Evidence is emerging that the interaction between male seminal fluid and female tissues promotes fertility, pregnancy, and health of offspring. This includes the acceleration of ovulation in a species known as a spontaneous ovulator, the domestic pig. Earlier studies revealed that seminal plasma acts by a local mechanism in the female pig. The aim of the present study was to examine local short-term and mid-term effects of seminal plasma (SP) on mRNA expression of immunoregulatory genes and transcripts associated with follicle- and oocyte maturation. In the porcine animal model, effects on mRNA expression in the female tract and preovulatory follicles were examined. SP suppressed mRNA expression of Prostaglandin-Endoperoxide Synthase 2 (PTGS2) ipsilateral to the infused uterine horn which was associated with a lower presence of immune cells in the uterine epithelium and lower PTGS2 immunoreaction. Depending on the sampling time (2 h vs. 17 h) and hormonal status, SP altered significant correlative relations of mRNA expression between PTGS2 and the transcripts Tumor Necrosis Factor Alpha, Tumor Necrosis Factor Alpha-Induced Protein 6 and Pentraxin 3 in uterus, granulosa and cumulus cells. A modulatory effect of SP on the oocyte gene network comprising eight oocyte transcripts was observed: uterine exposure to SP induced positive correlations (r >0.08, p<0.05) of maturation promoting factors among each other and with cumulus cells on the side of the treated horn. In conclusion, SP orchestrates the gene network regulating the bidirectional communication between oocytes and surrounding somatic cells. The modulation of the immune-cytokine network of the female reproductive system could contribute to the previously reported SP-induced acceleration of ovulation in the porcine species.

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Antimicrobial resistance is a steadily increasing problem and poses a serious threat to global public health. Therefore, it is highly necessary to advance the development of novel antimicrobial compounds and semen preservation strategies. The aim of this study was to evaluate a low temperature, antibiotic-free preservation procedure using Androstar Premium (ASP) extender (Minitüb) with antimicrobial lipopeptides. Firstly, seven lipopeptides in two concentrations (1 × minimum inhibitory concentration (MIC)/2 × MIC) were tested on their sperm-compatibility at 17 °C. Two lipopeptides, C16-KKK-NH 2 and C16-KKKK-NH 2 , did not negatively affect sperm quality and were further evaluated for their efficiency of bacterial growth inhibition at 5 °C. Besides an overall diminution of colony forming units, both peptides showed a reduction of bacterial subcultures (n = 103) with a decrement in Gram-positive rods from 65 (ASP w/o supplements) to 39/52 (ASP w/ C16-KKK-NH 2 /C16-KKKK-NH 2), in Gram-positive cocci from 21 to 9/10 and in Gram-negative species from 17 to 8/5 total subcultures. Furthermore, lipopeptides revealed activity towards selected bacteria of potential concern in artificial insemination like Trueperella pyogenes, Alcaligenes faecalis, Pseudomonas aeruginosa (not C16-KKK-NH 2), Pasteurella sp., Providencia stuartii, Escherichia coli (not C16-KKKK-NH 2) and Streptococcus porcinus (not C16-KKKK-NH 2). Consequently, both tested lipopeptides are promising candidates for alternative antibiotic-free preservation techniques of boar semen. Spreading antimicrobial resistance is a growing problem in human and veterinary medicine, poses a serious threat to global public health 1 and should therefore be addressed in a multifaceted approach from all areas of concern, including the industry branch of artificial insemination (AI) in pigs. It has been proven, that unhindered bacterial growth has detrimental effects on several sperm quality characteristics 2 and a transmission of potential pathogens to the inseminated sow has to be prevented in order to minimize negative effects on fertility and litter size 3. Semen collection is a non-sterile process and AI doses are conventionally stored at temperatures between 16 and 18 °C to prevent cold shock injuries to the highly sensitive boar spermatozoa 4. The combination of moderate temperatures and the use of nutrient-rich semen extenders promotes bacterial growth of psychrophilic and mesophilic species 5. As a countermeasure, antibiotics like gentamicin sulphate are routinely added to boar semen preserved for AI (Council Directive, European Union, 90/429/EEC). A steadily growing portion of contaminant bacteria isolated from AI doses are resistant to antibiotics commonly used as additives in semen extenders 6 , threatening animal welfare. This has far-reaching consequences for public health because of the natural backflow of parts of the rather large-volumed AI doses after insemination 7 ,

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