# Commission Implementing Regulation (EU) 2015/1375 of 10 August 2015 laying down specific rules on official controls for Trichinella in meat (Codification) (Text with EEA relevance), Division I: Complete pools (100 g of samples at a time)

Extent: E+W+S+N.I.

(a)16 ± 0,5 ml of hydrochloric acid is added to a 3 litre beaker containing 2,0 litre of tap water, preheated to 46 to 48 °C; a stirring rod is placed in the beaker, the beaker is placed on the preheated plate and the stirring is started.
(b)10 ± 0,2 g of pepsin or 30 ± 0,5 ml liquid pepsin is added.
(c)100 g of samples collected in accordance with point 2 is chopped in the blender.
(d)The chopped meat is transferred to the 3 litre beaker containing the water, pepsin and hydrochloric acid.
(e)The mincing insert of the blender is immersed repeatedly in the digestion fluid in the beaker and the blender bowl is rinsed with a small quantity of digestion fluid to remove any meat still adhering.
(f)The beaker is covered with aluminium foil.
(g)The magnetic stirrer must be adjusted so that it maintains a constant temperature of 44 to 46 °C throughout the operation. During stirring, the digestion fluid must rotate at a sufficiently high speed to create a deep whirl without splashing.
(h)The digestion fluid is stirred until the meat particles disappear (approximately 30 minutes). The stirrer is then switched off and the digestion fluid is poured through the sieve into the sedimentation funnel. Longer digestion times may be necessary (not exceeding 60 minutes) in the processing of certain types of meat (tongue, game meat, etc.).
(i)The digestion process is considered satisfactory if not more than 5 % of the starting sample weight remains on the sieve.
(j)The digestion fluid is allowed to stand in the funnel for 30 minutes.
(k)After 30 minutes, a 40 ml sample of digestion fluid is quickly run off into the measuring cylinder or centrifuge tube.
(l)The digestion fluids and other liquid waste are kept in a tray until reading of the results is completed.
(m)The 40 ml sample is allowed to stand for 10 minutes. 30 ml of supernatant is then carefully withdrawn by suction to remove the upper layers and leave a volume of not more than 10 ml.
(n)The remaining 10 ml sample of sediment is poured into a larval counting basin or petri dish.
(o)The cylinder or centrifuge tube is rinsed with not more than 10 ml of tap water, which has to be added to the sample in the larval counting basin or petri dish. Subsequently, the sample is examined by trichinoscope or stereo-microscope at a 15 to 20 times magnification. Visualisation using other techniques is allowed, provided examination of positive control samples has been shown to give an equal or better result than traditional visualisation methods. In all cases of suspect areas or parasite-like shapes, higher magnifications of 60 to 100 times must be used.
(p)Digests are to be examined as soon as they are ready. Under no circumstances should examination be postponed until the following day.
Where the digests are not examined within 30 minutes of preparation, they must be clarified as follows. The final sample of about 40 ml is poured into a measuring cylinder and allowed to stand for 10 minutes. 30 ml of the supernatant fluid is then removed, leaving a volume of 10 ml. This volume is made 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 no more than 10 ml for examination in a petri dish or larval counting basin. The measuring cylinder is washed with no more than 10 ml of tap water and these washings are 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 is poured into a measuring cylinder and made up to 40 ml with tap water and then the procedure described in this Section is followed. The procedure can be repeated 2 to 4 times until the fluid is clear enough for a reliable reading.

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Source: https://www.legislation.gov.uk/eur/2015/1375/annex/I/chapter/I/division/1/division/3/division/I (the official version)
Page: https://openlegislation.uk/eu-regulation-2015-1375/annex-I/chapter-I/division-1/division-3/division-I
Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives.