Differences in boar sperm head shape and dimensions recorded by computer-assisted sperm morphometry are not related to chromatin integrity

Our aim was to reduce heterogeneity and microbiological contamination risk on small ruminant semen cryopreservation by replacement of fresh egg yolk by pasteurized powdered egg yolk, assessing simultaneously the effect of the donor age (1 year v. 2 years old) on sperm head morphometry of fresh and thawed sperm. Briefly, fresh ejaculates from 8 rams and from 8 bucks were collected in autumn during 2 consecutive years. Immediately after collection, ejaculates from each species were mixed in equal quantities, and pooled semen was centrifuged twice (600 g for 10 min). Then the pellet was split into 2 aliquots and resuspended in an extender containing 15% (v/v) of fresh or powdered egg yolk supplemented with 5% glycerol in a Tris-based medium. Afterward, sperm samples were refrigerated at 5°C for 4 h before being frozen in nitrogen liquid vapours. Buck and ram sperm-head morphometry was analysed by computer-assisted sperm analysis (ISAS®) on fresh and thawed sperm previously stained with Diff Quick®, and the data were analysed using ANOVA (mean ± s.e., n = 6). From ram sperm studies, no differences were found between fresh and post-thaw sperm, neither between egg yolk-based extenders or donor ages, showing the following mean values of head length (8.4 ± 0.0 μm), width (4.9 ± 0.0 μm), area (34.2 ± 0.1 μm2), perimeter (23.4 ± 0.1 μm), ellipticity (1.7 ± 0.0), elongation (0.2 ± 0.0), rugosity (0.8 ± 0.0), and regularity (0.9 ± 0.0). Likewise buck semen did not show significant differences on sperm-head dimensions after cryopreservation, only on head-shape parameters as ellipticity, elongation, and regularity between fresh and thawed sperm from 2-year-old donors, independently of egg yolk used as cryoprotectant. However, the age of the buck had a significant effect on all assessed morphometric parameters in fresh and thawed sperms, except regularity (Table 1). Our results suggest that, from a morphometric point of view, the powdered egg yolk can be used satisfactory on small ruminant sperm cryopreservation, but in goats, the head changes due to the donor age should be considered. Table 1.Effect of fresh v. powdered egg yolk (EY) in the extender and donor age on fresh and thawed buck-sperm morphometry Research supported by INIA (RZ2009–00008–00–00), Generalitat de Catalunya (2009SGR0621 and CUR-DIUE), and FSE and Fundación Carolina.

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58 EFFECT OF MELATONIN IMPLANT ON BLANCA de RASQUERA BUCKS DURING SPRING ON SPERM MORPHOMETRY BEFORE AND AFTER CRYOPRESERVATION

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab58 ◽

2015 ◽

Vol 27 (1) ◽

pp. 122

Author(s):

A. Tabarez ◽

W. García ◽

M. J. Palomo

Keyword(s):

Egg Yolk ◽

Sperm Head ◽

Head Width ◽

Computer Assisted ◽

Sperm Cryopreservation ◽

Sperm Analysis ◽

Know How ◽

Before And After ◽

Sperm Morphometry ◽

Computer Assisted Sperm Analysis

In order to improve sperm cryopreservation throughout the year and accelerate the process of preservation of this Catalonian goat breed in extinction danger, we proposed to assess the effect of melatonin implant application in Blanca de Rasquera males during spring on sperm head morphometry of fresh and thawed sperm. Therefore 8 bucks of 30 months old approximately were divided into 2 groups. In one of the groups, 2 melatonin implants (Melovine®, CEVA) were inserted into bucks 60 days before starting the collection of semen, and the other group was kept untreated. Briefly, fresh ejaculates from each group of 4 bucks were collected in spring, immediately mixed in equal quantities, and centrifuged twice (600 g for 10 min). Then the pellet was resuspended in a Tris-based medium containing 15% (v/v) of powdered egg yolk supplemented with 5% glycerol. Afterward, sperm samples were refrigerated at 5°C for 4 h before being frozen in LN vapour. Buck sperm head morphometry was analysed by computer-assisted sperm analysis (ISAS®) on fresh and thawed sperm previously stained with Diff Quick®. Data were analysed by GLM multivariate procedure (IBM SPSS, 2011; mean ± s.e., n = 6), showing significant differences among treatments in all the morphometric parameters except head perimeter and rugosity (Table 1). Our results suggest that melatonin application in bucks increases the ellipticity and elongation of fresh and thawed sperm, meanwhile the cryopreservation process reduces both parameters. Likewise melatonin implants increase significantly the head length only on thawed sperms as cryopreservation process increases the head width, area in sperms from implanted males and regularity only in sperms from nonimplanted bucks. These head changes on fresh and thawed sperm morphometry should be deeply investigated in order to know how they could affect sperm cryosurvival and fertility. Table 1.Effect of melatonin implant on Blanca de Rasquera bucks during spring on morphometry of fresh and thawed sperm This research was supported by INIA (RZ2009–00008–00–00), Generalitat de Catalunya (2009SGR0621 and CUR-DIUE), and FSE and Fundación Carolina.

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Predictive Capacity of Boar Sperm Morphometry and Morphometric Sub-Populations on Reproductive Success after Artificial Insemination

Animals ◽

10.3390/ani11040920 ◽

2021 ◽

Vol 11 (4) ◽

pp. 920

Author(s):

Vinicio Barquero ◽

Eduardo R. S. Roldan ◽

Carles Soler ◽

Jesús L. Yániz ◽

Marlen Camacho ◽

...

Keyword(s):

Reproductive Success ◽

Litter Size ◽

Artificial Insemination ◽

Sperm Head ◽

Computer Assisted ◽

Operating Characteristics ◽

Head Shape ◽

Roc Curve Analysis ◽

Predictive Capacity ◽

Shape And Size

The aim of the study was to compare the morphometric features of sperm head size and shape from the Pietrain line and the Duroc × Pietrain boar crossbred terminal lines, and to evaluate their relationship with reproductive success after artificial insemination of sows produced from crossbreeding the York, Landrace and Pietrain breeds. Semen samples were collected from 11 sexually mature boars. Only ejaculates with greater than 70% motility rate and < 15% of abnormal sperm were used for artificial inseminations (AI) and included in the study. Samples were analyzed using an ISAS®v1 computer-assisted sperm analysis system for eight morphometric parameters of head shape and size (CASA-Morph). Sub-populations of morphometric ejaculates were characterized using multivariate procedures, such as principal component (PC) analysis and clustering methods (k-means model). Four different ejaculate sub-populations were identified from two PCs that involved the head shape and size of the spermatozoa. The discriminant ability of the different morphometric sperm variables to predict sow litter size was analyzed using a receiver operating characteristics (ROC) curve analysis. Sperm head length, ellipticity, elongation, and regularity showed significant predictive capacity on litter size (0.59, 0.59, 0.60, and 0.56 area under curve (AUC), respectively). The morphometric sperm sub-populations were not related to sow litter size.

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Season of collection and sperm head shape impacts expression of CARHSP and FTL from motile-rich boar sperm

Agri Gene ◽

10.1016/j.aggene.2017.10.002 ◽

2018 ◽

Vol 7 ◽

pp. 1-6 ◽

Cited By ~ 4

Author(s):

L.A. Rempel ◽

M.M. Krautkramer ◽

T.M. Loether ◽

J.J. Parrish ◽

J.R. Miles

Keyword(s):

Sperm Head ◽

Head Shape ◽

Boar Sperm

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309 MORPHOMETRICAL ANALYSIS OF THE LAMA GLAMA SPERM HEAD

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab309 ◽

2010 ◽

Vol 22 (1) ◽

pp. 310

Author(s):

C. I. Casaretto ◽

D. Lombardo ◽

S. Giuliano ◽

M. Gambarotta ◽

M. I. Carretero ◽

...

Keyword(s):

Artificial Insemination ◽

Companion Animals ◽

Sperm Head ◽

Computer Assisted ◽

Equivalent Circle Diameter ◽

Curve Length ◽

Length Width ◽

Sperm Morphometry ◽

Lama Glama ◽

Circle Diameter

During the last decades, the interest in breeding South American Camelids has increased, not only as companion animals but also for their high- quality fiber. Although several studies have been carried out on artificial insemination in Lama glama, this technique has not been widely applied in reproductive programs, principally due to the difficulty in collecting raw semen from males and the lack of knowledge about freezing/thawing techniques and semen characteristics. The aim of the present study was to objectively characterize llama sperm morphometry by a computer-assisted system, thereby increasing the knowledge on male llama physiology, leading to further developement of reproductive biotechnologies such as artificial insemination. Five semen samples were obtained by electroejaculation from each of 8 males, 6- to 10-year-old llamas of proven fertility. Smears were prepared from each sample and stained with Tinción 15® (Biopur S.R.L., Rosario, Argentina) and observed at x 1000 magnification. Images of sperm heads were captured by a Leica DC180 camera (Leica Microsystems Co., Wetzlar, Germany), obtaining 200 images from each sample. Binary images were obtained and area, length, width, equivalent circle diameter, curve length, curve width, perimeter, convex perimeter, roundness, and elongation were measured using QWin Plus (Leica Microsystems Co.). A total of 8005 sperm heads were measured. Descriptive statistics of the complete population was performed, with the following results obtained (mean ± SD): area (μm2) 20.09 ± 0.6, length (μm) 6.6 ± 0.3, width (μm) 4.14 ± 0.1, equivalent diameter ((μm) 5.06 ± 0.1, curve length (μm) 5.8 ± 0.3, curve width (μm) 3.48 ± 0.3, perimeter (μm) 18.54 ± 0.1, convex perimeter (μm) 17.34 ± 0.3, roundness 1.28 ± 0.04, and elongation 1.6 ± 0.01. Coefficients of variation were between 0.47 and 8.72%. A design considering the male as a fixed factor and the ejaculate as a nested factor was used for the purpose of identifying differences in morphometry between ejaculates of the same male and/or between males. Normality was tested using the Kolmogorov test. Significant differences between ejaculates of some males were found for curve length, curve width, perimeter, roundness, and elongation (P < 0.05). There were no intra-male differences for sperm head area, length, width, equivalent circle diameter, and convex perimeter. Of the parameters, there were significant differences between males for sperm area, length, equivalent circle diameter, and convex perimeter (P < 0.05). The differences found in sperm morphometry confirm the great polymorphism observed when subjectively evaluating llama semen morphology and make the establishment of a single pattern of normal llama sperm morphometry impossible.

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Semi-automatic morphometric analysis of normal boar sperm head

Animal Reproduction Science ◽

10.1016/j.anireprosci.2016.03.045 ◽

2016 ◽

Vol 169 ◽

pp. 112 ◽

Cited By ~ 1

Author(s):

A.M. Syanda ◽

G. Boe-Hansen ◽

N. Satake

Keyword(s):

Morphometric Analysis ◽

Sperm Head ◽

Boar Sperm

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Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and on developmental ability in vitro and in vivo after intracytoplasmic sperm head injection

Zygote ◽

10.1017/s0967199406003935 ◽

2007 ◽

Vol 15 (1) ◽

pp. 15-24 ◽

Cited By ~ 38

Author(s):

M. Nakai ◽

N. Kashiwazaki ◽

A. Takizawa ◽

N. Maedomari ◽

M. Ozawa ◽

...

Keyword(s):

Dna Fragmentation ◽

Freeze Drying ◽

Chelating Agents ◽

Sperm Head ◽

Control Group ◽

Sperm Dna Fragmentation ◽

Freeze Dried ◽

Boar Sperm

SUMMARYSuccessful offspring production after intracytoplasmic injection of freeze-dried sperm has been reported in laboratory animals but not in domesticated livestock, including pigs. The integrity of the DNA in the freeze-dried sperm is reported to affect embryogenesis. Release of endonucleases from the sperm is one of the causes of induction of sperm DNA fragmentation. We examined the effects of chelating agents, which inhibit the activation of such enzymes, on DNA fragmentation in freeze-dried sperm and on the in vitro and in vivo developmental ability of porcine oocytes following boar sperm head injection. Boar ejaculated sperm were sonicated, suspended in buffer supplemented with (1) 50 mM EGTA, (2) 50 mM EDTA, (3) 10 mM EDTA, or (4) no chelating agent and freeze-dried. A fertilization medium (Pig-FM) was used as a control. The rehydrated spermatozoa in each group were then incubated in Pig-FM at room temperature. The rate of DNA fragmentation in the control group, as assessed by the TUNEL method, increased gradually as time after rehydration elapsed (2.8% at 0 min to 12.2% at 180 min). However, the rates in all experimental groups (1–4) did not increase, even at 180 min (0.7–4.1%), which were all significantly lower (p < 0.05) than that of the control group. The rate of blastocyst formation after the injection in the control group (6.0%) was significantly lower (p < 0.05) than those in the 50 mM EGTA (23.1%) and 10 mM EDTA (22.6%) groups incubated for 120–180 min. The average number of blastocyst cells in the 50 mM EGTA group (33.1 cells) was significantly higher (p < 0.05) than that in the 10 mM EDTA group (17.8 cells). Finally, we transferred oocytes from 50 mM EGTA or control groups incubated for 0–60 min into estrous-synchronized recipients. The two recipients of the control oocytes became pregnant and one miscarried two fetuses on day 39.The results suggested that fragmentation of DNA in freeze-dried boar sperm is one of the causes of decreased in vitro developmental ability of injected oocytes to the blastocyst stage. Supplementation with EGTA in a freeze-drying buffer improves this ability.

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A mutation study of sperm head shape and motility in the mouse: lessons for the clinic

Andrology ◽

10.1111/andr.300 ◽

2014 ◽

Vol 3 (2) ◽

pp. 174-202 ◽

Cited By ~ 23

Author(s):

P. de Boer ◽

M. de Vries ◽

L. Ramos

Keyword(s):

Sperm Head ◽

Head Shape

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The effects of inbreeding on sperm morphometry of captive-bred endangered mammals

Canadian Journal of Zoology ◽

10.1139/cjz-2016-0291 ◽

2017 ◽

Vol 95 (8) ◽

pp. 599-606 ◽

Cited By ~ 3

Author(s):

M. Lawrence ◽

G. Mastromonaco ◽

K. Goodrowe ◽

R.M. Santymire ◽

W. Waddell ◽

...

Keyword(s):

Captive Breeding ◽

Negative Relationship ◽

Tail Length ◽

Poor Quality ◽

Sperm Head ◽

Head Width ◽

Canis Rufus ◽

Sperm Tail ◽

Sperm Morphometry ◽

Red Wolves

Captive breeding is used for the conservation of endangered species, but inbreeding can result when a small number of founders are used to establish populations. Inbreeding can reduce the proportion of normal sperm in an ejaculate, but may also have effects on sperm size and shape (morphometry). We investigated the effects of inbreeding on sperm morphometry of black-footed ferrets (Mustela nigripes (Audubon and Bachman, 1851)) and red wolves (Canis rufus Audubon and Bachman, 1851) from captive breeding programs to determine if more inbred males produced sperm of poor quality (bulky head, small midpiece, short tail). We measured sperm head length, head width, midpiece length, midpiece width, and tail length on 10 sperm from each male of both species. A negative relationship between variation in sperm tail length and inbreeding coefficient (f) was found in black-footed ferret, suggesting that more inbred individuals will have reduced genetic and phenotypic variation. Analyses indicated a negative relationship between sperm head width and f and a positive relationship between sperm tail length and f in red wolf, suggesting that more inbred male red wolves could have faster sperm. These results indicate that inbreeding affects functionally important aspects of sperm morphometry, but that these effects may not be entirely negative.

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