Sperm Quality Assessment in Honey Bee Drones

The quality of honey bee drone semen is relevant in different contexts, ranging from colony productivity to pathology, toxicology and biodiversity preservation. Despite its importance, considerably less knowledge is available on this subject for the honey bee when compared to other domestic animal species. A proper assessment of sperm quality requires a multiple testing approach which discriminates between the different aspects of sperm integrity and functionality. Most studies on drone semen quality have only assessed a few parameters, such as sperm volume, sperm concentration and/or sperm plasma membrane integrity. Although more recent studies have focused on a broader variety of aspects of semen quality, some techniques currently used in vertebrates, such as computer-assisted sperm analysis (CASA) or multiparametric sperm quality testing, still remain to be developed in the honey bee. This may be attributed to the particular sperm morphology and physiology in this species, requiring the development of technologies specifically adapted to it. This article reviews the present knowledge of sperm quality in honey bee drones, highlighting its peculiarities and proposing future lines of research.

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14 MORPHOLOGICAL AND FUNCTIONAL PARAMETERS OF ENDANGERED BERMEYA GOAT BREED SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv20n1ab14 ◽

2008 ◽

Vol 20 (1) ◽

pp. 87

Author(s):

C. O. Hidalgo ◽

A. Rodríguez ◽

C. Díez ◽

D. Martín ◽

M. Carbajo ◽

...

Keyword(s):

Semen Quality ◽

Sperm Concentration ◽

Membrane Integrity ◽

Analysis Data ◽

Computer Assisted ◽

Genetic Stock ◽

Term Survival ◽

Sperm Analysis ◽

Functional Parameters ◽

Fresh Semen

The Bermeya goats are an endangered autochthonous breed distributed in the north of Spain. To ensure their genetic diversity and long-term survival, morphological and functional parameters of the semen must be known in order to preserve the current genetic stock in a germplasm bank. The aim of this work was to establish basic characteristics and post-thaw survival of Bermeya goat's semen obtained by electro-ejaculation, that is not well described in the literature. The semen was collected by electro-ejaculation from 7 bucks, 1 to 7 years old, twice per week, for 9 weeks (n = 83). Fresh semen was evaluated for volume (V), concentration (C), motility, morphology, functional integrity of the sperm (spz) membranes (hypoosmotic swelling test; HOST), and acrosome integrity rate (NAR). Individual and progressive sperm motility were analyzed by means of a computer-assisted sperm analysis system (CASA: SCA 2002�, Microptic, Barcelona, Spain) immediately after dilution with the extender at 37�C, and after cooling to 4�C; five fields per sample (diluted to 204 � 106 spz mL–1) were evaluated under a phase contrast microscope (100�). The NAR and morphological abnormalities of sperm head, midpiece, tail, and cytoplasmic droplets were determined by counting 100 spz under 1000�. For freezing, ejaculates with at least 80% motile spz were diluted at 32�C with Krebs-Ringer solution containing 20% egg yolk and 14% glycerol to a final concentration of 400 � 106 spz mL–1, cooled to 4�C for 90 min, aspirated into 0.25-mL plastic straws (IMV�, L'Aigle, France), frozen at 7 cm above liquid nitrogen (LN2) phase for 10 min, and then plunged into the LN2. Straws were thawed in a water bath at 39�C for 30 s for post-thaw survival analysis. Data were analyzed by the GLM and FREQ procedures (SAS; SAS Institute, Inc., Cary, NC, USA) and expressed as means � standard error. Fresh semen characteristics were: V = 1.7 � 0.1 mL; C = 2619 � 106 � 153 spz mL–1; total and progressive motility were 89.0 � 2.1% and 66.9 � 2.1%, respectively. Percentages of head abnormalities were 4.8 � 0.5; midpiece: 3.8 � 0.7; tail: 4.7 � 1.0; cytoplasmic droplets: 8.3 � 0.7; intact acrosome: 91.8 � 0.6; and membrane integrity: 49.2 � 2.1. At 4�C, the % of total motile spz was 62.6 � 1.6, and the post-thaw survival rate was 46.3 � 1.5. There were only individual differences (P < 0.001) between bucks on sperm concentration, head abnormalities, and cytoplasmic droplets. In conclusion, our results indicate that semen quality is related to each individual animal and that electro-ejaculation allows collection of semen of satisfactory quality to use as fresh and for cryopreservation. However, the validity of our results for possible future sperm banking of endangered Bermeya goats semen must be confirmed by field trials.

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23 Sperm quality of Pure Spanish stallions is affected by inbreeding coefficient and age

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab23 ◽

2020 ◽

Vol 32 (2) ◽

pp. 137

Author(s):

Y. Pirosanto ◽

M. Valera ◽

A. Molina ◽

J. Dorado ◽

S. Demyda-Peyrás

Keyword(s):

Sperm Motility ◽

Inbreeding Depression ◽

Negative Correlation ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Concentration ◽

Inbreeding Coefficient ◽

Semen Parameters ◽

Computer Assisted

Inbreeding depression, a genetic condition produced by the mating of close-related individuals, has been associated with a reduction of fertility in several species. However, a loss in sperm quality was also associated with age. In horses, the few existing reports have described a tendency of both parameters to produce a negative effect on sperm quality. However, those reports were performed using a subjective evaluation of sperm motility. In the present study, a total of 692 ejaculates from 86 Pure Spanish stallions (PRE), aged between 3 and 22 years, were evaluated using a computer-assisted methodology to determine the effect of inbreeding in four semen parameters: free-gel volume (V), sperm concentration (C, by haemocytometer), and total (TM) and progressive (PM) sperm motility (by Spermvision sperm class analyser; Minitube). The inbreeding coefficient (F) was estimated using 300 000 PRE pedigree records approximately (minimum pedigree depth, eight equivalent complete generations; range, between 1 and 30.1%). Stallion, age, ejaculate, and season of semen collection were the variables included in the statistical model (general linear model), with ejaculate and season being the variables with a major effect (by variance components analysis). Our results showed that sperm concentration (r=−0.18; P<0.0001) and volume (to a lesser extent) were reduced with advancing age, both showing a major decline after 15 years of age. To the contrary, sperm motility was not affected by age of the stallion. We also found a negative correlation between the inbreeding coefficient and ejaculate volume (r=−0.14; P<0.001), with a marked decrease seen when F was between 7 and 20%. Also, a negative correlation was observed in PM (r=−0.08; P<0.05), although to a lower extent. Conversely, C and TM were not affected by inbreeding depression (P>0.05). In conclusion, our results demonstrated that high levels of inbreeding can compromise severely the sperm quality of the PRE stallion, which, subsequently, may have a negative influence on fertility. Ongoing studies using genomic data will help to detect genetic variants associated with stallion semen quality and how it is influenced by inbreeding in specific genomic regions.

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Validation of the Turkey Semen Cryopreservation by Evaluating the Effect of Two Diluents and the Inseminating Doses

Animals ◽

10.3390/ani10081329 ◽

2020 ◽

Vol 10 (8) ◽

pp. 1329

Author(s):

Michele Di Iorio ◽

Giusy Rusco ◽

Roberta Iampietro ◽

Lucia Maiuro ◽

Achille Schiavone ◽

...

Keyword(s):

Sperm Quality ◽

Sperm Concentration ◽

Membrane Integrity ◽

In Vivo Test ◽

Sperm Analysis ◽

Fertilizing Ability ◽

Vitro Analysis ◽

Hatching Rates

This study was designed to test the fertilizing ability of cryopreserved turkey semen, and here, two experiments were performed: an in vitro analysis to assess the effects of Tselutin and Lake diluents and an in vivo test to determine the fertility and hatching rates by also studying the feat of three insemination doses (250, 400 and 600 × 106 sperm/hen). Pooled semen samples were diluted with Tselutin or Lake extender which contained 20% of dimethylsulfoxide and 1 mM of Ficoll at final sperm concentration of 3 × 109 sperm/mL. Thereafter, semen was packaged into straws and frozen on liquid nitrogen. The post-thaw sperm quality was evaluated considering motility (computer-aided sperm analysis—CASA system) and membrane integrity (flow cytometry). Significantly higher values of progressive motility and some kinetic parameters in semen frozen with Lake were found. When we compared the extenders in vivo, no significant effects were detected, whilst sperm concentration significantly affected both fertility and hatching rates, with the best results obtained with the sperm concentration of 400 × 106 sperm/hen. From the results obtained, it emerged that the extender type only affected sperm motility characteristics, not the fertilizing ability of frozen-thawed semen, while inseminating dose markedly affected fertility and hatching rates.

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180 ASSESSMENT OF BULL SEMEN QUALITY LOADED IN NEW SensiTemp STRAWS USING SEMEN AND IN VITRO PRODUCTION TECHNOLOGIES

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab180 ◽

2016 ◽

Vol 28 (2) ◽

pp. 221

Author(s):

D. Le Bourhis ◽

S. Camugli ◽

P. Salvetti ◽

L. Schibler ◽

E. Schmitt

Keyword(s):

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Semen Analysis ◽

Membrane Integrity ◽

Computer Assisted ◽

Cleavage Rate ◽

Fluid Medium ◽

And Control

SensiTemp, a new in vitro maturation (IMV) bull straw concept, presents the advantage of colour changing while the straw is thawed. The colour of frozen straws is blue and straws start to become white when the temperature reaches 33°C, with a complete change of colour at 37°C. The objective of this study is to assess sperm quality after thawing of semen frozen in SensiTemp from 2 bulls, by analysing, in experiment 1, sperm motility and membrane integrity using computer-assisted semen analysis (CASA) and flow cytometry (FC), and, in experiment 2, the in vitro embryo production (IVP) using IVP technologies [IVM, IVF, and in vitro culture (IVC)]. The ejaculates of 2 bulls, selected during preliminary experiments on high in vitro fertility, were harvested at CIA L’Aigle, France, and split ejaculates were frozen in experimental (SensiTemp) and conventional (control) straws. In experiment 1 after thawing semen from the 2 types of straws (5 pooled straws each; 2 replicates), motility was assessed using the IVOS CASA system (Hamilton Thorne Inc., Beverly, MA, USA) and membrane integrity was evaluated through FC with Cytosoft software (Millipore-Guava Technologies Inc., Hayward, CA, USA). In experiment 2, IVF was used to evaluate the non-toxicity of SensiTemp and control straws. Cumulus-oocyte complexes (COC; n = 1178; 4 replicates) collected from slaughterhouse ovaries were matured in IVM medium (TCM-199 with bicarbonate, Sigma-Aldrich, Saint Quentin Fallavier, France; 10 µg mL–1 FSH-LH, Reprobiol, Liège, Belgium; and 10% FCS, Thermo Fisher, Illkirch, France) for 22 h. After fertilization, presumptive zygotes of each group (SensiTemp and control for each bull) were cultured in synthetic oviduct fluid medium (SOF, Minitube, Tiefenbach, Germany) with 1% estrous cow serum (ECS) and 0.6% BSA (Sigma-Aldrich, France) up to 8 days. All cultures were conducted at 38.5C in 5% CO2, and 5% O2. The cleavage and blastocysts rates were evaluated on Days 3 and 7, respectively, for each group. Embryo quality was recorded on Day 7 according to the IETS evaluation. Data from each bull were analysed separately using the chi-squared test (P < 0.05). In experiment 1, neither sperm motility from bull 1 (61.2 and 60.5%) and bull 2 (66.2 and 66.5%) nor membrane integrity from bull 1 (58.6 and 52.2%) and bull 2 (61.0 and 61.9%) were different between SensiTemp and control, respectively. Results from experiment 2 showed no difference (P > 0.05) in cleavage rate between SensiTemp and control for the 2 bulls: 92.1 and 91.7% for bull 1 and 94.2 and 94.6% for bull 2 respectively. The blastocysts rate on Day 7 did not differ (P > 0.05) among groups (47.5, 47.1 and 51.3, 50.4% for SensiTemp and control bull 1 and bull 2, respectively) nor the quality of embryos retrieved in the different groups: 25.4, 23.3, and 30.8, 29.6% in grade 1 embryo for SensiTemp and control bull 1 and bull 2, respectively. Those results demonstrate, in vitro, that the new SensiTemp straws were non-toxic and did not affect the semen quality after thawing nor did the SensiTemp straws affect the ability of sperm cells to fertilize oocytes and produce 8-day-old embryos.

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How to Increase Post-Thaw Semen Quality in Poor Freezing Stallions: Preliminary Results of the Promising Role of Seminal Plasma Added after Thawing

Animals ◽

10.3390/ani9070414 ◽

2019 ◽

Vol 9 (7) ◽

pp. 414 ◽

Cited By ~ 2

Author(s):

Jiří Šichtař ◽

Filipa Bubeníčková ◽

Jitka Sirohi ◽

Ondřej Šimoník

Keyword(s):

Plasma Membrane ◽

Seminal Plasma ◽

Semen Quality ◽

Membrane Integrity ◽

Control Group ◽

Computer Assisted ◽

Kinematic Parameters ◽

Prolonged Incubation ◽

Sperm Analysis ◽

Spermatozoa Motility

The aim of this study was to evaluate the effect of the addition of two types of seminal plasma (SP) after thawing on the functional characteristics of frozen–thawed (F–T) spermatozoa of poor freezing stallions during prolonged incubation periods. Seminal plasma from stallions with 35–40% (standard seminal plasma, (S-SP)) and 60–70% (above standard seminal plasma, (A-SP)) progressively motile spermatozoa after thawing was used. The motility, kinematic parameters (Computer Assisted Sperm Analysis), distribution of spermatozoa into subpopulations, integrity (carboxyfluorescein diacetate/propidium iodide staining), and functionality (hypo-osmotic swelling (HOS) test) of the spermatozoa plasma membrane were evaluated after thawing (T0) and after 30 min (T30) of incubation at 37 °C. There was no effect of SP addition on spermatozoa motility, but there was a significant positive effect on the kinematic parameters at T0 and T30. The addition of SP significantly increased the percentage of spermatozoa in the fast subpopulation at T0 as well as at T30. Plasma membrane integrity was not affected by the treatment, but functionality significantly decreased by 5% compared to the control group when samples were incubated for 30 min with A-SP. In conclusion, generally, the post-thaw addition of seminal plasma positively affected the post-thaw quality of semen from poor freezing stallions.

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Modified hypoosmotic swelling test for the assessment of boar and bull sperm sensitivity to cryopreservation

Acta Veterinaria Brno ◽

10.2754/avb201483040313 ◽

2014 ◽

Vol 83 (4) ◽

pp. 313-319 ◽

Cited By ~ 4

Author(s):

Petra Přinosilová ◽

Věra Kopecká ◽

Jaroslava Hlavicová ◽

Monika Kunetková

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Membrane Resistance ◽

Freezing Process ◽

Sperm Analysis ◽

Hypoosmotic Swelling Test ◽

Sperm Plasma Membrane ◽

Viable Sperm ◽

Sperm Membrane

Routine methods for the evaluation of sperm quality are not sufficiently useful to determine the sensitivity of sperm cells to cold shock. The aim of our preliminary study was to determine whether the sperm plasma membrane integrity evaluated by modified hypoosmotic swelling test based on simple hypoosmotic swelling test (HOS test) and eosin-nigrosin staining could be helpful in predicting the degree of boar sperm survivability during semen cryopreservation. Ejaculates collected from 24 boars and 20 bulls were used in the experiment. Fresh ejaculates were evaluated by routine sperm analysis and a modified HOS test, and subsequently frozen. Sperm cryosurvivability was defined as the percentage of motile spermatozoa that survived the freezing process. A higher percentage of sperm was recovered after the thawing of semen with a higher percentage of HOS-positive and eosin-negative sperm (P < 0.01). Both indicators were found to be correlated (r = 0.707 and r = 0.705, respectively; P < 0.01). Moreover, the percentage of HOS-positive and eosin-negative sperm was similar to the percentage of viable sperm after thawing as determined by traditional eosin-nigrosin staining in boars (50.90 ± 9.88% and 49.31 ± 11.63%, respectively) and bulls (55.91 ± 10.34% and 55.63 ± 6.64%, respectively) and both indicators showed a positive correlation (r = 0.558 and r = 0.504, respectively; P < 0.05). In conclusion, based on the obtained results, we can assume that the modified HOS test can detect differences in sperm membrane resistance which allows assessment of semen quality from the perspective of its further use, e.g. cryopreservation.

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Effect of Macrophages in Semen on Sperm Quality

10.21203/rs.3.rs-100361/v1 ◽

2020 ◽

Author(s):

Gangxin Chen ◽

Beihong Zheng

Keyword(s):

Cell Membrane ◽

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Concentration ◽

Membrane Integrity ◽

Significant Negative Correlation ◽

High Group ◽

Cross Sectional ◽

Cell Membrane Integrity

Abstract Background: This was a cross-sectional study in China, we analyzed the levels of macrophages(Mφ) in semen. The study evaluated the influence of the levels of Mφ in semen on sperm quality.Methods: The subjects were 78 males between 25 and 35 years old. The samples were divided into a low group(Mφ<6×105/ml) and a high group (Mφ>6×105/ml). Evaluation included consideration of the influencing factors of male semen quality, macrophage concentration, sperm motility, morphology, membrane integrity DNA fragmentation index (DFI), anti-sperm antibodies (AsAb), IL-10, and IL-12 in semen.Results: There was no difference in the physical or chemical indices of the semen, sperm concentration, AsAb, IL-10, or IL-12 between the two groups (P>0.05). The percentage of sperm forward motility (PR%), the rate of normal sperm shape, and the integrity of cell membranes in the low group were higher than those in the high group (P<0.05), while the percentage of sperm inactivity (IM%), the rate of sperm head deformity, the rate of deformity in the neck and middle segment, the sperm malformation index (SDI), the abnormal sperm index (TZI), and the sperm DFI in the low group were lower than those in the high group (P<0.05). The concentration of Mφ in the semen was linearly correlated with sperm concentration, sperm PR%, IM%, sperm normal shape rate, head deformity rate, neck and middle deformity rate, SDI, TZI, sperm DFI, and sperm cell membrane integrity (P<0.05), but there was no linear correlation with IL-10 or IL-12 (P>0.05).Conclusions: The concentration of Mφ in semen had no significant correlation with semen volume or sperm concentration, but it did have a significant negative correlation with sperm motility, sperm morphology, cell membrane integrity, and DNA breakage rate. There was no significant correlation with the concentration of IL-10 or IL-12.

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12 COMPUTER-ASSISTED SPERM ANALYSIS OF FRESH EPIDIDYMAL CAT SPERMATOZOA AND THE IMPACT OF COOLED STORAGE (4°C) ON SPERM QUALITY

Reproduction Fertility and Development ◽

10.1071/rdv20n1ab12 ◽

2008 ◽

Vol 20 (1) ◽

pp. 86

Author(s):

M. Filliers ◽

T. Rijsselaere ◽

P. Bossaert ◽

V. De Causmaecker ◽

J. Dewulf ◽

...

Keyword(s):

Reference Values ◽

Sperm Quality ◽

Membrane Integrity ◽

Computer Assisted ◽

Epididymal Sperm ◽

Sperm Analysis ◽

Computer Assisted Sperm Analysis ◽

The Impact ◽

Motility Parameters

Feline epididymal sperm is commonly used for in vitro fertilization. It also yields the opportunity to conserve genetic material from valuable males that suddenly die. Epididymal sperm quality parameters vary considerably among laboratories, implicating the need for objective evaluation methods. The aim of the present study was to describe reference values of computer-assisted sperm analysis (CASA) parameters of fresh epididymal cat sperm and to assess the effect of prolonged cooled storage (4�C) on various sample characteristics. Epididymides obtained from tomcats after routine orchiectomy (2–4 pairs/replicate) were sliced to release spermatozoa. The sperm suspension was placed on a 2-layer gradient and, after centrifugation, the sperm pellet was recovered. In Experiment 1 (20 replicates), sperm motility parameters were assessed immediately after retrieval (T0) using the Hamilton Thorne analyzer Ceros 12.1 (HTR; Hamilton Thorne Biosciences, Beverly, MA, USA). In Experiment 2, fresh (T0) sperm samples (4 replicates) were evaluated for motility parameters (HTR), acrosomal status (FITC-Pisum sativum agglutinin staining), morphology (eosin/nigrosin (E/N) staining), and membrane integrity (E/N and SYBR�-14-propidium iodide staining; Molecular Probes, Inc., Eugene, OR, USA). After addition (1:2) of a Tris-glucose-citrate diluent containing 20% egg yolk, samples were cooled and reassessed on Days 1 (T1), 3 (T3), 5 (T5), 7 (T7), and 10 (T10). Results were analyzed in a mixed linear model, with replicate as random factor and time as fixed effect (S-PLUS 7.0; Insightful Corp., Seattle, WA, USA). Results of Experiment 1 were as follows (mean � SD): motility (MOT): 80.8% � 23.5; progressive motility (PMOT): 69.9% � 23.2; velocity average pathway (VAP): 98.7 µm s–1 � 24.2; velocity straight line (VSL): 89.3 µm s–1 � 25.4; velocity curved line (VCL): 134.8 µm s–1 � 31.9; amplitude lateral head (ALH): 4.3 µm � 2.0; beat cross frequency (BCF): 34.6 Hz � 7.0; and straightness (STR): 89.6% � 6.6. In Experiment 2, MOT, PMOT, VAP, VSL, VCL, BCF, and the percentage of normal spermatozoa showed a decrease over time (P < 0.05) compared to fresh samples, starting from T1, T3, T5, T7, T5, T3, and T1, respectively. In contrast, STR, ALH, membrane integrity, and the percentage of acrosome-intact spermatozoa were not affected (P > 0.05) by cooled storage. To summarize, we have presented a set of reference values for CASA-parameters of fresh, epididymal cat spermatozoa. Cooled storage impaired most motility parameters and lowered the percentage of normal spermatozoa, but did not influence membrane integrity or acrosomal status. The effect of cooled storage on DNA fragmentation of sperm and its subsequent influence on in vitro embryo development require further investigation.

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88 SELECTION OF A CRYOSURVIVAL SPERM POPULATION DOES NOT IMPROVE THE IN VITRO PENETRATING ABILITY OF FROZEN - THAWED BOAR SPERMATOZOA

Reproduction Fertility and Development ◽

10.1071/rdv20n1ab88 ◽

2008 ◽

Vol 20 (1) ◽

pp. 124

Author(s):

M. L. Perals ◽

M. A. Gil ◽

E. M. Garcia ◽

J. Sanchez-Osorio ◽

J. M. Vázquez ◽

...

Keyword(s):

Sperm Quality ◽

Membrane Integrity ◽

Molecular Probes ◽

Computer Assisted ◽

Immature Oocytes ◽

Sperm Analysis ◽

Boar Spermatozoa ◽

Casa System ◽

Selection Of

Boars can be classified as good or bad sperm freezers according to their sperm cryosurvival. Different sperm selection techniques, such as PureSperm� (PS; MidAtlantic Diagnostics, Inc., Mount Laurel, NJ, USA), have been developed to improve functional competence of spermatozoa. The aim of this experimental study was to assess the ability of PS for improving the in vitro penetrating ability of frozen–thawed boar spermatozoa from good and bad sperm freezers. The sperm-rich fractions from two boars, good (Boar A) and bad (Boar B) freezers, were extended in a lactose/eggyolk/ glycerol/Equex Stem (Noba Chemical Sales, Inc., Scituate, ME, USA) mixture (1 � 109 sperm mL–1), dispensed into 0.5-mL straws, and frozen using a programmable cell freezer. After thawing (1.200�C min–1), semen from each boar was split into two aliquots of 500 µL. One aliquot was used as the control. The second was placed into a tube of PS gradient (90%/45%) and centrifuged at 425g for 20 min; the pellet re-suspended in 1 mL of BTS and re-centrifuged at 320g for 10 min (PS sample). Control and PS samples were diluted in supplemented TCM-199 (TCMm; Roca et al. 1998 Reprod. Fertil. Dev. 10, 479–485) at 200 � 106 sperm mL–1. Sperm survival (SV) was assessed afterTCMm dilution according to progressive sperm motility (PSM, %) using a computer-assisted sperm analysis (CASA) system (ISAS�), and plasma and acrosome membrane integrity (PMI; %) by flow cytometry (SYBR�-14/PE-PNA/PI; Molecular Probes, Leiden, The Netherlands). A homologous in vitro penetration (hIVP) assay, using immature oocytes (20 oocytes/2 mL TCMm supplemented with caffeine and calcium lactate), was used to assess sperm penetrating ability (Martinez et al. 1993 Theriogenology 40, 547–557). A total of 960 immature oocytes were inseminated (200 � 103 sperm/oocyte) in 3 batches. After 18 h of co-incubation at 39�C under 5% CO2 in air, the oocytes were washed, mounted on slides, fixed with ethanol:acetic acid (3:1, v/v) for 48 h, stained with 1% lacmoid, and examined under a phase contrast microscope (�400). Oocytes with swollen or unswollen heads of sperm found in the vitellus were considered as penetrated. Sperm penetrability ability (SPA) was assessed according to penetration rate (PR) and the mean number of sperm per oocyte (S/O). Data were analyzed using a PROMIXED model and expressed as mean � SEM. Boar A showed better (P ≤ 0.01) results for both SV and SPA parameters than boar B, independent of sperm treatment. PureSperm improved (P ≤ 0.05) PSM and PMI in both boar A (control v. PS: 48.0 � 5.8 v. 66.5 � 3.6 and 63.1 � 7.7 v. 88.4 � 1.3, respectively) and boar B (12.3 � 1.2 v. 22.2 � 3.7 and 44.3 � 3.5 v. 58.7 � 7.0, respectively). However, no differences (P ≥ 0.05) were observed in PR and S/O in either boar A (71.2 � 3.4 v. 78.3 � 3.1 and 5.0 � 0.4 v. 5.2 � 0.4, respectively) or boar B (34.3 � 3.6 v. 37.3 � 3.9 and 1.5 � 0.1 v. 1.5 � 0.1, respectively). In conclusion, under our laboratory conditions, PureSperm selection improves sperm quality but not in vitro penetrating ability of frozen–thawed spermatozoa of both good and bad sperm freezers. This work was supported by CICYT (AGF2005-00760), Madrid, Spain.

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Assessment of semen quality in infertile dogs using computer-assisted sperm analysis by the Hamilton-Thorne Semen Analyser

Bulletin of the Veterinary Institute in Pulawy ◽

10.2478/bvip-2013-0074 ◽

2013 ◽

Vol 57 (3) ◽

pp. 429-432 ◽

Cited By ~ 4

Author(s):

Anna Domosławska ◽

Sławomir Zduńczyk ◽

Wojciech Niżański ◽

Tomasz Janowski

Keyword(s):

Semen Quality ◽

Prostate Gland ◽

Sperm Concentration ◽

Quality Parameters ◽

Computer Assisted ◽

Normal Morphology ◽

Sperm Analysis ◽

History Of ◽

Possible Cause ◽

Motility Parameters

Abstract Semen quality parameters of infertile and fertile dogs were compared. Sperm concentration and semen motility parameters were measured by the Hamilton-Thorne Semen Analyser IVOS 12.3. The spermatozoal morphology and the percentage of live spermatozoa were examined microscopically. Forty-six dogs of various breeds were examined. Twenty dogs had a conception failure within last year. These dogs had a history of prior normal fertility. Twenty six fertile dogs served as control. All animals underwent a clinical examination as well as ultrasonography. Sperm concentration was significantly lower in infertile dogs than in fertile dogs. For most determined motility parameters (MOT, PMOT, VAP, VSL, VCL, BCF, RAPID, STATIC) significant differences between infertile and fertile dogs were found. The percentage of spermatozoa with normal morphology also significantly differed between both groups. Ultrasonography of the prostate gland and testes revealed no pathological conditions. The testicular degeneration was assumed to be a possible cause of infertility in these dogs. The present study showed that the most sperm CASA motility parameters were significantly lower in infertile dogs in comparison to the fertile ones, and confirmed the usefulness of the Hamilton-Thorne Semen Analyser for a quick and objective analysis of sperm concentration and motility in dogs.

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