This study focused on finding new information regarding the assessment of pig saliva cortisol samples in terms of practical effects of the sampling, sample storage conditions, and their laboratory analysis. The study was divided into two experiments. The first experiment was focused on finding the effect of sampling time on cortisol concentrations in pig saliva. The second experiment was focused on determining the effect of storage conditions on the value of salivary cortisol. Before the initiation of the study, we tested which one of the commercially available ELISA kits would be the most suitable for our experiments. Simultaneously, we carried out a pre-study to evaluate the effect of relocation and change in the housing type on the concentration of salivary cortisol in gestating sows. The samples were obtained by oral cavity swabbing, using a standard cotton swab. In the first study, piglets were examined at the age of 4 ± 1 days, and breeding management routine procedures were used as a stress factor. In the second study, the piglets were examined immediately after weaning (at 28 ± 2 days of age). The Cortisol EIA kit was found to be statistically more accurate and thus a more suitable ELISA kit for our experiment. Analysis of the relocation effect and the effect of change in the housing type showed that relocation does not seem to be a stress factor for gestating sows as no significant changes were observed in salivary cortisol concentration (P > 0.5); however, the change in the housing type lead to a significant increase in salivary cortisol (P < 0.001). In the first study, we determined using the ELISA method that the most significant difference occurred in 40 min (P < 0.01), which suggests that the best time for a sampling in order to assess salivary cortisol concentration is 40 min after stress induction by routine procedures. The conclusion of the second study was that in the monitored period of 60 h (P < 0.05), cortisol concentration decreased depending on the storage temperature. The decrease started between 48 and 60 h which showed that cortisol is stable in the saliva sample for at least 48 h. These findings will be further applied in our following studies focused on assessment of salivary cortisol concentration after stress induction.
A variety of stressful situations commonly occur on dairy farms which can impair the well-being of the animals. The aim of this study was to analyse the concentration of cortisol in the saliva of dairy goats and on the basis thereof to determine the degree of stress experienced by them in relation to selected situations on farms. The following situations were selected as stressful: first visit to the milking parlour; weaning off; loading and transport; deworming; and the disruption of social hierarchy. We examined 344 samples from 100 animals using cotton swabs for the saliva collection. Commercially available ELISA kits (Cortisol EIA Kit, BosterBio, California, USA) which can detect cortisol in the saliva of all animal species, were used for the analysis. During the first visit to the milking parlour, weaning off, deworming and disruption of social hierarchy there was a significant (P < 0.05) increase in cortisol concentrations compared to the basal values. For loading and transport there was a highly significant (P < 0.01) increase in cortisol concentrations compared to the basal values. Although these situations are inevitable on farms, efforts should be made to eliminate them as much as possible due to the stress the animals experience in them.
Soy is considered an allergen under Regulation No. 1169/2011 of the European Parliament and of the Council, which mandates the labelling of soy allergen on food packaging. This study is focused on detecting soy in food products using two kinds of methods. The first method, enzyme-linked immunosorbent assay (ELISA), was performed using two commercial kits (Veratox for Soy Allergen; Neogen, USA). The second method, polymerase chain reaction (PCR), was carried out with two different deoxyribonucleic acid (DNA)-isolating kits and DNA isolation via the cetyltrimethylammonium bromide (CTAB) method. A total of 57 samples of food were tested, including 45 samples of animal origin and 12 samples of plant origin. The results were compared with information on the packaging. From the group of samples that contained soy according to the packaging (12 pieces), 9 samples were found to be positive by ELISA method, 10 samples by the CTAB method and 11 samples by GeneSpin. On the other hand, from the group of samples that should not contain soy according to the packaging (30 pieces), the presence of soy in 2 samples was detected by ELISA. No significant difference was found between the examined methods. Results show that the situation on the market is satisfactory and that only products declared as containing traces of soy appear to be problematic.
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