Influence of antibiotics on bacterial load and sperm parameters during short-term preservation of collared peccary semen

Abstract

Studies on semen and sperm cells are critical to develop assisted reproductive technologies for the conservation of the collared peccary. The objective of the study was to compare the effect of different antibiotics on the bacterial load and sperm quality during short-term storage of peccary semen. Fresh semen samples from 10 males were extended in Tris-egg yolk or Tris-Aloe vera supplemented with streptomycin-penicillin (SP; 1 mg/mL -1000 IU/mL or 2 mg/mL -2000 IU/mL) or gentamicin (30 µg/mL or 70 µg/mL) before storage at 5°C. Bacterial load and sperm motility, membrane integrity and function, mitochondrial activity, and morphology, were evaluated at different time points for 36 h. The SP and gentamicin treatments concentration inhibited (p < 0.05) bacterial growth for 36 h regardless of the extender. Compared to the other treatments, Tris-egg yolk plus 70 µg/mL gentamicin maintained the sperm parameters for longer, including total motility (41.9 ± 6.1%) at 24 h, and membrane integrity (58.3 ± 2.1%) at 36 h. In contrast, the highest SP concentration in both extenders impaired sperm membrane integrity at 36 h (p < 0.05). For the liquid storage of collared peccary semen, it therefore is recommended to use Tris extender supplemented with egg yolk and gentamicin (70 µg/mL).

Investigating the need for antibiotic supplementation to the extender used for semen cryopreservation in collared peccaries

“…In the other hand, the penicillin is a β lactam that acts by inhibiting the synthesis of cell wall of gram-positive bacteria (19). Even if this is the first time that these antibiotics were used of the cryopreservation of peccary sperm, they were previously proven to provide efficient control of bacterial load during peccary semen chilling for 36 h. At this point, the streptomycinpenicillin combination was able to reduce the bacterial load to zero in various samples (12).…”

“…For the microbiological analysis, an aliquot of 100 µL of each semen sample was inoculated in 900 µL of sterile 0.85% saline, obtaining a dilution of 10-1, followed by a serial dilution up to 10-5. Aliquots of 100 µL of each dilution were sown, with the aid of a Drigalski loop, on the surface of Petri dishes containing Plate Count Agar (Hi Media, Mumbai, India), all in duplicate and incubated in a bacteriological incubator (Fanem LTDA, São Paulo, Brazil) at 37 • C for 24-48 h. Colonies were then counted on each plate and the number of microorganisms was expressed in Colony Forming Unit-CFU/mL multiplied by the inverse of each dilution (12).…”

“…Leica. Kista, Sweden), whose spermatozoa with a blue head (H-342) were judged to contain an intact membrane and those with a full head or partially red-labeled (PI) were considered to contain a non-intact membrane, and a red-labeled midpiece were considered having a mitochondrial function (12). The binding ability of spermatozoa was investigated using the hen egg perivitelline membrane binding assay, as previously validated for peccaries (15).…”

“…For freezing, the ejaculates were diluted in Tris plus glycerol (3%) and egg yolk (20%), and separated into three aliquots that were kept without antibiotics (Control group) or supplemented with gentamicin (70 µg/mL) or streptomycin/penicillin (1 mg/mL/1000 IU/mL), as previously reported for peccary semen refrigeration (12). Final dilution resulted in a 100 x 10 6 sperm/mL concentration.…”

Kojic acid inhibits pig sperm apoptosis and improves capacitated sperm state during liquid preservation at 17°C

“…Additionally, KA can improve the state of capacitated sperm by facilitating the process of oxidative phosphorylation, ATP level, and tyrosine oxidative phosphorylation (Figure 9). prevent the quality degradation of animal semen caused by bacterial contamination (Oplinger & Wagner, 2015;Santos et al, 2021).…”