Use of manual stimulation for collection of semen from an atactic stallion unable to mount

A 9-year-old atactic breeding stallion was trained to ejaculate, with only manual stimulation, while standing on the ground. Ejaculates obtained yielded fertile semen with morphologic and motility characteristics within the range for normal stallions. This method extended the breeding life of a stallion unable to mount a live or dummy mare or to ejaculate into an artificial vagina while standing on the ground.     

采精室的建立

本文从公猪的训练到器具的使用,介绍了巴西的一个育种群从人工授精公猪采精着手的实践经验. 在2007年末,巴西南部的Suinosul农场与当地的-个猪肉加工厂签订了-份协议,以保证750头F1代母猪的正常生产,并将22 kg活重的仔猪出售给加工厂,在其它养猪场进行育肥.这一协议给农场带来了重大变化.尽管农场从1978年起即开始饲养种畜出售给其他独立经营的农场,但一直采用自然交配的方式进行繁育.为满足新情况的需要,我们决定对终端公猪采用人工授精(AI)的方法进行配种.     

Semen collection in the ostrich (Struthio camelus) using a dummy or a teaser female

1. We investigated the feasibility of training male ostriches to ejaculate into an artificial cloaca (AC) using a teaser female or a dummy. 2. Male ostriches develop desirable behaviour patterns that allow them to be trained to the teaser and the dummy female. Training success can be high as both methods rely on natural stimulation and voluntary ejaculation into the AC provided suitable individuals are identified. 3. The key factors to successful training appear to be: the teaser method - temperament of male ostriches, the crouching behaviour of the female ostriches for the semen collector and acceptance of the collector by the male; dummy method - courtship of humans, learning ability of ostriches to habituate and then to mount a dummy. 4. Both methods are reliable, yield ejaculates of high quality and give reproducible results. The ejaculate volume was 1.09 +/- 0.13 ml, the concentration of spermatozoa 4.21 +/- 0.27 x 10(9)/ml, the total number of spermatozoa 4.67 +/- 0.62 x 10(9) and motility 4.3 +/- 0.1. 5. Commercial ostrich production that relies on natural mating can take advantage of those methods to develop artificial insemination technologies.     

Evaluation of a portable device for assessment of motility in stallion semen

In horse breeding, quality assessment of semen before insemination is often requested. Non‐laboratory‐based techniques for objective analysis of sperm motility are thus of interest. The aim of this study was evaluating a portable device for semen analysis (Ongo sperm test) and its comparison with computer‐assisted semen analysis (CASA). Semen was collected from 10 stallions, diluted to 100, 50 and 25 × 10⁶ sperm/ml and analysed for total (TM) and progressive motility (PM). The final sperm concentration influenced total motility analysed by Ongo (p < 0.05) which was higher at 100 × 10⁶ sperm/ml when compared to 25 × 10⁶ sperm/ml (p < 0.05) but not when compared to 50 × 10⁶ sperm/ml (n.s.). Sperm concentration did not influence total motility when assessed by SpermVision (n.s.). Agreement between methods was evaluated by correlation analysis and Bland–Altman plot. Intra‐assay variation of Ongo was 5.2% ± 3.0 for TM and 6.9% ± 3.4 for PM. Correlation between Ongo and CASA was r = 0.79, 0.88 and 0.83 for 100, 50 and 25 × 10⁶ sperm/ml for TM, and r = 0.87, 0.89 and 0.87 for PM, respectively (all p < 0.001). At the 100 and 25 mio/ml dilutions, the difference between the two systems deviated significantly from 0, while no such bias existed at the 50 mio/ml dilution (TM Ongo 85.0%, CASA 82.3%; PM Ongo 64.1%, CASA 66.1%). The 95% confidence interval was 19.9%, 18.9% and 19.2% ± mean for TM and 20.7%, 17.4% and 20.3% ± mean for 100, 50 and 25 × 10⁶ sperm/ml, respectively. In conclusion, Ongo sperm test sperm motility data were strongly correlated with data obtained by CASA. In addition, at a concentration of 50 × 10⁶ sperm/ml values measured with both systems were close to identical. At this concentration, which is recommended in equine AI, Ongo and CASA can be used interchangeably.     

A comparison of electro-ejaculation with the artificial vagina for ram semen collection

Nine fertile three-year-old Romney rams were ejaculated both by means of electro-ejaculation and by the artificial vagina. The ram repeatabilities of semen volume, concentration, total sperm number, percentage normal sperm and wave motion score of ejaculates collected by electro-ejaculation were 0.17, 0.08, 0.12, 0.61 and 0.23 respectively compared with the repeatabilities of ejaculates collected by the artificial vagina of 0.28,0.79, 0.56,0.72 and 0.54. The repeatability results showed that it was possible to rank rams on the quantity of sperm produced in a single ejaculate obtained by an artificial vagina, but not on the quantity obtained by electro-ejaculation. The correlation co-efficients between the ram means for the two techniques were for volume 0.05, concentration 0.06, total sperm number -0.21, percentage normal sperm 0.73, and wave motion score 0.35. Only the percentage normal sperm coefficient was significant.     

Intracellular calcium chelating agent (BAPTA‐AM) aids stallion semen cooling and freezing–thawing

This study aimed to investigate the effects of different concentrations of 1,2‐bis‐(o‐aminophenoxy)‐ethane‐N,N,N0 N0‐tetraacetic acid, tetra‐acetoxymethyl ester (BAPTA‐AM), an intracellular calcium chelating agent, on stallion semen cooling and freezing–thawing. After collection, semen was extended (1:1 v/v) on a skim milk‐based extender, centrifuged and resuspended at 400 million/ml into cooling or freezing extenders containing 0, 5, 25, 50, 100 and 200 μΜ BAPTA‐AM. Motility parameters were assessed after cooling in Equitainer at 5°C for 12, 24, 48, 72 and 120 hr and after freezing–thawing. In addition, mitochondrial membrane potential, intracellular ATP, reactive oxygen species and malondialdehyde concentrations were measured in cryopreserved‐thawed semen. Cooled stored (48 hr) semen containing 50 μΜ BAPTA‐AM and control extender (0 μΜ BAPTA‐AM) was used to assess fertility. Inclusion of 50 μΜ BAPTA‐AM resulted in superior sperm motility parameters during cooled storage when compared to other groups (p < 0.05). Furthermore, semen cryopreserved in extender containing 50 μΜ BAPTA‐AM showed increased intracellular ATP and mitochondrial membrane potential, whereas reactive oxygen species and malondialdehyde were increased after thawing for all groups (p < 0.05). Addition of 50 μΜ BAPTA‐AM to cooling extender resulted in similar pregnancy rates to the control group (75% vs. 73.6%, respectively; p > 0.05). In conclusion, the addition of BAPTA‐AM to semen extenders aided stallion semen cryopreservation in a dose‐dependent manner. Furthermore, the cooling extender supplemented with 50 μΜ BAPTA‐AM could be used to prolong the sperm motility during cooling without apparently compromising fertility. Field trials should be conducted to assess fertility of cryopreserved stallion semen with BAPTA‐AM.     

Effect of presence or absence of antibiotics and use of modified single layer centrifugation on bacteria in pony stallion semen

Bacteria contaminate semen during collection and handling. The objective of this study was to identify the bacteria in pony stallion semen, the effects of antibiotics included in commercial semen extenders (lincomycin and spectinomycin) and the effect of modified single layer centrifugation (MSLC), on bacterial load. Ejaculates from six pony stallions, 3 ejaculates per animal, were extended in EquiPlus extender either with or without antibiotics. Aliquots were processed by MSLC to form four treatment groups: control and MSLC with antibiotics (CA and SA, respectively) and control and MSLC without antibiotics (CW and SW, respectively). Bacteriological examinations were carried out within 2 hr. Thirty‐one species of bacteria were isolated from one or more ejaculates, with Corynebacterium spp. being the most frequently detected. Corynebacterium spp. were present in all ejaculates. The MSLC resulted in a significantly lower total bacterial count than controls (CA vs. SA, p < 0.001; CW vs. SW, p < 0.0001).