Introduction to new guidelines for validation of methods to examine visually recognisable substances.

Visual examination of visually recognisable substances, including microscopy, focus on targets or contaminants such as particles of animal origin, plant seeds, spore bodies of moulds, sclerotia, packaging material, microplastic and 'Besatz' (everything that differs from the norm). The two principal results are counts (numbers) and weights for macroscopic methods, or presence/absence for microscopic methods. The level of detection equals at least the size of one unit, usually with a weight exceeding 1 mg, which is in the range of parts per million (ppm). These parameters do not follow a normal distribution but Poisson (counts), lognormal (weights) or binomial (Booleans) distributions, with effect on the interpretation of validation parameters. As for other domains, examination methods for visual monitoring need to be properly validated and quality control during actual application is needed. In most cases procedures for validation of visual methods are based on principles adopted from other domains, such as chemical analysis. A series of examples from publications show inconsistent or not correct implementations of these validation procedures, which stress the need for dedicated validation procedures. Identification of legal ingredients and composition analysis in the domain of visual examination relies on the expertise of the laboratory staff, therefore validation of a method usually includes the validation of the expert. In the view of these specific circumstances, a Guidance for quality assurance and control of visual methods has been developed, which are being presented and discussed in this paper. The general framework of the Guidance is adopted from ISO standards (17023, 17043, 13528). Part 1 of the Guidance includes the general background, theory and principles. Part 2 presents the actual validation procedures with experimental designs and equations for calculating the relevant parameters, and can be used as blueprint for a SOP in a quality management system. An EURL and NRL network for physical hazards is strongly recommended.

The effects of feed naturally contaminated with Fusarium mycotoxins on the thymus in suckling piglets.

In this study, feed naturally containing Fusarium mycotoxins was fed to gilts during the perinatal period, and the effects on the thymus were investigated in one-week-old piglets. Twenty gilts were divided into equal control (0.26 mg deoxynivalenol, DON) and experimental (5.08 mg DON, 0.09 mg zearalenone and 21.61 mg fusaric acid per kg of feed) groups. One suckling piglet from each litter (n = 20) was sacrificed at one week of age to obtain thymus samples for further analysis. The cortex to medulla ratio of the thymus was morphometrically analysed using NIS Elements BR (Nikon) software. Paraffin-embedded thymus sections were stained to quantify apoptosis (with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling - TUNEL method), cellular proliferation (Ki-67) and macrophages (MAC 387). The results showed that the thymus cortex (P = 0.023) to medulla (P = 0.023) ratio was significantly lower in the experimental group. The number of apoptotic cells (cortex, P = 0.010, medulla, P = 0.001) and the number of proliferating cells in the thymus cortex (P = 0.001) and medulla (P < 0.001) were significantly higher in the experimental group. Our results indicate that feeding Fusarium mycotoxins to a parent animal during the perinatal period induces significant alterations in the thymus of one-week-old piglets, which indicates an immunosuppressive effect in piglets.