The annual proficiency test for the detection of animal proteins in animal feed of the IAG -International Association for Feeding stuff Analysis, Section Feeding stuff Microscopy was organized by Wageningen Food Safety Research, The Netherlands. The proficiency test was intended to provide the participants information on their performance of the implementation of the monitoring methods as well as to gather information about the current practices in the application of the microscopic method. The current 2021 version of the IAG proficiency test for animal proteins addressed all analytical sections of the methods for microscopy and PCR as published in Annex VI of Regulation (EC) 152/2009 together with accompanying SOPs. Regulation (EU) 2020/1560, in force from 16 th November 2020, introduced labelling information as extra parameter for decision on actions in the procedure, and changed the maximum determination cycles to two.The samples used in the proficiency test contained salmon meal (2%), ruminant processed animal protein (PAP, 0.1%) and milk powder (5%). A fourth sample was left blank. The matrix was artificially produced, mimicking a ruminant feed, for avoiding any nonintentional contamination with ruminant DNA. The labels of the samples did not show an indication of the composition nor of the intended target animal.A total of 41 participants subscribed to the proficiency test animal proteins. One participant did not submit their results and two submitted PCR results only, leaving 38 sets for microscopic evaluation. 16 sets of ruminant PCR results were submitted as well. The organisation and evaluation of the test and its results followed the Quality Guidance for Visual Research in Feed and Food. MicroscopyAll participants were requested to determine the presence or absence of land animal and/or fish, to indicate the type of material found and to describe the method used to achieve these results. Participants made a choice to follow either the old or the new protocol. Their choice to apply a second determination cycle would correctly match either one of these choices in all cases. Three participants made incorrect interpretations of the encountered number of particles (e.g. "suspect" for zero particles, "present" after a positive PCR result without any microscopical observation), and two participants did not report a final conclusion. Therefore, all evaluations were based on the actual number of particles reported by all participants.Incorrect positive results (positive deviations) were expressed in a specificity score and incorrect negative results (negative deviations) were expressed in a sensitivity score. An optimal score is 1.0. The results are analysed in two ways: numbers below threshold (between 1 and 5 particles per determination cycle inclusive) have been considered positive (complying to the zero tolerance) and as alternative considered as negative (for matching the official evaluation).The sensitivity for both basal ingredients were good (sample D, ruminant PAP: 0.97; sample B, salmon meal: 0.95). T...
of the method or a lack of expertise. This, because the labels provide a reminder to look for milk powder in the samples that contained it, providing statistics for the sensitivity of the method, and in samples that do not contain milk powder, providing statistics for the specificity of the method, whilst removing inattentiveness as an obfuscating factor. PCRThe ruminant material in the MBM sample (sample B) was correctly detected by all participants, one false positives was reported in the fish meal sample (sample C), all participants correctly detected ruminant material in the milk powder sample (sample D), and all participants correctly reported the absence of ruminant material in the blank (sample A). ConclusionsVery good or excellent results were achieved for the detection of processed animal proteins. The situation for detection of other ingredients, exemplified by the presence or absence of milk powder, needs considerable improvement. It is recommended to consider to reduce the scope of the current legally implemented method to PAPs and find adjusted protocols for other types of ingredients. Training for identification of these different particles need to be organised.
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