Evaluation of cryopreserved boar semen after supplementation sericin form silkworm (Bombyx mori) in semen extender

Boar cryopreserved semen is scarcely used for artificial insemination due to its quality which is largely reduced by membrane lipid peroxidation. This present study was designed to improve the post‐thawed boar semen quality by determining the optimal level of sericin supplementation (antioxidants) in semen extender. Five levels of sericin supplementation between 0% and 1% (w/v) were examined. Semen was frozen by the liquid nitrogen vapor method, thawed slowly at 5°C for 5 min, and used for the evaluation of sperm quality. The results indicated 0.5%–1% sericin supplementation was more effective on maintenance of sperm viability, acrosome integrity, and mitochondrial functions during freezing–thawing. Moreover, 0.75% sericin supplementation was most protective toward total sperm motility and sperm progressive motility. Additionally, 0.25%–0.75% sericin supplementation significantly suppressed increases in the index of lipid peroxidation. In conclusion, 0.75% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved boar semen. However, further research using AI will be necessary to demonstrate that this indication can be applied to the production of offspring in the farms.     

The effect of apramycin on bacterial contamination and the survival of boar semen during a short preservation period

264 ejaculate samples taken from seven boars were tested for the bacteriostatic action of apramycin and for its effect on some sperm quality parameters during 96-hour storage. Apralan soluble powder, produced by Elanco Products Limited, was used in the trials at the concentrations of 2.5 to 500 mg per 1000 ml of diluent. The preparation was evaluated according to sperm motility during storage, according to the concentration pattern of hydrogen ions, morphological changes in spermatozoa, thermoresistance test, bacteriological tests and other examinations. The concentrations of 200 to 250 mg of apramycin in combination with 500 000 i. u. of G-penicillin potassium salt had a statistically significantly more favourable effect on the survival of boar sperm, as compared with the combination of streptomycin with penicillin.     

Assessment of bacterial contamination of three multidose ophthalmic solutions

OBJECTIVE: To investigate the frequency of bacterial contamination of multidose proparacaine hydrochloride, tropicamide, and eyewash bottles used in veterinary ophthalmology examination rooms during normal operating procedures. METHODS: Three representative bottles each of proparacaine hydrochloride, tropicamide, and purified water eye wash were opened at the same time, numbered, and placed into small animal examination rooms. Doctors, students and technicians who were using the solutions were blinded to the study. Aerobic cultures were obtained at the time of opening (time 0), at 1 week (time 1), and at 2 weeks after opening (time 2) the bottles. The sites cultured included a drop of each solution, the inside of the bottle cap, the tip of the bottle, and the bottle threads and medication residue found in these threads. RESULTS: Aerobic cultures of tropicamide and proparacaine had no growth of bacteria from any of the evaluated sites. Staphylococcus epidermidis was cultured from the tip of one bottle of eye wash after 1 week. CONCLUSIONS: We conclude that proparacaine, tropicamide, and eyewash solutions used in veterinary examination rooms are adequately preserved for use in ophthalmic examination. They have a low level of bacterial contamination 1 or 2 weeks after opening when used and stored according to the recommendations of the product manufacturers and previous studies.     

Supplementation of salvianic acid A to boar semen extender to improve seminal quality and antioxidant capacity

The purpose of this test was to investigate the effect of salvianic acid A (SAA, CAS No. 76822‐21‐4) on the quality of boar semen during liquid storage at 17°C. The effects of different concentrations of SAA on semen quality and antioxidant capacity were analyzed. Boar semen was diluted with Beltsville Thawing Solution (BTS) containing different concentrations (0, 15, 30, 45, 60, 75 μM of SAA). During the storage period, sperm activity was measured every 24 hr, and plasma membrane integrity, acrosome integrity, total antioxidant capacity (T‐AOC), malondialdehyde (MDA) content, and catalase (CAT) activity were measured at 0, 1, 3, and 5 days. The results from our study suggest that different concentrations of SAA have different effects on semen preservation. Semen samples supplemented with SAA showed reduced effects of oxidative stress on sperm compared to the control samples. Supplementation of 30 μM of SAA significantly improved sperm motility, plasma membrane integrity, acrosome integrity, and antioxidant capacity. However, the addition of SAA to the extender was scarcely beneficial to the improvement of results of artificial insemination with boar semen after liquid preservation. Further studies are necessary in order to demonstrate that SAA has good effects on the liquid preservation of semen.     

An investigation of the bacterial contamination of small animal breathing systems during routine use

OBJECTIVE: To investigate the need for sterilization of anaesthetic breathing systems to prevent cross-infection between animals due to the re-use of anaesthetic circuit tubing. STUDY DESIGN: Prospective microbiological study. METHODS: Bacteriology samples were taken from 37 sterile breathing systems, each used for 1 day, at two sampling sites (one proximal and one distal to the animal). The number of patient connections, cumulative anaesthesia time, culture results, number of colony-forming units and the number of different species were recorded. Secondly, four sterile breathing systems were used for 2 months under routine conditions and sampled every 2 weeks and the same parameters recorded. Finally, the inner surfaces of four sterile breathing systems were inoculated with a known load of canine oropharyngeal bacteria. Bacteriology samples were taken at 1 minute, 1 hour and 1 day post-deposition. The number of colonies identified was compared with the initial load. RESULTS: Only a very small number of micro-organisms were isolated and these were generally organisms of low pathogenic potential. The proximal site was found to be significantly more colonized than the distal site (p < 0.001). Neither longer daily connection time (p = 0.54), nor a higher number of connections (p = 0.81) increased the incidence of proximal site colonization. Over the 2-month study period, the bacterial population did not increase. There was no correlation between cultures isolated from successive samples taken from the same tubing. There was rapid loss of viability of the micro-organisms deliberately inoculated onto the tubing surface: the number of colonies isolated from the breathing system after 1 minute was significantly lower than in the inoculum (p = 0.042). CONCLUSIONS AND CLINICAL RELEVANCE: Sterile anaesthesia breathing systems were colonized by environmental micro-organisms of low pathogenicity. Although long-term survival of recognized pathogens in a dry environment is still possible, the use of a bacterial filter or a sterilized breathing system for routine veterinary anaesthesia cannot be supported by current evidence.     

Effects of sulfanilamide on boar sperm quality,bacterial composition,and fertility during liquid storage at 17°C

Sulfanilamide (SA) is an effective broad‐spectrum antibacterial agent in human and veterinary medicine. The purpose of this study was to evaluate the effects of SA on boar sperm quality during liquid storage at 17°C and determine the optimal concentration of SA and its effects on bacterial growth, microbial composition, and maternal fertility. Boar ejaculates were diluted with a basic extender, containing different concentrations of SA, and stored in a 17°C incubator for 6 days. The sperm motility, plasma membrane integrity, and acrosome integrity were measured daily. The results showed that when the concentration of SA was 0.02 g/L, the sperm quality parameters were significantly higher than those of all other treatment groups (p < .05). We also monitored the bacterial growth and compared the differences in the microbial species between the 0.02 g/L SA group and the control by 16S rDNA sequencing. The results revealed that some bacteria, such as Staphylococcus and Pseudomonas, were considerably lower in the 0.02 g/L SA group than in the control group (p < .05). In addition, preserved semen was used for artificial insemination, and results showed that 0.02 g/L SA group had a higher litter size, and its pregnancy rate was 92.5%.     

Peptide derivatives of dermaseptin S4 in fresh bovine semen for bacterial contamination control: Physicochemical and structural characterization,antibacterial potency,and effects on red blood and sperm cells

The objectives of this study were to examine the physicochemical and structural properties of peptide derivatives of dermaseptin S4, investigate their detrimental effects on red blood and sperm cells and ascertain their antibacterial potency to control bacterial contaminants in fresh bovine semen. The dermaseptin S4 peptide derivatives used in this study were K4S4, S4(5–28), S4(5–28)a, K20S4(5–28), K4S4(1–16)a, K4S4(1–15)a and K4S4(1–15). Peptides K4S4, S4(5–28)a, K20S4(5–28), K4S4(1–15)a and K4S4(1–16)a, with a higher positive charge, were the most potent against the bacterial strains tested, with the lowest minimum inhibitory concentration (MIC), whereas S4(5–28) and K4S4(1–15), with a lower positive charge, showed the highest MIC (p < .01). Haemolysis percentage depended on peptide concentration (p < .01). The K4S4 was the most powerful haemolytic peptide, showing the highest haemolysis percentage at all peptide concentrations (p < .01). In contrast, S4(5–28), S4(5–28)a, K20S4(5–28) and K4S4(1–15) were not able to produce 50% cell lysis up to 100 µM (p < .01). All peptides reduced sperm motility in a dose-dependent manner when used in concentrations from 16 to 64 μM (p < .01). The highest reduction was seen due to K4S4 activity, and the lowest reductions of sperm motility were observed due to K4S4(1–16)a and K4S4(1–15)a activity (p < .01). Hence, we can conclude that K4S4(1–16)a and K4S4(1–15)a at a concentration of approximately 15 µM are the most promising peptides as antibacterial agents in fresh bovine semen, because at this concentration, they showed the most potent antibacterial activity against evaluated strains without significant effects on haemolysis or a reduction in sperm motility.     

Changes in the predicted function of the rumen bacterial community of Japanese Black beef cattle during the fattening stages according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses

We investigated changes in the predicted functions of the rumen bacterial community in Japanese Black beef cattle during fattening. Nine cattle were fed a high-concentrate diet during the early, middle, and late fattening stages consecutively (10–14, 15–22, and 23–30 months of age, respectively). The rumen fluid and solid samples collected at each stage were subjected to sequencing analyses. The sequencing results were clustered and classified into operational taxonomic units (OTUs). Representative sequences and a raw counting table for each OTU were submitted to the Piphillin website. The predicted functions were revealed by the Kyoto Encyclopedia of Genes and Genomes database as the ratio of the total sequence. In the early stage, “Biosynthesis of secondary metabolites” was significantly higher in the fluid fraction than in the solid fraction. “Two-component system” in the middle stage was significantly lower and “Purine metabolism” in the late stage was significantly higher in the fluid fraction than those in the solid fraction. The fluid fraction was significantly correlated with acetic acid, propionic acid, and bacterial metabolism, such as “Biosynthesis of secondary metabolites” and “Sugar metabolism.” Moreover, the solid fraction was correlated with “Purine metabolism” and “Biosynthesis of secondary metabolism”. These results suggest that the rumen bacterial community in Japanese Black beef cattle adapts to changes in rumen conditions by altering their functions in response to a long-term high-grain diet.