Intensity and Duration of Vibration Emissions during Shipping as Interacting Factors on the Quality of Boar Semen Extended in Beltsville Thawing Solution.

Vibration emissions during the transport of boar semen for artificial insemination (AI) affect sperm quality. In the present study, the common influence of the following factors was investigated: vibrations (displacement index (Di) = 0.5 to 6.0), duration of transport (0 to 12 h) and storage time (days 1 to 4). Normospermic ejaculates were collected from 39 fertile Pietrain boars (aged 18.6 ± 4.5 months) and diluted in a one-step procedure with an isothermic (32 °C) BTS (Minitüb) extender (n = 546 samples). Sperm concentration was adjusted to 22 × 106 sperm·mL-1. Extended semen (85 ± 1 mL) was filled into 95 mL QuickTip Flexitubes (Minitüb). For transport simulation on day 0, a laboratory shaker IKA MTS 4 was used. Total sperm motility (TSM) was evaluated on days 1 to 4. Thermo-resistance test (TRT), mitochondrial activity (MITO) and plasma membrane integrity (PMI) were assessed on day 4. Sperm quality dropped with increasing vibration intensity and transport duration, and the effect was enhanced by a longer storage time. A linear regression was performed using a mixed model, accounting for the boar as a random effect. The interaction between Di and transport duration significantly (p < 0.001) explained data for TSM (-0.30 ± 0.03%), TRT (-0.39 ± 0.06%), MITO (-0.45 ± 0.06%) and PMI (-0.43 ± 0.05%). Additionally, TSM decreased by 0.66 ± 0.08% with each day of storage (p < 0.001). It can be concluded that boar semen extended in BTS should be transported carefully. If this is not possible or the semen doses are transported a long way, the storage time should be reduced to a minimum.

Relations between the time of ovulation and fecal estrogen concentration in sows.

Artificial insemination (AI) is the most important biotechnology in pig reproduction. To achieve the best possible fertility results, appropriate timing of the insemination is essential. The optimal time for AI is 12 h before to 4 h after ovulation. This time-frame, unlike in estrus, is not recognizable through external indicators. It would, therefore, be beneficial to find simple and economical methods that support manual estrus checks and are able to determine the time of ovulation more accurately. On this basis, starting 80 h after weaning, 14 DanBred sows (parity: 5.2 ± 2.4) were checked for ovulation via ultrasound scans every 8 h over a period of 72 h. Additionally, rectal fecal samples were taken and analyzed for their estrogen concentration to assess possible relations to ovulation time. On average, sows ovulated 121 ± 10 h after weaning and 16 ± 9 h after onset of heat. There was a prominent drop in fecal estrogen levels 4 h before ovulation when compared to almost all other points in time (before ovulation: 20 h (P = 0.056), 12 h (P = 0.006); after ovulation: 4 h and 12 h (P < 0.001)). There are, however, significant differences in the sow-individual fecal estrogen concentrations for which several influencing factors must be considered.

Comparison of NUCLEOCOUNTER, ANDROVISION with Leja chambers and the newly developed ANDROVISION eFlow for sperm concentration analysis in boars.

Exact analysis of sperm concentration in raw and diluted semen is of major importance in swine artificial insemination, as sperm concentration is one of the most important characteristics of an ejaculate determining the value of the ejaculate and the productive life of the boar. The study compares different methods for sperm concentration analysis in raw and diluted boar semen: NUCLEOCOUNTER SP-100, the ANDROVISION with Leja chambers and the new ANDROVISION eFlow system. The Concordance Correlation Coefficient (CCC) between NUCLEOCOUNTER and ANDROVISION eFlow was 0.955 for raw (n = 185 ejaculates) and 0.94 for diluted semen (n = 109 ejaculates). The CCC between NUCLEOCOUNTER and ANDROVISION with Leja chambers was 0.66. A Bland-Altman plot of split-sample measurements of sperm concentration with NUCLEOCOUNTER and ANDROVISION eFlow showed that 95.1% of all measurements lay within ± 1.96 standard deviation. The coefficients of variance were 1.6 ± 1.3%, 3.6 ± 3.6% and 7.3 ± 6.3% for NUCLEOCOUNTER, ANDROVISION eFlow and ANDROVISION with Leja chambers in diluted semen, respectively. NUCLEOCOUNTER and ANDROVISION eFlow are comparable tools to measure the concentration of raw and diluted boar semen. In comparison to ANDROVISION with Leja chambers, concentration analyses of diluted semen using NUCLEOCOUNTER or ANDROVISION eFlow show a higher repeatability within and a higher concordance between the methods.

Boar Semen Shipping for Artificial Insemination: Current Status and Analysis of Transport Conditions with a Major Focus on Vibration Emissions.

In the modern pig reproduction system, artificial insemination (AI) doses are delivered from AI centers to sow farms via logistics vehicles. In this study, six breeding companies in three countries (Brazil, Germany, and the USA) were interviewed about their delivery process. It was found that there is currently no comprehensive monitoring system for the delivery of semen. The entire process "shipping of boar semen" was documented using Business Process Model and Notation (BPMN). Although it is not currently known which vibrations occur at all, it is suspected that vibration emissions affect the quality of boar semen. For this reason, a prototype of a measuring system was developed to calculate a displacement index (Di), representing vibration intensities. Vibrations were analyzed in standardized road trials (n = 120) on several road types (A: smooth asphalt pavement, B: rough asphalt pavement, C: cobblestone, and D: dirt road) with different speeds (30, 60, 90, 120, and 150 km/h). A two-way ANOVA showed significant differences in mean Di, depending on road surface and speed as well as an interaction of both factors (p < 0.001). A field study on a reference delivery from a German AI center to several sow farms indicated that 33% of the observed roads are in good quality and generate only a few vibrations (Di ≤ 1), while 40% are of a moderate quality with interrupted surfaces (Di = 1−1.5). However, 25% of the roads show markedly increased vibrations (Di ≥ 1.5), as a consequence of bad conditions on cobblestones or unpaved roads. Overall, more attention should be paid to factors affecting sperm quality during transport. In the future, an Internet of Things (IoT) based solution could enable complete monitoring of the entire transport process in real time, which could influence the courier's driving behavior based on road conditions in order to maintain the quality of the transported AI doses.

Detection and characterization of Lactobacillus spp. in the porcine seminal plasma and their influence on boar semen quality.

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.