus between birds. But these species do not

bite humans.

Cx. modestus

however will bite both man and

birds and thus act as an important bridge vector between

wild birds and humans during outbreaks.

Cx. modestus

was discovered in a densely populated residential area

just south of Copenhagen with local biting rates on hu-

mans reaching up to one per minute in the late afternoon

during peak season. The species was however found to be

confined to just a single small breeding site on the beach.

It is important to note that Usutu and West Nile virus were

never identified in Denmark, but migrating birds have

been found seropositive when returning to Denmark in

the spring. The large spread of West Nile virus and Usutu

virus within Europe the last 15 years and the presence of

seropositive migrating birds highlights why we need to be

aware of the presence of a bridge vector like

Cx. modestus

in Denmark.

In 2013, the invasive exotic mosquito

Aedes japonicus

became established in Hannover just 200 km from the

Danish border. Screening of water samples from private

gardens in southern Jutland that where hatched to adult

mosquitoes in the laboratory at the National Veterinary

Institute, did not reveal this species in southern Denmark

in 2014. But the rapid spread in Holland and Germany

suggests that it may eventually spread across the border

to Denmark.

Screening of pools of tick nymphs from Danish forests

using a newly developed DNA screening chip revealed

two new

Borrelia

pathogens in

Ixodes ricinus

[5].

Borrelia

spielmanii

and

Borrelia miyamotoi

which is different from

known European

Borrelia

species as it causes a relapsing

type of fever in humans. The screening also found the pa-

rasites

Babesia venatorum

and

Babesia divergens. Babesia

divergens

is a well know infection in Danish cattle, but

Figure 6.1 Biting midges

(Obsoletus

sp.) per day on cattle herds, May-October, 2012-2014. Monitoring starting

July, 2012

700

600

500

Biting midges per day

400

300

200

100

30. Apr

14. May

25. May

11. June

25. June

9. July

2012

23. July

6. Aug

2013

20. Aug

3. Sept

2014

17. Sept

1. Oct

15. Oct

29. Oct

Source: National Veterinary Institute, technical University of Denmark.

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Vectorborne zoonoses

this is the first time it is identified in Danish ticks. It is not

known if

Babesia venatorum

causes disease in humans.

Additionally, the screening confirmed the already known

presence of

Borrelia burgdorferi, Borrelia garinii, Borrelia

afzelii, Borrelia valaisiana, Anaplasma phagocytophilum

and

Rickettsia helvetica.

Further the screening found a wi-

despread presence of

Neoehrlichia mikurensis

(figure 6.2).

Neoerlichia mikurensis

was newly discovered in Denmark

[6] and has been associated with severe human disease in

Sweden [7].

References

1. Becker N, Jöst H, Ziegler U, Eiden M, Höper D,

Emmerich P, et al. (2012). Epizootic Emergence of Usutu

Virus in Wild and Captive Birds in Germany. PLoS ONE

7(2): e32604.

2. Tappe D, Plauth M, Bauer T, Muntau B, Dießel L,

Tannich E, Herrmann-Trost P (2014). A case of autochtho-

nous human

Dirofilaria

infection, Germany, March 2014.

Euro Surveill. 2014;19(17):pii=20790.

3. Bødker R, Klitgård K, Byriel DB, Kristensen B (2014).

Establishment of the West Nile virus vector,

Culex mode-

Figure 6.2 Neoehrlichia mikurensis in Denmark

stus,

in a residential area in Denmark. Journal of Vector

Ecology, 39(2), 1-3.

4. Golding N, Nunn MA, Medlock JM, Purse BV, Vaux

AGC, Schäfer SM (2012). West Nile virus vector

Culex

modestus

established in southern England. Parasites &

Vectors 2012, 5:32, 1-5.

5. Michelet L, Delannoy S, Devillers E, Umhang G,

Aspan A et al. (2014). High-throughput screening of tick-

borne pathogens in Europe. Front. Cell. Infect. Microbiol.

4:103, 1-13.

6. Fertner ME, Mølbak L, Boye Pihl TP, Fomsgaard A,

Bødker R (2012). First detection of tick-borne “Candidatus

Neoehrlichia mikurensis”

in Denmark 2011. Euro Surveill.

17(8):pii=20096.

7. Welinder-Olsson C, Kjellin E, Vaht K, Jacobsson

S, Wenneras C (2010). First case of human ”Candidatus

Neoehrlichia mikurensis”

infection in a febrile patient

with chronic lymphocytic leukemia. J Clin Microbiol.

48(5):1956-9.

a) From each site 15 pools of 15 nymphs each was screened for tick borne pathogens, here

Neoehrlichia mikurensis.

The color

indicates how many pools were positive at each site.

Source: National Veterinary Institute, Technical University of Denmark.

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7. Status on national and EU targets

By Gudrun Sandø ([email protected]) and Pernille C.S. Tillisch ([email protected])

In Denmark, action plans and programs on zoonoses

have been in place for more than 25 years. The first plan

targeted

Salmonella

in the broiler production and was

developed as a response to an increase in the number of

human cases related to eating chicken meat. Since then,

plans have been developed for

Salmonella

in pigs and pork,

Salmonella

in layers (eggs),

Campylobacter

in broilers and

Dublin in cattle and beef.

All plans have been outlined in cooperation between

industry, research institutes and authorities, and are

followed by a technical working group and a steering

committee. This ensures progress, that new knowledge is

incorporated in the plans, and an assessment of achieve-

ment of targets.

At EU level, harmonized surveillance programs and

common targets have been set for the broiler and laying

egg production.

An overview on the status on the targets can be seen

in Table 7.1.

7.1 National targets

The first action plan for

Campylobacter

was initiated in

2008 and followed by the second plan in 2013 that covers

the period until the end of 2016. The plan is targeting

Campylobacter

in broilers and chicken meat as well as other

sources and food. Targets have been set for the reduction

of the prevalence of

Campylobacter

positive flocks and of

the relative risk related to eating chicken meat. At flock

level the target is a 20% sum of reductions in 2011-2013

and 2014-2016. A reduction of 9.4% was obtained from

2011-2013, and the target for 2014-2016 is a reduction of

11%. At slaughterhouse level the target is a reduction of the

relative risk by 25% in 2014 and by 50% in 2016 compared

to 2013. A reduction of 28% was obtained in 2014.

The first action plan for reduction of

Salmonella

the broiler production was developed by the industry in

1988. An official plan of action was adopted in 1996 and

included the table-egg production as well. Both action

plans have been adjusted several times over the years.

The target was from the beginning an eradication of

Salmonella

from the broiler production. Today there is a

zero-tolerance of

Salmonella

in Danish produced broiler

meat. Meat from

Salmonella

positive flocks must be heat

treated, and broiler meat tested positive for

Salmonella

after slaughter cannot be marketed as fresh meat. In the

table-egg production, Denmark has achieved special

guarantees for

Salmonella

in the EU (for further infor-

mation see Annual Report 2012, Chapter 7). Both plans

are now focused on maintaining the low prevalence.

In the pig production, the fifth

Salmonella

action plan

was adopted at the end of 2013 and runs until the end of

2017. It continues the measures from former plans and

points at new measures as well (See Annual Report 2013

for more information). The target is set for the prevalence

Salmonella

positive carcasses at the slaughterhouse level

and is a maximum of 1% positive carcasses. The target

must be achieved in 2014 and maintained throughout the

period. At the end of 2014 the prevalence was just below

1%, and the target for 2014 was reached (Figure 7.1). The

trend in number of human

Salmonella

cases attributed

to the pig reservoir in the Salmonella source account is

evaluated annually. Since the large outbreaks in 2008-

2010, the annual number of cases in outbreaks from do-

mestic pork ranged between 0 and 65, and the estimated

number of sporadic cases ranged between 63 and 120,

resulting in an increasing overall number (Figure 7.2).

The first action plan for eradication of

Dublin

in cattle and beef was adopted in 2008 and has been

changed several times since then. In recent years, the

program has been tightened as the number of posi-

tive herds has been declining (for further information

see Annual Report 2013, Chapter 5). The target is to

eradicate

Dublin in cattle herds by the end of 2016.

7.2 EU targets

Based on the results of baseline studies in flocks of

poultry (Gallus

gallus

and turkeys), harmonised regulation

on targets and surveillance in the poultry production has

been laid down by the Commission.

In 2010, the EFSA published a scientific opinion[1] on

monophasic

Typhimurium-like strains in which it was

concluded that the monophasic

Salmonella

strains with

the antigenic formula

1,4,[5],12:i:- should be treated

equal to

Typhimurium. Subsequently EU regulations on

target and micribiological criteria set on

Salmonella

have

included these types as well.

The EU target for breeding and fattening turkey flocks

is laid down in Regulation (EC) No 1190/2012. According

to this regulation the target is maximum 1% flocks positive

for

Typhimurium and

Enteritidis. In Denmark, no

turkey flocks were positive with

Salmonella

in 2014 (Ap-

pendix Table A12).

In breeding flocks of

Gallus gallus,

the target according

to Regulation (EC) No 200/2010 is maximum 1% adult

flocks positive for

Typhimurium, including the mono-

phasic

1,4,[5],12:i:- strains,

Enteritidis,

Hadar,

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Figure 7.1.

Salmonella

in pork based on swab samples from carcasses, estimated prevalence

, 2001-2014

2.5

% positive samples

2.0

1.5

1.0

0.5

0.0

2001

2003

2005

2007

2009

2011

2013

a) Percent positive single samples, based on occurrence of

Salmonella

in pooled and single samples (see Appendix Table A5).

Source: Danish Veterinary and Food Administration.

Figure 7.2. Estimated number of human cases of salmonellosis and foodborne

Salmonella

outbreaks due to Danish pork,

2006-2014

Number of estimated human cases

400

300

200

100

2006

2007

2008

2009

2010

2011

2012

2013

2014

Danish pork (estimated sporadic)

Danish pork (outbreaks)

CI95% Lower

Source: Annual Report on

CI95% Upper

Zoonoses in Denmark from 2006 to 2014, Danish Zoonoses Centre, National Food Institute.

Infantis and

Virchow. In the legislation, no distinction

is made between breeding flocks from the table egg and

broiler production lines. In Denmark, three breeding

flocks from the broiler production were positive with

Salmonella.

The flocks were infected with

Typhimurium

DT41,

1,4,12:i:- DT120 and

Bareilly. In total 1.3%

of breeding flocks were positive for target

Salmonella

seotypes (Appendix Table A9 and A11), and the target

has not been reached.

The EU baseline study on table egg laying flocks car-

ried out in 2004 showed large differences in the prevalence

between Member States. Therefore, Member States specific

targets, according to Regulation (EC) No 517/2011, are

set either as an annual 10-40% reduction of positive adult

flocks dependent on the prevalence of positive adult flocks

in the Member State the previous year or a maximum of 2%

adult flocks positive. The target is set for

Typhimurium,

including the monophasic S. 1,4,[5],12:i:- strains, and

Enteritidis. For Denmark, the target is a maximum of 2%

adult flocks positive for the target serotypes. The preva-

lence in Denmark has been below 2% since 2004. In 2014,

two flocks (0.6%) were positive, both with target serotypes

(one flock with

Enteritidis FT21 and one flock with

Typhimurium DT40) (Appendix Table A9).

In broiler flocks of

Gallus gallus,

the target is maximum

1% flocks positive for

Enteritidis and

Typhimurium,

including the monophasic

1,4,[5],12:i:- strains, and is

laid down in Regulation (EC) No 200/2012. Denmark has

had intensive

Salmonella

control programs since the 1990’s

and the target of 1% was reached in 2000. In 2014, 0.7% of

broiler flocks were positive with

Salmonella,

and 0.4% of

the flocks were positive with target serotypes (Appendix

Table A11).

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Status on targets

Table 7.1. Status on targets for Campylobacter and Salmonella,

2014

National Action Plans

Target

Campylobacter

in broilers 2013-2016

Reduction in prevalence of positive

Flocks at farm

flocks: 20% (sum of reductions in two

periods: 2011-2013 and 2014-2016)

Fresh meat at slaughterhouse Reduction of the relative human risk

(RR) compared to the level in 2013

2014: RR reduced by 25%

2016: RR reduced by 50%

Salmonella

in poultry

Laying hen flocks of

Gallus

gallus

Carcasses at slaughterhouse

Initially eradication, later a reduction

strategy in the table egg production

Initially eradication, later a reduction

strategy in the broiler production

Zero-tolerance in Danish broiler meat.

Max. 1%

Salmonella

at carcass level in

2014-2017

No considerable increase in number of

estimated human cases from pigs in the

Danish

Salmonella

source account

Status

A reduction of 9.4% was obtained

for the period 2011-2013

A reduction of 28% was obtained in

2014 compared with 2013

0.6% (two flocks) (Table A9-A10)

Eggs from positive flocks are de-

stroyed or heat treated

1.4% positive batches (Table A11)

Positive batches are heat treated

Salmonella

in pigs 2014-2017

Carcasses at slaughterhouse

Estimated human cases from

pigs

a) Data from 2013 and 2014 cannot be compared due to a change in sampling per 1 January, 2014 from AM testing of sock samples

7-10 days before slaughter to cloacal swabs at the point of slaughter. The target has therefore been changed to cover two periods:

2011-2013 and 2014-2016. The target is an overall 20% reduction (the sum of percent reductions in the two periods).

b) Data from 2013 has been agreed as the baseline since 2012-data are not comparable with data from 2013 and onwards due to a

necessary improvement in the collection of data.

c) Supplementary to EU-regulations.

d) See Table A36 for explanation of the herd levels .

e) Including the monophasic strains

1,4,[5],12:i:-.

Source: Danish Veterinary and Food Administration.

Salmonella

Dublin in cattle 2012-2016

Herds at farm

Eradication of

Dublin in all herds by 6.3% of milk-producing herds and

2016., i.e. all herds in level 1

by the end 2.9% of non-milk producing herds

of 2016

are in level 2 or 3 (07.01.2015) (Table

A17)

EU Regulations

Regulation (EC) No. 1190/2012

Breeding and fattening turkey Max. 1% positive for

Enteritidis and No positive flocks (N=10)

flocks

Typhimurium

(Table A12)

Regulation (EC) No. 200/2010

Breeding flocks of

Gallus gallus

Max. 1% adult flocks positive for

2.0% (3 flocks) (Table A9 and A11)

Typhimurium ,

Enteritidis,

Hadar, 1.3% (2 flocks) with target serovars

Infantis and

Virchow

Regulation (EC) No. 1168/2006

Laying hen flocks of

Gallus

MS specific targets, for Denmark:

0.6% (two positive flocks with target

gallus

Max. 2% adult flocks positive for

serovars) (Table A9)

Typhimurium and

Enteritidis

Regulation (EC) No. 646/2007

Broiler flocks of

Gallus gallus

Max. 1% positive

Typhimurium

and 0.7% (26 positive flocks) (Table A11)

Enteritidis

0.4% (14 flocks) with target serovars

Overall 0.99% at carcass level in

2014 (Table A15)

In total 172 cases attributed to

Danish pork in 2014 (CI95%: 126-

214) vs. 128 cases in 2013 (CI95%:

86-163).

(Chapter 1, Figure 7.2, and Table

A1)

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International topica

8. International topics

By Inge-Lis Kyllesbæk Andersen ([email protected]) and Hanne Rosenquist ([email protected])

8.1

Trichinella

In June 2014 an amendment to the EU Regulation on

Trichinella

came into force. This new provision included

among other things a new sampling regime for

Trichinella

and introduction of the concept of “Controlled housing

conditions”. Slaughter pigs, sows and boars, which are

kept under controlled housing conditions, are exempted

testing. Free range pigs, horses, wild game (e.g. wild boar)

and other species, susceptible to

Trichinella

must be tested.

8.2 Ebola and imported foods

In 2014, an outbreak of Ebola virus in West Africa,

primarily in the countries Guinea, Liberia and Sierra

Leone, increased dramatically in size, and by the end of

2014 more than 20.000 confirmed, probable, and suspected

cases of Ebola virus disease with around 8000 deaths were

reported by WHO (http://apps.who.int/ebola/en/current-

situation/ebola-situation-report). Due to the seriousness

of the outbreak, WHO has declared the outbreak a Public

Health Emergency of International Concern.

Ebola is transmitted through direct contact with blood

or other bodily fluids from infected people. Concerns have

been raised about the risk of transmission through illegal

import of bush meat from Africa into Europe, since several

animal species, mainly primates and fruit bats, have been

found to harbor Ebola virus, and illegal import of bush

meat from Africa into e.g. France has shown to be large

(estimated to around five tonnes per week in personal bag-

gage through Paris Roissy-Charles de Gaulle airport) [1].

The risk from handling and consuming bush meat was

assessed by EFSA [2]. EFSA concluded that the potential

for introducing and transmitting Ebola virus via bush meat

to Europe is currently categorised as low risk. However,

due to lack of data and knowledge, which results in very

high uncertainty, the exact risk could not be estimated.

In conclusion, the low risk is explained by (i) the limited

number of outbreaks confirmed to date in Africa in spite of

the routine consumption of bush meat on that continent,

(ii) the handling of bush meat in Europe not involving

high risk practices such as hunting and butchering, and

(iii) the assumed low overall consumption of bush meat

in Europe. The probability of transmission of Ebola via

bush meat to Denmark is considered to be lower than for

Europe in general. This is because illegal import of bush

meat to Denmark is considered to be negligible. Further,

the border control is aware of the risk.

EFSA and ECDC have also assessed the risks related to

dogs and cats having been in contact with people infected

with Ebola virus [3]. In Europe, this event is assumed to be

very rare. If it happens, the probability of the pet becoming

infected may range from very low to high. A similar range

for the probability (low-high) was assessed for human ex-

posure to the virus through infected pets, as it will depend

on the specific circumstances. EFSA and ECDC therefore

recommend that the risk in each case is assessed jointly by

veterinary and public health authorities.

8.3 References

1. Chaber A-L, Allebone-Webb S, Lignereux Y, Cun-

ningham AA and Rowcliffe JM (2010). The scale of illegal

meat importation from Africa to Europe via Paris. Con-

servation Letters, 3, 317-323.

2. EFSA (European Food Safety Authority) (2014). An

update on the risk of transmission of Ebola virus (EBOV)

via the food chain. EFSA Journal;12(11):3884, 25 pp.

3. EFSA (European Food Safety Authority) (2014). Risk

related to household pets in contact with Ebola cases in

humans. EFSA Journal;12(12):3930, 12 pp.

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9. Surveillance and control

programmes

The collaboration on zoonoses between national and

regional authorities, the industry and non-governmental

organizations in Denmark is presented in Figure 9.1. Ac-

cording to the Danish legislation, 41 infectious diseases are

notifiable in Denmark. An overview of the notifiable and

non-notifiable human and animal diseases, presented in this

report, is provided in Appendix Table A30 and Table A31,

respectively, including reference to the relevant legislation.

9.1 Surveillance of human disease

Information on human cases due to zoonotic pathogens

presented in this report is reported to Statens Serum Institut

through different channels depending on the disease:

•

Notifiable through the laboratory surveillance system:

Salmonella, Campylobacter, Yersinia,

Verotoxin-produ-

cing

E. coli

(VTEC) and

Listeria.

•

Individually notifiable zoonotic pathogens:

Chlamy-

dia psittacci

(ornithosis),

Leptospira

(Weils disease),

Mycobacterium,

Bovine Spongieform Encephalopathy

(BSE) prions (var. Creutzfeldt-Jakob Disease), Vero-

toxin-producing

E. coli

(VTEC) and Lyssavirus (rabies).

•

Non-notifiable zoonotic pathogens:

Brucella, Crypto-

sporidium, Echinococcus

and

Trichinella.

In Denmark, the physicians report individually notifiable

zoonotic diseases to the Danish Health and Medicines Aut-

hority and the Department of Infectious Disease Epidemio-

logy at Statens Serum Institut. Physicians send specimens

from suspected cases to one of the clinical microbiology

laboratories depending on the geographical region. Positive

cases diagnosed by a clinical microbiological laboratory are

reported through the laboratory surveillance system to the

Unit of Gastrointestinal Infections at Statens Serum Institut.

The laboratories must report positive results to Statens Se-

rum Institut within one week. Furthermore, all

Salmonella

and VTEC isolates are sent to the reference laboratory at

Statens Serum Institut for further sero- and genotyping.

The results are recorded in the Register of Enteric Pathogens

maintained by Statens Serum Institut. Cases are reported as

episodes, i.e. each patient-infectious agent combination is

only recorded once in any six-month period. Overviews of

results from the Register of Enteric Pathogens are presented

as follows:

•

All laboratory confirmed human cases are presented in

Appendix Table A2.

•

VTEC O-group distribution in humans is presented in

Appendix Table A3.

•

The

Salmonella

serovar and MLVA distribution is pre-

sented in Appendix Table A5-A8.

Figure 9.1. Overview of the monitoring and outbreak investigation network for reporting infectious pathogens in humans,

animals, foodstuffs and feedstuffs in Denmark, 2013

Ministry of Food,

Agriculture and Fisheries

Danish Veterinary and Food

Administration

Ministry of Health

Danish Health and Medicines

Authority

General Practioners

& Hospitals

Central Outbreak

Management Group

Statens Serum Institut (SSI)

Clinical Microbiology

Industry

Ministry of Higher Education and

Science

Tecnical University of Denmark

Ministry of the

Environment

Danish Environmental

Protection Agency

Non-governmental

Organisation

National Food Institute

Danish Zoonosis

Centre

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9.2 Outbreaks of zoonotic gastrointestinal

infections

In Denmark, local and regional foodborne outbreaks

are typically investigated by the Food Control Offices

collaboration with the medical officers at the Danish Health

and Medicines Authority, and the regional clinical micro-

biology laboratories. Larger regional and national outbreaks

are investigated by Statens Serum Institut, the National Food

Institute, Technical University of Denmark and the Danish

Veterinary and Food Administration in collaboration. These

institutions may also aid in the investigation of local outb-

reaks. Representatives from these institutions meet regularly

in the Central Outbreak Management Group to discuss

surveillance results, compare the reported occurrence of

zoonotic agents in animals, food and feedstuffs with that in

humans, react upon outbreak alerts and coordinate investi-

gations. The formal responsibility of food- or waterborne

outbreak investigations is divided between three ministeries

based on the outbreak source: the Ministry of Health for

infectious diseases; the Ministry of Food, Agriculture and

Fisheries for foodborne and animal related diseases; and the

Ministry of the Environment (along with the municipalities)

for waterborne diseases.

Outbreaks may be detected in various ways. Individuals

who experience illness related to food intake in settings

such as restaurants or work place cafeterias may report

these incidents directly to the Food Control Office. General

practitioners and hospitals are obliged to report all suspected

water- and foodborne infections to the Danish Health

and Medicines Authority and to Statens Serum Institut.

Further, clusters of cases may be noted in local laboratories

or identified through the laboratory surveillance system of

gastrointestinal bacterial infections or subtyping of bacterial

isolates from patients, both at Statens Serum Institut.

A list of verified outbreaks (not including household

outbreaks) reported to the Food- and waterborne Outbreak

Database (FUD) is presented in Appendix Table A4 and

some of the outbreaks from 2014 are outlined in Chapter 2.

9.3 Surveillance and control of animals and

animal products

Salmonella

surveillance and control programmes for

poultry, pigs and cattle are presented in Appendix Tables

A32-A37. Sample analysis is performed at authorised private

laboratories, the Danish Food and Veterinary Administrati-

ons laboratory, the National Food Institute and the National

Veterinary Institute at the Technical University of Denmark.

Salmonella

isolates are forwarded to the National Food

Institute for serotyping, some isolates are also phage- and

genotyped as well as tested for antimicrobial resistance. An

overview of the methods used for subtyping is presented in

Appendix Table A38.

Overviews of results from surveillance and control of

Salmonella

are presented as follows:

•

Results from the table egg production are presented in

Appendix Tables A5-A7 and A9-A10.

•

Results from the broiler production are presented in Ap-

pendix Tables A5-A7, A11 and A18.

•

Results from the duck and turkey productions are pre-

sented in Appendix Table A5-A6, A12 and A18.

•

Results from the pig production are presented in Appen-

dix Tables A5-A6, A15, A18 and Figures A1-A3.

•

Results from the cattle production are presented in Ap-

pendix Tables A5-A6, A8, A16-A17 and Figure A4.

•

Results from the feeding stuff production are presented

in Appendix Tables A19-A20.

•

Results from the rendering plants are presented in Ap-

pendix Table A21.

•

Results based on suspicion of diseases in pets, zoo animals

and wild life are presented in Appendix Table A23-A24.

Overviews of results from monitoring and control of

Campylobacter

are presented as follows:

•

Results from the broiler production are presented in Ap-

pendix Tables A13-A14 and A18.

•

Results based on suspicion of diseases in pets, zoo animals

and wild life are presented in Appendix Table A23-A24.

Pig and cattle carcasses are screened for

Mycobacterium

and

Echinococcus

during meat inspection at the slaughter-

house. Although Denmark is assigned as a region where

the risk of

Trichinella

in domestic swine is negligible, all

slaughter pigs are still examined for

Trichinella

at slaughter

as well as wild boars, and horses slaughtered for human con-

sumption. In addition, boars and bulls are tested for

Brucella

and bulls are tested for

Mycobacterium

at semen collection

centres. All positive results for notifiable infectious diseases

are reported to the Danish Veterinary and Food Admini-

stration. Results are presented in Appendix Table A15-A16.

Results from the surveillance for Bovine Spongiform

Encephalopathy (BSE) in cattle, and Transmissible Spongi-

form Encephalopathy (TSE) in sheep/goat are presented in

Appendix Tables A25-A27.

Results from the monitoring of

Coxiella burnetii

fever) in cattle are presented in Appendix Table A16.

Results based on suspicion of diseases with

Chlamydia

_________________________________________________________________________________________________________

1) The Danish Veterinary and Food Administration (DVFA) is one authority but operates from more locations throughout the country.

To be able to distinguish the locations the terms DVFA is used synonymous with the location in Glostrup and Food Control Office

followed by the location synonymous with the location in question.

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Surveillance and control programmes

psittacci, Cryptosporidium, Trichinella,

classical rabies and

European Bat

Lyssavirus

in zoo animals, pets and wild life

are presented in Appendix Table A23-A24.

9.4 Official testing of zoonotic pathogens in

foodstuffs

In Denmark, control of zoonotic microorganisms in

foodstuffs is mainly carried out as projects which are co-

ordinated at the central level of the Danish Veterinary and

Food Administration. Sampling and testing are carried out

with the following purposes:

•

To verify that food business operators comply with mi-

crobiological criteria laid down in the legislation

•

To verify the microbiological safety of food for which no

microbiological criteria are laid down at EU Community

level.

•

To monitor the effect of established risk management

procedures in order to evaluate if these provide the desired

results or need to be reconsidered.

•

To generate data for the preparation of risk profiles and

risk assessments to support microbial risk management

•

To discover emerging problems with microbiological

contaminants.

Appendix Table A28 provides information on the cen-

trally coordinated studies conducted in 2014.

For further information consult the website of the Da-

nish Veterinary and Food Administration, www.fvst.dk.

New legislation on

Salmonella

and

Campylobacter

in broilers and

Salmonella

in pigs and pork

By Gudrun Sandø ([email protected])

The changes of Order no. 1512 of 13/12/2013, regulating the control of

Salmonella

and

Campylobacter

in poultry,

include

•

Change of testing for Campylobacter.

12 cloacal swabs from 24 animals from each flock are to be analyzed in

one pool instead of, as in 2010-2013, sock samples in the flock.

•

Salmonella

control in ducks is no longer included in the programme, as no action is taken on isolation of

Salmonella

in ducks, and the prevalence is known to be high.

•

The Salmonella

control after slaughter now includes more samples at the small slaughterhouses.

The changes of Order no. 1183 of 06/11/2014, regulating the control of

Salmonella

in pigs and pork, include

•

A new way to calculate the index, that divides the herds into the categories 1,2 or 3 (footnote g) Table A37).

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Appendix

Trends and sources in human salmonellosis

Table A1. Estimated no. of reported human cases and percentage of cases per major food source, travel or outbreaks,

2012-2014

2014

Source

Domestic pork

Domestic beef

Domestic table eggs

Domestic broilers

Domestic ducks

Imported pork

Imported beef

Imported broilers

Imported turkey

Imported duck

Travels

Unknown source

Outbreaks,

unknown source

Total

Estimated no. of

reported cases

(95 % credibility

interval

)

172 (126-214)

25 (20-31)

33 (22-47)

22 (1-69)

No data

13 (0-44)

3 (0-7)

33 (14-53)

22 (11-34)

538 (528-549)

216

(

178-252)

1,122

Percen-

tage of

reported

cases

15.4

2.2

3.0

2.0

1.1

0.2

2.9

2.0

48.0

19.2

4.0

2013

Estimated no. of

reported cases

(95 % credibility

interval

)

128 (86-163)

21 (4-58)

17 (7-31)

9 (0-21)

30 (15-50)

(13-33)

12 (2-26)

7 (0-18)

No data

458 (445-471)

228 (191-264)

204

1,136

Percen-

tage of

reported

cases

11.3

1.9

1.5

0.8

2.6

2.0

1.1

0.6

40.3

20.1

18.0

2012

Estimated no. of

reported cases

(95 % credibility

interval

)

110 (84-139)

85 (72-100)

15 (1-35)

10 (1-23)

3 (0-10)

11 (4-20)

21 (3-47)

13 (1-28)

22 (13-34)

539 (527-550)

332 (293-369)

1,198

Percen-

tage of

reported

cases

8.0

7.1

1.3

0.8

0.2

0.9

1.8

1.1

1.6

45.0

27.7

4.3

a) The model is based on a Bayesian framework which gives 95% credibility intervals.

b) No samples from imported turkey meat were found positive for

Salmonella

in 2014.

c) No samples from domestic broiler meat were found positive for

Salmonella

in 2012 and 2013.

d) No data from imported beef in 2013. The number of cases attributed to this source was modelled from previous years’ data.

Source: Danish Zoonosis Centre, National Food Institute.

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Appendix

Human disease and outbreak data

Table A2. Zoonoses in humans, number of laboratory-confirmed cases, 2009-2014

a) Not notifiable, hence the incidence cannot be calculated.

b) Notifiable.

Typhimurium and the monophasic

1,4,[5],12:i:- strains.

d) Data presented are from one laboratory (Statens Serum Institut) only, representing a proportion of the Danish population. The

proportion of the population represented varies from year to year, thus results from different years are not comparable. Testing for

these pathogens is carried out only if specifically requested on the submission form.

e) The cases were imported.

f) Includes 19 cases verified by PCR only (see Table A3).

g) The nation-wide neonatal screening of congenital toxoplasmosis stopped in 2007

Source: Statens Serum Institut.

Table A3.

VTEC

O-group distribution in humans

, 2014

Zoonotic pathogen

Bacteria

Brucella abortus/melitensis

a,d

Campylobacter coli/jejuni

Chlamydia psittaci

Leptospira

spp.

Listeria monocytogenes

Mycobacterium bovis

Salmonella

total

Enteritidis

Typhimurium

b,c

Other serotypes

VTEC total

O157

Other O-groups or non-typeable

Yersinia enterocolitica

Parasites

Cryptosporidium

spp.

a,d

Echinococcus multilocularis

a,e

Echinococcus granulosus

a,e

Trichinella

spp.

a,e

Viruses

Lyssavirus

Incidence

per 100,000

inhabitants

2014

67.1

0.3

0.2

1.6

0.02

19.9

4.8

7.6

4.4

0.7

3.4

7.7

Reported no. of cases

2014

3,782

1,122

268

427

248

192

432

2013

3,766

1,136

346

337

453

186

163

345

2012

3,728

1,198

242

415

541

190

154

291

2011

4,068

1,166

293

386

487

224

197

224

2010

4,035

1,598

388

521

689

184

159

192

2009

3,352

2,129

600

767

762

165

141

238

O-group

O157

O103

O26

O146

O145

O117

Number of episodes

Continued in the next column

O-group

O91

O-rough

Other O-groups or non-typeable

Isolates total

Confirmed by PCR only

Notified

Total

Number of

episodes

229

280

a) All O-groups that resulted in five or more episodes are listed.

b) The cases are reported through the notification system, isolates or DNA not available for verification.

Source: Statens Serum Institut.

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Appendix

Table A4. Food- and waterborne disease outbreaks

reported in the Food- and Waterborne Outbreak Database (FUD)

(n=60), 2014

Pathogen

Bacillus cereus

Campylobacter

Clostridium perfringens

E. coli

(not typed)

E. coli

(AEEC) O171:H19

Histamine

L. monocytogenes,

MLST224

d,e

L. monocytogenes,

MLST391

L. monocytogenes,

MLST399

L. monocytogenes,

MLST6

Lectines

Norovirus

Patients

No. of

laboratory

patients

confirmed

391

147

153

430

134

Setting

Restaurant

Private party

Sports event

Restaurant

Catering

Restaurant

National

Hospital

National

Canteen

Institution

Restaurant

School

Shop

Restaurant

Conference Center

Restaurant

Canteen

Restaurant

Private party

Restaurant

Institution

Canteen

Restaurant

Canteen

Restaurant

Private party

Canteen

Source

Composite meal

Chicken (imp/dk)

Chicken

Composite meal

Buffet meal

Herbs (imp)

Composite meal

Fish

Cold cuts

Unknown

Composite meal

Fish

Dried beans

Buffet meal

Composite meal

Strawberries (imp)

Composite meal

Unknown

Buffet meal

Composite meal

Oysters (imp)

Composite meal

Rasberries (imp)

Buffet meal

Composite meal

Buffet meal

Composite meal

Buffet meal

Oysters (imp)

Buffet meal

Composite meal

FUD

no.

1403

1437

1375

1420

1417

1400

1394

1393

1390

1359

1418

1387

1401

1373

1376

1384

1385

1407

1439

1435

1433

1432

1419

1412

1411

1405

1404

1402

1398

1397

1389

1388

1383

1371

1369

1367

1365

1364

1363

1362

1353

1352

Continued on the next page

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Appendix

Table A4. Food- and waterborne disease outbreaks

reported in the Food- and Waterborne Outbreak Database (FUD)

(n=60), 2014 (Continued from previous page)

Pathogen

Agona

S. Enteritidis, MLVA0206

c,g

Enteritidis, MLVA0017

Enteritidis, MLVA0019

Infantis

Salmonella

1,4,5,12:i:-, MLVA0201

Typhimurium, MLVA1788

Salmonella

1,4,12:i:-, MLVA0007

Salmonella

1,4,5,12:i:-, MLVA1277

Salmonella

1,4,5,12:i:-, MLVA0008

Salmonella

1,4,5,12:i:-, MLVA0334

Shigella sonnei

Unknown

VTEC O103:H2,

eae

and

ehxA, vtx1a

VTEC O157:H-, eae,

vtx1a, vtx2a

VTEC O157:H7, eae,

vtx1a, vtx2a

Yersinia enterocolitica

O:3, biotype 4

Total

Patients

No. of

laboratory

patients

confirmed

2,209

295

Setting

National

Restaurant

Regional

National

Shop

National

Sports event

Private party

Canteen

Restaurant

National

Institution

Restaurant

National

Source

Unknown

Chicken (imp)

Eggs

Chicken (imp)

Unknown

Pork

Unknown

Pork

Minced beef

Unknown

Pork

Sugar snaps (imp)

Composite meal

Unknown

Beef

Unknown

FUD

no.

1317

1327

1391

1379

1380

1370

1372

1410

1368

1374

1378

1446

1408

1431

1377

1392

1409

1354

Note: (imp)= imported product. (imp/dk)= uncertain origin of product.

a) In addition to the above mentioned outbreaks, 1 household outbreak (FUD 1350) with 2 persons involved was registered in 2014.

The cause of the otbreak was possible presence of histamine in tunaseaks.

b) Outbreak FUD No 1317 - Additional 13 cases had onset in 2013. However, these cases were not reported in Annual Report 2013.

c) Outbreak FUD No 1327 referred to in Chapter 2 concerning, in total, 12 cases of

Salmonella

Enteritidis, MLVA0206 was already

mentioned in the report for 2013. Seven of these cases are therefore not presented in this table.

d) Multi-Locus Sequence Type (see chapter 3).

e) Four cases in FUD No. 1373 had onset in 2014.

f) Cold smoked fish.

g) MLVA profiles for the most common human MLVA-types can be found in tables A6, A7 and A8.

Source: Food- and waterborne Outbreak Database (FUD).

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Appendix

Monitoring and surveillance data

Table A5. Top 15 (humans) serotype distribution (%) of

Salmonella

from humans, animals, carcasses and meat, 2014.

N=number of culture positive units

Human

cases

Serotype

Enteritidis

1,4,[5],12:i:-

Typhimurium

Infantis

Dublin

Stanley

Newport

Virchow

Agona

Kentucky

Chester

Paratyphi B var Java

Oranienburg

Derby

Muenchen

Others

Unknown

Total

Pig

Pork

Beef

N=11

80.8

9.1

100

Layer

Broiler

flocks

N=26

3.8

30.8

19.2

26.9

3.8

15.4

100

batches

N=6

66.7

16.7

100

N=2

50.0

100

Imported meat (batches)

Pork

N=20

31.8

22.7

4.5

22.7

13.6

4.5

100

Beef

Broiler

Ducks

N=4

25.0

50.0

100

N=9

8.3

25.0

8.3

25.0

33.3

100

N=24

4.2

41.7

20.8

33.3

100

animals batches batches flocks

N=1,122 N=170 N=108

23.9

20.5

17.6

3.4

1.9

1.7

1.6

1.4

1.0

0.9

0.8

18.0

2.3

100

22.4

18.2

1.2

55.9

2.4

100

22.2

15.7

6.5

45.4

1.9

8.3

100

a) One isolate per serotype per unit is included, thus the number of isolates may exceed the number of units. Thus, in 2014 more

isolates were included from imported pork and broiler meat.

b) Isolates collected from coecum samples taken randomly at slaughter. Where more than one

Salmonella

positive pig with different

serotypes was randomly selected from a herd, one pig per serotype was included.

c) Sampling of pork carcasses at slaughterhouses according to the surveillance programme (Table A37).

d) Data are from sampling of beef carcasses at slaughter houses according to the surveillance programme (10 positive samples) (Ta-

ble A36) and from a centrally coordinated study (one positive sample) (see section 9.4 and Table A28 for description).

e) Sampling of production flocks prior to slaughter according to surveillance programmes (Tables A33).

f) Sampling of broiler meat (neck skin) at slaughterhouses according to the surveillance programme (4 positive batches) (Table A33),

and case-by-case control of Danish meat (2 positive batches).

g) Case-by-case control of imported meat. One batch of imported pork with three serotypes:

Typhimurium (3),

1,4,[5],12:i:- (2)

and

Derby (1). One batch of imported broiler meat with four serotypes:

Paratyphi B var Java (1),

1,4,12:b:- (1),

6,7:r:- (1)

and

Infantis (1). For further information regarding case-by-case control programme and Annual Report on Zoonoses in Den-

mark, 2007.

h) Centrally coordinated study (see section 9.4 and Table A28 for description).

Source: Danish Veterinary and Food Administration, Statens Serum Institut, and National Food Institute.

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Appendix

Table A6. Top 10 (humans) MLVA

distribution (%) of

Salmonella

Typhimurium including the monophasic S. 1,4,[5],12:i:-

from humans, animals, carcasses and imported meat, 2014. N= number of isolates

MLVA type

STTR 9|5|6|10|3 Danish

3|12|9|NA|0211 0007

3|14|9|NA|0211

3|13|9|NA|0211

3|12|10|NA|0211

3|13|10|NA|0211

3|12|17|NA|0211

3|14|8|12|0311

3|14|12|NA|0211

3|14|10|NA|0211

Other

Total

0201

0008

0005

0192

1277

1690

0334

0126

Human

cases

N=423

9.2

7.8

5.7

4.3

3.8

2.8

2.1

1.9

1.4

56.7

100

Pork

batches

N=40

2.5

12.5

5.0

7.5

2.5

70.0

100

Layer

flocks

N=1

100

Broiler

flocks

N=13

15.4

69.2

100

Broiler

batches

N=3

33.3

66.7

100

Imported meat (batches)

Pork

N=11

20.0

6.7

66.7

100

Beef

N=1

100

Ducks

N=11

100

5|19|NA|NA|0201 1788

For footnotes a-h see Table A5.

i) The isolates are analysed for the following loci:

STTR9|STTR5|STTR6|STTR10|STTR3 and the results are reported in the

same order in the table. ”Danish” is the Danish MLVA-number for the MLVA profile. ”NA”=

locus missing.

Source: Danish Veterinary and Food Administration, Statens Serum Institut, and National Food Institute.

Table A7. Top 10 (humans) MLVA

distribution (%) of

Salmonella

Enteritidis from humans, animals and imported meat, 2014.

N= number of isolates

Table A8. Top 10 (humans) MLVA

distribution (%) of

Salmo-

nella

Dublin from humans, carcasses and imported meat,

2014. N= number of isolates

Broi-

MLVA type

Human Layer

ler

cases flocks flocks

SE 1|5|2|9|3 Danish N=268 N=1 N=1

4|10|5|3|1 0019

27.7

100

3|10|7|2|2

4|9|5|3|1

4|11|5|3|1

4|11|4|3|1

3|11|7|2|2

4|9|8|2|2

3|10|8|2|2

3|13|9|2|2

4|12|5|3|1

Other

Total

0004

0017

0020

0034

0029

0206

0005

0022

0031

16.4

15.4

12.3

7.7

7.2

4.1

3.6

3.1

2.6

27.2

100

Imported

meat (batch)

Broiler

N=1

100

Human Beef

cases batches

SE1|SE2|SE5|SD1 Danish N=21

N=9

2|3|14|4

0043

14.3

2|4|11|6

Other

Total

0104

14.3

71.4

100

MLVA type

i,j

Imported

meat (batch)

Beef

N=1

100

For footnotes a-h see Table A5.

i) The isolates are analysed for the following loci:

SE1|SE2|SE5|SD1 and the results are reported in the same or-

der in the table. ”Danish” is the Danish MLVA-number for the

MLVA profile.

j) Kjeldsen MK, Torpdahl M, Campos J, Pedersen K, Nielsen

EM (2014). Multiple-locus variable-number tandem repeat

analysis of

Salmonella enterica subsp. enterica

serovar Dublin. J

Appl Microbiol. 116(4):1044-54.

k) One isolate of each MLVA-type.

Source: Danish Veterinary and Food Administration, Statens

Serum Institut, and National Food Institute.

For footnotes a-h see Table A5.

i) The isolates are analysed for the following loci:

SE1|SE5|SE2|SE9|SE3

and the results are reported in the same order in the table. ”Danish”

is the Danish MLVA-number for the MLVA profile.

Source: Danish Veterinary and Food Administration, Statens Serum

Institut, and National Food Institute.

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Appendix

Table A9. Occurrence of

Salmonella

in the table egg production

, 2005-2014

Rearing period

(parent flocks)

2005

2006

2007

2008

2009

2010

2011

2012

2013

2014

Positive

Adult period

(parent flocks)

Positive

Pullet-rearing flocks

355

289

326

258

253

225

195

197

173

150

Positive

Table egg layer flocks

655

565

510

508

454

455

410

359

373

347

Positive

a) See Tables A32 and A34 for description of the surveillance programmes.

b) One flock positive with

Enteritidis PT21, and one flock positive with

Typhimurium DT40. For information on MLVA

types see Tables A6-A7.

Salmonella

was not detected in grandparent flocks during rearing period (7 flocks).

Salmonella

was not detected in grandparent flocks during adult period (8 flocks).

Source: Danish Agriculture and Food Council, and Danish Veterinary and Food Administration.

Table A10. Occurrence of

Salmonella

in the table egg layer flocks sorted by type of production, 2005-2014

Deep litter

2005

2006

2007

2008

2009

2010

2011

2012

2013

2014

217

185

155

151

133

117

109

101

108

Positive

Free range

Positive

178

164

146

145

130

136

130

136

137

125

Organic

Positive

175

148

146

135

110

157

131

Battery

Positive

a) One flock positive with

Typhimurium DT40. For information on MLVA types see Table A6.

b) One flock positive with

Enteritidis FT21. For information on MLVA types see Table A7.

Source: Danish Agriculture and Food Council, and Danish Veterinary and Food Administration.

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Appendix

Appendix C

Table A11. Occurrence of

Salmonella

in the broiler production

, 2005-2014

Rearing period

(parent flocks)

2005

2006

2007

2008

2009

2010

2011

2012

2013

2014

214

190

152

146

140

126

114

123

128

121

Positive

Adult period

(parent flocks)

185

282

258

293

225

200

213

183

152

131

Positive

Broiler flocks

Positive

4,034

3,621

3,703

3,845

3,767

3,773

3,795

3,448

3,498

3,470

Slaughterhouse

(flocks/batches)

1,174

875

884

518

375

346

306

368

288

277

Positive

a) See Tables A32-A33 for description of the surveillance programmes.

b) In 2003-2005, only one flock per house was registered per year although there may have been more than one flock in the house,

however all flocks were sampled according to the surveillance programme.

c) From 2006, data cover only samples taken following the

Salmonella

programme. Verification samples taken once a week by pro-

ducers of poultry meat approved to market

Salmonella-free

poultry meat are not included. Collection of verification samples started

in the middle of 2005.

d) From 2008, all AM positive flocks are heat treated at slaughter. Sampling is now carried out as verification of the AM results of

the negative flocks.

e) Both isolates were

Typhimurium DT120, one MLVA 3|7|11|15|0311 and one with both 3|7|11|15|0311 and 3|7|12|14|0311. For

more information on

Typhimurium MLVA types see Table A6.

Typhimurium DT41 MLVA 2|12|12|8|0212 (1),

1,4,12:i:- DT120 MLVA 3|15|8|NA|0211 (1),

Bareilly (1). For more infor-

mation on

Typhimurium MLVA types see Table A6.

Typhimurium (DT170 (2), DT41 (2), RDNC (1)),

Enteritidis (PT1B (1)),

1,4,5,12:i:- (DT193 (7), Not phage typed (1)),

Senftenberg (1),

Infantis (7),

Indiana (1),

Newport (1),

Manhattan (1),

Tennessee (1).

1,4,5,12:i:- and

1,4,12:i:- DT193 (1),

1,4,5,12:i:- DT193 (2),

Infantis (1).

Salmonella

was not detected in grandparent flocks during rearing period (13 flocks).

Salmonella

was not detected in grandparent flocks during adult period (6 flocks).

Source: Danish Agriculture and Food Council, and Danish Veterinary and Food Administration.

Table A12. Occurrence of

Salmonella

in turkey and duck flocks, 2006-2014

Duck flocks

2006

2007

2008

2009

2010

2011

2012

2013

2014

266

108

% pos

80.5

64.7

63.5

56.5

58.1

49.0

20.3

Turkey flocks

% pos

10.0

4.2

2.6

3.6

a) See Table A35 for description of the surveillance programme for turkey flocks. The two

major turkey and duck slaughterhouses in Denmark closed down in 2004 and 2007, respec-

tively. Therefore, most commercially reared duck and turkey flocks are transported abroad

for slaughter.

Since 20/09/2013 samples from ducks were no more taken.

Source: Danish Agriculture and Food Council.

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Appendix

Table A13. Occurrence of

Campylobacter

in broiler flocks, 2005-2014

UKLART om N er swabs/socks eller flocks

Cloacal swabs at slaughter

2005

2006

2007

2008

2009

2010

2011

2012

2013

2014

4,952

4,522

4,527

4,950

4,591

3,474

% pos

30.4

30.8

26.8

26.3

29.4

27.7

Sock samples at farm

3,132

3,379

3,376

3,508

% pos

16.5

14.4

11.6

13.1

a) See Tables A33 for description of the surveillance programmes. In 2014 the sampling method changed back from boot swabs col-

lected in the stable 7-10 days before slaughter to cloacal swabs at slaughter according to Regulation no. 1512 of 13/12/2013.

Source: Danish Agriculture and Food Council, Danish Veterinary and Food Administration, and National Veterinary Institute,

Technical University of Denmark (until 2009).

Table A14. Occurrence of

Campylobacter

in non-heat treated broiler meat at slaughter and retail

, 2012-2014

Chilled broiler meat (samples)

At slaughter

Denmark

2012

Conventional

Organic/free-range

In total

2013

Conventional

Organic-free-range

In total

2014

Conventional

Organic/free-range

In total

1,044

870

927

108

% pos

21.5

28.2

90.3

25.7

75.0

521

849

884

Denmark

% pos

9.7

12.1

42.9

17.8

154

170

208

At retail

Import

% pos

28.2

12.8

71.1

31.9

a) Centrally coordinated studies (see Table A28 and section 9.4 for description). Limit of quantification: 10 cfu/g.

b) The prevalence is calculated as a mean of quarterly prevalences., except orgnanic/free-range results.

c) Leg-skin samples only.

d) Included are 238 leg-skin samples, prevalence = 24,4%.

Source: National Food Institute.

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Appendix

Figure A1. Serological surveillance of Salmonella in breeding and multiplying pigs

based on monthly testing of blood

samples, 2009-2014

% positive breeding- and

multiplying pigs

2009

2010

% positive

2011

2012

2013

2014

% positive, moving avg for 12 month

a) For more information about the surveillance programme, see Table A37.

Source: Danish Agriculture and Food Council.

Figure A2. Serological surveillance of

Salmonella

in slaughter pigs

, 2009-2014. Percentage of seropositive meat juice

samples (first sample per herd per month)

% positive slaughter pigs

2009

2010

% positive

2011

2012

2013

2014

% positive, moving avg. for 12 month

a) For more information about the surveillance programme, see Table A37.

b) The peaks in January 2010 and August 2011 were due to data transfer problems. The reason for the increase in late summer

2012 is unknown.

Source: Danish Agriculture and Food Council.

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Appendix

Appendix C

Appendix

Table A15. Occurrence of zoonotic pathogens in pigs and pork in Denmark, 2014

Herds

Zoonotic pathogen

At farm

Brucella abortus

Leptospira

At slaughterhouse (slaughter pigs)

Salmonella

spp.

c,d

Salmonella

spp.

c,f

(slaughtering >50 pigs/month)

Salmonella

spp.

c,f

(slaughtering 50 or less pigs/month)

Salmonella

spp.

c,h

Trichinella

spp.

Mycobacterium bovis

Echinococcus granulosis/multilocularis

6,761

337

Pos

Animals/Samples

20,163

121

17,250

537

801

18,250,233

18,407,939

Pos

173

% pos

5.0

0.98

1.30

21.6

a) Including samples from boars (examined at pre-entry, every 18 month, and prior to release from semen collection centres)

(17,007 samples), samples collected in connection with export (3,011), import (no samples this year) or diagnostic samples (145

samples). 5-8 ml blood samples were analysed using either the SAT, RBT, CFT or ELISA methods.

b) Sampling is based on suspicion of leptospirosis due to increased abortions or other reproductive problems in a herd. Samples are

investigated using immunoflourescence techniques.

c) See Table A37 for describtion of the

Salmonella

surveillance programme.

d) Data are from December 2014. Slaughter pig herds monitored using serological testing of meatjuice samples collected at slaugh-

ter.

e) Includes herds belonging to

Salmonella

level 2 and 3 only.

f) Swab samples from four designated areas of the half-carcass were collected at the slaughterhouse after min. 12 h chilling. Sample

size is 4x100 cm

. Samples from five animals were pooled, except at slaughterhouses where 50 pigs or less were slaughtered per

month, in which case samples were analysed individually.

g) When estimating the prevalence of

Salmonella,

both the loss of sensitivity and the probability of more than one sample being

positive in each pool are taken into consideration. A conversion factor has been determined on the basis of comparative studies, as

described in Annual Report 2001.

h) Coecum samples are randomly collected from slaughter pigs at slaughter.

i) Samples collected from slaughter pigs at slaughter were examined using the method described in Directive 2075/2005/EEC. In

2007, Denmark achieved official status as region with negligible risk of

Trichinella,

according to EU Regulation (EC) No 2075/2005.

j) Slaughter pigs were examined by meat inspectors at slaughter.

Source: Danish Veterinary and Food Administration, National Veterinary Institute and National Food Institute, Technical Univer-

sity of Denmark.

Figure A3.

Salmonella

in pork, monitored at slaughterhouses

, 2009-2014

3.0

% positive samples

2.5

2.0

1.5

1.0

0.5

0.0

2009

2010

% positive

2011

2012

2013

2014

% positive, moving avg. for 12 month

a) For more information about the surveillance programme, see Table A37.

Source: Danish Veterinary and Food Administration.

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Appendix

Table A16. Occurrence of zoonotic pathogens in cattle and beef in Denmark, 2014

Herds

Zoonotic pathogen

At farm

Brucella abortus

Mycobacterium bovis

b, c

Coxiella burnetii

At slaughterhouse

Salmonella

spp.

(slaughtering >50 cattle/month)

Salmonella

spp.

(slaughtering 50 or less cattle/month)

Mycobacterium bovis

b, h

VTEC O157

Echinococcusus granulosis/multilocularis

228

Pos

Animals/Samples

1,452

1,150

168

5,350

969

485,147

Pos

% pos

0.2

0.4

a) Denmark has been declared officially brucellosis free since 1979. The last outbreak was recorded in 1962. Including samples from

bulls (examined at pre-entry, every year, and prior to release from semen collection centres) (1,197 samples), samples collected in

connection with export (206), import (43) or diagnostic samples (6). 5-8 ml blood samples were analysed using either the SAT, RBT,

CFT or ELISA methods.

b) Denmark has been declared officially tuberculosis free since 1980. The last case of TB in cattle was diagnosed in 1988.

c) Analysis using the interdermal tuberculin test. Including samples from bulls (examined at pre-entry, every year, and prior to

release from semen collection centres) and samples collected in connection with export.

d) Bulk tank milk samples taken for diagnostic testing and analysed using an ELISA method.

e) Serum samples taken for diagnostic testing (41 samples, 6 pos), export (85 samples, no pos), import (2 samples, no pos) and

breeding (40 samples, no pos) and analysed using an ELISA method. An additional 9 samples from placenta was analysed using the

FISH method, none were positive.

f) See Table A36 for describtion of the surveillance programme. Swab samples from four designated areas of the half-carcass were

collected at the slaughterhouse after min. 12 h chilling. Sample size is 4x100 cm

. Samples from five animals were pooled, except at

slaughterhouses where 50 cattle or less were slaughtered per month, in which case samples were analysed individually.

g) When estimating the prevalence of

Salmonella,

both the loss of sensitivity and the probability of more than one sample being

positive in each pool are taken into consideration. A conversion factor has been determined on the basis of comparative studies, as

described in Annual Report 2001.

h) Slaughtered cattle were examined by the meat inspectors at slaughter.

i) Caecal content are tested from one animal per herd, collected at slaughter (DANMAP programme). A 25 g faecal sample from one

slaughter calf per herd is examined using overnight enrichment, immunomagnetic separation method and plating on CT-SMAC

plates for O157.

Source: Danish Veterinary and Food Administration, Danish Agriculture and Food Council, National Veterinary Institute, and

National Food Institute, Technical University of Denmark.

Figure A4.

Salmonella

in beef, monitored at slaughterhouses

, 2009-2014

1.0

% positive samples

0.5

0.0

2009

2010

% positive

2011

2012

2013

2014

% positive, moving avg. for 12 month

a) For more information about the surveillance programme, see Table A36.

Source: Danish Veterinary and Food Administration.

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Appendix

Table A17 Cattle herds in the

Dublin surveillance programme

, January 2014

Non-milk

producing herds

Salmonella

Dublin level

Level 1

Total

Level 2

On the basis of milk samples

On the basis of blood samples

Probably

Salmonella

Dublin free

Titer high in blood- or milk samples

Contact with herds in level 2 or 3

Other causes

Level 3

Total

Total number of herds

a) See Table A36 for description of the surveillance programme.

Source: Seges, Cattle.

Milk producing

herds

93.7

4.9

0.6

0.2

0.6

6.3

3,205

169

216

3,421

3,205

14,070

241

416

14,486

97.1

0.6

1.7

0.6

2.9

Salmonellosis, official supervision

Non

Salmonella

Dublin free

Table A18 Results from the intensified control of

Salmonella

and

Campylobacter

in fresh meat based on case-by-case

risk assessments 2014

Batches

tested

Campylobacter

Danish

Imported

Salmonella

Danish

Imported

Pork

Broiler

Pork

Broiler

Turkey

144

105

153

155

Broiler

124

149

No. of

batches

positive

No. of batches

deemed unsafe

based on a risk

assessment

Batches deemed

unsafe based on

other criteria

Mean

prevalence

in batches

b,d

47.1

46.3

26.6

9.4

28.1

Mean relative

human risk

in batches

c,d

3.5

4.5

16.8

5.6

13.3

a) Microbiological criteria specified in regulation (EC) No 2073/2005 as amended. For Danish broiler meat there is a zero-tolerance

for

Salmonella

and all positive batches must be heat treated before being put on the marked (Order no. 1512 of 13/12/2013).

b) The

Salmonella

prevalence in each batch is based on the proportion of positive pooled samples (12 pools per batch) and number

of subsamples per pool. Only results for batches subjected to risk assessment have been included.

c) Calculated as the risk relative to a batch of the same size with a mean prevalence (weighted average in Danish and imported meat)

Campylobacter

or of a

Salmonella

type with an average impact to cause human infection.

d) In 2014, a lower limit for when batches contaminated with

Campylobacter

were sent for risk assessment was introduced. Therefore

these figures are not comparable to previous years.

Source: Danish Veterinary and Food Administration, and National Food Institute.

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AppendixC

Table A19. Feed business operators own sampling of

Salmonella

in compound feeds, feed processing and feed material

(batch-based data), 2012-2014

2014

Feed processing plants (process control)

Ordinary inspections - clean zone

Ordinary inspections - unclean zone

Compound feed, farm animals

Feed materials, farm animals

Transport vehicles, clean zone/hygiene samples

Transport vehicles, unclean zone/hygiene samples

7,557

456

858

1,656

1,143

235

Positive

2013

Positive

2012

Positive

7,132

577

375

1,295

973

255

7,105

736

316

1,369

884

259

a) Presence of

Salmonella

in compound feed is indirectly monitored by environmental samples collected during feed processing.

b) Predominantly soy bean meal and rapeseed cake.

c) Samples from transport vehicles (hygiene samples) prior to loading of feed compounds.

Falkensee,

Idikan,

Isangi,

Rissen,

Tennessee,

Agona,

Derby,

Falkensee,

Putten,

Rissen,

1,4,5,12:i:-.

Cubana,

Havana,

Infantis,

Livingstone,

Liverpool,

Mbandaka,

Senftenberg,

Tennessee,

13,23:-:-.

Derby.

Derby,

Rissen.

Source: Danish Veterinary and Food Administration and the feed business operators.

Table A20. Control of

Salmonella

in compound feeds, feed processing and feed material (batch-based data), 2011-2014

2014

N Positive

Feed processing plants (process control)

Ordinary inspections

Feed materials, farm animals

402

333

2013

N Positive

2012

N Positive

311

2011

N Positive

377

a) Presence of

Salmonella

in compound feed is indirectly monitored by environmental samples collected during feed processing.

Companies are sampled one to four times per year.

b) Primarily findings of

Salmonella

in the dirty zone.

c) Predominantly soy bean meal and rapeseed cake.

Infantis (3 dirty, 1 clean),

Rissen (1 dirty, 1 clean),

13,23:-:- (1 clean),

Agona (1 clean),

Falkensee (1 dirty),

Senften-

berg (1 dirty).

Infantis (2),

Idikan (1),

Mbandaka (1).

Source: Danish Veterinary and Food Administration.

Table A21

Salmonella

in three categories of meat and bone meal by-products not intended for human consumption

2014

Category of

processing plant

1+2

By-products of this material cannot be used for

feeding purposes

By-product of this material may be used for feed for

fur animals

By-products from healthy animals slaughtered in a

slaughterhouse. Products of these may be used for

petfood

and for feed for fur animals

Total

Own-check samples

210

450

Positive

Product samples

1,717

Positive

a) Regulation No. 1774 of 03/10/2002.

b) For cats and dogs. Only by-products from pigs are used in this pet food.

Source: Daka Denmark A/S.

725

1,786

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Appendix

Table A22. Pathogens in batches

of ready-to-eat vegetables, herbs and fruits

, 2014

Salmonella

Type of sample

Vegetables

Baby corn

Cucumber

Pepper

Salad/leafy green

Sprouts

Sugar peas

Tomato

Other vegetables

Herbs

Parsley

Spearmint

Spring onions

Other herbs

Fruit and berries

Apples and pears

Grapes

Rasberries

Strawberries

Other berries

Other fruits

Total

153

Pos

Campylobacter

153

Pos

153

E. coli

>100 cfu/g

a) Five samples per batch.

b) Centrally coordinated study (See section 9 for description) to control and investigate

Salmonella, Campylobacter

and

E. coli

Danish and imported ready-to-eat vegetables, sprouts and herbs.

c) 1 batch of babycorn from Thailand.

d) 1 batch of baby leaves from Denmark.

e) 1 batch of azuki sprouts from Denmark.

f) Including aubergine, broccoli, spinach, zucchini.

g) Including chives, oregano, thyme, dill.

h) 1 batch of dill from Italy.

i) Batches with >100 cfu/g in one or more samples.

Source: Danish Veterinary and Food Administration.

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Appendix

Table A23. Occurrence of zoonotic pathogens in pets and zoo animals in Denmark

, 2014

Pet animals

Dogs

Zoonotic pathogen

Salmonella

spp.

Chlamydia psittaci

Cryptosporidium

spp.

Lyssavirus

(classical)

European Bat

Lyssavirus

Pos

Cats

Pos

Others

Pos

Zoo animals

Mammals &

reptiles

Pos

Birds

Pos

a) All samples are analysed based on suspicion of disease, and does not reflect the country prevalence.

Dublin.

c) Two chimpansees, one alpaca, one reindeer, one camel, one tiger, one zebra (four of these animals were tested due to

export).

d) Three ring-tailed lemurs, two chimpansees, one alpaca, one tiger, one zebra, one antilope, one monkey, one seacow (three

of these animals were tested due to export).

Source: National Veterinary Institute, Technical University of Denmark, and Danish Veterinary and Food Administration.

Table A24. Occurrence of zoonotic pathogens in wild and farmed wildlife in Denmark

, 2014

Farmed wildlife

Wild boar

Zoonotic pathogen

Salmonella

spp.

Campylobacter

spp.

Chlamydia psittaci

Cryptosporidium

spp.

Echinococcus multilocularis

Trichinella spp

Lyssavirus

(classical)

European Bat

Lyssavirus

482

Pos

Minks &

chincillas

Pos

Wildlife

Mammals

477

Birds

Pos

446

a) All samples are analysed based on suspicion of disease or risk based and does not reflect the country prevalence, except for

animals analysed for

Echinococcus multilocularis.

These animals are collected as part of a survey.

Typhimurium (1),

Derby (1),

Enteritidis (1),

1,4,12:i:- (1),

1,4,5,12:i:- (1).

c) 41 badgers.

Typhimurium (1),

Newport (1), not serotyped (2)

e) 10 starlings

f) 3 mice, 24 raccoon dogs, 59 roe deer, 13 seals.

g) 1 raccoon dogs and 5 roe deer.

h) 344 foxes, 20 badgers, 112 raccoon dogs, 1 raccoon.

i) 7 foxes and 2 raccoon dogs.

j) In 2007, Denmark achived official status as region with negligible risk of

Trichinella,

according to EU regulation (EC) No 2075/2005.

k) Samples reported from slaughterhouses. An additional 133 samples were reported from the laboratory. Double registrations

from slaughterhouses and laboratory may occur.

l) 50 mammals from the sea, 46 minks, 53 raccoon dogs, 273 foxes, 1 raccoon and 23 badgers.

m) 16 bats, 3 foxes, 2 badgers, 1 marten, 1 mink, 1 roe deer, 1 seal.

Source: National Veterinary Institute, Technical University of Denmark, and Danish Veterinary and Food Administration.

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Appendix

Table A25. The Bovine Spongiform Encephalopathy (BSE) surveillance programme

for cattle, 2014

Type of surveillance

Active surveillance

Healthy slaughtered animals

Risk categories:

Emergency slaugthers

Slaughterhouse antemortem inspection revealed suspi-

cion or signs of disease

Fallen stock

Animals from herds under restriction

Passive surveillance

Animals suspected of having clinical BSE

Total

1,122

20,392

21,559

Positive

a) According to the EU Regulation (EC) 999/2001 as amended, Commission Decision 2009/719/EC as amended and Danish Order

no. 878 of 01/07/2013 as amended.

b) Samples (brain stem material) are tested using a IDEXX technique or Prionics-Check PrioStrip. Confirmatory testing is carried

out using histopathology or immunohistochemistry. Further confirmation on autolysed material is performed at the European

Union TSE reference laboratory.

Source: National Veterinary Instistute, Technical University of Denmark, and Danish Veterinary and Food Administration.

Table A26. The Transmissible Spongiform Encephalopathy (TSE) surveillance programme

for sheep and goats, 2014

Type of surveillance

Active surveillance

Fallen stock (>18 months)

Animals from herds under restriction

Passive surveillance

Animals suspected of having clinical TSE

Total

701

703

Positive

a) According to the EU Regulation (EC) 999/2001 as amended, Commission Decision 2009/719/EC as amended and Danish Order

no. 1288 of 20/12/2011 as amended.

b) Samples (brain stem material) are tested using a IDEXX technique or Prionics-Check PrioStrip. Confirmatory testing is carried

out using histopathology or immunohistochemistry. Further confirmation on autolysed material is performed at the European

Union TSE reference laboratory.

Source: National Veterinary Institute, Techncal University of Denmark, and Danish Veterinary and Food Administration.

Table A27. Distribution

(%) of prion protein genotype of sheep randomly selected, 2014

Genotype

NSP 1

NSP 2

NSP 3 (ARQ/ARQ)

NSP 3 (Other)

NSP 4

NSP 5

Total

ARR/ARR

ARR/AHQ, ARR/ARH, ARR/ARQ

ARQ/ARQ

AHQ/AHQ, AHQ/ARH, AHQ/ARQ, ARH/ARQ, ARH/ARH, ARQ/ARH,

ARH/AHQ, ARQ/AHQ

ARR/VRQ

ARH/VRQ, ARQ/VRQ, VRQ/VRQ, AHQ/VRQ

Sheep

n=100

27.0

26.0

31.0

10.0

1.0

5.0

100

a) The genotypes were grouped in the NSP classification system according to their different susceptibility:

NSP 1: Genetically most resistant, NSP 2: Genetically resistant, NSP 3: Genetically little resistance,

NSP 4: Genetically susceptible, and NSP 5: Genetically highly susceptible.

Source: National Veterinary Institute, Technical University of Denmark, and Danish Veterinary and Food Administration.

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Appendix

Table A28. Centrally coordinated studies conducted in 2014

Title of project

No. of

samples

927

108

248

Pathogen surveyed

Campylobacter

spp.

Campylobacter

spp.

Campylobacter

spp.

Further information

Appendix Table A14

Data are being processed

Appendix Table 14

Appendix Table A18

Data are being processed

Campylobacter

spp. in fresh, chilled

Danish broiler meat (conventional)

Campylobacter

spp. in fresh, chilled

Danish broiler meat (organic)

Campylobacter

spp. in fresh, chil-

led and frozen Danish and imported

broiler meat (processed meat)

Intensified control for

Salmonella

spp.

7,622

Campylobacter

spp.,

and

Campylobacter

in fresh Danish

(639 batches)

Salmonella

spp.

and imported meat (poultry and pig)

Official verification of microbiological

criteria (EU 2073/2005)

ESBL in Danish poultry production

Import control - processed products

of animal origin (not fish)

Import control - fish, fish products

and bivalve molluscan shellfish

Import control - food of non animal

origin

Listeria monocytogenes

in cold smo-

ked halibut

Listeria monocytogenes, Salmonella

spp.,

Escherichia coli,

staphylococci in

fish goods from Greenland

Microbiological classification of mus-

sel production areas in Denmark

MRSA in pigs in the top of the

breeding pyramid

MRSA in slaughterpig herds

Pathogens in Danish and imported

ready-to-eat vegetables

DANMAP - Antibiotic resistance in

poultry, pigs and cattle, and in Danish

and imported broiler, beef and pork meat

Salmonella

Dublin in beef

Salmonella

in animal feed

Salmonella

in pigs at slaughter

Salmonella

spp. and

Escherichia coli

raw, frozen scallop from Greenland

Salmonella

spp. antibiotic resistance in

fresh, chilled and frozen imported beef

and duck meat incl. ESBL in duck meat

2440

Listeria monocytogenes, Salmo-

nella

spp., staphylococci,

Escherichia coli,

Aerobic colony

count,

Enterobacteriaceae

ESBL

Listeria monocytogenes, Salmo-

nella

spp., staphylococci

Listeria monocytogenes, Sal-

monella

spp.,

Escherichia coli,

staphylococci

114

100

215

100

350

1,008

815

1,505

375

409

804

364

Data are being processed

Salmonella

spp. (herbs)

Data are being processed

Norovirus (frozen straberries)

Hepatitis A (frozen strawberries)

Listeria monocytogenes

Salmonella

spp.,

Listeria mo-

nocytogenes, Escherichia coli,

staphylococci

Data are being processed

Salmonella

spp.,

Escherichia coli

Data are being processed

Methicillin resistant

Staphylococ-

Data are being processed

cus aureus

Methicillin resistant

Staphylococ-

Data are being processed

cus aureus

Salmonella

spp.,

Campylobacter

spp.,

Escherichia coli

Campylobacter

spp.,

Enterococ-

cus faecium, E. faecalis, ESC

Escherichia coli

Appendix Table A22

Results are presented in the

2014 DANMAP report

Salmonella

spp.,

Escherichia coli,

Data are being processed

Enterobacteriaceae,

enterococci

Salmonella

spp.

Salmonella

spp.

Results are published on the

DVFA website www.fvst.dk

(In Danish)

Appendix Tables A5 and

A15

Salmonella

spp.,

Escherichia coli

Data are being processed

Salmonella

spp., ESC

Escheri-

chia coli

Appendix Tables A5-A6 and

a) Results are published on the DVFA website www.fvst.dk (in Danish).

Source: Danish Veterinary and Food Administration.

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Appendix

Table A29.

Listeria monocytogenes

in Danish produced ready-to-eat (RTE) foods

, 2014

Samples analysed by a

qualitative method

Batches

Food category

Cheese, RTE

Milk and dairy products, RTE

Products made from broiler

meat, RTE

Sampling place

At processing

Pos

Single

samples

120

170

175

125

284

330

Pos

Samples analysed by a

quantitative method

Batches

Pos

Single

samples

110

295

245

Pos

Products made from other poul-

At processing

try meat, RTE

Products made from pork, RTE

Products made from beef, RTE

Fruit, RTE

Vegetables, RTE

Fish and Fishery products, RTE

Shellfish and products thereoff,

RTE

Other RTE products

At processing

a) Samples are collected by the local food control offices according to EU Regulation (EC) No 2073/2005.

Listeria monocytogenes

present in a 25 g sample of the product.

c) Five samples from each batch, analysed individually.

Source: Danish Veterinary and Food Administration.

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Appendix

Monitoring and surveillance programmes

Table A30. Overview of notifiable and non-notifiable human diseases presented in this report, 2014

Patogen

Bacteria

Brucella

spp.

Campylobacter

spp.

Chlamydophila psittaci

(Ornithosis)

Listeria monocytogenes

Leptospira

spp.

Mycobacterium bovis/ tuberculosis

Coxiella burnetii

Salmonella

spp.

VTEC

Yersinia enterocolitica

Parasites

Cryptosporidium

spp.

Echinococcus multilocularis

Echinococcus granulosus

Trichinella

spp.

Viruses

Lyssavirus

(Rabies)

Prions

BSE/Creutzfeld Jacob

Notifiable

1979

1980

1993

1980

1905

1979

2000

1979

1964

1997

Notification route

Laboratory

Physician

Physician (and laboratory

)

Laboratory

Physician and laboratory

Laboratory

Physician (via telephone)

Physician

a) Danish order no. 277 of 14/04/2000. Cases must be notified to Statens Serum Institut.

b) The regional microbiological laboratories report confirmed cases.

c) The physician report individually notifiable infections.

d) The laboratories voluntarily report confirmed cases.

Source: Statens Serum Institut.

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Appendix

Table A31. Overview of notifiable and non-notifiable animal diseases presented in this report, 2014

Patogen

Bacteria

Brucella

spp.

Cattle

Sheep and goats

Pigs

Campylobacter

spp.

Chlamydophila psittaci

Birds and poultry

Listeria monocytogenes

Leptospira

spp. (only in

production animals)

Mycobacterium bovis/tuber-

culosis

Cattle

Coxiella burnetii

Salmonella

spp.

Cattle

Swine

Poultry

VTEC

Yersinia enterocolitica

Parasites

Cryptosporidium

spp.

Echinococcus multilocularis

Echinococcus granulosus

Trichinella

spp.

Viruses

Lyssavirus

(Rabies)

Prions

TSE

Sheep and goats

BSE

Cattle

Notifiable

1920

OBF in 1979

ObmF in 1995

No cases since

1999

1920

2003

EU legislation

Danish legislation

Decision 2003/467/EC

Directive 2003/99/EC

Order no 305 of 3/5 2000

Order no. 739 of 21/8 2001

Order no. 205 of 28/3 2008

Order no. 871 of 25/8 2011

Act no. 432 of 09/06/2004

1920

OTF in 1980

2005

1993

Decision 2003/467/EC

Order no. 1417 of 11/12 2007

Act no. 432 of 09/06/2004

Order no. 954 of 10/07/2013

Order no. 404 of 08/05/2012

Order no. 1512 of 13/12/2013

2004

1993

1920

Council Directive 64/433/EC Act no. 466 of 15/05/2014

Regulation 2075/2005/EC

Order no. 412 of 28/05/2008

Order no. 330 of 14/04/2011

yes

Regulation 999/2001/EC

(as amended)

Regulation 999/2001/EC

(as amended)

Order no. 1288 of 20/12/2011

yes

Order no. 878 of 01/07/2013

(as amended)

a) Clinical cases, observations during the meat inspection at the slaughterhouse, positive blood samples or finding of agents are

notifiable.

b) Officially Brucellosis Free (OBF) according to Council Directive 64/432/EC as amended and Commision Decision 2003/467/EC.

No cases in since 1962.

c) Officially

Brucella melitensis

Free (ObmF) according to Council Directive 91/68/EC and Commision Decision 2003/467/EC.

Never detected in sheep or goat.

d) Officially Tuberculosis Free (OTF) according to Council Directive 64/432/EC as amended and Regulation (EC) No 1226/2002,

and Commission Decision 2003/467/EC. No cases in since 1988 or in deer since 1994.

e) Only clinical cases notifiable.

f) Denmark was recognized as a country with neglible risk for BSE at World Organisation for Animal Health (OIE) general session

in May 2011.

Source: Danish Veterinary and Food Administration.

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Appendix

Table A32.

Salmonella

surveillance programme for the rearing flocks and adult flocks of the grandparent and parent

generation of the broiler and table egg production, 2014

Time

Rearing flocks

Day-old

a,b

Samples

taken

Per

delivery

Material

Grandparent generation

5 transport crates from one delivery:

crate liners (>1 m

in total) or swab

samples (>1 m

in total). Analysed as

one pool

Material

Parent generation

5 transport crates from one delivery:

crate liners (>1 m

in total) or swab

samples (>1m

in total). Analysed as

one pool

2 pairs of boot swabs (analysed as one

pooled sample) or 1 faeces sample of

60 g

2 pairs of boot swabs (analysed as one

pooled sample) or 1 faeces sample of

60 g

2 pairs of boot swabs (analysed as one

pooled sample). Cage birds: 60x1 g

samples of fresh droppings. Analysed

as one pool

2 pairs of boot swabs (analysed as one

pooled sample) or 1 faeces sample of

60 g

Parent generation

Hatcher basket liners from 5 baskets

(>1 m

in total) or 10 g of broken eggs-

hells from each of 25 hatcher baskets

(reduced to 25 g sub-sample). Analy-

sed as one pool

Wet dust samples. Up to four hatchers

of the same flock can be pooled

2 pairs of boot swabs (analysed as one

pooled sample) or 1 faeces sample of

60 g

5 pairs of boot swabs (analysed as two

pooled samples), or 1 faeces sample

consisting of 2x150 g

5 pairs of boot swabs (analysed as two

pooled samples), 2 dust samples (250

ml) and 5 birds (analysed for antimi-

crobial substances)

1st & 2nd week

b, c

Per unit

4th week

a,c

Per unit

5 pairs of boot swabs (analysed as two

pooled samples), or 1 faeces sample

consisting of 2x150 g

2 pairs of boot swabs (analysed as one

pooled sample). Cage birds: 60x1 g

samples of fresh droppings. Analysed

as one pool

5 pairs of boot swabs (analysed as two

pooled samples), or 1 faeces sample

consisting of 2x150 g

Grandparent generation

Hatcher basket liners from 5 baskets

(>1 m

in total) or 10 g of broken egg-

shells from each of 25 hatcher baskets

(reduced to 25 g sub-sample). Analy-

sed as one pool

8th week

b,c

Per unit

2 weeks prior to

moving

a,d

Adult flocks

Every two weeks

a,b

(Every 16th week)

Per unit

Per flock

After each hatch

Every week

Per hatch Wet dust samples. Up to four hatchers

of the same flock can be pooled

Per unit

0-4 weeks after

moving, 8-0 weeks

before slaughter

b,d

After positive

findings

b,d

Per unit

5 pairs of boot swabs (analysed as two

pooled samples), or 1 faeces sample

consisting of 2x150 g

5 pairs of boot swabs (analysed as two

pooled samples), 2 dust samples (250

ml) and 5 birds (analysed for antimi-

crobial substances)

Per unit

a) Sampling requirements set out by Regulation (EC) No 2160/2003.

b) Samples collected by the food business operator.

c) Sampling requirements set out by Order no 952 of 10/07/2013.

d) Samples collected by the Danish Veterinary and Food Administration.

e) When eggs from a flock exceed the capacity of one incubator, each incubator should be sampled as described.

Source: Danish Veterinary and Food Administration.

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Appendix

Table A33.

Salmonella

and

Campylobacter

surveillance programme for the broiler flocks, 2014

Time

Salmonella

15 - 21 days before slaughter

a,c,d

7 - 10 days before slaughter

b,e

After slaughter

b,c

Samples taken

Per flock

Per batch

Material

5 pairs of boot swabs. Analysed individually

300x1 g neck skin, analysed in pools of max. 60 grams.

Sampling size depends on whether the slaughterhouse

slaughters only AM-negative flocks or AM-negative as well

as AM-positive flocks

12 cloacal swabs from 24 animals, analysed in one pool

Campylobacter

After slaughter

Per flock

a) Sampling requirements set out by Regulation (EC) 2160/2003.

b) Sampling requirements set out by Order no. 1512 of 13/12/2013 replacing 1105 of 18/09/2013 replacing 1462 of 16/12/2009.

c) Samples collected by the food business operator.

d) Once a year, one pair of socks is collected by the Danish Veterinary and Food Administration.

e) Samples are collected by a representative of the slaughterhouse, laboratorium or the Danish Veterinary and Food Administration.

f) For flocks to be slaughtered outside Denmark 1 pair of boot swabs is collected by the owner 10 days before slaughter at the latest.

g) If the flock is slaughtered over several days, the last batch is sampled.

Source: Danish Veterinary and Food Administration.

Table A34.

Salmonella

surveillance programme for the pullet-rearing, table egg layer and barnyard/hobby flocks in the

table egg production, 2014

Time

Pullet-rearing

Day-old

a,c

Samples taken

Per delivery

Material

5 transport crates from one delivery: Crate liner (> 1 m

total) or swab samples (> 1 m

in total) (Analysed as one

pooled sample)

5 pairs of boot swabs (analysed as two pooled samples) or

5 faeces samples of 60 gram

5 pairs of boot swabs (analysed as two pooled samples) or

5 faeces samples of 60 gram. 60 blood samples (serology)

2 pairs of boot swabs (analysed as one pooled sample) or 1

faeces sample consisting of 2x150 g. 250 ml (100 g) dust or

a dust sample by a cloth of min. 900 cm

2 pairs of boot swabs (analysed as one pooled sample) or 1

faeces sample consisting of 2x150 g.

5 pairs of boot swabs (analysed as two pooled samples) or

5 faecal samples consisting of 60 gram each

5 pairs of boot swabs (analysed as two pooled samples) or

5 faeces samples consisting of 60 gram each

4 weeks old

a,c

2 weeks before moving

a,b

Per flock

Table egg layers (Production for certified packing stations)

24 weeks old

a,c

Per flock

Every 2 weeks from age 20

weeks

a,c,d, e

Per flock

After positive serological findings

Per flock

After positive findings of other

Per flock

serotypes than

Enteritidis,

Hadar,

Infantis,

Virchow or

Typhimurium including the mo-

nophasic strains

1,4,[5],12:i:-

Barnyard and hobby flocks

Every 18 weeks

a,c,g

Per flock

Egg samples

a) Sampling requirements set out by Order no 1260 of 15/12/2008, replaced by Order no. 953 of 10/07/2013 and no. 1134 of

27/09/2013.

b) Samples collected by the Danish Veterinary and Food Administration.

c) Samples collected by the food business operator.

d) According to Regulation (EC) 2160/2003 sample collection must be carried out every 15 weeks as a minimum.

e) Until 01/10/2013 sampling every 9

week only.

f) Voluntary for hobby flocks.

g) For flocks with 30 birds or less: No testing if only delivered to a well-known circle of users.

Source: Danish Veterinary and Food Administration.

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Appendix

Table A35.

Salmonella

surveillance programmes for turkey flocks, 2014

Time

Turkey production

Max. 21 days before slaughter

a,b

Samples taken

Per flock

Material

2 pairs of boot swabs. Analysed

individually

a) Sampling requirements set out by Regulation (EC) 584/2008.

b) Samples collected by the food business operator or the local food control offices.

Source: Danish Veterinary and Food Administration.

Table A36.

Salmonella

surveillance programme

for the cattle production, 2014

No. of samples

Milk producing herds

4 samples distributed over 18

months

10 samples

Non-milk producing herds

1 sample every 180 days at

slaughter

Samples taken

Bulk tank samples

Blood samples

Purpose/Comment

Calculation of herd level

If the owner wants a herd moved from

level 2 to 1

Calculation of herd level

Consecutive negative samples required

for level 1

Slaughterhouses slaughtering more than

200 cattle per day

Slaughterhouses slaughtering more than

200 cattle per month but 200 or less

cattle per day

Slaughterhouses slaughtering 50-200

cattle per month

Slaughterhouses slaughtering less than

50 cattle per month

Blood samples

4-8 samples depending on herd Blood samples

size

Beef carcasses at the slaughterhouse

5 samples daily, pooled into

one analysis

5 samples per 200 slaughtered

cattle, pooled into one analysis

5 samples every 3

month,

pooled into one analysis

1 sample every 3

month

Swab samples from 4 designated areas

after 12 hours chilling (4x100cm

)

Swab samples from 4 designated areas

after 12 hours chilling (4x100cm

)

Swab samples from 4 designated areas

after 12 hours chilling (4x100cm

)

Swab samples from 4 designated areas

after 12 hours chilling (4x100cm

)

a) Order no. 886 of 02/07/2014 as ammended. In 2013 and 2014, the programme for eradication of

Salmonella

Dublin from the

Danish cattle production was intensified. This implies regionalisation of the country according to prevalence and compulsory era-

dication plans in Level 2 herds.

b) Herd levels based on serological testing (blood and milk):

Level 1: Herd assumed free of infection based on bulk milk samples (milk producing herd) or blood samples (non-milk

producing herd or milk producing herd assumed free from infection),

Level 2: Herd not assumed free of infection,

Level 3: Herd infected based on culture and clinical signs.

c) No samples are taken, if the herd has been tested for

Dublin within the last 180 days or 8 samples have been tested within the

last 24 months.

d) Number of samples equals total number of animals in the herd minus 2 (max. 8 animals, min. 4 animals).

Source: Danish Agriculture and Food Council, and Danish Veterinary and Food Administration.

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Appendix

Table A37.

Salmonella

surveillance programme

for the pig production, 2014

Time

Breeding and multiplier herds

Every month

Samples taken

10 blood samples per

epidemiological unit

Purpose/Comment

Calculation of

Salmonella-index

based on

the mean seroreaction from the last three

months with more weight to the results

from the more recent months (1:3:6)

Clarify distribution and type of infection

in the herd

Clarify distribution and type of infection

in the herd, and possible transmission

from sow herds to slaughter pig herds

Max. twice per year

Sow herds

When purchaser of piglets is

assigned to level 2 or 3,

max. twice per year

Herds positive with

Typhimu-

rium,

Infantis,

Derby and

Choleraesuis are considered posi-

tive for the following 5 years

Slaughter pigs, herds

At slaughter

Herds with

Salmonella-index

or above: Pen-faecal samples

Pen-faecal samples

No samples are collected from

Reduce repeated sampling in positive

the herd during the 5 year period herds infected with a persistent serotype

when the herd is considered po-

sitive, unless the herd is proven

negative

Meat juice, 60-100 samples per

Calculation of slaughter pig index based

herd per year. Herds in RBOV : on the mean proportion of positive

one meat juice sample per month samples from the last three months with

most weight to the result from the most

recent month (1:1:3)

. Assigning herds

to level 1-3 and assigning herds to risk-

based surveillance (RBOV)

e,g

Coecum samples, avg. 73 samp-

les per month, 12 months per

year

Swab samples from 4 designa-

ted areas after 12 hours chilling

(4x100cm

)

Swab samples from 4 designa-

ted areas after 12 hours chilling

(4x100cm

)

Swab samples from 4 designa-

ted areas after 12 hours chilling

(4x100cm

)

Swab samples from 4 designa-

ted areas after 12 hours chilling

(4x100cm

)

Random collection of samples for mo-

nitoring of the distribution of serotypes

and antimicrobial resistance.

Slaughterhouses slaughtering more than

200 pigs per day

Slaughterhouses slaughtering more

than 200 pigs per month or 200 or less

pigs per day

Slaughterhouses slaughtering more

than 50 pigs per month or less than

200 pigs per month

Slaughterhouses slaughtering less than 50

pigs per month

Slaughter pigs, animals

At slaughter

Pork carcasses at the slaughterhouse

5 samples daily, pooled into one

analysis

5 samples per 200 slaughtered pig,

pooled into one analysis

5 samples every 3

month,

pooled into one analysis

1 sample every 3

month

a) Sampling requirements set out by Order no. 1280 of 4/12/2014.

b) Herds with index above 10 have to pay a penalty for each pig sold.

c) The herd owner must inform buyers of breeding animals about the infection level and type of

Salmonella.

d) These serotypes are primarily spread by live trade, and are known to persist in herds.

Typhimurium includes the monophasic

1,4,[5],12:i:-.

e) RBOV: risk-based surveillance in herds with a slaughter pig index of zero (no positive samples in the previous three months) the

sample size is reduced to one sample per month. Increasing seroprevalence from level 1 to level 3.

f) Since November 2014: based on the proportion of seropositive samples from the last three months. Both the number of seroposi-

tive and total number of samples are weighted with most weight to samples from the most recent month (1:1:5).

g) Pigs from herds with highest level of infection (Level 3) must be slaughtered under special hygienic precautions.

h) Centrally coordinated study (Table A28)

Source: Danish Veterinary and Food Administration.

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Appendix

Table A38. Typing methods used in the surveillance of foodborne pathogens in Denmark, 2014

Methods

Salmonella enterica

Serotype

Phage type

Human

All

None

Food

All

Few

Typhimurium and

Enteritidis

Almost all isolates

Animal

All

Few

Typhimurium and

Enteritidis, all isolates from

poultry

Almost all isolates

Typhimurium

Ente-

ritidis and

Dublin for the

Salmonella

source account,

outbreak investigations and

research

Outbreak investigations

Some for outbreak investiga-

tion and research

Antimicrobial

resistance

MLVA

All

Salmonella

except

Enteritidis

Typhimurium

Enteri-

Typhimurium

Ente-

tidis and

Dublin

ritidis and

Dublin for the

Salmonella

source account,

outbreak investigations and

research

Outbreak investigations

Some for outbreak investiga-

tion and research

PFGE

WGS

Campylobacter coli/jejuni

Antimicrobial

resistance

FlaA-SVR

MLST, WGS

VTEC

Serotype

Virulence profile

PFGE

WGS

Listeria

Serogroup

PFGE

WGS

Yersinia enterocolitica

O-group

All isolates send to SSI

None

a) Including the monophasic strains

1,4,[5],12:i:-.

Source: Statens Serum Institut, and Danish Zoonosis Laboratory, National Food Institute.

Isolates from 3 districts for

DANMAP surveillance

Outbreak investigations

Outbreaks investigations,

research

All

Outbreak investigations

All

For DANMAP surveillance

Only for DANMAP

purposes and the case-by-case surveillance purposes

program

Outbreak investigations

None

All

All (O157)

Outbreak investigations

None

All

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Population and slaughter data

Table A39. Human population, 2014

Age groups (years)

0-4

5-14

15-24

25-44

45-64

65+

Source: Statistics Denmark, 1 January 2015.

Males

153,001

340,301

373,323

713,218

752,353

478,818

2,811,014

Females

145,367

323,978

356,621

701,478

748,946

572,311

2,848,701

Total

298,368

664,279

729,944

1,414,696

1,501,299

1,051,129

5,659,715

Total

Table A40. Number of herds/flocks, livestock and animals slaughtered, 2014

Herds/flocks

Slaughter pigs (>30 kg)

Cattle

Broilers

Layers (excl. barnyard)

Turkeys

Sheep & lambs

Goats

Horses

7,012

18,577

556

214

6,975

3,179

Livestock (capacity)

6,471,342

1,563,933

n/a

3,240,000

360,670

142,558

20,821

Number slaughtered

18,407,939

485,147

102,941,054

3,825

80,475

1,451

1,328

Source: The Central Husbandry Register and Danish Veterinary and Food Administration.

Table A41. Number of farms in the broiler production, 2014

No. of holdings

Rearing period (grandparent)

Adult period (grandparent)

Rearing period (parent)

Adult period (parent)

Hatcheries

Broilers

234

No. of houses/flocks

141

556

Livestock (capacity)

50,000

90,000

250,000

700,000

n.a.

Source: Danish Veterinary and Food Administration and Danish Agriculture and Food Council.

Table A42. Number of farms in the table egg production, 2014

No. of holdings

Rearing period (grandparent)

Adult period (grandparent)

Rearing period (parent)

Adult period (parent)

Hatcheries

Pullet-rearing

Layers (excl. Barnyard)

154

No. of houses/flocks

214

Livestock (capacity)

50,000

70,000

20,000

50,000

1,050,000

3,240,000

Source: Danish Veterinary and Food Administration, and Danish Agriculture and Food Council.

Annual Report on Zoonoses in Denmark 2014

MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU

Annual Report on Zoonoses in Denmark 2014

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Contributing institutions:

Danish Zoonosis Centre

National Food Institute

Technical University of Denmark

Mørkhøj Bygade 19

DK - 2860 Søborg

Tel: +45 40 21 53 77

E-mail: [email protected]

www.food.dtu.dk

Statens Serum Institut

Artillerivej 5

DK - 2300 København S

Tel: +45 3268 3268

E-mail: [email protected]

www.ssi.dk

The Danish Veterinary and Food Administration

Stationsparken 31-33

DK - 2600 Glostrup

Tel: +45 7227 6900

E-mail: [email protected]

www.fvst.dk

The National Veterinary Institute

Technical University of Denmark

Bülowsvej 27

DK - 1790 Copenhagen V

Tel: +45 3588 6000

E-mail: [email protected]

www.vet.dtu.dk

Danish Agriculture and Food Council

Axelborg, Axeltorv 3

DK - 1609 Copenhagen V

Tel: +45 3339 4000

E-mail: [email protected]

www.lf.dk

Annual Report on Zoonoses in Denmark 2014

MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU MOF, Alm.del - 2015-16 - Bilag 148: Annual Report on Zoonoses in Danmark 2014, fra DTU

National Food Institute

Technical University of Denmark

Mørkhøj Bygade 19

DK - 2860 Søborg

T: 35 88 70 00

F: 35 88 70 01

www.food.dtu.dk

ISSN: 1600-3837