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Bisphenol A (BPA) is a highly-produced organic compound of anthropogenic origin with a wide-range use and is ubiquitously present in both living organisms and the environment. A previously published analytical method for testing of the free (aglycone) BPA in foodstuffs was simplified and optimized for sheep muscle tissue, kidney and liver, by using only a single MISPE purification step allied with HPLC-FLD analysis. The recovery rates and RSD values over a concentration range of 1–10 µg/kg were in the range of 67‒86% and 3‒34%, respectively, while linearity in the matrix, represented by the r 2 , was ≥0.999. LOD values were 0.5‒1 µg/kg and LOQ values were 1 µg/kg. The analytical method used is a contribution to the field of veterinary toxicology and food testing and also proved to be applicable for other food-producing animal species, e.g. pigs, poultry and freshwater fish. The MISPE sorbent material for testing of BPA was reusable, with up to five re-use cycles without significant loss in performance characteristics. The paper reports on a rapid determination of free (aglycone) BPA in tissues of food-producing animals by HPLC-FLD and presents a substantial improvement of analytical performance with regard to time- and cost-savings, as well as environmental protection. MISPE is an advanced analytical technology, and the results proved that in a single and simplified SPE step it had good performance characteristics such as selectivity, recovery and precision, allied with low LOD and LOQ values. It was proved that MISPE BPA cartridges could be used at least five times without significant loss of methodological recovery and precision.

Bisphenol A (BPA), an endocrine-disrupting chemical and environmental pollutant, has been reported by many researchers to induce male reproductive toxicity in different experimental models. In this study, we investigated whether long-term exposure for two months to 25 µg/kg body weight (low dose) of BPA affects spermatogenesis or sperm quality in young Istrian Pramenka rams exposed via diet. We evaluated body and testicular weights, histopathology of testes and epididymides, and sperm analyses, and compared these parameters between the group of treated rams and the control group of rams. Although there were some differences between the two groups, these differences were not large or statistically significant. The only statistically significant difference was the lower epithelial height of seminiferous tubules in treated rams, compared to control rams. In addition to assessing toxicity, BPA concentrations in the blood plasma of treated rams were determined after the first administration, and the toxicokinetic parameters of total BPA were calculated. In this study, no major signs of altered reproduction in rams were detected.

Testicular histopathology is considered the most sensitive and reliable method to detect the effects of chemicals on sperm production. To carry out a sensitive examination of testicular histopathology and interpret the changes require knowledge of spermatogenic stages. Spermatogenic staging based on acrosome development during spermiogenesis is conventionally performed in animal species routinely used for research and toxicity testing. In contrast, small ruminants, such as sheep and goats, are rarely used as animal models to evaluate toxicity in male reproductive organs. To the best of our knowledge, a comparable spermatogenic staging system in rams has not yet been fully characterised. Hence, this study aimed to adapt the existing spermatogenic staging based on acrosome development in bull testes to fit the seminiferous epithelium cycle of ram testes. The results show that spermatogenic staging based on acrosome development in bull testes can, with slight modifications, be efficiently used for the staging of ram testes.

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