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Simple Summary: In the rabbit species, the use of frozen semen could play a twofold role; on the one hand, it could bring many benefits to commercial breeding operations that rely on artificial insemination, and on the other hand, it would enable the conservation of biodiversity through the establishment of semen cryobanks. One of the less explored aspects of cryopreserving rabbit semen involves establishing the ideal sperm concentration in straws. Achieving this goal is crucial for minimizing the variability in outcomes and accurately determining the number of sperm delivered during artificial insemination procedures. This study sought to determine the ideal sperm concentration in straws for cryopreserving rabbit semen, with the broader goal of improving and standardizing the freezing protocol developed by our research group over the past decade. Our study provided a comprehensive analysis of how different sperm concentrations within straws (ranging from 15 to 75 million sperm) influenced the critical stages of the cryopreservation process, starting from initial dilution with the extender, through to cooling and equilibration, and culminating in the thawing phase. This investigation shed light on the role of the sperm concentration when determining the overall success of rabbit semen cryopreservation. This study aimed to investigate the impact of sperm concentrations on the in vitro quality of cryopreserved rabbit semen. The semen pools (n = 8, from 80 donors) were split into five aliquots with final sperm concentrations of 15, 25, 35, 55, and 75 × 106 per straw. The sperm motility parameters (CASA system) and membrane integrity (flow cytometric analysis) were both evaluated at various stages of the cryopreservation process: fresh semen dilution, cooling, equilibration, and immediately after and 30 min post-thawing. The results indicated the significant influence of the sperm concentration on the total motility (TM) and progressive motility (PM), with a consistent decline in all sperm variables over the time points. Notably, the semen with a final concentration of 15 × 106 exhibited a higher TM and PM after cooling and equilibration. The post-thawing quality (TM, PM) was higher (p < 0.05) in the mid-range sperm concentrations of 25 × 106 (49.9% and 19.7%) and 35 × 106 (46.2% and 19.7%) compared to the other concentrations. This study demonstrated that the sperm concentration per straw played a significant role in specific phases of the cryopreservation process. These findings contribute valuable insights for refining and standardizing the cryopreservation protocol for rabbit semen, emphasizing the importance of the sperm concentration. [ABSTRACT FROM AUTHOR]

Simple Summary: The aim of this study was to evaluate the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Phyto-L (Pro Tech s.r.l.), a commercial product containing organosulfur compounds (OSCs) such as propyl propane thiosulfonate (PTSO) from Allium spp., on 108 enteropathogenic E. coli (EPEC) strains responsible for colibacillosis in rabbits. Bacterial suspensions with a charge of 108 CFU/mL were tested with different concentrations (20, 10, 5, 2.5, 1.25, 0.6, 0.3, and 0.15 μL/mL) of Phyto-L. To evaluate MBC values, bacterial suspensions corresponding to the MIC and above the MIC were plated on Tryptic Soy agar (TSA) without Phyto-L. The MICs of the tested strains corresponded to 1.25 μL/mL (37/108-34.3%), 2.5 μL/mL (70/108-64.8%), and 5 μL/mL (1/108-0.9%). The MBCs were 1.25 μL/mL (15/108-13.9%), 2.5 μL/mL (46/108-42.6%), 5 μL/mL (9/108-8.3%), 10 μL/mL (20/108-18.5%), 20 μL/mL (8/108-7.4%), and higher than 20 μL/mL (10/108-9.3%). Based on the results obtained, Phyto-L has antibacterial activity on EPEC strains. Therefore, in field applications, Phyto-L should be useful in limiting the E. coli load in the rabbit gut, preventing the occurrence of colibacillosis. Moreover, considering that 104–105 CFU/g of feces is the charge of E. coli normally present in the intestinal contents of rabbits under physiological conditions, it is possible that lower dosages than those found in this study may be effective in preventing the disease in rabbit farms. Colibacillosis, caused by enteropathogenic Escherichia coli (EPEC), is one of the most common diseases in rabbit farms, resulting in economic losses due to mortality and decrease in production. Until recently, antimicrobials were used to both treat and prevent disease on livestock farms, leading to the possible risk of antimicrobial resistance (AMR) and the selection of multidrug-resistant (MDR) bacteria. Therefore, interest in alternative control methods, such as the use of natural substances, has increased in the scientific community. The aim of this study was to evaluate the antimicrobial efficacy of Phyto-L (Pro Tech s.r.l.), a product containing organosulfur compounds (OSCs) such as propyl propane thiosulfonate (PTSO) from Allium spp., against 108 strains of E. coli isolated from rabbits with colibacillosis from 19 farms. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Phyto-L were assessed. Bacterial suspensions with a charge of 108 CFU/mL, corresponding to those found in the rabbit gut under pathologic conditions, were tested with different concentrations from 20 to 0.15 μL/mL of Phyto-L. For each strain, the MIC and concentrations above the MIC were plated on Tryptic Soy agar (TSA) without Phyto-L to assess the MBCs. MIC and MBC values ranged from 1.25 to 5 μL/mL and 1.25 to 20 μL/mL, respectively, depending on the strain tested. The data showed an interesting antibacterial activity of Phyto-L against EPEC strains. Therefore, this product could be effective in preventing colibacillosis in field application, especially considering that 104–105 CFU/g of feces is the amount of E. coli usually found in the gut contents of rabbits under physiological condition. [ABSTRACT FROM AUTHOR]