Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by plants as a chemical defense against herbivores. Plants containing PAs are widely distributed in almost all geographical regions posing a risk of honey contamination. To provide safety of honey and decrease the potential risk for the consumers, a sensitive method based on gas chromatography-mass spectrometry enabling determination of a content of 1,2-unsaturated PAs in honey was developed. Honey samples were purified on MCX cartridges, and PAs were eluted with a solvent mixture consisting of ethyl acetate, methanol, ammonia, and triethylamine. Subsequently, 1,2-unsaturated alkaloids were reduced to their common backbone structures and derivatized with heptafluorobutyric anhydride. The method was validated according to SANTE 2015. All received parameters are in consistence with the document requirements as recovery ranged from 73.1 to 93.6%. The repeatability and reproducibility were calculated as relative standard deviation and ranged from 3.9 to 8.6% and from 10.6 to 17.8%, respectively. The limit of quantification was determined as 1 μg kg −1. Good linearity of the method was obtained with the coefficient of determination R 2 > 0.99. The method was applied to 40 Polish and 14 Asian honey sample analyses.

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The aim of this study was to perform an in-house validation of multiplex PCR method for C. botulinum detection in food and feed samples. The study was carried out on food and feed matrixes artificially contaminated by spores of C. botulinum reference strains. The following characteristic parameters for qualitative detection were estimated: limit of detection expressed as LOD 50 according to the Spearman-Kärber formula, specificity, sensitivity, and accuracy according to the PN-EN ISO 16140:2004. The validated method showed high specificity. Specific PCR products were revealed only for DNA obtained from samples contaminated with C. botulinum spores. PCR inhibition was observed, especially during examination of contaminated feed. The calculated LOD 50 for feed was nearly 10 times higher than for food. The implemented method enables to obtain test results during 3 d without timeconsuming process of isolation and proving the ability of strains to produce botulinum toxins.

The aim of this study was to assess the possibility of genetically modified DNA transfer from feed containing RR soybean or/and MON810 maize to animal tissues, gut bacterial flora, food of animal origin, and the fate of GM DNA in the animal digestive tract. The experiment was carried out on broilers, laying hens, pigs and calves. All animals were divided into four groups: I -control group (non-modified feed), II -GM soybean group (non-modified maize, RR soybean), III -GM maize group (MON810 maize, non-modified soybean), and IV -GM maize and soybean group (MON810 maize, RR soybean). Samples of blood, organs, tissues, digesta from the gastrointestinal tract, and eggs were analysed for the presence of plant species specific genes, and transgenic sequences of CaMV 35S promoter and NOS terminator. PCR amplifications of these GM sequences were conducted to investigate the GM DNA transfer from feed to animal tissues and bacterial gut flora. In none of the analysed samples of blood, organs, tissues, eggs, excreta and bacterial DNA were plant reference genes or GM DNA found. A GM crop diet did not affect bacterial gut flora as regards diversity of bacteria species, quantity of particular bacteria species in the animal gut, or incorporation of transgenic DNA to the bacteria genome. It can be concluded that MON810 maize and RR soybean used for animal feeding are substantially equivalent to their conventional counterparts. Genetically modified DNA from MON810 maize and RR soybean is digested in the same way as plant DNA, with no probability of its transfer to animal tissues or gut bacterial flora.

IntroductionSilage quality deteriorates with Clostridium spp. contamination, and if consumed, such silage jeopardises herd health and productivity. Minimising its occurrence reduces economic and animal welfare risks. The study investigated the influence of environmental and technological determinants on the Clostridium genus’ occurrence in silage.Material and MethodsAnalyses were conducted on 305 silage samples directly collected from farms located in all Polish provinces. Cultures and isolates were evaluated phenotypically and examined for occurrence of Clostridium spp., particularly C. perfringens and C. botulinum using PCR techniques. The results were statistically analysed using the ᵡ2 test for continuous and Student’s t-test for non-continuous values.ResultsThe most influential effect on Clostridium spp. occurrence is exerted by factors potentially associated with primary production, like the type of fertilisation and the contamination level of the ensiled feed material. Clostridium spp. was detected in 232 (76%) samples, and C. perfringens strains, predominantly toxinotype A, in 79 (26%). C. botulinum occurrence was not detected.ConclusionsDeterioration of silage by clostridia could be prevented by a properly conducted ensiling process with the addition of starter cultures, but the presence of spores mainly depends on primary production and the extent of contamination of the feed material.

The aim of this study was an examination of 240 multifloral honey samples collected from Polish apiaries to determine Clostridium botulinum occurrence. Honey was collected from apiaries directly after the extraction process. Samples were inoculated by using the dilution and centrifugation method. Suspected isolates were examined by using mouse bioassay, polymerase chain reaction (PCR), and real-time PCR methods. C. botulinum type A and B strains were detected in 5 of 240 examined honey samples (2.1%). Bacterial strains were also detected that were phenotypically similar to C. botulinum but that did not exhibit the ability to produce botulinum toxins and did not show the presence of the botulinum cluster (ntnh and bont genes) or expression of the ntnh gene. The methods used in the examination, especially the expression analysis of ntnh gene, enabled specific analysis of suspected strains and could be used routinely in environmental isolate analyses of C. botulinum occurrence.

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IntroductionThe influence of feeding genetically modified MON 810 hybrid maize on the growth and haematological and biochemical indices of rats was tested.Material and MethodsTwo conventional (non-GM) and two test (MON 810) lines of maize were used in semi-purified diets at the level of 40% w/w. The non-GM I, MON 810 I, non-GM II, and MON 810 II maize lines were near-isogenic. A total of 40 male 6-week-old Wistar-derived rats were assigned to four equal feeding groups corresponding to the four maize lines for 16 weeks. Overall, health, body weight gain, clinical pathology parameters, gross changes, and appearance of tissues were compared between groups.ResultsThere were no statistically significant differences in the weight gain or relative organ weights of rats, but there were some non diet-related histopathological changes in the liver, kidneys, and spleen. Except for creatinine level, no diet-related effects were observed in haematology or most of the biochemical indices. Transgenic DNA of MON 810 maize was not detected in the tissues or faeces nor in the DNA of E. coli isolated from the rectum digesta of rats given transgenic feeds. In our experiment, various metabolic indices of rats fed non-GM diets or genetically modified (MON 810) maize for 16 weeks were similar. No adverse nutrition-related health effects were detected.ConclusionMON 810 maize seems to be as safe as the conventional maize lines.