Abstract
The presence of pathogenic bacteria in ejaculates has been a topic in boar semen preservation over the last decades. Since little information is available on commensal bacteria in boar semen, the aim of the present study was to identify commensal lactobacilli in fresh cryopreserved boar semen and to examine their influence on boar semen quality. Therefore, 111 boar ejaculates were investigated for the presence of Lactobacillus species. Thirty samples (27%) contained viable Lactobacillus species (e.g. L. amylovorus, L. animalis, L. reuteri and Weisella minor). L. animalis and L. buchneri DSM 32407 (isolated from the bovine uterus) qualified for further examinations based on their growth rate in six antibiotic-free boar semen extenders. After a 120 min short-term incubation with an antibiotic-free BTS-extender, progressive motility was diminished (P = 0.001) upon addition of 105 and 106 colony forming units (CFU/mL) L. animalis. The supplementation with L. buchneri DSM 32407 had no significant (P > 0.05) influence on sperm quality during short-term co-incubation. After 168 h long-term co-incubation, motility analysis revealed a negative (P = 0.026) impact of 105 CFU/mL L. buchneri DSM 32407. A concentration- and storage-dependent effect is particularly obvious (P < 0.001) using 106 CFU/mL L. buchneri DSM 32407. Most notably, the thermo-resistance (TRT) for 106 CFU/mL L. buchneri DSM 32407 (P = 0.001) was inferior to BTS with and without gentamicin after 72 and 168 h of semen co-incubation. The supplementation of 105 CFU/mL L. buchneri DSM 32407 impaired progressive motility to a lesser extent. The percentage of mitochondrially active spermatozoa after 96 h (P = 0.009) and membrane-intact spermatozoa after 168 h (P < 0.001) was lower when 106 CFU/mL L. buchneri DSM 32407 were suspended compared with all other groups. Finally, the addition of L. buchneri DSM 32407 to BTS-extended boar semen had no competitive effect on the total amount of bacteria 48 h after co-incubation. In summary, the present study demonstrated that there are Lactobacillus species present in the porcine seminal plasma, which can be cultivated using standard procedures. However, long-term co-incubation of lactic acid bacteria with spermatozoa had a negative influence on spermatozoa.
Highlights
In the swine industry, artificial insemination (AI) is a common reproductive technology to apply spermatozoa to the female reproductive tract
Ejaculates from 81 (73%) mature Pietrain boars out of a total number of 111 collected samples were culture-negative under selective conditions for lactobacilli
Bacteria, which grow in sufficient amount for subsequent experiments, were subjected to a PCR using phylogenetic 16S rDNA primers
Summary
Artificial insemination (AI) is a common reproductive technology to apply spermatozoa to the female reproductive tract. Due to the natural bacterial content of fresh boar semen, measures to control bacterial growth during storage of insemination doses are necessary. This reduces the risk of potential bacterial contamination of the uterus. All ejaculates collected from healthy donors contain bacteria stemming from natural occurrence within the male reproductive tract. Bacterial load in raw semen commonly ranges between 104 and 106 CFU/mL [1]. In addition to animal origins, bacteria from the environment may contaminate semen during collection or processing in the laboratory [2]. Consequences of bacterial contamination predominantly reside in loss of sperm motility, and induction of sperm agglutination and membrane damage, resulting in poor fertility and high economic losses in sow herds [3,4]
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