OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Trichinellosis
Istituto Superiore di Sanità
Department of Infectious, Parasitic
and Immunomediated Diseases
Viale Regina Elena 299,
00161 Rome,
ITALY
Tel.:
+39 06 4990 2304
Fax:
+39 06 4990 3561
e-mail address:
website:
edoardo.pozio@iss.it
www.iss.it/site/Trichinella/index.asp
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Edoardo Pozio
Name(including Title and
Position) of OIE Reference
Expert:
Edoardo Pozio
Name (including Title and
Position) of writer of this report
(if different from above):
Annual reports of OIE Reference Centres, 2011
1
Trichinellosis
Part I: Summary of general activities related to the disease
The Istituto Superiore di Sanità (ISS) is the leading technical and scientific public body of the Italian National
Health Service. Its activities include research, control, training and consultation in the interest of public health
protection. Two sections of the Department of Infectious, Parasitic and Immunomediated Diseases, namely the
Section of gastroenteric and tissue parasitic diseases and the Section of vector borne diseases and international
health, are working in the field of parasites and parasitic diseases. The OIE reference laboratory of trichinellosis is
appointed at the Section of gastroenteric and tissue parasitic diseases.
1.
2.
Test(s) in use/or available for the specified disease/topic at your laboratory
Test
For
Specificity
ELISA
Antibody
pig
ELISA
Antibody
human
Western blot
Antibody
pig
Western blot
Antibody
human
PCR
Parasite identification
species
Total
2,945
Production and distribution of diagnostic reagents
Type of reagent
Amount supplied nationally
(including for own use)
Amount supplied to other
countries
Control positive pig sera
2 mL
23 mL
Antigens for 30,400 tests
2.4 mg
35.6 mg
Reference larvae
6
9
Meat samples for proficiency tests
495
414
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
a)
Establishment and maintenance of a network with other OIE Reference Laboratories
designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results
Two ring trials have been organised:
-
2
to detect Trichinella larvae in meat samples. Twenty-eight labs from 26 countries of the European Union and
2 outside the EU (Serbia, Switzerland) participated. The scope was to evaluate the performance of national
reference laboratories for Trichinella to detect the parasites in the meat samples for the maintenance of the
accredited method or to validate the method. Ten samples (nine spiked with Trichinella larvae and one
Annual reports of OIE Reference Centres, 2011
Trichinellosis
negative) of 100 g each of pork and/or horse meat were sent to each lab. The outcome was quite good. In
fact, only few labs failed to recognise as positive some of the samples with a low number of larvae.
-
To identify the species/genotype of Trichinella larvae. Then samples containing single or pool of larvae were
sent to 12 EU labs and 2 labs outside the EU (Serbia, Switzerland). The outcome was quite good. In fact,
only few labs failed to identify some larvae.
An interlaboratory test evaluation has been organized to validate a commercial kit to detect Trichinella
infection in meat samples by a latex agglutination test. The kit has been evaluated by five international labs
with positive outcomes.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
None.
5.
Research and development of new procedures for diagnosis and control
The aim of the study was to identify the most frequently Trichinella specific antigens recognized by pig sera on
excretory/secretory antigens (ESA) by western blot (wb), in order to define an unequivocal pattern of positive sera.
To this end, 210 pig sera were tested by wb using ESA: 1) 30 from experimentally Trichinella infected pigs; 2)
90 from naturally Trichinella infected pigs for which the worm burden was known; and 3) 90 ELISA crossreacting sera (CRS) from Trichinella non-infected pigs. All sera from naturally and experimentally infected pigs
reacted with the three band pattern of 48–72 Kd (first band from 48 Kd to 55 Kd, second band from 59 Kd to
63 Kd and third band from 64 Kd to 72 Kd). No one of the 90 pig CRS recognised the three band pattern.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
Trichinella britovi focus of the Sardinia island, Italy
Up to 2005, the Island of Sardinia (Italy) of the Mediterranean basin has been considered free of Trichinella sp. in
domestic and wild animals and in humans. In 2005, two human outbreaks occurred for the consumption of pork
from free ranging pigs infected by Trichinella britovi in the Orgosolo village. The epidemiological investigation
revealed the presence of four infected pigs (0.6%), but no infection was detected in wild boars and foxes hunted in
the island. A passive introduction of the parasite from the neighbouring island of Corsica (France) was suspected.
In January 2011, a new human outbreak of trichinellosis occurred in the same village of Orgosolo for the
consumption of pork from pigs free ranging in the same municipality. The epidemiological investigation revealed
the presence of 8 T. britovi infected animals (2.8%). The free-ranging pigs originating from municipalities
neighbouring Orgosolo tested always negative. The discovery of T. britovi in Corsica in 2004 and its circulation
among free ranging pigs and wild animals up to the recent years seem to be related.
Trichinella spp. infections in different host species of an endemic district of Serbia
During the 2009–2010 period, muscle samples were collected from 167 wild animals: 94 wild boars (Sus scrofa),
57 red foxes (Vulpes vulpes), 13 golden jackals (Canis aureus), 3 wolves (Canis lupus) and 116,398 domestic
pigs. Trichinella spp. infection was detected in 302 (0.26%) domestic pigs, in 11 (11.7%) wild boars, 7 (12.3%)
red foxes, 7 (53.8%) golden jackal, and 3 (100%) wolves. Trichinella spiralis and Trichinella britovi were the only
two species identified in the isolates as single or mixed infections. The results point out the circulating of
Trichinella species by a domestic or a sylvatic cycle, the transmission between these two cycles, and the role of
some host species as reservoirs of T. spiralis or T. britovi or of both species in Serbia.
Trichinella pseudospiralis in wild boars (Sus scrofa) of the Czech Republic
In the Czech Republic, 121,185 wild boars (Sus scrofa) were hunted in 2009 and 96,232 of them were tested for
Trichinella by artificial digestion. In December 2010 and January 2011, Trichinella sp. larvae were detected in
three wild boars hunted in the eastern part of the Czech Republic. The larvae were identified as Trichinella
pseudospiralis. All T. pseudospiralis-positive wild boars had similar weight (around 35 kg) and were shot at the
same baited site by the same hunter. A common origin of the infection in the three wild boars has been
hypothesized. These findings support the tendency of a more frequent detection of the non-encapsulated species
T. pseudospiralis in Europe, which probably is related to the increased number of tested wild boars and to the use
Annual reports of OIE Reference Centres, 2011
3
Trichinellosis
of the artificial digestion instead of the less sensible trichinoscopy to detect these zoonotic parasites in meat
samples.
Molecular identification of nematode larvae different from those of the Trichinella genus detected by muscle
digestion
From 2008 to 2011, 15 nematodes isolated by artificial digestion from muscle tissues of different animal species
tested for Trichinella, have been analysed (about 1% of those analysed as Trichinella spp. in the same period).
Eight of these isolates have been successfully identified at the species, genus or family level by using four
molecular markers (12S mtDNA; COI; 18S rDNA; ITS1). Nematode larvae isolated from birds (Falco
tinnunculus; Circus cyaneus) and a pig, were identified as Toxocara cati; larvae from Meles meles were identified
as Toxocara canis; larvae from a domestic pig were identified as belonging to a free living nematode of the genus
Panagrolaimus; larvae from a wild boar (Sus scrofa) were identified as belonging to the Metastrongylus genus;
and larvae from a rough-legged buzzard (Buteo lagopus) were identified as belonging to the superfamily
Filarioidea. The recovery of nematodes different from those of the genus Trichinella during routine meat
inspection, seems to be not so unusual showing the need to inform technicians in charge for the analysis about the
possibility of false positives, and the need for a molecular based identification system allowing a reliable and
quick response.
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
The Reference Laboratory for Trichinellosis has been accredited by the Italian accreditation body ACCREDIA
according to UNI EN/ISO 17025:2005, since 2006, and the accreditation was confirmed for 2011–2012 following
the audit carried out on 7–8 July, 2011. Laboratory animals infected with Trichinella strains are maintained in an
animal house under biosefty measures.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
The reference lab has answered specific queries of European Commission and Italian Government on the Chapter
8.13 Infection with Trichinella spp.
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
Four persons from Estonia, Italy and Latvia have been trained on Molecular identification of Trichinella species
and on Digestion methods to detect Trichinella larvae in meat samples.
Establishing an OIE Reference laboratory for Trichinella at the Botswana national veterinary laboratory for the
southern Africa development community. Unfortunately, after the agreement signature in 2010, nobody from
Botswana answered the emails which were sent to them to organize their visit to Italy according to the project.
Only at the end of 2011, Dr Mercy W. Kawonga [mkawonga@gov.bw] sent an answer, but no other actions
occurred.
10. Provision of diagnostic testing facilities to other OIE Member Countries
a) Trichinella larvae identification at the species level by a multiplex PCR
4
OIE Member Country
No. of tests
Argentina
10
Bulgaria
65
Croatia
20
Czech Rep
10
Estonia
240
France
5
Annual reports of OIE Reference Centres, 2011
Trichinellosis
OIE Member Country
No. of tests
Germany
85
Iran
10
Ireland
10
Italy
100
Latvia
335
Lithuania
325
Norway
65
Poland
1250
Romania
250
Serbia
30
Spain
15
South Africa
5
Sweden
75
Zimbabwe
5
Total
2945
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
A two-day workshop was organised in Rome on May 2011, on: 1. presentation and discussion of the results of the
proficiency tests among European laboratories on the digestion method to detect Trichinella larvae in pork and/or
horse meat; 2. presentation and discussion of epidemiological data on Trichinella in humans and animals;
3. Organization of PT to detect Trichinella larvae. Seventy-five people from 30 European countries attended the
meeting. The workshop was organised by the European Union Reference Laboratory for Parasites. The meeting
Agenda, presentations and report can be downloaded from:
www.iss.it/crlp/wops/index.php?lang=2&tipo=25&anno=2011.
A tree-day meeting entitled ‘Expert Workshop on Quality Assurance in Trichinella Inspection and Testing
Programs’ was organised in Rome on November 2011. The meeting was organized by this OIE lab and the
International Commission of trichinellosis (ICT) and it was in part funded by OECD. Sixteen people from
11 countries of Europe, America, Asia and Australasia attended the meeting. The final document will be published
on the web site of ICT.
13. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
Communications at the 13th International Conference on Trichinellosis, Changchun, China, 1-6 August, 2011:
Pozio E, Bandino E, Murgia MC, Soddu M, Interisano M, Amati M, Marucci G, Gomez Morales MA. An
updating on the Trichinella britovi focus of the Sardinia island, Italy.
Gómez Morales MA, Amati M, Ludovisi A, Schimmel H, Ramirez Villaescusa A, Pozio E. Towards the first
international pig reference serum with anti-Trichinella antibodies: isochronous studies on the reference swine
serum candidates.
Gómez Morales MA, Amati M, Ludovisi A, Zivojinovic M, Blaga R, Ribicich M, Pozio E. Is possible to define
the pattern of Trichinella sp. infection for human and pig sera by western blot?
Annual reports of OIE Reference Centres, 2011
5
Trichinellosis
Ludovisi A, La Grange LJ, Gómez Morales MA, Amati M, Pozio E. Development of a serological test to detect
the humoral immune response against Trichinella zimbabwensis in Nile crocodiles (Crocodylus niloticus).
Pozio E. The opportunistic nature of Trichinella - exploitation of new geographies and habitats.
Pozio E., Krivokapich S, Marucci G. Cross-breedings between Trichinella T12 and the other encapsulated
Trichinella species suggest its reproductive isolation.
Marucci G., Interisano M, La Rosa G, Pozio E. Molecular identification of nematode larvae different from those of
the Trichinella genus detected by muscle digestion.
Marucci G., Interisano M, Gomez Morales MA, Glawischnig W, Claes M, Kärssin A, Zakrisson G, Pozio E.
Validation of a latex agglutination test for the detection of Trichinella infections in pigs.
Zivojinovic M, Sofronic-Milosavljevic Lj, Cvetkovic J, Pozio E, Interisano M, Radojicic S,.Kulusic Z. Trichinella
spp. infections in different host species of an endemic district of Serbia.
Koudela B1, Harna J, Račka K, Pozio E. Trichinella pseudospiralis in wild boars (Sus scrofa) of the Czech
Republic.

Scientific publications in peer-reviewed journals
Murrell KD, Pozio E. 2011. Worldwide occurrence and impact of human trichinellosis, 1986-2009. Emerg Infect
Dis 17:2194-202.
Reichard MV, Tiernan KE, Paras KL, Interisano M, Reiskind MH, Panciera RJ, Pozio E. 2011. Detection of
Trichinella murrelli in coyotes (Canis latrans) from Oklahoma and North Texas. Vet Parasitol. 182:368-71.
Romano F, Motta A, Melino M, Negro M, Gavotto G, Decasteli L, Careddu E, Bianchi C, Bianchi DM, Pozio E.
2011. Investigation on a focus of human trichinellosis revealed by an atypical clinical case: after wild-boar (Sus
scrofa) pork consumption in northern Italy. Parasite 18:85-7.
Alban L, Pozio E, Boes J, Boireau P, Boué F, Claes M, Cook AJ, Dorny P, Enemark HL, van der Giessen J, Hunt
KR, Howell M, Kirjusina M, Nöckler K, Rossi P, Smith GC, Snow L, Taylor MA, Theodoropoulos G, Vallée I,
Viera-Pinto MM, Zimmer IA. 2011. Towards a standardised surveillance for Trichinella in the European Union.
Prev Vet Med 99:148-60.
Merialdi G, Bardasi L, Fontana MC, Spaggiari B, Maioli G, Conedera G, Vio D, Londero M, Marucci G, Ludovisi
A, Pozio E, Capelli G. 2011. First reports of Trichinella pseudospiralis in wild boars (Sus scrofa) of Italy. Vet
Parasitol 178:370-3.
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6
Annual reports of OIE Reference Centres, 2011