Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Boar semen can tolerate rapid cooling rates prior to freezing

Jorge D. Juarez A B , Inma Parrilla A , Juan M. Vazquez A , Emilio A. Martinez A and Jordi Roca A C

A Department of Medicine and Animal Surgery, Faculty of Veterinary Science, University of Murcia, E-30100 Murcia, Spain.

B Department of Livestock Science, Universidad Nacional Agraria de la Selva, Tingo María, Huánuco, Perú.

C Corresponding author. Email: roca@um.es

Reproduction, Fertility and Development 23(5) 681-690 http://dx.doi.org/10.1071/RD10216
Submitted: 6 September 2010  Accepted: 27 January 2011   Published: 17 May 2011

Abstract

Two experiments were performed in the present study that demonstrated that boar spermatozoa are capable of surviving rapid cooling rates within a range of 15–5°C before freezing. Boar ejaculates diluted in Beltsville thawing solution (BTS) (1 : 1, v/v) were held at 17–20°C and shipped over a 24-h time period from two AI centres to a cryobiology laboratory, where they were pooled (Experiment 1) or cryopreserved individually (Experiment 2) using a standard 0.5-mL straw freezing protocol. The effects of cooling before freezing were assessed after thawing through the objective evaluation of sperm motility and flow cytometric analysis of membrane integrity, acrosomal status, changes in membrane lipid architecture monitored by merocyanine and annexin V binding and intracellular production of reactive oxygen species. In Experiment 1 (six replicates), two semen pools (five ejaculates per pool) were cooled from 15 to 5°C at rates of 0.08, 0.13, 0.40 and 1.50°C min–1. These cooling rates did not result in any significant differences (P > 0.05) in any of the post-thaw sperm assessments, even in thawed samples incubated under capacitation conditions. In Experiment 2, three individual ejaculates from 16 boars were slowly (0.08°C min–1) or rapidly (1.5°C min–1) cooled before freezing. A consistent interboar variability (P < 0.01) was detected, which was independent of the cooling rate used. Cooling rate only significantly influenced (P < 0.05) sperm assessments in four of 16 boars, which exhibited slightly higher percentages of motile cells and intact plasma and acrosomal membranes in the samples that had been cooled slowly. These findings demonstrate that boar spermatozoa undergoing cryopreservation can withstand rapid cooling rates before freezing.

Additional keywords: flow cytometry, freezability, spermatozoa.


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