309 MORPHOMETRICAL ANALYSIS OF THE LAMA GLAMA SPERM HEAD

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.

74 EFFECT OF POWDERED EGG YOLK IN THE EXTENDER AND DONOR AGE ON FRESH AND THAWED SPERM MORPHOMETRY IN SMALL RUMINANTS

2015 ◽  
Vol 27 (1) ◽  
pp. 130
Author(s):  
M. J. Palomo ◽  
W. Garcia ◽  
A. Tabarez
Keyword(s):  
Small Ruminant ◽  
Small Ruminants ◽  
Egg Yolk ◽  
Sperm Head ◽  
Computer Assisted ◽  
Donor Age ◽  
Mean Values ◽  
Sperm Analysis ◽  
Sperm Morphometry ◽  
Ram Sperm

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.

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

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.

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

Animals ◽  
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.

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

2007 ◽  
Vol 68 (2) ◽  
pp. 196-203 ◽  
Author(s):  
F. Saravia ◽  
I. Núñez-Martínez ◽  
J.M. Morán ◽  
C. Soler ◽  
A. Muriel ◽  
...  
Keyword(s):  
Sperm Head ◽  
Computer Assisted ◽  
Head Shape ◽  
Boar Sperm ◽  
Sperm Morphometry ◽  
Chromatin Integrity

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

2017 ◽  
Vol 95 (8) ◽  
pp. 599-606 ◽  
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.

84 EVALUATION OF BULL SEMEN AT 1 H VS. 48 H OF POST-FROZEN STABILIZATION TIME

2006 ◽  
Vol 18 (2) ◽  
pp. 150
Author(s):  
G. M. Brogliatti ◽  
G. Larraburu ◽  
R. Cavia ◽  
M. E. Carini
Keyword(s):  
Liquid Nitrogen ◽  
Artificial Insemination ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Stabilization Time ◽  
Bull Semen ◽  
Straight Line ◽  
Semen Extender ◽  
Lateral Head

The process of cryopreservation of bull semen in liquid nitrogen at −196°C is usually carried out after 3 to 6 h of refrigeration at 4°C post-collection. To guarantee the quality of the final product, the frozen straws are evaluated after cryopreservation. The seminal samples are usually stabilized during 48 h before being analyzed (Hafez, Reproduction and Artificial Insemination in Animals, 1989); this would retard the possible commercialization. The objective of the present study was to determine motility parameters and viability of semen doses stabilized by 1 h or more than 48 h in liquid nitrogen at −196°C. A total of 122 ejaculated from 23 different adult bulls (Angus, Brangus, Braford, and Hereford) were evaluated in an artificial insemination center between January and April 2005. The semen was diluted in a semi-defined semen extender (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, France). Parameters of velocity average path (VAP, μm/s), velocity straight line (VSL, µm/s), amplitude lateral head (ALH, µm), linearity (LIN, %), percentage of rapid cells (RAPID, %), and viability (VIA, %) were determined by Computer Assisted Semen Analysis (CASA, HTM-ceros 12.1, Berkeley, CA, USA). The obtained results were analyzed statistically with T Student and are summarized in Table 1. The results indicate that there is no difference in the velocity of the spermatozoa evaluated 1 h or 48 h post-frozen. There is no difference in VAP, VSL, movement of amplitude lateral head (ALH), or linearity (LIN). The percentage of viable spermatozoa was not affected in either group. Statistical analysis indicates that there is no difference (P > 0.05) in any of the evaluated parameters. The results demonstrate that spermatic motility and viability of frozen bull semen could be evaluated before 48 h post-frozen. This allows reduction of the time between freezing and evaluation and immediate availability of the bull straws. Table 1. Parameters of motility and viability at 1 h vs. 48 h of post-frozen stabilization time This research was supported by Centro Genético Bovino EOLIA S.A.

231 SPERM SUBPOPULATIONS IN THE KOALA (PHASCOLARCTOS CINEREUS) EJACULATE

2009 ◽  
Vol 21 (1) ◽  
pp. 213
Author(s):  
N. Satake ◽  
S. D. Johnston ◽  
W. V. Holt
Keyword(s):  
Sperm Motility ◽  
Sperm Head ◽  
Computer Assisted ◽  
Multivariate Pattern Analysis ◽  
Phascolarctos Cinereus ◽  
Non Linear ◽  
Group 3 ◽  
Group 2 ◽  
Head Morphology ◽  
Multivariate Pattern

Koala semen contains a heterogeneous mixture of sperm morphotypes, mainly attributable to extreme degree of shape variability displayed by the hooked sperm head. By analogy with other species, we anticipate that the morphotypes may exhibit correspondingly different sperm-motility behaviors, largely caused by the differences in hydrodynamic interactions with the suspending media. This trend has been shown in human spermatozoa where motility behavior was demonstrably correlated with the sperm head morphology (Overstreet et al. 1981). In this study, we have investigated the heterogeneity of koala sperm motility profiles in semen in an effort to determine whether distinct sperm subpopulations within ejaculates are recognizable by the use of computer-assisted sperm motility analysis. Ejaculates from 5 males were collected by electroejaculation, then diluted and transported in Tris-citrate-glucose (TCG) diluent. Spermatozoa were washed through a 35–60% Percoll gradient to separate seminal plasma and the majority of the prostatic bodies from spermatozoa. Spermatozoa from the washed pellet were then diluted in TCG at 35°C, incubated for 10 min, and video recorded using a negative phase ×10 objective. Sperm motion parameters were then analyzed using the Hobson sperm tracker (Hobson Vision Systems, UK: Holt et al. 1996 J. Androl. 17, 587–596). Multivariate pattern analysis (PATN; CSIRO Australia; Abaigar 1999 Biol. Reprod. 60, 32–41) was used to distinguish 3 sperm subgroups, consistently shown in each ejaculate, within the data (1936 tracks × 6 kinetic parameters; VCL, VAP, MAD, BCF, ALH, LIN). After group allocation by PATN, all parameters showed significant differences between each of the groups (P < 0.0001). Group 1, approximately 25% of the sperm tracks, showed profiles of spermatozoa with fast, non-linear motility (VCL 106.88 ± 28.15; BCF 3.23 ± 3.81; LIN 14.08 ± 10.20). Group 2, approximately 27% of sperm tracks, showed profiles of fast, linear motility (VCL 63.92 ± 13.50; BCF 7.90 ± 3.42; LIN 28.10 ± 12.15). Group 3, 48% of sperm tracks, showed profiles of slow, non-linear or circular patterns of motility (VCL 39.05 ± 11.92; BCF 0.02 ± 0.35; LIN 5.15 ± 4.88). The recognition of 3 clearly identifiable subgroups supports our hypothesis that heterogeneity of sperm motility patterns exists within koala ejaculates. These may be a reflection of the heterogeneity in sperm-head morphotypes in koala semen, but that remains to be investigated in more detail. The clear distinctions between these groups, and the observation that all 3 subpopulations exist in each of the ejaculates, also suggest that the spermatozoa exhibit functional differences, possibly related to biochemical or maturational status. Many thanks to Dr. Michael Pyne and Dr. Vere Nicholson and their teams and animals at Currumbin Wildlife Sanctutary and Dreamwolrd QLD for all their help and support for the collection of samples.

144 Dynamic of Sperm Subpopulations in Red Deer Capacitated Samples

2018 ◽  
Vol 30 (1) ◽  
pp. 212
Author(s):  
M. Ramón ◽  
M. Iniesta-Cuerda ◽  
A. Martín-Maestro ◽  
P. Peris-Frau ◽  
I. Sánchez-Ajofrín ◽  
...  
Keyword(s):  
Red Deer ◽  
Lateral Displacement ◽  
Semen Analysis ◽  
Sperm Head ◽  
Change Points ◽  
Computer Assisted ◽  
Change Over Time ◽  
Flagellar Beat ◽  
Time Table ◽  
Over Time

An ejaculate is a mixture of sperm subpopulations (SP) with varying motility characteristics. Moreover, males with high percentages of fast and linear-moving sperm have high rates of fertility (Ramón et al. 2003 Biol. Reprod. 89, 110). The objective was to assess dynamics, over time, of SP of capacitated red deer sperm. Thawed sperm were selected with 45/90% Percoll, diluted at 10 × 106 sperm mL−1 in SOF plus 10% oestrous sheep serum and incubated for 2 h at 38.5°C under 5% CO2. Sperm motility was assessed by computer-assisted semen analysis at 1, 5, 15, 30, 45, 60, and 120 min and 24 h. Sperm were classified as described previously (Martínez-Pastor et al. 2005 Biol. Reprod. 72, 316-327) and the evolution of SP during capacitation was characterised with piece-wise regression that identified change points. Five sperm SP were identified based on velocity according to an actual path (VCL), velocity according to a straight path (VSL), velocity according to the average, smoothed path (VAP), linearity (LIN), straightness (STR), wobble (WOB), amplitude of lateral displacement of sperm head (ALH), and frequency of the flagellar beat (BCF). Sperm in SP1 were fast, linear sperm with high ALH; they corresponded to capacitated sperm. In contrast, SP5 were slow, non-linear sperm, with low ALH (Table 1). The dynamics of each SP differed over time was different along the time. Percentages of SP1, SP4, and SP3 were significantly decreased at 60, 90, and 100 min, whereas percentages of SP2 and SP5 did not change over time. This study was consistent with previous reports that kinematic sperm characteristics change over time. Table 1.Sperm subpopulations (SP) based on kinematic end points.

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