EN   EN 
 
 
 EUROPEAN  
COMMISSION   
Brussels, XXX   
SANTE/11094/2021 ANNEX CIS 
(POOL/G4/2021/11094/11094 -EN 
ANNEX CIS.docx)  
[…](2021) XXX  draft  
ANNEX  
 
ANNEX  
to the  
Commission Implementing Regulation (EU) .../...  
amending Annex VI to Commission Regulation (EC) No 152/2009 as regards the method 
of analysis for the detection of terrestrial invertebrate constituents for the official 
control of feed   EN 1  EN ANNEX  
 
Annex VI to Regulation (EC) No 152/2009 is amended as follows:  
 
(1) point 1 is replaced by the following:   
‘1. PURPOSE AND SCOPE  
The determination  of constituents of animal origin in feed shall be performed by light 
microscopy or polymerase chain reaction (PCR) in accordance with the provisions laid down 
in this Annex.  
These two methods make it possible to detect the presence of constituents of anim al origin in 
premixtures , feed materials and compound feed. However, they do not make it possible to 
calculate the amount of such constituents in premixtures, feed materials and compound feed. 
Both methods have a limit of detection below 0 ,1 % (w/w).  
The P CR method makes it possible to identify the taxonomic group of constituents of animal 
origin present in premixtures, feed materials and compound feed.  
These methods shall apply for the control of the application of the prohibitions laid down in 
Article 7(1 ) of Regulation (EC) No 999/2001 of the European Parliament and of the Council*, 
Annex IV to that Regulation and Article  11(1) of Regulation (EC) No 1069/2009  of the 
European Parliament and of the Council**. 
Depending on the type of feed being tested, these methods may be used, within one single 
operational protocol, either on their own or combined together in accordance with the 
standard operating procedures ( ‘SOPs ’) established by the EU reference laboratory for animal 
proteins in feedingstuffs (EURL -AP) and published on its website***.  
______________  
* Regulation (EC) No 999/2001  of the European Parliament and of the Council of 
22 May 2001 laying down rules for the prevention, control and eradication of certain 
transmissible spongiform encephalopathies (OJ L 147 31.5.2001, p. 1).  
** Regulation (EC) No 1069/2009 of the European Par liament and of the Council of 
21 October  2009 laying down health rules as regards animal by -products and derived 
products not intended for human consumption and repealing Regulation (EC) No 
1774/2002 (Animal by -products Regulation) (OJ L 300 14.11.2009, p.  1). 
*** https://www.eurl.craw.eu/legal -sources -and-sops/method -of-reference -and-sops/  .’; 
 
(2) point 2.1 is replaced by the following : 
‘2.1. Light microscopy  
2.1.1.  Principle  
The constituents of animal origin which may be present in premixtures, feed 
materials and compound feed  sent for analysis are identified on the basis of typical 
and microscopically identifiable characteristics like muscle fibres and other meat 
particles, cartilage, bones, horn, hair, bristles, invertebrates cuticular fragments, 
insect tracheal structures, blood, milk globules, lactose crystals, feathers, egg shells, 
fish bones and scales.  EN 2  EN Microscopic examinations shall be performed after preparati on of samples  by 
sedimentation . 
Sample s shall be subject to a sedimentation s tep as follows :  
(a) for the detection of constituents of animal origin other than terrestrial invertebrates , a 
single Tetrachloroethylene ( TCE ) sedimentation step as detailed in point 2.1.3.4.3 ;   
(b) for the detection of constituents of terrestrial invertebrate s, a double Petroleum ether 
/ Tetrachloroethylene ( PE/TCE ) sedimentation step as detailed in point 2.1.3.4.4.  
2.1.2.  Reagents and equipment  
2.1.2.1 . Reagents  
 2.1.2.1.1.  Concen trating agent   
– Tetrachloroethylene  (specific gravity 1 ,62) 
– Petroleum ether  (PE) boiling point 40 -60 °C (specific gravity 0 ,65) 
 2.1.2.1.2.  Staining reagent  
– Alizarin Red solution (dilute 2 ,5 ml 1M hydrochloric acid in 100 
ml water and add 200 mg Alizarin Re d to this solution) . 
 2.1.2.1.3.  Mounting media  
– Lye (NaOH 2 ,5 % w/v or KOH 2 ,5 % w/v) . 
– Glycerol (undiluted, viscosity: 1490 cP) or a mounting medium 
with equivalent properties for non -permanent slide preparation . 
– Norland ® Optical Adhesive 65 (viscosity: 1200 cP) or a resin with 
equivalent properties for permanent slide preparation . 
 2.1.2.1.4.  Mounting media with staining properties  
– Lugol solution (dissolve 2 g potassium iodide in 100 ml water and 
add 1  g iodine while frequently shaking) . 
– Cystine reagent (2 g lead acetate, 10 g NaOH/100 ml water) . 
– Fehling’s reagent (prepared before use from equal parts (1/1) of 
two-stock solutions A and B : solution A (dissolve 6 ,9 g copper (II) 
sulphate pentahydrate in 100 ml water ); solution B (dissolve 34 ,6 g 
potassium s odium tartrate tetrahydrate and 12 g NaOH in 100 ml 
water) . 
– Tetramethylbenzidine/Hydrogen peroxide (dissolve 1 g 3,3’,5,5’ 
tetramethylbenzidine (TMB) in 100 ml glacial acetic acid and 150 
ml water. Before use, mix 4 parts of this TMB solution with 1 part 
3% hydrogen peroxide) . 
 2.1.2.1.5.  Rinsing agents  
– Ethanol ≥ 96 % (technical grade) . 
– Acetone (technical grade) . 
 2.1.2.1.6.  Bleaching reagent  
– Commercial sodium hypochlorite solution (9 - 14 % active 
chlorine) . EN 3  EN 2.1.2.2. Equipment  
– Analytica l balance with an ac curacy of 0, 001 g . 
– Grinding equipment: knife or rotor mill. If a rotor mill is used, mill sieves ≤ 0 ,5 
mm shall be prohibited . 
– Sieves with square meshes of 0, 25 mm and 1 mm width. With the exception of 
sample pre -sieving, the diameter of the sieves shall  not exceed 10 cm to avoid 
loss of materials. Calibration of sieves is not required . 
– Conical glass separation funnel with a content of 250 ml with Teflon or ground 
glass stopcock at the base of the cone. Stopcock opening diameter shall be 
≥ 4mm. Alternativel y, for single TCE sedimentation only, a conical bottomed 
settling beaker may be used provided the laboratory has demonstrated that 
detection levels are equivalent to that obtained using the conical glass 
separation funnel . 
 
 
Separation funnel  
 
– Stereomicroscope covering at least a 6 ,5x to 40x final magnification range . 
– Compound microscope covering at least a 100x to 400x final magnification 
range with transmitted light bright field. Polarised light, differential 
interferential contrast can additionally be used . 
– Standard laboratory glassware.  
– Equipment for slide preparation: classical microscope slides, hollow slides, 
coverslips (20x20 mm), tweezers, fine spatula . 
– Laboratory oven . 
– Centrifuge.  
– Filter paper: qualitative cellu lose filter (pore s ize 4 -11 µm).  
 
2.1.3. Sampling and sample preparation  
2.1.3.1.  Sampling  
A representative sample, taken in accordance with Annex  I shall be used.  
2.1.3.1.1.  Sample drying  
EN 4  EN Samples with a moisture content > 14 % shall be dried prior to handling 
in accordance with Annex III.  
2.1.3.1.2.  Sample pre -sieving  
In order to collect information on possible environmental contamination 
of the feed, it is recommended to pre -sieve at 1 mm pelleted feeds and 
kernels and to subsequently prepare, analyse, and report separately  on the 
two resulting fractions, which must be considered as distinct samples.  
2.1.3.2.  Precautions to be taken  
In order to avoid laboratory cross -contamination, all reusable equipment shall be 
carefully cleaned before use. Separation funnel pieces shall b e disassembled before 
cleaning. Separation funnel pieces and glassware shall be pre -washed manually and 
then washed in a washing machine. Sieves shall be cleaned by using a brush with 
stiff synthetic hairs. A final cleaning of sieves with acetone and compr essed air is 
recommended after sieving of fatty material like fishmeal.  
2.1.3.3.  Preparation of samples consisting of fat or oil  
The following protocol shall be followed for the preparation of samples consisting of 
fat: 
– If the fat is solid, it shall be war med in a n oven until it is liquid.  
– By using a pipette, 40 ml of fat or oil shall be transferred from the bottom of 
the sample to a centrifugation tube.  
– The sample shall be centrifuge d during 10 min at 4000 r.p.m. . 
– If the fat is solid after centrifugation, it shall be warme d in an oven until it is 
liquid.  
– The centrifugation  shall be repeated  during 5 min at 4000 r.p.m.  
– By using a small spoon or a spatula, one half of the decanted impurities shall 
be transferred to microscopic slides for examination. Glycerol  is recommended 
as mounting medium . 
– The remaining impurities shall be used for preparing the sediment as described 
in the point 2.1.3.4.3 , first indent . 
The same protocol, with the exception of the first and fourth indent s, shall be applied 
for the prepara tion of samples consisting of oil.  
2.1.3.4.  Preparation of samples other than fat or oil  
2.1.3.4.1.  Sub-sampling and grinding : at least 50 g of the sample shall be sub -
sampled for analysis and subsequently ground.  
2.1.3.4.2.  Preparation of raw material : a portion of at least 5 g of the ground sub -
sample shall be pr epared. It shall be sieved at 0, 25 mm and the two 
resulting fractions shall be examined.  
2.1.3.4.3.  Single TCE sedimentation  for the detection of constituents  of animal 
origin other than terrestr ial invertebrates . 
– Extraction and preparation of the sediment:  
A portion of 10 g (accurate to 0, 01 g) of the ground sub -sample 
shall be transferred into the separation funnel or conical bottomed EN 5  EN settling beaker and 50 ml of TCE shall be added. The portion  
transferred into the funnel shall be limited to 3 g in case of fishmeal 
or other pure animal products, mineral ingredients or premix tures 
which generate more than 10  % of sediment. The mixture shall be 
vigorously shaken for at least 30 s and at least 50 m l more of TCE 
shall be added cautiously while washing down the inside surface of 
the funnel to remove any adhering particles. The resulting mixture 
shall be left to stand for at least 5 min before the sediment is 
separated off by opening the stopcock.  
If a conical bottomed settling beaker is used then the mixture shall 
be vigorously stirred for at least 15 s and any particles adhering to 
the side of the beaker shall be carefully washed down the inside 
surface with at least 10 ml of clean TCE . The mixture sh all be left 
to stand for 3 min and then stirred again for 15 s and any particles 
adhering to the side of the beaker shall be carefully washed down 
the inside surface with at least 10 ml of clean TCE . The resulting 
mixture shall be left to stand for at leas t 5 min and then the liquid 
fraction is removed and discarded by careful decanting, taking care 
not to lose any of the sediment.  
The sediment shall be collected on a filter paper placed into a 
funnel to allow the separation of the remaining TCE while avoid ing 
fat deposition into the sediment. The sediment shall be dried. It is 
recommended to subsequently weigh the sediment (accurate to 
0,001 g) to control the sedimentation step. Lastly, the sediment 
shall be sieved at 0 ,25 mm and the two resulting fractions  shall be 
examined, unless sieving is not deemed necessary.  
– Extraction and preparation of the flotate:  
After recovery of the sediment with the method described above, 
two phases shall  remain in the separation funnel: a liquid one 
consisting of TCE and a s olid one made of floating material. This 
solid phase is the flotate and shall be recovered by pouring off 
completely TCE from the funnel by opening the stopcock. By 
inverting the separation funnel, the flotate shall be transferred into 
a large petri dish a nd air dried in a fu mehood. It shall be sieved at 
0,25 mm and the two resulting fractions shall be examined.  
– Use of staining  reagents  
In order to facilitate the correct identification of the constituents of 
animal origin, the operator may use staining reag ents during the 
sample preparation in accordance with guidelines issued by the 
EURL -AP and published on its website.  
In case Alizarin Red solution is used to colour the sediment, the 
following protocol shall apply:  
- The dried sediment shall be transferred i nto a glass test tube 
and rinsed twice with approximately 5 ml of ethanol (each time 
a vortex of 30 s shall be used, the solvent shall be let settle 
about 1 min 30 s and poured off).  EN 6  EN - the sediment shall be bleached by adding at least 1 ml sodium 
hypochlorite solution. The reaction shall be allowed to continue 
for 10 min. The tube shall be filled with water, the sediment 
shall be let settle 2 -3 min, and the water and the suspended 
particles shall be poured off gently.  
- the sediment shall be rinsed t wice more with about 10 ml of 
water (a vortex shall be used for 30 s, let settle, an d pour off the 
water each time).  
- 2 to 10 drops of the Alizarin Red solution shall be added and 
the mixture shall be vortexed. The reaction shall be let occur for 
30 s and t he coloured sediment shall be rinsed twice with 
approximately 5 ml ethanol followed by one rinse with acetone 
(each time a vortex of 30 s shall be used, the solvent shall be let 
settle about 1 min and poured off).  
- the coloured sediment shall be dried.  
2.1.3.4.4.   Double PE/TCE sedimentation  for the detection of terrestrial invertebrate 
constituents.   
All steps shall be realised in a conical glass separation funnel of 250 ml 
as described in point 2.1.2.2 , fourth indent .  
– A portion of 10 g (accurate to 0, 01 g ) of the ground sub -sample 
shall be transferred into the separation funnel and submitted first to 
a single TCE sedimentation as described in point 2.1.3.4.3 
including the recovery of the sediment on a filter paper placed on a 
funnel. This sediment may be u sed as the one  obtained from point 
2.1.3.4.3.  
– The small volume of TCE drained together with the sediment shall 
be transferred into a graduated cylinder. By opening the stopcock of 
the separation funnel the graduated cylinder has to be filled further 
until obtaining 30 ml of TCE. Once this volume is achieved, the 
stopcock shall be closed.  
– This collected volume of TCE shall be substituted by adding a 
volume  of 30 ml of petroleum ether  boiling point 40-60 °C into the 
separation funnel. The content of the separ ation funnel shall be 
mixed thoroughly to obtain a 30 % PE/70 % TCE mixture (with a 
density of approximately 1 ,26 g.cm-3). Allow the material to settle 
down for 10 min. Two new fractions will segregate: a second 
sediment and a final flotate (< 1, 26 g.cm-3). The second sediment is 
to recover in a petri dish (or a filter paper placed on a funnel) by 
opening the stopcock until only a few solvent mixture and the final 
flotate remain in the sedimentation funnel. The remaining liquid 
and the final flotate  shall be collected separately on a filter paper 
placed on a funnel . The wall of the sedimentation funnel shall be 
rinced with a flush of PE to collect all material from the final 
flotate. The final flotate should be allowed to dry. The final fl otate 
shall  be sieved at 0, 25 mm and the two resulting fractions shall be 
examined for the detection of terrestrial invertebrate constituents, 
unless sieving is not deemed necessary.  EN 7  EN 2.1.4.  Microscopic examination  
2.1.4.1. Slide preparation  
Microscopic slides shall be prepared from the sediment and, depending on operator's 
choice, from either the flotate or the raw material. When appropriate , for the 
detection of terrestrial invertebrate constituents only, slides shall also be prepared 
from the final flotate obtained  as described in point 2.1.3.4.4. The two resulting 
fractions (the fine and the coarse one) shall be prepared. Test portions of fractions 
spread on slides shall be representative of the whole fraction.  
A sufficient number of slides shall be prepared in ord er to ensure that a complete 
examination protocol as laid down in point 2.1.4.2 can be carried -out. 
Microscopic slides shall be mounted with the adequate mounting medium in 
accordance with the SOP established by the EURL -AP and published on its website. 
The slides shall be covered with coverslips.  
2.1.4.2. Observation flowchart for the detection of animal particles in compound feed, feed 
material and premix tures . 
The prepared microscopic slides shall be observed in accordance with the 
obser vation flowcharts  in Diagrams 1 and 2.  
The microscopic observations shall be conducted using the compound microscope on 
the sediment and, depending on the operator’s choice, either on the flotate or on the 
raw material. Additionally, for the detection of terrestrial invert ebrate constituents, 
observations shall also be conducted on the final flotate obtained as described in 
point 2.1.3.4.4 in accordance with D iagram 3. The stereomicroscope may be used in 
addition to the compound microscope for the coarse fractions. Each sli de shall be 
screened entirely at various magnifications. Precise explanations on how to use the 
flowcharts are detailed by a SOP established by the EURL -AP and published on its 
website.  
The minimum numbers of slides to be observed at each step of the obser vation 
flowcharts shall be strictly respected unless the entire fraction material does not 
permit to reach the stipulated slide number, for instance when no sediment is 
obtained. No more than 6 slides per determination shall be used for recording of the 
number of particles.  
When additional slides are prepared using a more specific mounting medium with 
staining properties, as described  in point 2.1.2.1.4, on the flotate or the raw material 
to further characterise structures (e.g. feathers, hairs, muscle or blood particles) , 
which have been detected on slides prepared by other mounting media, as described  
in point 2.1.2.1.3, the number of particles shall be counted based on a number of 
slides per determination  not exceeding 6, including the additional slides with a more 
specific mounting medium. The additional slides prepared from the final flotate 
obtained , as described in point 2.1.3.4.4 , for the detection of terrestrial invertebrate 
constituents shall not be considered for the identification of other nature s (terrestrial 
vertebrates and fish).  
In order to facilitate the identification of the particles' nature and origin, the operator 
may use support tools like decision support systems, image libraries and reference 
samples.  
Diagram 1  EN 8  EN Observation flowchart after single TCE sedimentation for the detection of 
animal particles other than from terrestrial invertebrates in compound feed, 
feed material and premix tures for the first determination  
 
 
 
Diagram 2  
Observation flowchart after single TCE sedimentation for the detection of 
animal particles other than from terrestrial invertebrates in compound feed, 
feed material and premix tures for the second determination  
EN 9  EN  
 
EN 10  EN Diagram 3  
Observation flowchart after double PE/TCE sedimentation for the detection of 
terrestr ial invertebrates’ constituents in compound feed, feed material and 
premix tures 
 
 
 
 
2.1.4.3.  Number of determinations  
Determinations shall be performed on different sub -samples of 50 g each.  
If, following the first determination carried out in accordance with the obse rvation 
flowchart in Diagram 1, or D iagram 3 when appropriate, no animal particles are 
detected, no additional determination shall be  necessary and the result of the analysis 
shall be reported using the wording  set out  in point 2.1.5.1.  
If, following the first determination carried out in accordance with the obse rvation 
flowchart in D iagram 1, one or more animal particles of a given nature (i.e. terrestrial 
vertebrate s or fish) are detected and the nature of the particles found confirms the 
declared content of the sample, no second determination shall be  necessary. If the 
number of the animal particles of a given nature detected during this first 
determination is higher than 5, the result of the analysis shall be reported per animal 
nature usi ng the wording  set out  in point 2.1.5.3. Otherwise, the result of the analysis 
shall be reported per animal nature using the wording  set out  in point 2.1.5.2.  
If, following the first determination carried out in accordance with the observation 
flowchart in  Diagram 3, more than 5 particles of terrestrial invertebrates are detected, 
no second determination shall be  necessary and the result of the analysis shall be 
reported using the wording  set out  in point 2.1.5.3 for this nature.  
EN 11  EN In all other cases, including when no declaration of content has been provided to the 
laboratory, a second determination shall be carried out from a new sub -sample. If, 
following the second determination carried out in accordance with the observation 
flowc hart in Diagram 2, or Diagram 3 when appropriate, the sum of the animal 
particles of a given nature detected over the two determinations is higher than 10, the 
result of the analysis shall be reported per animal nature using the wording  set out  in 
point 2. 1.5.3. Otherwise, the result of the analysis shall be reported per animal nature 
using the wording  set out  in point 2.1.5.2.  
2.1.5.  Expression of the results  
When reporting the results, the laboratory shall indicate on which type of material 
the analysis has been carried  out (sediment, flotate, final flotate or raw material). The 
reporting shall clearly indicate how many determinations have been carried  out and if 
sieving of the fractions prior to slide preparation, in accordance with point 2.1.3.4.3, 
first indent, third  paragraph, or point 2.1.3.4.4, third indent, was not performed.  
The laboratory report shall at least contain information on the presence of 
constituents derived from terrestrial vertebrates and from fish.  
The different situations shall be reported in the following ways.  
2.1.5.1.  No animal particle of a given nature is detected:  
– “As far as was discernible using a light microscope, no particle derived from 
terrestrial vertebrates was detected in the submitted sample. ” 
– “As far as was discernib le using a light microscope, no particle derived from 
fish was detected in the submitted sample. ” 
– “As far as was discernible using a light microscope, no particle derived from 
terrestrial invertebrates was detected in the submitted sample. ” 
2.1.5.2.  Between 1 and 5 animal particles of a given nature are detected when only one 
determination has been performed, or between 1 and 10 particles of a given nature 
are detected in case of two determinations (the number of detected particles is below 
the decisi on limit established in the SOP of the EURL -AP and published on its 
website):  
When only one determination has been performed:  
– “As far as was discernible using a light microscope, no more than 5 particles 
derived from terrestrial vertebrates were detected in the submitted sample. The 
particles were identified as … [bone, cartilage, muscle, hair, horn , other (please 
specify as appropriate) ]. This low level presence is below the decision limit 
established for this microscopic method. ” 
– “As far as was discernibl e using a light microscope, no more than 5 particles 
derived from fish were detected in the submitted sample. The particles were 
identified as … [fishbone, fish scale, cartilage, muscle, otolith, gill , other  
(please specify as appropriate) ]. This low level  presence is below the decision 
limit established for this microscopic method .” 
– “As far as was discernible using a light microscope, no more than 5 particles 
derived from terrestrial invertebrates were detected in the submitted sample. 
The particles were i dentified as … [cuticle fragments, mouthparts, muscles, 
tracheal structures , other (please specify as appropriate) ]. This low level 
presence is below the decision limit established for this microscopic method .” EN 12  EN When two determinations have been performed:  
– “As far as was discernible using a light microscope, no more than 10 particles 
derived from terrestrial vertebrates were detected over the two determinations 
in the submitted sample. The particles were identified as … [bone, cartilage, 
muscle, hair, horn , other (please specify as appropriate) ]. This low level 
presence is below the decision limit established for this microscopic method. ” 
– “As far as was discernible using a light microscope, no more than 10 particles 
derived from fish were detected over the tw o determinations in the submitted 
sample. The particles were identified as … [fishbone, fish scale, cartilage, 
muscle, otolith, gill , other (please specify as appropriate) ]. This low level 
presence is below the decision limit established for this microscopic method .” 
– “As far as was discernible using a light microscope, no more than 10 particles 
derived from terrestrial invertebrates were detected over the two 
determinations in the submitted sample. The particles were identified as … 
[cuticle fragme nts, mouthparts, muscles, tracheal structures , other (please 
specify as appropriate) ]. This low level presence is below the decision limit 
established for this microscopic method .” 
Additionally:  
– In case of sample pre -sieving, the laboratory report shall me ntion in which 
fraction (sieved fraction, pelleted fraction or kernels) the animal particles have 
been detected insofar as the detection of animal particles only in the sieved 
fraction may be the sign of an environmental contamination.  
– When only animal par ticles which cannot be categorised as either terrestrial 
vertebrates or fish are detected (e.g. muscle fibres), the report shall mention 
that only such animal particles were detected and that it cannot be excluded 
that they originate from terrestrial verte brates.  
2.1.5.3.  More than 5 animal particles of a given nature are detected when only one 
determination has been performed, or more than 10 particles of a given nature are 
detected in case of two determinations:  
When only one determination has been performed:  
– “As far as was discernible using a light microscope, more than 5 particles 
derived from terrestrial vertebrates were detected in the submitted sample. The 
particles were identified as … [bone, cartilage, muscle, hair, horn , other (please 
specify  as appropriate) ].” 
– “As far as was discernible using a light microscope, more than 5 particles 
derived from fish were detected in the submitted sample. The particles were 
identified as … [fishbone, fish scale, cartilage, muscle, otolith, gill , other 
(please specify as appropriate) ].” 
– “As far as was discernible using a light microscope, more than 5 particles 
derived from terrestrial invertebrates were detected in the submitted sample. 
The particles were identified as … [cuticle fragments, mouthparts, muscles , 
tracheal structures , other (please specify as appropriate) ].” 
When two determinations have been performed:  
– “As far as was discernible using a light microscope, more than 10 particles 
derived from terrestrial vertebrates were detected over the two determi nations EN 13  EN in the submitted sample. The particles were identified as … [bone, cartilage, 
muscle, hair, horn , other (please specify as appropriate) ].” 
– “As far as was discernible using a light microscope, more than 10 particles 
derived from fish were detected o ver the two determinations in the submitted 
sample. The particles were identified as … [fishbone, fish scale, cartilage, 
muscle, otolith, gill , other (please specify as appropriate) ].” 
– “As far as was discernible using a light microscope, more than 10 particles 
derived from terrestrial invertebrates were detected over the two 
determinations in the submitted sample. The particles were identified as … 
[cuticle fragments, mouthparts, muscles, tracheal structures , other (please 
specify as appropriate) ].” 
Additionally:  
– In case of sample pre -sieving, the laboratory report shall mention in which 
fraction (sieved fraction, pelleted fraction or kernels) the animal particles have 
been detected insofar as the detection of animal particles only in the sieved 
fractio n may be the sign of an environmental contamination.  
– When only animal particles which cannot be categorised as either terrestrial 
vertebrates or fish are detected (e.g. muscle fibres), the report shall mention 
that only such animal particles were detected and that it cannot be excluded 
that they originate from terrestrial vertebrates. ’