1977L0096 — EN — 05.06.2003 — 009.001 — 1
This document is meant purely as a documentation tool and the institutions do not assume any liability for its contents
►B COUNCILDIRECTIVE
of21December 1976
on the examination for trichinae (trichinella spiralis) upon importation from third countries of
freshmeatderivedfromdomesticswine
(77/96/EEC)
(OJ L 26, 31.1.1977, p. 67)
Amended by:
Official Journal
No page date
►M1Council Directive 81/476/EEC of 24 June 1981 L 186 20 8.7.1981
►M2Council Directive 83/91/EEC of 7 February 1983 L 59 34 5.3.1983
►M3Commission Directive 84/319/EEC of 7 June 1984 L 167 34 27.6.1984
►M4Council Regulation (EEC) No 3768/85 of 20 December 1985 L 362 8 31.12.1985
►M5Commission Directive 89/321/EEC of 27 April 1989 L 133 33 17.5.1989
►M6Commission Directive 94/59/EC of 2 December 1994 L 315 18 8.12.1994
►M7Council Regulation (EC) No 807/2003 of 14 April 2003 L 122 36 16.5.2003
Amended by:
►A1Act of Accession of Greece L 291 17 19.11.1979
►A2Act of Accession of Austria, Sweden and Finland C 241 21 29.8.1994
(adapted by Council Decision 95/1/EC, Euratom, ECSC) L 1 1 1.1.1995
▼B
COUNCILDIRECTIVE
of21December 1976
ontheexaminationfortrichinae(trichinellaspiralis)uponimporta-
tion from third countries of fresh meat derived from domestic
swine
(77/96/EEC)
THE COUNCIL OF THE EUROPEAN COMMUNITIES,
Having regard to the Treaty establishing the European Economic
Community,
Having regard to Council Directive 72/462/EEC of 12 December 1972
on health and veterinary inspection problems upon importation ofbovine animals and swine and fresh meat from third countries (
1), as
last amended by Directive 75/379/EEC (2), and in particular Article 21
thereof,
Having regard to the proposal from the Commission,
Whereasin Directive 72/462/EEC the Council provided in Article 21
for the laying down of the method and proceduresrequired fordetecting the presence of trichinae in fresh pigmeat;
Whereasthe application of Directive 72/462/EEC will not have the
desired effects as long as disparities exist between the Member Statesasto the guaranteesrequired in res pect of the detection of trichinaeupon importation of fresh meat from third countries; whereas it is
therefore necessary to lay down Community arrangements in this field;
Whereas, in order to protect consumer health, it is necessary that fresh
pigmeat be systematically subjected to an examination using methods
recognized aseffective, in order to eliminate meat containing trichinae;
Whereaswhen the examination iscarried out in the exporting third
country, it must be carried out in slaughterhouses which comply with
certain conditionsand which contain, in particular, a s creening labora-tory provided with suitable equipment;
Whereas in order to be able to distinguish the meat samples examined
from those not examined it is necessary to provide for the affixing of aspecial mark to meat which has been examined with a negative result;
Whereas there should be a procedure establishing close and effective
cooperation between the Commission and the Member States for asses-sing the advisability of permitting establishments in the third countriesto carry out thisexamination or to work on the meat examined;whereas there should also be a procedure for bringing the technicalprovisions relating in particular to the examination methods, the
requirementsconcerning the s creening laboratoriesand the procedure
for marking examined meat into line with technical progress and withexperience acquired;
Whereas the Member States should be allowed to admit fresh meat
which hasnot been s creened for trichinae in the exporting thirdcountry, provided that thismeat, undergoestreatment by freezing;which ensures that any trichinae which may be present are renderedharmless, either in the exporting third country, or in the Member State
for which the meat isintended; whereasthistreatment mus t neverthe-
less be carried out according to certain well defined procedures andestablishments fulfilling certain conditions,1977L0096 — EN — 05.06.2003 — 009.001 — 2
(1) OJ No L 302, 31. 12. 1972, p. 28.
(2) OJ No L 172, 3. 7. 1975, p. 17.
▼B
HAS ADOPTED THIS DIRECTIVE:
Article 1
The definitions used in this Directive are those contained in Directive
72/462/EEC.
Moreover:
(a) ‘fresh meat’ means fresh meat of domestic swine;
(b) ‘examination’ refersto the examination to detect the pres ence of
trichinae in fresh meat.
Article 2
1. In order to be admitted to intra-Community trade, fresh meat
originating in third countries which contains skeletal muscles (striatedmuscles) shall be examined under the supervision and responsibility ofan official veterinarian.
2. The examination shall be carried out in accordance with one of
the methodsprovided for in Annex I, on the whole carcas e or, failingthis, on each half carcase, quarter carcase or piece to be imported intothe Community.
3. The examination shall take place in a slaughterhouse approved in
the exporting country in accordance with Article 4 of Directive 72/462/
EEC and authorized to carry out thisexamination in accordance withArticle 4 of thisDirective.
4. The examination shall take place before the health marking
provided for in Chapter X of Annex B to Directive 72/462/EEC.
5. If it is not possible to carry out the examination in the exporting
country, the Member State for which the fresh meat is intended mayauthorize itsimportation provided that the examination iscarried outwithin itsterritory at the time of the public health ins pection providedfor in Article 24 (2) of Directive 72/462/EEC, at an inspection post
within the meaning of Article 27 (1) (b) of that Directive.
6. (a) If the outcome of the examination isnegative, the fres h meat
shall be marked immediately after the examination, in accor-dance with Annex III.
(b) In the case of ink stamping, use shall be made of a colorant
within the meaning of Article 17 (3) of Directive 72/462/EEC.
Article 3
1. By way of derogation from Article 2, the Member State for which
it isintended may authorize the exemption from examination of fres hmeat from certain third countries or parts of such countries, provided itisfrozen in accordance with the provis ionsof Annex IV.
2. This treatment shall be carried out in an establishment situated in
the exporting third country, and described in Article 4 (1).
Freezing in the exporting third country must be the subject of certifica-
tion by the official veterinarian on the health certificate accompanyingthe meat, asreferred to in Article 22 (3) of Directive 72/462/EEC.
3. If the treatment hasnot been carried out in the exporting third
country, it must be carried out at an inspection post as described inArticle 2 (5).
Freezing in a Member State must be the subject of certification by the
official veterinarian on the certificatesaccompanying the meat, as
referred to in Article 25 of Directive 72/462/EEC.
Article 4
1. The authorization for a slaughterhouse to carry out the examina-
tion and of a cutting plant to cut up or bone meat which hasundergonesuch examination, or the authorization for an establishment to carry out1977L0096 — EN — 05.06.2003 — 009.001 — 3
▼B
the freezing treatment referred to in Article 3, shall be decided on in
accordance with the procedure laid down in Article 9. In addition tothe requirementsof Article 4 of Directive 72/462/EEC, account s hall
be taken of the guaranteesgiven in res pect of compliance with this
Directive and, in the case of slaughterhouses, of:
(a) the presence of the rooms and apparatus necessary for carrying out
the examination;
(b) the qualifications of the personnel responsible for carrying out the
examination.
Authorization shall be granted to a slaughterhouse and cutting plant
only where the competent authoritiesof the third country concernedhave officially recognized that the slaughterhouse and cutting plant
are in a position to satisfy the conditions laid down in Article 5 and
in Annex III; also, in the case of a slaughterhouse, that it has a labora-tory which complieswith the conditionslaid down in Annex II,Chapter I, and which is in a position to satisfy the requirements of theother chaptersof Annex II and thos e of Annex I.
Authorization shall be granted to an establishment to carry out the
freezing treatment only if the competent authoritiesof the third countryconcerned have officially recognized that the establishment is in a posi-tion to satisfy the conditions laid down in Annex IV.
2. On the list(s) referred to in Article 4 (4) of Directive 72/462/EEC,
a special indication shall be inserted against the names of the establish-mentswhich have been granted an authorization within the meaning ofparagraph 1.
Article 5
1. In slaughterhouses which have been granted an authorization in
accordance with Article 4, swine the meat of which is intended for
the Community must be slaughtered in different rooms or in the
absence thereof at different times from swine the meat of which is notintended for the Community, unless the meat of such swine is exam-ined in accordance with the same procedure.
2. The cutting and boning of meat which hasundergone an examina-
tion with negative results and is intended for the Community must becarried out in cutting plants, in accordance with Article 4.
In these cutting plants, the cutting and boning of such meat must be
carried out in different roomsor in the abs ence thereof, at differenttimesfrom meat which isnot intended for the Community, unles sthemeat isexamined in accordance with the s ame procedure.
Article 6
The inspections in third countries provided for in Article 5 of Directive
72/462/EEC must also verify whether the present Directive is beingapplied.
Article 7
The Member States shall draw up and communicate to the Commission
the list of the inspection posts referred to in Article 2 (5) at which:
— the examination,
— the freezing referred to in Article 3,
may be carried out.
They shall ensure that these posts have the equipment necessary for
carrying out the operationsin ques tion.
Article 8
Using the procedure laid down in Article 9, decisions may be taken to
adapt or add to the Annexeshereto in the light of technological devel-1977L0096 — EN — 05.06.2003 — 009.001 — 4
▼M2
▼M2
opments; such decisions must ensure that health guarantees are main-
tained at their present standard.
Article 9
1. The Commission shall be assisted by the Standing Committee on
the Food Chain and Animal Health set up by Article 58 of Regulation
(EC) No 178/2002 (1).
2. Where reference ismade to thisArticle, Articles5 and 7 of Deci-
sion 1999/468/EC (2) shall apply.
The period laid down in Article 5(6) of Decision 1999/468/EC shall be
set at three months.
3. The committee shall adopt its rules of procedure.
Article 11
The Member Statess hall bring into force on 1 January 1979 at the
latest the laws, regulations and administrative provisions needed forcompliance with thisDirective. They s hall forthwith inform theCommission thereof.
Article 12
This Directive is addressed to the Member States.1977L0096 — EN — 05.06.2003 — 009.001 — 5
(1) OJ L 31, 1.2.2002, p. 1.
(2) OJ L 184, 17.7.1999, p. 23.▼M7
▼M1
▼B
▼B
ANNEX I
METHODSOFEXAMINATIONFORTRICHINAE
I. TRICHINOSCOPIC EXAMINATION
(a)Apparatus
An incandescent lamp trichinoscope with 50 × and 80 to 100 × magni-
fication.
A pressure glass consisting of two glass plates — one of which is
divided into equal fields — small curved scissors, small forceps, a
knife for cutting specimens, small numbered containers for storing the
specimens separately, a dropping pipette, a glass of acetic acid and a
glass of potassium hydroxide solution for brightening any calcifications
or softening dried meat.
(b)Takingofspecimens
In the case of whole carcases, at least one specimen of the size of a
hazelnut isto be taken from both diaphragm pillarsat the trans ition
of the sinewy part.
If there is only one diaphragm pillar, one specimen the size of two
hazelnuts is to be taken. In the absence of both diaphragm pillars,
two specimens approximately the size of a hazelnut are to be taken
from the rib part or the breastbone part of the diaphragm or, as the
case may be, from the lingual muscle or the jaw muscle or from the
abdominal muscles.
For pieces of meat, from each piece take three samples of skeletal
muscle, containing little fat, if possible the size of a hazelnut, and
taken from different points, where possible near to bones or tendons.
(c)Method
If both diaphragm pillars are present the trichinae inspector must cut,
from each of the above specimens taken from a whole carcase, seven
piecesthe s ize of an oat-kernel making 14 in all; if only one
diaphragm pillar is present, 14 pieces, from different places and if
possible from the transition to the sinewy part; he must then compress
them between the glass plates in such a way that normal print can be
clearly read through the slide preparation. If the flesh of the specimens
to be examined is dry and old, the preparations must be softened in a
mixture of one part potassium hydroxide solution to about two parts
water for 10 to 20 minutes before pressing.
If, in the case of whole carcases, specimens have to be taken from the
rib part or the breastbone part of the diaphragm, the lingual muscle or
jaw muscle or the abdominal muscles, then 14 pieces the size of an
oat-kernal are to be cut from each specimen, i.e. a total of 28.
From each of the samples taken from pieces of meat, the trichinae
inspector must cut four pieces the size of an oat-kernal, making 12
piecesin all.
The trichinoscopic examination must be carried out in such a manner
that each preparation is scanned slowly and carefully. If the trichino-
scopic examination reveals suspect areas, the nature of which cannot
be definitely ascertained even with the most powerful magnification
of the trichinoscope, these must be checked by microscope.
The microscopic examination should be carried out in such a manner
that each preparation is scanned slowly and carefully at a magnifica-
tion of 30 to 40 ×.
In the case of an uncertain result, the examination must be continued
on a further number of specimens and slide preparations, if necessary
with the aid of higher magnifications, until the information required is
obtained. The trichinoscopic examination must be carried out for at
least three minutes.
The trichinoscopic examination must be carried out for at least six
minutes in the case of substitute specimens taken from the rib part or
breastbone part of the diaphragm pillars, the lingual muscle or the jaw
muscle or the abdominal muscles.1977L0096 — EN — 05.06.2003 — 009.001 — 6
▼B
The minimum time fixed for the examination doesnot include the time
necessary for sample-taking and for making the preparations.
As a general rule, the trichinoscopic examiner should not inspect more
than 840 piecesa day, though by way of exception he may ins pect up
to 1 050.
II. THE ARTIFICIAL DIGESTION METHOD
(a)Apparatusandmaterial
— knife for taking samples,
— small numbered containers, with closures, for storing samples, if
necessary until repetition of the examinations,
— incubator,— 2 to 3 litre glass funnel with stand, a connecting hose in rubber,
clamps for fastening the connecting hose,
— a plastic sieve (approximately 18 cm in diameter, and approxi-
mately 1 mm mesh),
— gauze,— a small tapering tube with a sealed end,— a glass block dish,— a meat mincer,
— a stereo-microscope (magnification 15 to 40 ×) with a suitable light
source,
— digestive liquid made up as follows:
10 g of pepsin (80 u/g FIP: Fédération internationale de phar-
macie), 5 ml HCl (at least 37 %) made up to a litre with tap-water.
(b)Takingofspecimens
1. In the case of whole carcases a sample of at least 20 g to be taken
from a diaphragm pillar at the transition to the sinewy part. In the
absence of diaphragm pillars a specimen of at least 20 g to be taken
from the rib part or the breastbone part of the diaphragm or from
the lingual muscle or the jaw muscle or the abdominal muscles.
2. For pieces of meat, a sample of at least 20 g of skeletal muscle to
be taken, containing little fat and where possible near to bones or
tendons.
(c)Method
For the examination of a collective specimen from 10 pigs, a 10 g
specimen shall be prepared from each individual 20 g sample. The
remaining 10 g shall be kept for additional single-specimen examina-
tion should this be necessary.
10 specimens, each weighing 10 g, shall be combined into a collective
specimen; it shall be minced in a meat mincer (with 2 mm perfora-
tions) and placed loosely in a sieve lined with a layer of gauze. The
sieve shall then be placed in a funnel connected by a length of rubber
hose to a small tapered tube with a sealed end; the funnel shall be
filled from the edge with digestive liquid until the matter for analysis
is completely covered. The proportion of matter for analysis to diges-
tive liquid must be approximately 1:20 to 1:30.
After 18 to 20 hoursof incubation at 37 to 39 ºC, the s mall tapered
tube shall be disconnected and removed. After carefully drawing off
the supernatant liquid, the sediment present in the tip of the tube is to
be carefully rinsed in a dish. Then to be examined for the presence of
trichinae with a stereo-microscope at 20 to 40 × magnification.
In the case of a positive or doubtful result from the examination of a
collective specimen, the remaining single specimens to be examined
individually after addition of a further 20 g from each pig, or in the
case of pieces of meat, the addition of 20 g taken from each piece,
according to (b) above.1977L0096 — EN — 05.06.2003 — 009.001 — 7
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III. METHOD USING THE ARTIFICIAL DIGESTION OF COLLECTIVE
SAMPLES
(a)Apparatusandreagents
— knife and tweezers for collecting specimens,
— meat mincer with 2 to 3 mm diameter perforations,
— a 3 litre Erlenmeyer flask with a rubber or cotton-wool plug,
— a conical separation funnel of 2 000 ml capacity,
— an ordinary A-base stand of approximately 28 cm length with an
80 cm stem,
— a ring, diameter approximately 10 to 11 cm which can be fixed to
the stand,
— a clamp with a flat vice (23 × 40 mm) which can be attached to the
stand by means of a double coupling,
— a sieve (mesh size 177 microns) with an external diameter of 11 cm
fitted with brass or stainless steel mesh,
— a funnel with an internal diameter of not less than 12 cm,
— 100 ml glass measuring cylinders,
— a stereo-microscope (magnification 15 to 40 ×) with a suitable light
source, or a trichinoscope with horizontal table for the compressor
with suitable light source,
— when using the trichinoscope: a larval counting basin is used which
may be described as follows:
the larval counting basin is made from 3 mm thick acrylic plates,
asfollows :
(i) the bottom of the basin to be 180 × 40 mm, marked off into
squares,
(ii) the sides to be 230 × 20 mm,
(iii) the end to be 40 × 20 mm. The bottom and the endss hould be
inserted between the sides, thus forming a basin with two
small handles in both ends. The upper side of the bottom
should be raised 7 to 9 mm from the base of the frame formed
by the sides and the ends. The parts should be fixed by using
glue appropriate for the material,
— a number of 9 cm diameter Petri dishes (when using stereo-micro-
scope) marked on their undersides into 10 × 10 mm square
examination areas using a pointed instrument,
— a number of 10 litre binsto be us ed when applying decontamina-
tion, such as formol treatment, to the apparatus and for the
remaining digestive juice in the case of positive results,
— concentrated (37 %) hydrochloric acid,
— pepsin strength: 1: 10 000 NF (US National Formulary)
corresponding to 1: 12 500 BP (British Pharmacopoea)
corresponding to 2 000 FIP (Fédération internationale de phar-
macie),
— a number of trayswhich can hold 50 s amplesof approximately 2 g
each,
— a balance accurate to 0,1 g.
(b)Collectionofspecimens
1. In the case of whole carcases, a specimen to be taken of approxi-
mately 2 g from a pillar of the diaphragm at the transition to the
sinewy part. In the absence of diaphragm pillars, a specimen of the
same size to be taken from the rib part of the breastbone part of the
diaphragm, from the lingual muscle or the jaw muscle or the
abdominal muscles.
2. For cuts of meat, a sample of approximately 2 g of skeletal muscle
to be taken, containing little fat and, where possible, near to bones
or tendons.
(c)Method
1. (i)Complete pools (100 samples at a time)
Approximately 1 g of sample shall be taken from each of the
100 individual samples from the pigs. The pooled sample is
put once through the mincer.
The minced meat shall be placed in the 3 litre Erlenmeyer flask
together with 7 g of pepsin, approximately 2 litres of tap-water1977L0096 — EN — 05.06.2003 — 009.001 — 8
▼M3
heated to about 40 to 41 ºC and 25 ml of concentrated hydro-
chloric acid. The mixture shall be shaken to dissolve the pepsin.
The pH of the solution will be about 1,5 to 2.
— For digestion, the Erlenmeyer flask shall be incubated at 40
to 41 ºC for approximately four hours. The flask shall be
regularly shaken during the time of incubation, at least
twice every hour.
— The digested solution shall be filtered through the sieve into
the conical 2 litre separation funnel and left undisturbed on
the stand for at least one hour.
— A total volume of approximately 45 ml should be run off
into a measuring cylinder and divided between three Petri
dishes, the bottoms of which should be marked off into
squares, with 15 ml in each dish.
— Each Petri dish shall be minutely examined for larval
trichinae under the stereo-microscope.
— Where larval counting basins are employed, the 45 ml shall
be distributed between two larval counting basins and
examined under the trichinoscope.
The larvae appear as identifiable organisms in the deposit
and often, when the water islukewarm, rolling and unrol-
ling movements of the ‘spiral’ may be observed.
— Digests should be examined as soon as they are ready.
Under no circumstances should examination be postponed
until the following day.
If the digests are unclear or not examined within 30 minutes
of their preparation, they should be clarified as follows. The
final sample of 45 ml is poured into a measuring cylinder
and allowed to stand for 10 minutes. At the end of this
time, 30 ml of supernatant fluid is removed by suction and
the remaining 15 ml ismade up to 45 ml with tap-water.
After a further settling period of 10 minutes, 30 ml of
supernatant fluid is removed by suction and the remaining
15 ml is poured into a Petri dish or larval counting basin
for examination. The measuring cylinder should be washed
with 10 ml of tap-water and these washings should be added
to the sample in the Petri dish or larval counting basin for
examination.
(ii)Pools of less than 100 samples
Up to 15 single samples could be added to a total pool of 100
samples and be examined together with these samples. If more
than 15 samples and less than 100 samples are examined, thedigestion fluid should be reduced proportionately.
2. In the case of a positive or doubtful result following the examina-
tion of a collective sample, a further 20 g sample should be taken
from each pig in accordance with (b) above. The 20 g samples from
five pigs should be pooled and examined by the method described
above. In thisway s amplesfrom 20 groupsof five pigswill be
examined. When trichinellae are detected in a pooled sample from
five pigs, further 20 g samples should be collected from the indivi-
dual pigs in the group and each should separately be examined
using the method described above.
IV. THE MECHANICALLY ASSISTED POOLED SAMPLE DIGESTION
METHOD/SEDIMENTATION TECHNIQUE
(a)Apparatusandreagents
— knife or scissors for cutting specimens,
— trays marked off with 50 squares each of which can hold samples
of approximately 2 g of meat,
— a Stomacher Lab-blender 3 500 Thermo model,
— plastic bags suitable for the Stomacher Lab-blender,
— conical separation funnels of 2 litre capacity, preferably fitted with
teflon safety plugs,
— stands, rings and clamps,
— sieves, mesh size 177 microns, external diameter 11 cm with stain-
less steel mesh,
— funnels with an internal diameter not less than 12 cm, to support
the sieves,1977L0096 — EN — 05.06.2003 — 009.001 — 9
▼M3
— 100 ml glass measuring cylinders,
— a 25 ml dispenser,
— beakersof 3 litre capacity,— spoon or glass rod for stirring the digestion fluid in the beaker,
— a plastic syringe and tube for suction,
— a measuring spoon for 6 g,— a thermometer accurate to ± 0,5 ºC within the range 1 to 100 ºC,— a vibrator, e.g. an electric shaver with the head removed,
— a relay which will switch on and off at one minute interval,
— a trichinoscope with a horizontal table or a stereo-microscope, with
a suitable light source,
— larval counting basin (when using trichinoscope): the larval
counting basin is made from 3 mm thick acrylic plates as follows:
(i) the bottom of the basin to be 180 × 40 mm, marked off into
squares,
(ii) the sides to be 230 × 20 mm,
(iii) the end to be 40 × 20 mm. The bottom and the endss hould be
inserted between the sides, thus forming two small handles in
both ends. The upper side of the bottom should be raised 7 to
9 mm from the base of the frame formed by the sides and the
ends. The parts should be fixed by using glue appropriate for
the material,
— a number of 9 cm diameter Petri dishes (when using stereo-micro-
scope) marked on their undersides into 10 × 10 mm square
examination areas using a pointed instrument,
— 17,5 % hydrochloric acid solution,
— pepsin strength 1: 10 000 NF (US National Formulary)
corresponding to 1: 12 500 BP (British Pharmacopoea)corresponding to 2 000 FIP (Fédération internationale de phar-
macie),
— a number of 10 litre binsto be us ed when applying decontamina-
tion, such as formol treatment, to the apparatus and for the
remaining digestive juice in the case. of positive results,
— a balance accurate to 0,1 g.
(b)Collectionofspecimens
1. In the case of whole carcases, a specimen to be taken of approxi-
mately 2 g from a pillar of the diaphragm at the transition to the
sinewy part. In the absence of diaphragm pillars, a specimen of the
same size to be taken from the rib part or the breastbone part of the
diaphragm, from the jaw muscle or the abdominal muscle.
2. For cuts of meat, a sample of approximately 2 g of skeletal muscle
to be taken, containing little fat and, where possible, near to bones
or tendons.
(c)Method
1.Digestion procedure
(i)Complete pools (100 samples at a time)
— The Stomacher Lab-blender 3 500 should be fitted with a
double plastic bag and the temperature control set at 40 to
41 ºC
— One and a half litresof water preheated to about 32 to 35 ºC
ispoured into the inner plas tic bag and the water heated to
40 to 41 ºC.
— 25 ml of 17,5 % hydrochloric acid isthen added to the
water in the Stomacher.
— 100 samples of approximately 1 g each (at 25 to 30 ºC)
taken from each of the individual samples, in accordance
with (b), are then added.
— 6 g pepsin is finally added. This order of addition should be
strictly adhered to in order to avoid decomposition of the
pepsin.
— The Stomacher isthen allowed to pound the content of the
bag for 25 minutes.
— The pastic bag is then removed from the Stomacher and the
digestion fluid is filtered through the sieve into a 3 litre
beaker.1977L0096 — EN — 05.06.2003 — 009.001 — 10
▼M3
— The plastic bag is washed with approximately 100 ml of
water, which is then used to rinse the sieve and finally
added to the filtrate in the beaker.
Up to 15 single samples could be added to a total pool of 100
samples and be examined together with these samples.
(ii)Smaller pools less than 100 samples
— The Stomacher Lab-blender 3 500 should be fitted with a
double plastic bag and the temperature control set at 40 to
41 ºC.
— A digestion fluid is prepared by mixing about one and a
half litresof water and 25 ml of 17,5 % hydrochloric acid.
6 g of pepsin is added and the whole mixed at a temperature
of 40 to 41 ºC. This order of addition should be strictly
adhered to in order to avoid decomposition of the pepsin.
— Of the digestion fluid, a volume corresponding to 15 ml per
gram of sample is measured (e.g. for 30 samples the volume
required is30 × 15 ml or 450 ml) and trans ferred to the
inner of the two plastic bags together with the meat samples
of approximately 1 g (at 25 to 30 ºC) taken from each of the
individual samples in accordance with (b).
— Water at a temperature of approximately 41 ºC ispoured
into the outer bag to a total volume in the two bagsof one
and a half litres.
— The Stomacher isthen allowed to pound the content of the
bag for 25 minutes.
— The plastic bag is then removed from the Stomacher and the
digestion fluid is filtered through the sieve into a 3 litre
beaker.
— The plastic bag is washed with approximately 100 ml of
water, which is then used to rinse the sieve and finally
added to the filtrate in the beaker.
2.Recovery of larvae by sedimentation
— Ice (300 to 400 g of ice flakes, scaly ice or crushed ice) is
added to the digestion fluid, bringing its volume up to about 2
litres. The digestion fluid is then stirred until the ice has melted.
In the case of smaller pools (see 1 (ii)), the amount of ice
should be reduced correspondingly.
— The chilled digestion fluid is transferred to a 2 litre separation
funnel, equipped with a vibrator in an extra clamp.
— Sedimentation for 30 minutes, during which time the sedimenta-
tion funnel isvibrated intermittently, i.e. one minute vibration
followed by one minute pause.
— After 30 minutes, a 60 ml sample of the sediment is quickly run
off into a 100 ml measuring cylinder. (The funnel is rinsed with
detergent solution after use).
— The 60 ml sample is allowed to stand for at least 10 minutes,
after which time the supernatant should be withdrawn by
suction, leaving a volume of 15 ml to be examined for the
presence of larvae.
— For suction, a disposable syringe can be used, equipped with a
plastic tube.
The length of the tube should be such that 15 ml will remain in
the measuring cylinder when the flanges of the syringe rest on
the cylinder'srim.
— The remaining 15 ml ispoured into a larval counting bas in or
two Petri dishes and examined using a trichinoscope or stereo-
microscope respectively.
— Digests should be examined as soon as they are ready. Under no
circumstances should examination be postponed until the
following day.
If the digests are unclear, or are not examined within 30 minutes
of their preparation, they should be clarified as follows. The
final sample of 60 ml is poured into a measuring cylinder and
allowed to stand for 10 minutes. At the end of this time, 45 ml
of supernatant fluid is removed by suction and the remaining
15 ml ismade up to 45 ml with tap-water. After a further
settling period of 10 minutes, 30 ml of supernatant fluid is
removed by suction and the remaining 15 ml is poured into a
Petri dish or larval counting basin for examination. The
measuring cylinder should be washed with 10 ml of tap-water1977L0096 — EN — 05.06.2003 — 009.001 — 11
▼M3
and these washings should be added to the sample in the Petri
dish or the larval counting basin for examination.
3. In the case of a positive or doubtful result, following the examina-
tion of a collective sample, a further 20 g sample should be taken
from each pig in accordance with (b) above. The 20 g samples from
five pigs should be pooled and examined by the method described
above. In thisway, s amplesfrom 20 groupsof five pigswill be
examined. When trichinellae are detected in a pooled sample from
five pigs, further 20 g samples should be collected from the indivi-
dual pigs in the group and each should separately be examined
using the method described above.
V. THE MECHANICALLY ASSISTED POOLED SAMPLE DIGESTION
METHOD/ ‘ON FILTER ISOLATION’ TECHNIQUE
(a)Apparatusandreagents
Those indicated in method IV (a).Supplementary equipment to the abovementioned:
— 1 litre Gelman funnel, complete with filter holder (diameter
45 mm),
— filter discs; the filter discs consist of:
a circular stainless steel mesh with an aperture of 35 microns (the
diameter of the disc should be 45 mm),
two rubber ringsmade of 1 mm thick rubber (the external diameter
should be 45 mm and the internal diameter 38 mm),
the circular mesh is placed between the two rubber rings and
bonded to them using a two-component glue suitable for the two
materials,
— an Erlenmeyer flask with a capacity of 3 litres and fitted with a
side tube for suction,
— a filter pump,
— plastic bags with a capacity of at least 80 ml,
— equipment for sealing the plastic bags,— rennilase, strength 1: 150 000 soxhlet units per gram.
(b)Collectionofspecimens
See method IV (b).
(c)Method
1.Digestion procedure
(i)Complete pools (100 samples at a time)
See method IV (c) (1) (i).
(ii)Smaller pools less than 100 samples
See method IV (c) (1) (ii).
2.Recovery of larvae by filtration
— Ice (300 to 400 g of ice flakes, scaly ice or crushed ice) is
added to the digestion fluid, bringing its volume up to about 2
litres.
In the case of smaller pools, the amount of ice should be
reduced correspondingly.
— The digestion fluid is then stirred until the ice has melted. The
chilled digestion fluid is then left for at least three minutes to let
the larvae coil.
— The Gelman funnel, fitted with a filter holder and filter disc, is
mounted on the Erlenymeyer flask connected to a filter pump.
— The digestion fluid is poured into the Gelman funnel and
filtered. Towards the end of filtration, the passage of the diges-
tion fluid through the filter can be assisted by applying suction
with the filter pump. Suction should cease before the filter
becomesdry, i.e. when 2 to 5 ml of fluid are left in the funnel.
— When all the digestion fluid has been filtered, the filter disc is
removed and placed in an 80 ml capacity plastic bag, together
with 15 to 20 ml of rennilase solution. The solution of rennilase
ismade by adding 2 g of rennilas e to 100 ml of tap-water.
— The plastic bag is sealed twice and placed in the Stomacher
between the inner and outer bag.1977L0096 — EN — 05.06.2003 — 009.001 — 12
▼M3
— The Stomacher isallowed to pound for three minutes , e.g. while
it isworking on a complete or incomplete pool.
— After three minutes, the plastic bag, complete with filter disc
and rennilase solution, is removed from the Stomacher and
opened with scissors. The liquid contents are poured into a
larval counting basin or Petri dish. The bag is washed out with
5 to 10 ml of water which isthen added to the larval counting
basin for examination by trichinoscope or to the Petri dish for
examination by stereo-microscope.
— Digests should be examined as soon as they are ready. Under no
circumstances should examination be postponed until the
following day.
NoteFilter discs should never be used when not completely clean.
Unclean discs should never be allowed to dry out.
Filter discs can be cleaned by leaving them in rennilase solution
overnight. Before use, they should be washed in fresh rennilase
solution using the Stomacher.
3. In the case of a positive or doubtful result, following the examina-
tion of a collective sample, a further 20 g sample should be taken
from each pig in accordance with (b) above. The 20 g samples from
five pigs should be pooled and examined by the method described
above. In thisway, s amplesfrom 20 groupsof five pigswill be
examined. When trichinellae are detected in a pooled sample from
five pigs, further 20 g samples should be collected from the indivi-
dual pigs in the group and each should separately be examined
using the method described above.
VI. THE MAGNETIC STIRRER METHOD FOR POOLED SAMPLE DIGES-
TION
(a)Apparatusandreagents
— knife and tweezers for cutting specimens,
— trays marked off into 50 squares each of which can hold samples of
approximately 2 g of meat,
— a Moulinette blender,— magnetic stirrers, with thermostatically controlled heating plate and
teflon coated stirring rods, approximately 5 cm long,
— conical separation funnels of 2 litre capacity,— stands, rings and clamps,
— sieves, mesh size 177 microns, external diameter 11 cm with stain-
less steel mesh,
— funnels with an internal diameter not less than 12 cm, to support
the sieves,
— beaker of 3 litre capacity,— measuring cylinders of approximately 50 ml capacity, or centrifuge
tubes,
— a trichinoscope with horizontal table or a stereo-microscope, with a
suitable light source,
— larval counting basin (when using trichinoscope): the larval
counting basin is made from 3 mm thick acrylic plates as follows:
(i) the bottom of the basin to be 180 × 40 mm, marked off into
squares,
(ii) the sides to be 230 × 20 mm,
(iii) the end to be 40 × 20 mm. The bottom and the endss hould be
inserted between the sides, thus forming two small handles in
both ends. The upper side of the bottom should be raised 7 to
9 mm from the base of the frame formed by the sides and the
ends. The parts should be fixed by using glue appropriate for
the material,
— a number of 9 cm diameter Petri dishes (when using stereo-micro-
scope) marked on their undersides into 10 × 10 mm square
examination areas using a pointed instrument,
— aluminium foil,— 25 % hydrochloric acid,— pepsin strength: 1: 10 000 NF (US National Formulary)
corresponding to 1: 12 500 BP (British Pharmacopoea)1977L0096 — EN — 05.06.2003 — 009.001 — 13
▼M3
corresponding to 2 000 FIP (Fédération internationale de phar-
macie),
— tap-water heated to 46 to 48 ºC,— a number of 10 litre binsto be us ed when applying decontamina-
tion, such as formol treatment, to the apparatus and for the
remaining digestive juice in the case of positive results,
— a balance accurate to 0,1 g.
(b)Collectionofspecimens
1. In the case of whole carcases, a specimen to be taken of approxi-
mately 2 g from a pillar of the diaphragm at the transition to the
sinewy part. In the absence of diaphragm pillars, a specimen of the
same size to be taken from the rib part or the breastbone part of the
diaphragm, from the jaw muscle or the abdominal muscles.
2. For cuts of meat, a sample of approximately 2 g of skeletal muscle
to be taken, containing little fat and, where possible, near to bones
or tendons.
(c)Method
1. (i)Complete pools (100 samples at a time)
— 100 samples of approximately 1 g each, taken from each of
the individual samples in accordance with (b), are chopped
in the Moulinette blender. The blender should be operated
three to four timesfor approximately one s econd each time.
— The chopped meat istrans ferred to a 3 litre beaker and
sprinkled with 10 g of pepsin. 2 litres of tap-water,
preheated to 46 to 48 ºC, ispoured into the beaker, together
with 16 ml of hydrochloric acid.
— The mincing insert of the Moulinette blender is immersed
repeatedly in the digestion fluid in the beaker to remove
any meat still adhering.
— The stirring rod is placed in the beaker and the beaker is
covered with aluminium foil.
— The beaker isplaced on the preheated heating plate of the
magnetic stirrer and the stirring process started. Before
starting the stirring process, the magnetic stirrer should be
adjusted so that it maintains a constant temperature of 44
to 46 ºC throughout the period of operation. During the stir-
ring process, the digestion fluid should rotate at a
sufficiently high speed to create a deep whirl without
splashing.
— The digestion fluid is stirred for 30 minutes, at the end of
which the stirrer is switched off and the digestion fluid is
poured through the sieve into the sedimentation funnel.
— The digestion fluid is allowed to stand in the funnel for 30
minutes.
— After 30 minutes, a 40 ml sample of digestion fluid is
quickly run off into the measuring cylinder or centrifuge
tube.
— The 40 ml sample is allowed to stand for 10 minutes, at the
end of which time 30 ml of supernatant is withdrawn by
suction, leaving a volume of 10 ml.
— The remaining 10 ml sample of sediment is poured into a
larval counting basin or Petri dish.
— Then the cylinder or centrifuge tube isrins ed with about
10 ml of tap-water which hasto be added to the s ample in
the larval counting basin or Petri dish. Subsequently, the
sample is examined by trichinoscope or stereomicroscope
respectively.
— Digests should be examined as soon as they are ready.
Under no circumstances should examination be postponed
until the following day.
If the digests are not examined within 30 minutes of their
preparation, they should be clarified as follows. The final
sample of about 40 ml is poured into a measuring cylinder
and allowed to stand for 10 minutes, after which time 30 mlof the supernatant fluid is removed leaving a volume of
10 ml. Thisvolume ismade up to 40 ml with tap-water.
After a further settling period of 10 minutes, 30 ml of the
supernatant fluid is withdrawn by suction leaving a volume
of 10 ml for examination in a Petri dish or larval counting1977L0096 — EN — 05.06.2003 — 009.001 — 14
▼M3
basin. The measuring cylinder should be washed with 10 ml
of tap-water and these washings should be added to the
sample in the Petri dish or the larval counting basin for
examination.
If the sediment is found to be unclear on examination, the
sample should be poured into a measuring cylinder and
made up to 40 ml with tap-water and then the above proce-
dure should be followed.
(ii)Pools of less than 100 samples
When needed, up to 15 samples of 1 g each may be added to a
total pool of 100 samples and examined together with these
samples according to (c) (1) (i). More than 15 samples should
be examined as a complete pool. For pools up to 50 samples,
the digestion fluid may be reduced to 1 litre.
2. In the case of a positive or doubtful result following the examina-
tion of a collective sample, a further 20 g sample should be taken
from each pig in accordance with (b) above. The 20 g samples from
five pigs should be pooled and examined by the method described
above. In thisway s amplesfrom 20 groupsof five pigswill be
examined. When trichinellae are detected in a pooled sample from
five pigs, further 20 g samples should be collected from the indivi-
dual pigs in the group and each should separately be examined
using the method described above.
VII. THE AUTOMATIC DIGESTION METHOD FOR POOLED SAMPLES
OF UP TO 35 g
(a)Apparatusandreagents
— Knife or scissors for cutting specimens,
— trays marked off with 50 squares each of which can hold samples
of approximately 2 g of meat,
— a Trichomatic 35 blender with filtration insert,— hydrochloric acid solution 8,5 % ± 0,5 weight,
— transparent polycarbonate membrane filters with a diameter of
50 mm and a pore size of 14 microns,
— pepsin strength 1: 10 000 NF (US National Formulary)
corresponding to 1: 125 000 BP (British Pharmacopoea)corresponding to 2 000 FIP (Federation Internationale de Phar-
macie),
— a balance, accurate to 0,1 g,
— tweezerswith a flat tip,
— a number of microscope slides with a side-length of at least 5 cm
or a number of at least 6 cm diameter Petri dishes marked on their
underside equipped into 10 × 10 mm large areas using a pointed
instrument,
— a (stereo-) microscope with transmitted light (magnification 15 to
60 times) or a trichinoscope with a horizontal table,
— a bin for collection of waste liquids,
— a number of 10 litre binsto be us ed when applying de-contamina-
tion, such as formol treatment, to the apparatus and for the
remaining digestive juice in the case of positive results.
(b)Collectionofspecimens
1. In the case of whole carcases a specimen to be taken of approxi-
mately 2 g from a pillar of the diaphragm at the transition to the
sinewy part. In the absence of diaphragm pillars, a specimen of the
same size to be taken from the rib part of the breastbone, part of the
diaphragm, from the jaw muscle or the abdomina muscle.
2. For cuts of meat, a sample of approximately 2 g of skeletal muscle
to be taken, containing little fat and, where possible, near to bones
or tendons.
(c)Method
1.Digestion Procedure
— Place the blender with filtration-insert, connect the waste tube
and lead the tube to the waste bin.1977L0096 — EN — 05.06.2003 — 009.001 — 15
▼M5
▼M5
— When the blender is switched on, the heat-up will start.
— Before start, the bottom valve, located below the reaction
chamber, should be opened and closed.
— Up to 35 samples of approximately 1 g each (at 25 to 30º C)
taken from each of the individual samples, in accordance with
point b, are then added. Make sure that larger pieces of tendons
are removed asthismay clot the membrane filter.
— Pour water to the edge of a liquid chamber connected to the
blender (approximately 400 ml).
— Pour about 30 ml hydrochloric acid (8,5 %) to the edge of the
smaller, connected liquid chamber.
— Place a membrane filter under the coarse filter in the filter
holder in the filter insert.
—►M67g◄of pepsin is added last. The order of addition
should be strictly adhered to in order to avoid decomposition
of the pepsin.
— Close the lids to the reaction- and liquid chambers.
— Select the period of digestion. Short digestion period (5
minutes) for pigs at normal age of slaughtering and extended
digestion time (8 minutes) for other samples.
— The automatic dispensing starts when the start button on the
blender isactivated and diges tion with following filtration will
proceed automatically. After 10 to 13 minutesthe proces sis
completed and stops automatically.
— The lid to the reaction chamber isopened once it ischecked that
the chamber isemptied. If there isfoam or remainsof diges tion
liquid in the chamber repeat the procedure according to point 4.
2.Recovery of larvae
— Dismount the filter holder and transfer the membrane filter to a
slide or a Petri dish.
— The membrane filter isexamined by meansof a micros cope or a
trichinoscope.
3.Cleaning of equipment
— In the case of a positive result, fill the reaction chamber in the
blender /twothirdwith boiling water. Ordinary tap-water ispoured into
the connecting liquid chamber until the lower level sensor is
covered. The automatic cleaning programme isthen carried
out. De-contaminate the filter-holder together with the
remaining equipment, for example by meansof formal treat-
ment.
— After the day'swork fill the liquid chamber in the blender with
water and carry out a standard programme.
4.Use of Membrane filters
Each polycarbonate membrane filter may be used no more than five
times. The filter should be turned between each use. In addition the
filter should be checked after each use for any damage which would
make it unsuitable for further use.
►M65.◄Method to be used when digestion is incomplete and
filtration cannot therefore be carried out
When the automatic process in the blender is carried out according
to point 1, open the lid to the reaction chamber and check whether
there isfoam or liquid remaining in the chamber. It thisisthe cas e,
carry out the following procedure:
— Close the bottom valve below the reaction chamber.
— Dismount the filter holder and transfer the membrane filter to a
slide or a Petri dish.
— Put a new membrane filter in the filter holder and mount the
filter holder.
— Fill water into the liquid chamber in the blender until the lower
level-sensor is covered.
— Carry out the automatic cleaning programme.
— After the cleaning programme hasbeen completed open the lid
to the reaction chamber and check for liquid remains.
— If the chamber is empty, dismount the filter holder and transfer
the membrane filter with a tweezer to a slide or a Petri dish.1977L0096 — EN — 05.06.2003 — 009.001 — 16
▼M6
▼M5
▼M5
— The two membrane filtersare examined according to point 2. If
the filterscannot be examined repeat the entire diges tion
process with extended digestion time according to point 1.
►M66.◄In the case of a positive or doubtful result, following the
result of a collective sample, a further 20 g sample should be taken
from each pig in accordance with point b above. These samples
shall be investigated individually according to the abovementioned
method.1977L0096 — EN — 05.06.2003 — 009.001 — 17
▼B
ANNEX II
CHAPTER I
CONDITIONSFOR THEAUTHORIZATIONOF TRICHINAE(TRICHI-
NELLASPIRALIS)LABORATORIES
1. Trichinae laboratories must be situated next to the swine slaughterhouses
and, if the establishment does not already possess such facilities to fulfil the
other requirements of the Directive on third countries, have at least the
following facilitiesavailable:
(a) a lockable room adequately equipped for the preparation of specimens;
its walls must be smooth and must be lined with a washable light-
coloured covering or paint up to a height of 2 m. A preparation room
must be provided for each method of examination used;
(b) an adequately equipped lockable examination room which can be
darkened when examination isperformed by trichinos cope;
(c) equipment providing adequate ventilation and, if necessary, air condi-
tioning equipment which ensures that the room temperature does not
exceed + 25 ºC;
(d) adequate natural or artificial lighting which doesnot alter colours ; direct
sunlight must be avoided;
(e) in the preparation room, adequate equipment for cleaning and disin-
fecting hands;
►M3(f)◄possibly, refrigerators in which to store meat samples;
(g) a washroom for cleansing and disinfecting examination equipment (e.g.
containers for samples, compressors, knives and scissors), with:
— a waterproof floor-covering which isrot-proof and eas y to clean and
disinfect,
— smooth walls which, up to a height of at least 2 m, are lined by a
washable and light-coloured covering or paint.
This provision need not be applied when using the methods indicated
under pointsII, III, IV, V, VI of Annex I provided the laboratoriesare
equipped with a large suitably plumbed sink;
►M3(h)◄changing rooms, wash basins and rest rooms and flush lava-
tories;
►M3(i)◄wash basins with hot and cold running water, provided with
cleansing and disinfecting materials and disposable towels;
►M3(j)◄watertight corrosion-resistant containers, with hermetically
sealed lids, for collecting the samples after examination, and so designed
asto prevent unauthorized removal of the contents ;
►M3(k)◄adequate supplies of hot and cold drinking water;
►M3(l)◄equipment for removing waste water which fulfils the condi-
tions for the authorization of slaughterhouses;
►M3(m)◄proper equipment for protection against pests (insects,
rodents, etc.).
CHAPTER II
REQUIREMENTS APPLICABLE TO STAFF, PREMISES, EQUIPMENT
ANDINSTRUMENTSINTRICHINAELABORATORIES
2. Absolute cleanliness is required at all times of laboratory staff, premises,
equipment and instruments:
(a) staff must wear clean working clothes and wash their hands several times
during working hoursand after each break;
(b) no animal may enter trichinae laboratories;
(c) equipment and instruments used must be kept clean and in good repair.
They must be carefully cleansed and disinfected several times during and
at the end of the day'swork.
3. Drinking water must be used for all purposes.1977L0096 — EN — 05.06.2003 — 009.001 — 18
▼M3
▼B
▼M3
▼B
▼M3
▼B
▼B
4. As regards health, staff taking meat samples for examination must comply
with the rulesin Annex B, Chapter IV (11) and (12) of Directive 72/462/
EEC.
5. The meat samples required for examination must be taken immediately after
slaughter and examined without delay in the trichinae laboratory of the
slaughterhouse.
These examinations are forbidden away from the slaughterhouse in which the
animalswere s laughtered.
6. To avoid fatigue and its consequences, inspection staff should be given short
breaksduring the working day.
CHAPTER III
REQUIREMENTSINRESPECTOFTRICHINOSCOPES
The construction and design of trichinoscopes must meet the following
minimum criteria:
1. Simple operation.2. High light intensity:
— accurate results must be obtainable even in a room which is not comple-
tely dark,
— a projector bulb of 100 W (12 V) must be used as the light source.
3. Adequate magnification:
— normal working magnification: 50 x,
— 80 to 100 x magnification for more precise assessment of objects not
clearly identifiable under normal working magnification.
4. Resolving power:
— a clear sharp picture of well-defined colour must be obtainable at each
magnification.
5. Switch mechanism:
— each change of magnification must be accompanied by automatic adjust-
ment of the brightness of the image.
6. Increase of contrast:
— the condensor must be provided with an iris diaphragm enabling the
contrast to be increased for the closer inspection of difficult cases,
— the iris diaphragm must be easy to operate (e.g. control lever on the
platform of the trichinoscope).
7. Easy focusing:
— rapid focusing by means of an adjusting ring,
— precise focusing by means of a control lever.
8. Voltage regulation:
— so that brightness may be adjusted as required.
9. One-way movement of the compressor:
— an automatic blocking mechanism must ensure that the compressor
movesin only one direction, to prevent unintentional dis placement.
10. Free view of the projector screen.
11. Projector screen:
— at least 54 cm in diameter,
— high reflecting capability,— durable,— can be dismantled,
— easy to clean.1977L0096 — EN — 05.06.2003 — 009.001 — 19
▼B
ANNEX III
MARKING OF MEAT WHICH HAS BEEN EXAMINEDFOR
TRICHINAE
1. Marking of the meat must be carried out under the responsibility of the offi-
cial veterinarian. For this purpose, he shall keep and maintain:
— the instruments intended for marking which he may hand over to the
assistant staff only at the time of marking and for the length of time
required for thispurpos e,
— the tags mentioned in paragraph 5. These tags shall be given to the assis-
tant staff at the time when they must be used and in the required number.
2. The mark must be round with a diameter of 2·5 cm. The following informa-
tion must appear on the mark in perfectly legible characters:
— towardsthe centre the capital letter ‘T’ with arms1 cm long and 0·2 cm
wide,
— under the letter ‘T’ one of the following sets of initials: CEE, EEG,
EWG, EØF, EEC ►A1, EOK◄or►A2ETY◄. The lettersmus t
be 0·4 cm high.
3. Carcases must be marked in ink or hot-branded on the inside of the thighs, in
accordance with paragraph 2.
4. Headsmus t be marked in ink or hot-branded, with a mark meeting the
requirementsof paragraph 2.
With the exception of cutsexempt from health marking by virtue of Annex
B, Chapter X (43) of Council Directive 72/462/EEC, those taken in cutting
plants from carcases marked in accordance with the rules must, where they
bear no stamp, be marked in accordance with paragraph 2 before the health
mark isaffixed.
The label provided for in the second subparagraph of the abovementioned
paragraph 43 must comply with the conditions of paragraph 6 below.
5. Marking may also be effected by means of a round tag. This tag, to be
affixed to each piece or to each carcase must not be reusable, must be made
of strong materials and must meet all hygiene requirements. The following
information must appear on the stamp seals in perfectly legible characters:
— towardsthe centre the capital letter ‘T’,
— under the letter ‘T’ one of the following sets of initials: CEE, EEG,
EWG, EØF, EEC ►A1, EOK◄or►A2ETY◄. The lettersmus t
be 0·2 cm high.
6. The label provided for in Annex B, Chapter X (44) of the Directive
mentioned in paragraph 4 above must, in addition to the health mark, bear a
clearly legible mark identical to that provided for in paragraph 2.1977L0096 — EN — 05.06.2003 — 009.001 — 20
▼B
ANNEX IV
THEFREEZINGOFMEAT
I.Method1
1. Meat brought in already frozen must be kept in this condition.
2. The technical equipment and energy supply of the refrigerating room,
must be such as to ensure that the temperature referred to under para-
graph 6 isreached very rapidly and maintained in all partsof the room
and of the meat.
3. Insulated packaging should be removed before freezing, except for meat
which hasalready reached throughout the temperature referred to under
paragraph 6 when it isbrought into the refrigeration room.
4. Consignments in the refrigeration room must be kept separately and
under lock.
5. The date and time when each consignment is brought into the refrigera-
tion room must be recorded.
6. The temperature in the refrigeration room must be at least − 25 ºC. It
should be measured with calibrated thermo-electric instruments and
continuously recorded. It may not be measured directly in the cold air
flow. The instruments must be kept under lock. The charts must include
the relevant numbers from the meat inspection register on importation
and the date and time of the commencement and completion of freezing,
and must be retained for one year after compilation.
7. Meat with a diameter or thickness of up to 25 cm must be frozen for at
least 240 consecutive hours, and meat with a diameter or thickness of
between 25 and 50 cm must be frozen for at least 480 consecutive hours.
Thisfreezing proces smay not be applied to meat which hasa larger
diameter or isthicker. The freezing time s hall be calculated from the
point when the temperature referred to in paragraph 6 isreached in the
freezing room.
II.Method2
The general provisions of paragraphs 1 to 5 of method 1 shall be
complied with, and the following time-temperature combinations
applied:
1. Meat with a diameter or thickness of up to 15 cm must be frozen
according to one of the following time-temperature combinations:
— 20 daysat − 15 ºC,
— 10 daysat − 23 ºC,— 6 daysat − 29 ºC.
2. Meat with a diameter or thickness of between 15 cm and 50 cm must be
frozen according to one of the following time-temperature combinations:
— 30 daysat − 15 ºC,
— 20 daysat − 25 ºC,— 12 daysat − 29 ºC.
The temperature in the refrigeration room must be no higher than the
level of the selected inactivation temperature. It should be measured
with calibrated thermoelectric instruments and continuously recorded. It
may not be measured directly in the cold air flow. The instruments must
be kept under lock. The chartsmus t include the relevant numbersfrom
the meat inspection register on importation and the date and time of the
commencement and completion of freezing, and must be retained for one
year after compilation.
III.Method3
Control of the temperature at the center of the meat pieces.
1. The following time-temperature combinationss hall be applied where the
temperature iscontrolled at the center of the meat piecesand the condi-
tionsin paragraphs2 to 6 are fulfilled:
— 106 hoursat − 18 ºC,1977L0096 — EN — 05.06.2003 — 009.001 — 21
▼M6
▼B
▼M6
▼M6
— 82 hoursat − 21 ºC,
— 63 hoursat − 23,5 ºC,
— 48 hoursat − 26 ºC,
— 35 hoursat − 29 ºC,— 22 hoursat − 32 ºC,— 8 hoursat − 35 ºC,— ½ hour at − 37 ºC.
2. Meat brought in already frozen must be kept in this condition.
3. Consignments in the refrigeration room must be kept separately and
under lock.
4. The date and time when each consignment is brought into the refrigera-
tion room must be recorded.
5. The technical equipment and energy supply of the refrigeration room
must be such as to ensure that the temperature referred to under para-
graph 1 isreached very rapidly and maintained in all partsof the meat.
6. The temperature should be measured with calibrated thermoelectric
instruments and continuously recorded. The probe of the thermometer
hasto be placed at the center of a calibrated piece of meat of a s ize no
smaller than the thickest piece of meat to be frozen. This calibrated piece
of meat should be placed at the least favorable site in the freezing room,
not close to the cooling equipment or directly in the cold air flow. The
instruments must be kept under lock. The charts must include the rele-
vant numbers from the meat inspection register on importation and the
date and time of the commencement and completion of freezing, and
must be retained for one year after compilation.1977L0096 — EN — 05.06.2003 — 009.001 — 22
▼M6
ANNEX V
INSPECTIONANDFREEZINGOFHORSEMEAT
1.Inspection
The inspection of horse meat has to be performed according to a digestion
method mentioned in Annex I with the following modifications:
— Specimens of at least 10 g to be taken from the lingual muscle or the jaw
muscle. In the absence of lingual muscle or jaw muscle a specimen of the
same size to be taken from a pillar of the diaphragm at the transition to
the sinewy part. The muscle should be clean of connective tissue and fat.
— A 5 g sample is digested for inspection if the artificial digestion of
collective samples according to Annex I, Nr. III to VII is applied. For
each digest, the total weight of muscle under examination must not
exceed 100 g for methodsIII, IV, V and VI in Annex I or 35 g for
method VII in Annex I.
— In the case of a positive result a further 10 g specimen must be taken for
a subsequent independent examination.
2.Freezing ofhorsemeat
To kill trichinae by freezing, horse meat must undergo cold treatment in
accordance with one of the methodsdes cribed in Annex IV.1977L0096 — EN — 05.06.2003 — 009.001 — 23