Improvement of llama (Lama glama) seminal characteristics using collagenase.

Anim Reprod Sci

Area de Física Biológica, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Chorroarín 280, 1427 Ciudad Autónoma de Buenos Aires, Argentina.

Published: March 2010

AI Article Synopsis

  • Llama semen traditionally has high structural viscosity and low sperm motility, which makes assisted reproductive techniques challenging.
  • Research evaluated how different dilutions and incubation times with H-TALP-BSA medium, with or without collagenase, affected semen quality for fertility procedures.
  • Results indicated that collagenase treatment improved sperm motility and separation from seminal plasma while enhancing functional sperm membranes, suggesting better prospects for assisted reproduction in llamas.

Article Abstract

Llama semen is characterized by great structural viscosity and minimal sperm progressive motility. These characteristics, inherent to South American Camelid ejaculates, have slowed down the development of assisted reproductive techniques in these species. The aim of the present research was to evaluate the effect of different combinations of dilutions and incubation time with H-TALP-BSA medium, with and without adding 0.1% collagenase, on the qualitative and quantitative semen characteristics, for its use in assisted fertility techniques. Ejaculates (n=8; r=3) were obtained using electroejaculation. Each ejaculate was evaluated and then split into four aliquots. Two of these were diluted 4:1 and 8:1 in 0.1% collagenase in H-TALP-BSA (treatments 1 and 3) and the other two 4:1 and 8:1 in H-TALP-BSA without collagenase (treatments 2 and 4). Treatments 1 and 2 were incubated 4 min at 37 degrees C while treatments 3 and 4 were incubated 8 min. All aliquots were centrifuged at 800 x g for 4 min immediately after incubation. Supernatants were pipetted to observe thread formation and pellets were re-diluted in H-TALP-BSA. Supernatants from samples treated with collagenase did not form a thread when pipetted, while the ones from samples that were not treated with the enzyme did. Only semen samples treated with collagenase showed progressive sperm motility, with averages over 40%. There were no significant differences (P>0.05) for the percentage of live spermatozoa and for the percentage of detached heads between raw and treated semen samples. Percentages of spermatozoa with functional membranes were significantly higher (P< or =0.05) in samples treated with collagenase than in raw semen and in samples incubated without collagenase. These results show that treating semen with 0.1% collagenase in H-TALP-BSA improves semen rheological properties while facilitates the separation of spermatozoa from seminal plasma in llama; it also promotes sperm progressive motility, while maintaining sperm membrane functionality and integrity. Consequently, this protocol could be used for in vitro llama embryo production with ejaculated spermatozoa.

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http://dx.doi.org/10.1016/j.anireprosci.2009.06.005DOI Listing

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