Essential oils possess a variety of biological properties (i.e., antioxidant, antibacterial, and cytotoxic) that could possibly be applied in reproductive medicine, but their effects on spermatozoa are still partially unknown. The aim of the study was to describe the effects of Thymbra capitata (L.) Cav. and Rosmarinus officinalis (L.) essential oils on the main morpho-functional parameters of swine spermatozoa. Essential oils were preliminary characterized by gas chromatography and added with emulsifiers to facilitate diffusion. Experimental samples were prepared by suspending a fixed number of spermatozoa in 5 mL of medium with 10 different concentrations of essential oil (0.2–2 mg/mL, at intervals of 0.2). After 3 h of incubation, samples were analyzed for pH, viability, objective motility, and acrosome status. Results showed that the effects of the essential oils are concentration-dependent and that R. officinalis is well tolerated up to 0.6 mg/mL. T. capitata impaired the spermatozoa starting from the lowest concentration, with complete spermicidal effect from 0.4 mg/mL. The patterns of damage, confirmed by SEM, were different and quite distinct. As expected, spermatozoa proved to be sensitive to external stimuli and capable of showing different functional patterns, providing interesting insights to the action/toxicity mechanisms. The results of the present work represent the first step towards the systematic characterization of the effects of these compounds on spermatozoa. This kind of studies are necessary to strengthen the idea of future applications of essential oils in the reproductive field due to their antioxidant, antibacterial, or spermicidal properties.

The growing interest towards essential oils stems from their biological capabilities that include antibacterial and antioxidant effects. Such properties may be extremely useful in the reproductive field; nonetheless essential oils show toxic effects that can lead to cell disruption. The present study aimed to evaluate and compare the effects of tea tree oil (TTO) and its principal component terpinen-4-ol (TER) on the morpho-functional parameters of swine spermatozoa. Experimental samples were prepared by suspending 15 × 107 spermatozoa in 5 mL of medium with different concentrations of the above-mentioned compounds: from 0.2 to 2 mg/mL at an interval of 0.2 for TTO, while TER concentrations were adjusted according to its presence in TTO (41.5%). After 3 h incubation at 16 °C, samples were analyzed for pH, viability, acrosome status, and objective motility. The results highlighted a concentration-dependent effect of TTO with total motility as the most sensitive parameter. TER was better tolerated, and the most sensitive parameters were related to membrane integrity, suggesting a different pattern of interaction. The study confirms the importance of evaluating the effects of natural compounds on spermatozoa before exploiting their beneficial effects. Spermatozoa seem to be good candidates for preliminary toxicological screenings in the light of their peculiar properties.

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Bifidobacterium lemurum sp. nov., from faeces of the ring-tailed lemur (Lemur catta)

The efficiencies of two published primer pairs (RSP13-RSP14 and RSP2-RSP21) were compared in reverse transcriptionpolymerase chain reaction (RT-PCR) to improve the diagnosis of Grapevine rupestris stem pitting-associated virus (GRSPaV), which has consistently been found associated with rupestris stem pitting (RSP), one of the most common infectious diseases of grapevine. Fifty-six grapevine accessions from different locations in Northern Italy were tested. Of these, 48 were positive with primers RSP13-RSP14 and 37 with primers RSP2-RSP21, suggesting that primer pair RSP13-RSP14 has a broader detection spectrum than primers RSP2-RSP21. The coat protein (CP) gene was also amplified by RT-PCR and the amplicons analysed by single-strand conformation polymorphism (SSCP) and sequencing. The results support the existence of wide variability within this virus, independent of cultivar or geographic origin. Sequence analysis of the 17 Italian GRSPaV isolates examined showed a molecular identity ranging from 78AE8 to 99AE7%, within which, four groups of sequence variants were recognized. Flexuous rod-shaped particles, approximately 800 nm in length, were detected in plant extracts from infected grapevines and were positively decorated with the specific antiserum As7-276 in immunosorbent electron microscopy (ISEM). This is the first report of GRSPaV particles in grapevines observed by transmission electron microscopy (TEM) in Italy.