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Correlation of CASA Velocity and Linearity Parameters With Sperm Mobility Phenotype in Turkeys
Abstract
Since all domestic turkeys are produced through artificial insemination, a measurable sperm characteristic that would be predictive of fertility would allow for the culling of poor males, resulting in improved reproductive efficiency. The sperm mobility test (SMT), which quantifies sperm penetration into an Accudenz solution, has been shown to correlate highly with fertilization potential of individual turkeys. Since this sire-selection test is based on the differences in sperm mobility between whole ejaculates from individual males, the objective of this study was to determine whether specific sperm velocity parameters would correlate with the SMT and to determine whether these characteristics could account for phenotypic differences in sperm mobility observed between males. The SMT was used to rank males within a flock (n = 110) in triplicate and to classify them into high, average, and low sperm mobility phenotypes on the basis of the sperm mobility index. Several sperm velocity parameters were evaluated for each male by a computer-aided sperm analysis (CASA) system, the Hobson Sperm Tracker. The types of measurements taken of 200 sperm tracks/ejaculate included the following: curvilinear velocity (VCL), average path velocity (VAP), straight-line velocity (VSL), linearity (LIN), beat-cross frequency (BCF), and mean angular displacement (MAD). Significant positive correlations were found between VSL, LIN, BCF, and sperm mobility, and a significant negative correlation was seen between MAD and sperm mobility. Subpopulations of sperm that had penetrated the Accudenz solution were isolated from each mobility phenotype and were analyzed by CASA, and significant correlations were again observed between VSL, LIN, BCF, and sperm mobility. We conclude that sperm velocity and linearity contribute to overall sperm mobility phenotype and are important characteristics of turkey sperm function. Key words: Motility, computer, spermatozoa.
... Perbedaan yang signifikan pada keenam bangsa terhadap parameter motilitas level 3 khususnya pada VCL, LIN, ALH berpengaruh terhadap kelompok spermatozoa yang mengalami hiperaktifasi, pada VAP, VSL, LIN berpengaruh terhadap motilitas progresif, VCL, ALH, BCF merupakan indikator vigor spermatozoa. Velocity dan Linearity memberikan kontribusi terhadap karakteristik penting fungsi spermatozoa (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005) (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005, Herwiyanti, 2004 LIN memberikan kontribusi terhadap karakteristik penting fungsi spermatozoa (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005, Herwiyanti, 2004 ...
... Perbedaan yang signifikan pada keenam bangsa terhadap parameter motilitas level 3 khususnya pada VCL, LIN, ALH berpengaruh terhadap kelompok spermatozoa yang mengalami hiperaktifasi, pada VAP, VSL, LIN berpengaruh terhadap motilitas progresif, VCL, ALH, BCF merupakan indikator vigor spermatozoa. Velocity dan Linearity memberikan kontribusi terhadap karakteristik penting fungsi spermatozoa (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005) (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005, Herwiyanti, 2004 LIN memberikan kontribusi terhadap karakteristik penting fungsi spermatozoa (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005, Herwiyanti, 2004 ...
... Perbedaan yang signifikan pada keenam bangsa terhadap parameter motilitas level 3 khususnya pada VCL, LIN, ALH berpengaruh terhadap kelompok spermatozoa yang mengalami hiperaktifasi, pada VAP, VSL, LIN berpengaruh terhadap motilitas progresif, VCL, ALH, BCF merupakan indikator vigor spermatozoa. Velocity dan Linearity memberikan kontribusi terhadap karakteristik penting fungsi spermatozoa (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005) (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005, Herwiyanti, 2004 LIN memberikan kontribusi terhadap karakteristik penting fungsi spermatozoa (King et al., 2000;Duty et al., 2004;Shibahara et al., 2003;Rounge, 2003;Suzuki et al., 2003;Katebi et al., 2005, Herwiyanti, 2004 ...
ABSTRAK Tujuan Penelitian ini adalah untuk mempelajari 18 parameter motilitas spermatozoa menggunakan metode CASA serta motilitas progresif secara visual dibandingkan ddengan metoda CASA. Materi adalah semen segar dari 8 bangsa sapi yaitu Limousin, Bali Madura, Simental, Brahman, dan Ongole. Sata dianalisa dengan analisis varian, sementara data motilitas progresif diuji t-test. Hasil menunjukkan bahwa motilitas progresif menggunakan CASA tidak berbeda nyata (P>0.05) diantara 6 bangsa sapi. Sementara itu parameter motilitas hiperaktif menggunakan CASA tidak berbeda nyata (P>0.05) kecuali on linier, non limear, juga parameter: DCL, DAP, DSL, VCL, VAP, VSL, LIN< STR< WOB, ALH, AOH adalah berbeda nyata (P<0.05) diantara bangsa sapi. Uji t pada motilitas progresif menunjukkan tidak ada beda nyata (P>0.05) antara CASA dengan visula. ABSTRACT The aims of this research were to determine 18 parameters spermatozoa motility using CASA method and to determine spermatozoa progressive motility using visual assessment and CASA. The material of this research were bull fresh semen from 6 breeds, there were Limmousin, Bali, Madura, Simmental, Brahman and Ongole which owned by National Artificial Insemination Center Singosari. Data were collected from CASA method of 18 parameters sperm motility and visual assessment method of progressive sperm motility. Data were analyzed by variance and factor analysis, and progressive motility using visual and CASA were analyzed by paired T-Test. The result showed that motility and progressive motility using CASA was no significant different (P>0.05) for six breeds. There were no significant different (P>0.05) on hyperactive parameter using CASA, but on linear, non linear, curve linear and cell detail parameter : DCL, DAP, DSL, VCL, VAP, VSL, LIN, STR, WOB, ALH, AOH were significant different (P<0.05) among breeds. Analysis using paired T-test of progressive motility assessment was no signifgicant different between CASA and visual microscopic (P>0.05).
... As is shown in (Table 3), non progressive motility (NP) type C percentage after DPP Administration for three months has increased (from 8.84% to 10.10%) but in non-significant value. This increment in the (NP) type C percentage strengthens the percentages of motility partitions are in a positive way towards improving the motility parameters [43][44][45].The immotile spermatozoa percentage (IM% Class D) has dropped significantly from 70.13% in the pretreatment measure to 51.52% in the post treatment (Table 3), which can be regarded as a good sign of general sperm motility improvement. ...
... A research foun d that there is significant positive correlations between sperm velocities and sperm mobility [68] and as the motility parameters improved in our current research DPP administration (Table 2 and 3) in oligoasthenozoospermic patients, so that we can conclude that sperm velocities improvement is a reflection for sperm motility improvement. Both the progressive linear motility (VSL) and the vigorous non-linear non-progressive motion (VCL) are of high importance [43], in our current study both increased and improved by the administration of DPP (Table 4) in oligoasthenozoospermic patients. Table 9, in both of the cases, DPP induced a non -significant differences in serum levels of ALP. ...
Medicinal plants are identified and used throughout human history; it has a great economic value especially in drugs discovery. Date palm pollen (DPP) is used traditionally in Sudan for treating sub-fertile male patients. Male infertility is heterogeneous group of disorders, most of them are idiopathic. This study is aimed to investigate the role of pharmaceutical preparation of DPP in amelioration of male sub fertility with detection of any possible adverse effects on the major body system functions, through blood picture, liver enzymes and kidney function. This study is a single group pretest-posttest experimental prospective comparative self-control. Sub-fertile men with Idiopathic oligoasthenozoospermia or azoospermia were received 500 mg capsules of DPP twice daily for three months after conducting their safety profiles to detect any toxic effects on hematological, hepatological and nephrological functions, Blood samples were taken from the patients for serum level of FSH (for azoospermic patients), FSH and Testosterone (for oligoasthenozoospermic patients). Finally, Semen sample have been obtained for computerized assisted semen analysis (CASA) report I and II. DPP administration induced significant increase (p≤0.001) in testosterone level (in oligoasthenozoospermic patients) and FSH level (in azoospermic patients). DPP induced significant changes (p≤0.001) towards improvement in the total and progressive sperm motility percentages measured in oligoasthenozoospermic patients by CASA dynamic analysis report I and II. The toxicological studies for DPP approved their safety use in human.
... The importance of VSL in the fertilizing capacity of spermatozoa has been observed in both bulls (14) and humans (15), and it was confirmed in the present study as both the lowest VSL and NRR values were obtained from Bull 3, in addition to the positive correlation acquired between these parameters. VSL, VAP, and VCL are all measurements of sperm velocity over specific paths, so these values indicate that sperm classified as having high mobility swim faster than those classified with lower mobility (16). Thus, the role of VSL in sperm transport may be during the passage through the female reproductive tract and penetration of the oocyte vestments (14). ...
... The highest positive interaction was detected between the VCL and VAP kinetic parameters, followed by the interaction between WOB and LIN. In rabbit and bull semen, in accordance with the present study, velocity parameters strongly correlated with progressive motility (13)(14)(15)(16)(17)(18), indicating that spermatozoa with forward motility and a straight linear path may cover more distance in a short period of time. In addition to that, a strong and highly significant correlation was found between straightness and linearity in bulls (13). ...
Prediction of male fertility with in vitro semen quality parameters remains a challenge for the bull industry. Fluorescein staining and Computer-Assisted Semen Analysis (CASA) ensure kinetically proper and functionally objective results to improve spermatological parameters control. Therefore, we aim to examine the CASA kinetic parameters and fluorescein staining of cryopreserved bull semen and its relationship with fertility. A total of ninety frozen straws from five Simmental bulls were evaluated. After thawing semen, motility, kinetic parameters and insemination doses were evaluated by CASA; fluorescein stainings were evaluated by using fluorescein microscope. 60th-day non-return rates (NRR) were recorded for determination of pregnancy rate. There were significant differences at motility and kinetic parameters among bulls. Bull 3 had the lowest percentage of NRR and VCL, VSL, BCF, ALH parameters (P<0.05). A positive correlation was detected between the pregnancy rate and some kinetic parameters. Besides, the correlation between the CASA parameters was determined as well. Therefore, we concluded that various kinetic parameters obtained with CASA software system algorithms and fluorescein stainings can be linked to fertility. However, further research is needed to acquire more precise link with fertility.
... Consequently, CASA is now widely used to assess sperm movement characteristics for a variety of species (rat, rabbit, pig, bull, broiler chicken, pigeon, turkey, human). Furthermore, significant correlations of CASA motility parameters with fertility have also been reported [16,17]. ...
... Overall, we did not detect any significant relationships between any sperm measurement (VCL, VSL, VAP, LIN, BCF, STR) and any measure of egg fertility. In contrast, studies with broilers and turkeys showed differences in VSL, LIN, BCF, and VAP between high and low sperm mobility males [17]. Moreover, VSL is reported to be associated with fertility in rats and boars [25,26] although, in humans, both positive and negative associations have been reported [16]. ...
This study investigated the relationship between sperm kinematics and egg fertility in Japanese quail in an attempt to identify a semen trait that could be used to predict male fertility. Males (n = 45) and females (n = 180) from five strains (A, B, C, D, E) were used. Ejaculates (n = 720) were collected from 8 to 38 weeks of male age. Semen volume and sperm concentration were recorded and sperm motility was analyzed using Sperm Class Analyzer (SCA®-CASA). At the time of ejaculate collection, males were allowed to mate with females in order to obtain egg fertility data: percent fertile eggs and the numbers of sperm (SpermOPVL) and sperm-holes (HolesIPVL) present on the egg perivitelline membranes. Sperm concentration was positively correlated with ejaculate volume (r = 0.35; P < 0.01) and percent fertile eggs (r = 0.08; P < 0.05). There were high correlations (P < 0.01) among the sperm kinematic parameters: percent motile (MOT%), percent rapid (Rapid%), percent progressive (PROG%), percent medium (Medium%), velocity curvilinear (VCL), velocity straight line (VSL), and velocity average path (VAP), linearity (LIN%), straightness (STR%) and beat cross frequency (BCF). The strains differed with respect to the correlations for sperm kinematics and egg fertility. For example, SpermOPVL was correlated (P<0.05) with VSL (r = 0.18), VAP (r = 0.18), and BCF (r = 0.23) for Strain A, with PROG% (r = 0.17) for Strain B, and with Medium% (r = 0.18) for Strain C. When the data were adjusted for the effects of strain and age, Medium% was correlated with HolesIPVL (r = 0.36; P< 0.01) and percent fertile eggs (r = 0.31; P< 0.01), whereas PROG% was correlated with SpermOPVL(r = 0.30; P < 0.01) and HolesIPVL (r = 0.30; P < 0.01). Males could be ranked into high and low fertility categories based on initial (i.e., Week 8) and the life-time (i.e., Weeks 8-38) data for sperm kinematics and egg fertility. Of the males classified as poorly fertile by Week 8 sperm kinematics, 30% were also confirmed as poor on the basis of life-time sperm kinematics. Of males classified as poorly fertile by Week 8 egg fertility, 47% were confirmed as poor on the basis of life time-data egg fertility. We concluded that sperm concentration, Medium%, PROG%, VAP, VSL and BCF are important determinants of egg fertility in quail, and that these relationships depend on genotype.
... Computerassisted sperm analysis (CASA) technology was introduced in the late 1980s ( Amann and Waberski 2014;Mortimer et al. 2015). With CASA, spermatozoal images are captured by a microscope camera, using the defined software to determine spermatozoa variables, such as motility and morphology ( King et al. 2000;Lu et al. 2014;Maroto-Morales et al. 2016;Santiago-Moreno et al. 2016). Flow cytometry (FC) can detect spermatozoa labelled by various specific fluorescent probes and rapidly calculates the relevant numbers of spermatozoa for assessing different functional characteristics (for example, plasma and acrosomal integrity, mitochondrial function and DNA status) ( Christensen et al. 2004;Cordelli et al. 2005;Graham and Mocé 2005;Mocé and Graham 2008;Partyka et al. 2010). ...
... Therefore, the WST-8 assay might be more efficient and reliable than the MTT assay. Several studies reveal that in vitro analysis of spermatozoa viability and motility are correlated with fertilising potential in vivo (Donoghue 1999;King et al. 2000;Blesbois et al. 2008;Hossain et al. 2011;Farah et al. 2013;Petrunkina and Harrison 2013). It is therefore important to measure spermatozoa viability and motility before artificial insemination in order to maximise fertility. ...
1. The results of spermatozoa assessment by the WST-8 (2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H-tetrazolium, monosodium salt) assay, flow cytometry (FC) or computer-assisted spermatozoa analysis (CASA) were compared.
2. Different live/killed ratios of cockerel semen were serially diluted to 120, 60, and 30 × 10⁶ cells/ml, and each sample was analysed by (1) WST-8 assay at 0, 10, 20, 30, 40, 50, 60 min, (2) viability with flow cytometry, and (3) motility with CASA.
3. The WST-8 reduction rate was closely correlated with spermatozoa viability and motility. The optimal semen concentration for the WST-8 assay was 120 × 10⁶ cells/ml, and the standard curves for spermatozoa viability and motility predictions respectively were yviability60 = 162.8x + 104.96 (R² = 0.9594) after 60 min of incubation and ymotility40 = 225.09x + 96.299 (R² = 0.8475) after 40 min of incubation.
4. It was conclused that the WST-8 assay is useful for the practical evaluation of cockerel spermatozoa viability and motility. Compared to FC and CASA, the WST-8 assay does not require expensive and complex instrumentation in the lab. Furthermore, one well of the WST-8 reaction can be used to predict spermatozoa viability and motility at the same time which all lead it to be efficient and economical for semen quality assessment.
... The slides were mounted on a heated stage (41 • C). The sperm dilutions, recording, and Hobson Tracker settings followed those described by King et al. (2000) and Kot lowska et al. (2007) for turkey spermatozoa. Semen samples were incubated for 5 min in the motility buffer (50 mM Tris buffer, pH 7.4, 120 mM NaCl, 10 mM glucose, 2 mM CaCl 2 ) at 39 • C. The motility buffer composition used in the present study was previously adapted for CASA analysis of turkey spermatozoa by King et al. (2000) and applied in our studies on turkey semen storage (Kot lowska et al., 2007;S lowińska et al., 2012. ...
... The sperm dilutions, recording, and Hobson Tracker settings followed those described by King et al. (2000) and Kot lowska et al. (2007) for turkey spermatozoa. Semen samples were incubated for 5 min in the motility buffer (50 mM Tris buffer, pH 7.4, 120 mM NaCl, 10 mM glucose, 2 mM CaCl 2 ) at 39 • C. The motility buffer composition used in the present study was previously adapted for CASA analysis of turkey spermatozoa by King et al. (2000) and applied in our studies on turkey semen storage (Kot lowska et al., 2007;S lowińska et al., 2012. A study of Lemoine et al. (2008) showed no induction of acrosome reaction of chicken semen in NaCl-TES buffer in the presence of Ca 2+ and BSA if buffer was not supplemented with the inner perivitelline layer. ...
Numerous studies have indicated that yellow semen syndrome (YSS) of turkey is associated with the production of low semen quality, resulting in reduced fertility and hatchability. It is unknown at present if the etiology of YSS also could be linked to low-molecular weight metabolites. The aim of this study was to examine the metabolome of white and yellow seminal plasma of turkeys. Two different metabolomics approaches, shotgun (direct infusion) and liquid chromatography-mass spectrometry (LC-MS), were employed to identify metabolites differentially abundant in yellow seminal plasma. Significant changes in the levels of 1549 and 2093 metabolites were detected in yellow vs. white seminal plasma using shotgun and LC-MS, respectively. Of these, 354 metabolites (189 increased and 165 decreased) after shotgun and 936 metabolites (363 increased and 573 decreased) after LC-MS were putatively identified using the Human Metabolome Database. Significantly differentiated metabolites were subjected to Ingenuity Pathway Analysis. Lipid metabolism, molecular transport, and nucleic acid metabolism were the top pathways that differentiated white and yellow seminal plasma. These data strongly suggest that disturbance of carbohydrate and lipid metabolism is characteristic for YSS. The abnormal metabolism of lipids may contribute to the numerous lipid vacuoles previously observed in the reproductive tracts of YSS males. An increased level of riboflavin in YSS may be responsible for yellow turkey semen pigmentation. A disturbance in thyroid hormone metabolism visible at protein and metabolic levels may be involved in YSS in turkey. The low quality of YSS may be linked with the presence of drug residues in the reproductive tract.
... In this sense, Froman (2003) proposes that spermatozoa with a high VSL value present a competitive advantage since they can remain for a longer period in the sperm storage tubules, a structure present in the reproductive apparatus of hens that allows the storage of spermatozoa for a variable time of approximately two weeks after artificial insemination or natural mating, which is a great advantage (Etches 1998). In turkeys, linearity is an important characteristic of sperm function (King et al. 2000) and this variable has also been shown to determine fertilisation success in other species such as Atlantic salmon (Gage et al. 2004), rats (Moore and Akhondi 1996), and boars (Holt et al. 1997). Likewise, in humans, VSL has been determined to be one of the only sperm variables that is significantly related to the natural pregnancy rate in subfertile couples (Garrett et al. 2003). ...
... It has been proposed that sperm have a more progressive pattern of motility when they have smaller (lower ALH) but more frequent (higher BCF) head movements (Papin et al. 2021). Furthermore, BCF has been positively correlated with highly motile sperm subpopulations (King et al. 2000); however, some other studies have linked this parameter to sperm hyperactivation and DNA fragmentation (Aghazarian et al. 2021), while others have concluded that the sensitivity of BCF for detecting adverse effects on sperm motion is unclear (Kato et al. 2001). Although advanced, automated semen analysis methods such as CASA offer many advantages including standardisation, speed, accuracy and objectivity, their parameters do not reliably predict fertility in equines (Whitesell et al. 2020). ...
... Furthermore, STR and LIN can also describe the swimming pattern, while BCF and VCL could be considered specific markers of the viability of sperm [141,151]. Meanwhile, the MAD index is a precious parameter of the male gametes' path deviation from linearity and is inversely correlated with the sperm mobility phenotype [152]. Lower values in all kinematic indices, even insignificant in BCF and WOB, alongside a reduction in the level of sperm forward motility, were recorded in Pb-treated animals, probably indicating that male gametes will not be Fig. 8 Tissue histopathological alterations and stereological analysis of seminiferous tubules in Pb-treated mice. ...
Background
Taurine (TAU) is the most abundant non‐protein amino acid in the central nervous system (CNS). However, the molecular mechanism of TAU in the CNS is still poorly understood. Meanwhile, disruption in mitochondrial dynamics is evident in CNS disorders. This study aimed to investigate the effect of TAU on mitochondrial dynamics.
Methods
TAU (0.25, 0.5 and 1% in drinking water) was administered to young mice for six months. Several memory/cognition parameters and indices of anxiety/depression were assessed. Meanwhile, various mitochondrial indices and the expression/activity of genes involved in mitochondrial biogenesis and dynamics (Akt, CREB, NRF1, TFAM, PGC‐1α, Mfn1, Mfn2, UCP2, PINK1, OPA1, Drp1 and Fis1) were examined.
Results
TAU significantly enhanced memory performance, suppressed anxiety and depression‐like behaviour, increased mitochondrial biogenesis/dynamics and improved mitochondrial indices. It should be mentioned that there was no significant difference between different concentrations of TAU in changing most brain mitochondrial dynamic biomarkers in the current study.
Conclusions
These findings offer more insights into the molecular mechanism for TAU's action in the CNS. However, there is a need for further research to confirm these effects in humans. Overall, this study suggests the potential application of TAU in various neurological disorders and the need for clinical studies on the effects of this amino acid in the brain.
... Fresh and cryopreserved semen samples were diluted to 1.0 × 10 9 sperm/mL in a solution consisting of 111 mM NaCl buffered with 50 mM N-tris-[hydroxymethyl]methyl-2 amino-ethanesulfonic acid (TES), pH 7.4, containing 25 mM glucose and 4 mM CaCl 2 [ 2 ]. An aliquot of each sperm suspension (0.060 mL) was carefully overlaid on 0.600 mL of pre-warmed Accudenz® solution at a 4 % concentration (w/v) [ 5 ], in a semimicro polystyrene disposable cuvette. Subsequently, they were incubated in a water bath at 41 °C for 5 min and placed in a spectrophotometer (Jasco V-570, Tokyo, Japan), where the absorbance was measured at a λ of 550 nm after 1 min. ...
Significant changes in the quality and metabolic profile of fresh turkey sperm as a result of both cryopreservation and reproductive age have already been individually confirmed in our previous studies. This new dataset adds a relevant piece to the tangled puzzle of changes in metabolite levels affecting cryopreserved turkey sperm quality, taking into consideration two different reproductive period ages. Fresh semen samples were collected at 44 and 56 weeks of age and exposed to the cryopreservation process. All fresh and frozen-thawed samples were subjected to analysis of mobility, viability and osmotic tolerance as parameters for evaluating the sperm quality, while NMR spectroscopy was used to assess the quantitative changes in water and lipid-soluble metabolites. Our results showed that the cryopreservation process significantly affected all of the measured qualitative parameters both at 44 and 56 weeks. Concerning the metabolic profile, a greater number of quantitative changes for both water and lipid-soluble components were found in frozen semen at 56 weeks than at 44 weeks of age. These data could contribute to identifying new strategies aimed at improving freezing procedures even as reproductive age increases.
... Improving the survival rate and motility of chilled poultry semen stored at 4-8 oC for a period of 48-72 h with a natural extender such as Tris Coconut-water orange juice extender (TCWOE) could be an effective way of increasing utilization of artificial insemination (AI) in the poultry breeding industry. The success and prospects of artificial insemination in poultry have been documented that they generally vary with species due to variation in semen quality in terms of functionality within and between species, males as well as ejaculates (Blanco et al. 2000;King et al. 2000;Yu et al. 2002;Blesbois et al. 2005;Chaveiro et al. 2006;Roca et al. 2006;Leahy and Gadella 2011). Ultimately, prospects of semen dilution with extender for artificial insemination varies with species, methods, and procedures for semen preservation and insemination. ...
... The notable exception was BCF, which was significantly highest when exposed to 20% EG. Although increased BCF appears to be directly related to motility in turkeys [70], its significance for evaluating sperm fertility or function is less clear in mammals and teleosts. Some reports have correlated decreased BCF to an overall lack of ATP and or sperm membrane damage [46], however, its inconsistent identification as a significant parameter makes it usefulness unclear. ...
abstract: Three experiments were performed to evaluate the effects of osmolarity, temperature, media and cryoprotectants on white spotted bamboo shark (Chiloscyllium plagiosum) spermatozoa. Semen was collected from 7 different males using a massage technique with the animals held in tonic immobilization with gills submerged. A Computer Assisted Sperm Analysis (CASA) system was used to assess sperm motility. Experiment one evaluated the effects of Hank’s balanced salt solution (HBSS) with different osmolarities (200, 400, 800 and 1000 mOsm Kg1) and incubation temperatures (5 °C and 24 °C [room temperature]) over time (6 h, 12 h and 24 h). Membrane integrity and morphology were degraded (P 0.05) sperm parameters were observed with HBSS 1000 and HBSS 800, respectively. In addition, TM, VAP, and curvilinear velocity (VCL) were all reduced (P
... Furthermore, STR and LIN can also describe the swimming pattern, while BCF and VCL could be considered specific markers of the viability of sperm [141,151]. Meanwhile, the MAD index is a precious parameter of the male gametes' path deviation from linearity and is inversely correlated with the sperm mobility phenotype [152]. Lower values in all kinematic indices, even insignificant in BCF and WOB, alongside a reduction in the level of sperm forward motility, were recorded in Pb-treated animals, probably indicating that male gametes will not be Fig. 8 Tissue histopathological alterations and stereological analysis of seminiferous tubules in Pb-treated mice. ...
Lead (Pb) is a highly toxic heavy metal. Pb exposure could adversely affect many organs, including the male reproductive system. Oxidative stress and mitochondrial impairment play a fundamental role in the pathogenesis of Pb-induced male reproductive system injury. Taurine (TAU) is abundantly found in mammalians body. The positive effects of TAU on the oxidative stress biomarkers and mitochondrial function have been reported. In the current study, the effects of TAU on Pb-induced reproductive toxicity is evaluated. Mice received Pb (20 mg/kg/day; gavage, 35 consecutive days). Then, sperm indices (quality and quantity) together with sperm kinetics, sperm mitochondrial parameters, testicular and sperm oxidative stress biomarkers, testis and plasma testosterone levels, and the expression of genes involved in the steroidogenesis process have been evaluated. Pb caused significant histopathological alterations and oxidative stress in male mice reproductive system and sperm. Moreover, significant mitochondrial function impairment was evident in sperm isolated from Pb-treated mice. Pb exposure also suppressed the expression of StAR, 17β-HSD, CYP11A, and 3β-HSD genes in the male gonad. It was found that TAU (500 and 1000 mg/kg) significantly improved oxidative stress biomarkers in both male gonads and gametes of Pb-treated mice. TAU also significantly restored sperm mitochondrial function and kinetics. The expression of genes involved in steroidogenesis was also higher in TAU-treated animals. These data suggest TAU as a safe and effective agent against Pb-induced reproductive toxicity. The effects of TAU on oxidative stress markers, mitochondrial function, and the steroidogenesis process seem to play a fundamental role in its protective properties.
... Improving the survival rate and motility of chilled poultry semen stored at 4-8 oC for a period of 48-72 h with a natural extender such as Tris Coconut-water orange juice extender (TCWOE) could be an effective way of increasing utilization of artificial insemination (AI) in the poultry breeding industry. The success and prospects of artificial insemination in poultry have been documented that they generally vary with species due to variation in semen quality in terms of functionality within and between species, males as well as ejaculates (Blanco et al. 2000;King et al. 2000;Yu et al. 2002;Blesbois et al. 2005;Chaveiro et al. 2006;Roca et al. 2006;Leahy and Gadella 2011). Ultimately, prospects of semen dilution with extender for artificial insemination varies with species, methods, and procedures for semen preservation and insemination. ...
Liquid storage of semen preservation for poultry species is deemed to be the only reliable and cost effective method compare to cryopreservation. This experiment was conducted to examine the preservation capacity of Tris coconut-water orange juice extender (TCWOE) on tom and cock semen for artificial insemination. TCWOE was prepared. Semen ejaculates were collected and pooled from five toms and cocks separately. The pooled semen from each species was divided into two portions and in one portion of each species the extender was added in the ratio of 1:3 (semen: extender), making a total of four treatments. Experimental design used was a complete randomized design consisting of three trials. Semen microscopic parameters like motility, viability, membrane integrity, and acrosome integrity were examined and recorded for freshly extended semen and preserved semen at 4-8oC every 12 h interval till 72 h. The motility and membrane integrity of freshly extended cock semen was significantly higher (P<0.05) compared to motility of extended tom semen and membrane integrity of un-extended tom semen. During storage up to 24 h, extended tom and cock semen showed significantly higher (P<0.05) motility and membrane integrity compare to un-extended semen. Extended cock semen had significant (P<0.05) higher percentage motility, livability and membrane integrity compared to un-extended and extended tom semen from 36 h to 72 h of storage. However, no significant difference (P>0.05) was observed for acrosome integrity. Conclusively, cock semen survived longer and had better results considerable for artificial insemination than tom semen up to 72 h storage. However, the extender was beneficial for storage of both species semen.
... 116 Furthermore, King and colleagues showed an inverse relationship between sperm mobility phenotype and MAD. 122 Pb-exposed mice, possibly indicating that the exposed spermatozoa will not be capable of transiting successfully throughout the female reproductive tract and finally reach the place of fertilization. 120 The insignificant improved motility-related path indices, such as linearity (LIN) and straightness (STR), along with significant higher VSL, VCL, and VAP (velocity indices) in the animals exposed to glycine (250 and 500 mg/kg) as compared with their treated groups only with (Pb) suggests an overall capability of this amino acid in reversing the deteriorating role of the toxic element on the swimming speed of spermatozoa. ...
Lead (Pb) is a highly toxic heavy metal widely dispersed in the environment because of human industrial activities. Many studies revealed that Pb could adversely affect several organs, including the male reproductive system. Pb-induced reproductive toxicity could lead to infertility. Thus, finding safe and clinically applicable protective agents against this complication is important. It has been found that oxidative stress plays a fundamental role in the pathogenesis of Pb-induced reprotoxicity. Glycine is the simplest amino acid with a wide range of pharmacological activities. It has been found that glycine could attenuate oxidative stress and mitochondrial impairment in various experimental models. The current study was designed to evaluate the role of glycine in Pb-induced reproductive toxicity in male mice. Male BALB/c mice received Pb (20 mg/kg/day; gavage; 35 consecutive days) and treated with glycine (250 and 500 mg/kg/day; gavage; 35 consecutive days). Then, reproductive system weight indices, biomarkers of oxidative stress in the testis and isolated sperm, sperm kinetic, sperm mitochondrial indices, and testis histopathological alterations were monitored. A significant change in testis, epididymis, and Vas deferens weight was evident in Pb-Mohammad Mehdi Ommati and Hassan Nategh Ahmadi contributed equally in this study.
... Tung et al. [20] measured STR values of 0.87 ± 0.02 for bovine sperm tracked for 2.81 s. Using this value we estimated the deflection angles for sperm movement, by simulating agent movement in a box and calculating their STR [20,35]. The agents were positioned in the middle of the chamber, restricting movement to z-direction. ...
Sperm migration in the female genital tract controls sperm selection and, therefore, reproductive success as male gametes are conditioned for fertilization while their number is dramatically reduced. Mechanisms underlying sperm migration are mostly unknown, since in vivo investigations are mostly unfeasible for ethical or practical reasons. By presenting a spatio-temporal model of the mammalian female genital tract combined with agent-based description of sperm motion and interaction as well as parameterizing it with bovine data, we offer an alternative possibility for studying sperm migration in silico. The model incorporates genital tract geometry as well as biophysical principles of sperm motion observed in vitro such as positive rheotaxis and thigmotaxis. This model for sperm migration from vagina to oviducts was successfully tested against in vivo data from literature. We found that physical sperm characteristics such as velocity and directional stability as well as sperm-fluid interactions and wall alignment are critical for success, i.e. sperms reaching the oviducts. Therefore, we propose that these identified sperm parameters should be considered in detail for conditioning sperm in artificial selection procedures since the natural processes are normally bypassed in reproductive in vitro technologies. The tremendous impact of mucus flow to support sperm accumulation in the oviduct highlights the importance of a species-specific optimum time window for artificial insemination regarding ovulation. Predictions from our extendable in silico experimental system will improve assisted reproduction in humans, endangered species, and livestock.
... Contrarily, sperm mobility has been demonstrated to have a significant correlation with paternity [50][51][52][53]. In fact, sperm mobility has been reported to be predictive of fertility [54,55]. In addition, it was demonstrated that a higher sperm concentration cannot counteract a lower sperm mobility in terms of the fertility rate [53,56], and according to Froman [52], every mobile sperm cell must be motile, but not every motile sperm cell is mobile. ...
The sperm of each avian species and breed have unique characteristics that render them more or less susceptible to the freezing–thawing process; therefore, a suitable cryopreservation protocol that is specific for the sperm of each type of bird is needed. In this context, little information about the common pheasant’s sperm is available. Therefore, the aim of this study was to test different parameters at each step of the process of freezing into pellets and thawing to detect the least deleterious parameter settings. Sixteen different protocols were tested by studying two levels in each of the four steps (dilution, equilibration at 5 °C, final dimethylacetamide concentration, and dimethylacetamide equilibration time) comprising the freezing process. The pheasant sperm exhibited a high susceptibility to the damage caused by freezing into pellets; however, the survival of the sperm reached 29%, and the greatest recovered mobility was 22%. The mobility of the sperm was affected by the dilution and the dimethylacetamide concentration, and the viability of the sperm was affected by the equilibration at 5 °C and the dimethylacetamide equilibration. The protocols that caused the least damage to the pheasant sperm were found to be those with higher dilution rates, 10 minutes of equilibration at 5 °C, and 6% dimethylacetamide equilibrated for 1 or 5 minutes. In the present study, we individualise some applicable parameters for certain critical steps of the freezing–thawing process; however, further investigations are needed in order to improve upon and complete a suitable protocol for the cryopreservation and thawing of pheasant sperm.
... Tung et al. [20] measured STR values of 0.87 ± 0.02 for bovine sperm tracked for 2.81 s. Using this value we estimated the deflection angles for sperm movement, by simulating agent movement in a box and calculating their STR [20,35]. The agents were positioned in the middle of the chamber, restricting movement to z-direction. ...
Sperm migration in the female genital tract controls sperm selection and, therefore, reproductive success as male gametes are conditioned for fertilization while their number is dramatically reduced. Mechanisms underlying sperm migration are mostly unknown, since in vivo investigations are mostly unfeasible for ethical or practical reasons. By presenting a spatio-temporal model of the mammalian female genital tract combined with agent-based description of sperm motion and interaction as well as parameterizing it with bovine data, we offer an alternative possibility for studying sperm migration in silico. The model incorporates genital tract geometry as well as biophysical principles of sperm motion observed in vitro such as positive rheotaxis and thigmotaxis. This model for sperm migration from vagina to oviducts was successfully tested against in vivo data from literature. We found that physical sperm characteristics such as velocity and directional stability as well as sperm-fluid interactions and wall alignment are critical for success, i.e. sperms reaching the oviducts. Therefore, we propose that these identified sperm parameters should be considered in detail for conditioning sperm in artificial selection procedures since the natural processes are normally bypassed in reproductive in vitro technologies. The tremendous impact of mucus flow to support sperm accumulation in the oviduct highlights the importance of a species-specific optimum time window for artificial insemination regarding ovulation. Predictions from our extendable in silico experimental system will improve assisted reproduction in humans, endangered species, and livestock.
... It was previously reported that high VCL, VSL, ALH, and LIN of spermatozoa are highly related to fertility rate in bull (Al-Qarawi et al., 2002;Perumal et al., 2011). High motile group showed that high VSL, VAP, VCL, LIN, and BCF compared to those of low motile group (King et al., 2000). In human, ALH of spermatozoa in pregnant group was significantly higher than that of non-pregnant group. ...
In this study, we examined the effect of a liposome-based extender (Optixcell) and a tris-citric egg-yolk extender (Triladyl) on the frozen-thawed spermatozoa characteristics and the calving rate. The percentages for the total motility of the frozen-thawed spermatozoa were similar in the Optixcell and Triladyl groups. However, among the motile spermatozoa with a straight line velocity (VSL) ≥ 25 µm/sec, the curvilinear velocity (VCL, µm/sec), VSL (µm/sec), average path velocity (VAP, µm/sec), amplitude of lateral head displacement (ALH, µm), beat cross frequency (BCF, Hz), and plasma membrane integrity of the frozen-thawed spermatozoa for the Optixcell group were significantly higher than those for the Triladyl group. Furthermore, the calving rate in the Optixcell group (79.0%) was higher than that of the Triladyl group (62.8%). However, the acrosomal membrane integrity of the frozen-thawed spermatozoa in the Optixcell and Triladyl groups was not significantly different. These results indicate that semen freezing with Optixcell improved the motility and plasma membrane integrity of frozen-thawed spermatozoa and the calving rate of Hanwoo cows (native Korean cattle). In conclusion, our results suggest that semen freezing with the liposome-based extender Optixcell is more efficient than with the tris-citric egg-yolk extender Triladyl for improved offspring production.
... Although the MOT did not change significantly during the infection, it was positively correlated with the fertilization rate. As shown in turkey (King et al., 2000), the sperm motility phenotype was not characterized by the percentage of motile sperm but by the sperm with certain movement patterns. Similarly, in our experiment, it can be stated that sperm motility was defined by movement pattern recorded as high BCF, which resulted in a higher hatching rate. ...
A controlled fish propagation is nowadays the fundamental prerequisite for well-managed aquaculture production. However, despite practices aimed at creating equal conditions during the spawning, the share of males giving offspring is highly diversified. In traditional pond aquaculture, the fish are exposed to various environmental factors that may affect their fertility through immune activation, among which parasites are one of the most influential. It is highly feasible that in conditions of sperm overload and short sperm-egg distance, the differences in fertilization success between males are related to sperm characteristics determined by male fitness. The present study aimed to investigate if mounting an immune response through parasitic infection affects common carp (Cyrpinus carpio L) sperm quality and reproductive success.
Mature carp males were experimentally infected with a dose of 1 × 10⁹ blood parasites Trypanoplasma borreli by intraperitoneal inoculation. Control fish were PBS-injected. Parasite number was determined at weekly intervals using a Bürker counting chamber. The level of parasitaemia developed within four weeks post-infection served as the criterion to classify each individual either as parasite susceptible or resistant. On day 24 post-infection, carp males were engaged in controlled spawning. Equal volumes of quantified semen, obtained by gentle abdominal massage, were used to fertilize the egg portions obtained from four females of different genetic origin. The calculated hatchability rate was used as the indicator of the males' reproductive success. The parasite-infected males demonstrated average hatchability rates of 14.18 ± 4.83%, while the control males demonstrated 11.29 ± 3.93% rates. Parasites induced a robust adaptive antibody response, but individual antibody levels were not associated with hatchability rate. A significantly higher number of spermatozoids with damaged DNA was detected in parasite-susceptible males than in the resistant ones. The 11-ketotestosterone and most CASA parameters did not differ between parasite-infected and control fish except beat cross frequency (BCF), which was higher in infected fish (57.45 ± 18 Hz in infected vs 43.64 ± 9 Hz in controls). This CASA parameter, alongside the percentage of motile spermatozoa (MOT), was positively associated with the hatchability rate.
Our study shows that in artificial spawning, a mild level of parasitaemia does not negatively influence common carp males' reproductive success. The only sperm characteristic altered by immune activation is BCF, which was correlated with hatchability in infected fish. However, the mechanism by which the activated immune system can alter BCF values and how this sperm parameter affects hatchability remains unknown and requires further investigation.
... Several sperm velocity parameters: curvilinear velocity (VCL), average path velocity (VAP) and straight-line velocity (VSL), were evaluated. Sperm velocities contribute to overall sperm mobility and are important characteristics of sperm function [14,15]. Methods: The study is a single group pretest-posttest experimental prospective comparative self-control; clinical trial research, it has been conducted in Sudan -Khartoum state at andrology clinic in the Reproductive Health Care Centre (RHCC), in the period from September till December 2013. ...
Background: The use of minerals in treatment of different diseases is as old as man himself. zinc is the most famous trace mineral related to male sexual function. Oligoasthenozoospermic subfertile patients were treated with zinc sulphate for three months.
Objectives: Aim of the research is to investigate the role of Zinc and if it affects the abnormalities of some semen parameters and to study the possible role of pharmaceutical preperations of zinc in amelioration of male subfertility as well as to assess the ability of Zinc to induce changes in the serum and semen zinc levels in addition to the levels of reproductive hormones (FSH and Testosterone).
Type of the study: The study is a single group pretest-posttest experimental prospective comparative self-control; clinical trial research.
Methods: The patients were tested before and after the treatment for semen analysis via Computerized Assisted Semen Analysis (CASA) dynamic analysis report I and II as well as for FSH and Testosterone hormonal levels, serum and semen zinc levels .
Results: Zinc administration induced a significant increase(p≤0.001) in FSH, Testosterone, serum and semen zinc level as well as in the total and progressive sperm motility percentages .
Conclusions: Zinc administration induced significant changes (p≤0.001) towards improvement in the total and progressive sperm motility percentages in oligoasthenozoospermic patients by CASA dynamic analysis report I and II.
... However, our CASA data analysis provides clear quantitative evidence that older males produce fewer motile sperm and that these sperm have poorer forward movement. The CASA parameters of sperm movement (VCL, VSL) have previously been reported to be positively correlated with sperm mobility and egg fertility in turkeys as well as quail (King et al., 2000;Hirano et al., 2001;Farooq et al., 2018). In humans also, age-related declines in %MOT, %Rapid and %PROG and other sperm kinematics are linked to lower egg fertilization rate (Abramsson, 1988;Sloter et al., 2006). ...
For successful breeding programs, it is important to quantify the useful period of a male's reproductive life and it is often done simply by measurement of semen quality. This information is lacking for Japanese quail so we tested whether there is a decline in ejaculate quality and sperm kinematics with age, and whether the decline varies among strains. Nine males (n = 9) from each of 5 strains (A, B, C, D and E) were subjected to 4 semen collections (n = 16 per male) at 8, 16, 26 and 36 weeks of age. Ejaculate volume, sperm concentration and total sperm per ejaculate were measured, and sperm kinematics were analysed using a Sperm Class Analyser (SCA®). There was a significant effect of age for ejaculate volume, total sperm per ejaculate and per cent medium sperm. The effect of the interaction between age and strain was significant for percent progressive motile sperm, percent rapid sperm, velocity curvilinear, velocity straight line, velocity average path, linearity, straightness and beat cross frequency. Ejaculate volume peaked at Week 26 in all strains, while peak values for sperm concentration and total sperm per ejaculate were observed at Week 16 for most strains. There were declines in percent motile sperm, progressive motile sperm and rapid sperm, and in velocity curvilinear velocity, velocity straight line and velocity average path, by Week 16 for most strains. Linearity declined by Week 26 in some strains, and all strains showed a significant decline in beat cross frequency by that age. In conclusion, the ability of CASA to detect age-related changes in sperm kinematics makes it a valuable tool for identifying the best males and thus improving quail flock fertility. It is essential that breeders understand that age affects both sperm production and sperm kinematics, and that the changes vary with strain.
... For avian species, a similar test has been established: penetration of a dense solution of Accudenz ® (Accurate Chemical & Scientific Corporation, Westbury, NY, USA) is used to determine the progressive motility of sperm [31] and is correlated to fertility in the chicken [32]. King et al. [33] were able to prove a correlation between the sperm mobility test and distinct CASA (computer assisted semen analysis)-parameters in turkeys (Melleagris gallopavo). However, all these tests contribute only to determining the ability of gametes to reach the site of fertilization. ...
The aim of this study was to adapt an inner perivitelline membrane (IPVM) test as an interspecies penetration assay for avian spermatozoa. The IPVM of different bird species was evaluated to test the penetrating ability of avian spermatozoa in an intra- and interspecies design. Isolation of the IPVM via acid hydrolysis was tested in pre-incubated chicken eggs and in six other avian species. The separation protocol was modified (time, acid concentration) to facilitate practicability. Separated membranes were evaluated with dark field microscopy for the presence of holes produced by penetrating spermatozoa. In chicken eggs, the influence of different membrane storage conditions was tested. In the penetration assay, the IPVM of chicken eggs was used as a model for fresh and frozen–thawed rooster sperm and for fresh spermatozoa of cockatiels and falcons. Results demonstrated that the time of egg-incubation had a significantly negative influence on the isolation ability of the IPVM (p < 0.0001). IPVM-separation was successful for a maximum of two days after preincubation. In the experiments with eggs from other avian species, results were heterogenous: there was no isolation in geese and cockatiels, 20% in the European kestrel, and 40% in pheasant, quail, and duck. In the penetration assay, holes were found in 100% of the IPVM of chicken eggs after incubation with native and frozen–thawed rooster semen and in 10% with fresh cockatiel semen. Falcon spermatozoa failed to produce visible holes. In conclusion, the IPVM of chicken eggs seems to be unsuitable to establish a functional sperm assay in other species tested but is suitable for quality evaluation of cryopreserved rooster sperm.
... We also confirmed results of previous studies in both turkey (King et al., 2000) and rooster ) that high-mobility sperm swim faster and more linearly under almost every metric examined. This result makes sense given that a sperm's ability to swim through Accudenz is influenced by its velocity and linearity. ...
The sperm mobility assay measures the ability of sperm to swim through a dense layer of Accudenz®, and the sperm mobility phenotype has been shown to predict fertility and other sperm performance traits in roosters and turkeys. In this study, we examined turkey sperm morphometry and rates of early embryonic death associated with high and low mobility semen. We also assessed whether the hypo‐osmotic stress test, which evaluates the structural integrity of the sperm plasma membrane, may be used as a faster and simpler assay for sperm mobility and viability. We confirmed previous work that found that high mobility sperm are faster and swim more linearly than low mobility sperm, and that mobility traits were repeatable within males. In contrast to previous studies, we did not find higher rates of fertility, but low mobility sperm was associated with higher rates of early embryonic death, though this trend was not significant. High mobility sperm had longer sperm heads, explained by longer nuclei, despite shorter acrosomes. Although these sperm were faster, midpiece length and flagellum length did not differ between high and low mobility sperm. Finally, mobility was not found to be associated with sperm performance in the hypo‐osmotic stress test.
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... 1998; King et al., 2000;Blebois et al., 2008 (Vantman, 1988). 닭의 경우, 정액 내 정자 농도가 평균 2∼5 × 10 9 /mL 정도 로 매우 높은 농도의 정액을 생산하므로 문제점을 야기할 수 있으며, 정액을 낮은 농도로 희석을 실시하면, 희석충격 에 의하여 운동성이 낮아지며 정자가 죽는 현상이 있음이 보고되어 있다 (Harrison et al., 1982). ...
... Since studies have reported this phenomenon, the motility of sperm during capacitation has been considered essential to fertilization. Computer-aided sperm analysis (CASA) has been used to obtain parameters of sperm motility [3], some studies have reported a correlation between those parameters of sperm motility and the rate of fertilization [4][5][6][7]. ...
... Analyses were done in duplicate; 100 sperm tracks were recorded from each side of the slide, for a total of 200 tracks/sample. Semen samples were incubated for 5 min in motility buffer (50 mM Tris buffer, pH 7.4, 120 mM NaCl, 10 mM glucose, and 2 mM CaCl 2 ) at 39 • C. The motility buffer composition used in the present study was previously adapted for CASA analysis of turkey spermatozoa by King et al. (2000) and applied in our studies on turkey semen storage (Kot lowska et al., 2007;S lowińska et al., 2012, 2013. The study of Lemoine et al. (2008) showed no induction of acrosome reaction of chicken semen in NaCl-TES buffer (applied in present study) in the presence of Ca 2+ and bovine serum albumin (BSA) if buffer was not supplemented with the inner perivitelline layer. ...
Yellow semen syndrome (YSS) is the most widely recognized problem among male turkeys. Yellow semen is of low quality and, when used for insemination, results in reduction of fertility and hatchability. Elevated level of serum albumin-like protein accession no. XP_003205725 is a characteristic feature of yellow seminal plasma suggesting albumin role in YSS pathology. However, knowledge regarding the expression of albumin in the reproductive tract in relation to YSS is very limited. The aim of this study was to identify albumin secretion and localization sites in the turkey reproductive tract in relation to YSS. Reproductive tract tissues and liver originating from turkeys producing white semen (WS) and YSS were used for analysis of albumin mRNA expression and its localization using immunohistochemistry. Moreover, albumin abundance in tissues, blood and seminal plasma was analyzed using two dimensional electrophoresis and western blot analysis. Albumin mRNA expression was found in all parts of the reproductive tract. Apart from the liver, the highest expression of albumin was found in the ductus deferens in YSS turkeys. The testicular spermatids, Leydig, and myoid cells and the epithelium of the epididymis and ductus deferens were the main secretion sites of albumin in the reproductive tract in turkeys. Higher albumin abundance was found in the reproductive tract and seminal plasma of YSS toms compared to WS toms. Our results demonstrated that germ cells from spermatocytes to spermatids, Leydig cells, and myoid cells synthesized and secreted albumin in turkey testis, and epithelial cells are the main secretion sites in epididymis and ductus deferens. Ductus deferens secretion of albumin seems to be mostly responsible for YSS. Over-secretion by the ductus deferens may be the main origin of albumin abundance in YSS semen. Knowledge regarding disturbances of albumin secretion in relation to YSS may be useful for future work on studies related to better understanding the molecular basis of YSS.
... Especially, group 3 was higher values than other groups in VCL, VSL, VAP, LIN parameters. VCL and VAP are evaluate of sperm velocity over specifical paths, so these values indicate that sperm classified as high mobility swim faster than those classified as lower mobility [21]. Thus, VSL's role in sperm transport may be during the passage through the female reproductive tract and penetration of the oocyte vestments [13,17]. ...
Background: Most part of ram spermatozoa membrane has unsaturated fatty acids (phospholipids). Membrane structure of cells is composed of double ordered phospholipid layers adorned with mosaic-like protein, glycoprotein and glycolipids. Sperm freezing protocols could be negatively affected on ram sperm motility, viability and acrosome integrity during cryopreservation. For these reasons, researchers were designed their topics has led to the search for effective antioxidant systems against peroxidative damage and spermatozoon dysfunction. There are three protective enzymatic systems against reactive oxygen species (ROS) damage in sperm. These include superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase / reductase cycles Catalase (CAT) is a hemo-protein in the enzyme tetramer structure. The aim of this study was to investigate the effects of trehalose, catalase and their combinations on ram sperm parameters after the cryopreservation/thawing process.Materials, Methods & Results: At the out of breeding season (March-May) seven rams (1-3 years of age) were used in this study. Ejaculates were collected by electro-ejaculator twice a week. Pooled ejaculates were at 37°C, divided into six aliquots, diluted with the Tris based extender containing Trehalose 25 mM (Group-1), Trehalose 50 mM (Group-2), Catalase 200 µg (Group-3), Catalase 400 µg (Group-4), Trehalose 50 mM + Catalase 400 µg (Group-5) and no anti-oxidant (control), respectively, were cooled to 5°C than frozen in 0.25 mL French straws on the nitrogen vapour and stored in liquid nitrogen. The extender supplemented with Group 1 (54.1 ± 1.53; 73.1 ± 4.37), 50 mM (58.3 ± 4.01; 63.1 ± 0.30) and Group 5 (56.6 ± 1.05; 58.3 ± 0.55) resulted in higher subjective motility in comparison to the control (40.0 ± 3.87; 40.5 ± 0.22) group respectively (P < 0.05). Besides, Group 1 (60.16 ± 4.39) and Group 5 (59.60 ± 2.21) led to higher CASA total motility when compared to control (44.40 ± 8.13) group (P < 0.05). Sperm progressive motility was better in Group 1 (20.57 ± 6.90) than the Group 3 (10.63 ± 3.59) [P < 0.05]. Casa kinetic parameters of catalase 200 (Group 3) was higher values than other groups in VCL, VSL, VAP, LIN parameters (P < 0.05). There were no statistically significant differences on the membrane integrity parameter between the groups (P > 0.05).Discussion: Freezing ram sperm is extremely difficult process when compared bull and dog semen. Previous studies showed that antioxidants which were adding into the ram freezing extender gave positive effects solely or combination. In this study similar results were taken at trehalose 25, 50 mM and trehalose 50 mM + catalase 400 µg except 200 and 400 µg catalase groups. These findings supported some researches but lots of them opposite of catalase results. Catalase is found semen and ameliorates the sperm parameters when adding the liquid storage. Also after diluted and equilibrated catalase groups motilities were better than the control group. During the freezing stage catalase efficiency has been restricted. On the other hand when it combined with the trehalose 50 mM, catalase activity was triggered. Trehalose acts on sperm as non-permanent had a protective action related both osmotic effect and specific interactions with membrane phospholipids. Our data suggest that solely Trehalose 50 mM or combination with Catalase 400 µg can be added to Tris based extender for improving the post-thawed sperm quality in ram semen.
... The slides were mounted on a heated stage (41 • C). Conditions of sperm dilutions, recordings, and Hobson Tracker settings followed those described by King et al. (2000) and Kot lowska et al. (2007) for turkey spermatozoa. Semen samples were incubated for 5 min in the motility buffer (50 mM Tris buffer, pH 7.4, 120 mM NaCl, 10 mM glucose, 2 mM CaCl 2 ) at 39 • C. Turkey semen was diluted 300 to 500 times to obtain 30 to 50 sperm on the screen at the start of motility. ...
The extensive use of artificial insemination in turkeys has led to the development of in vitro semen storage. However, fertility rates from liquid stored and frozen/thawed turkey semen are still unsatisfactory. The aim of the study was to assess spermatozoa viability, mitochondrial membrane potential (MMP), and reactive oxygen species production (ROS) in liquid stored and cryopreserved turkey semen with the use of flow cytometry. Moreover, motility and adenosine triphosphate (ATP) content in sperm were monitored at the same time to link flow cytometry data with sperm movement and energetics. Liquid storage led to a decrease in sperm motility (80.6 vs. 55.6%, for fresh and stored for 48 h), live sperm with an intact MMP (59.9 vs. 30.5% for fresh and stored for 48 h), and a 20-fold decrease in ATP content after 24 h of storage. A 3-fold increase in ROS+ sperm was observed after 48 h of storage (9.3 vs. 26.8% for fresh and stored for 48 h). Semen equilibration before cryopreservation affected only ATP content. However, freezing/thawing led to a dramatic decrease in all of the studied semen quality parameters. A 5-fold decrease in live sperm with intact MMP (59.8 vs. 11.9%) and a 7-fold increase in sperm ROS+ (10.8 vs. 74.4%) were recorded between fresh and frozen/thawed semen. The results strongly suggested that a significant loss of MMP and a disturbance in sperm ATP production during semen storage can be the main reason for the decline in sperm motility. The disturbance of mitochondria activity during storage seems to be associated with the increase in oxidative stress in turkey semen. Turkey sperm mitochondria also appear to be very sensitive to cryodamage. Diminished energy production in turkey spermatozoa, visible as the low percentage of sperm with an intact MMP and low level of ATP after freezing/thawing, which is associated with high ROS generation, could be responsible for the low fertilizing ability of cryopreserved turkey semen.
... Similar fertility distribution pattern (i.e. 30%, 46% and 22% as high, average and low fertility, respectively) has previously been reported for turkeys by mobility indexing (King, Holsberger, & Donghue, 2000) which suggests that categorization of quail sperm by 25% method could separate male population close to the normal pattern of fertility. In addition to that, the present study suggests that male fertility in quails can be explained by sperm motility parameters recorded by SCA, although future studies need to explore specific motility parameters that link closely to the fertility. ...
One of the basic steps in objective analysis of sperm motility is the subdivision of a motile sperm population into slow, medium and rapid categories based on their velocity. However, for CASA analysis of quail sperm, the velocity values for categorization of slow, medium and rapid sperm have not yet been standardized. To identify the cut-off values of “velocity curvilinear” (VCL) for quail sperm categorization, we captured and analysed 22,300 tracks of quail sperm using SCA®-CASA. The median and mean VCL values were 85 and 97 μm/s. To define the VCL cut-off values, we used two methods. In the first, we identified the upper (rapid sperm) and lower (slow sperm) cut-off values using: (i) median VCL ± 25% or ± 50% or ± 75% of median VCL value; (ii) first and third quartile values of VCL data (i.e. 25% cut-off setting); and (iii) 33% and 66% of VCL data. Among these settings, sperm categories and their corresponding motility characteristics recorded using the “25%” setting (i.e. slow ≤36 ≤ medium ≤154 ≤ rapid) were found the most realistic and coherent with male ranking by fertility. In the second method, we calculated heteroscedasticity in the total VCL data using PCA and the two-step clustering method. With this approach, the mean of the high and low clusters was 165 and 51 μm/s, respectively. Together, the mean from two methods suggested that, for SCA®-CASA categorization of quail sperm, sperm should be classed as “rapid” at VCL ≥160 μm/s and “slow” at VCL ≤45 μm/s.
... The importance of VSL for fertility assessment has also been observed in bulls (Gillan et al., 2008). VSL, VAP, and VCL are measures of sperm velocity over specific paths; these values indicate that sperm with high mobility swim faster than those with lower mobility (King et al., 2000). On the other hand, there was no significant effect of CLC on the control groups with respect to the kinetic parameters, when the age groups were evaluated separately. ...
The objective of the study was to determine the effect of cholesterol-loaded cyclodextrin (CLC) on the quality parameters of semen from Aksaray Malakli Shepherd dogs of different age groups. Forty-eight male dogs were divided into 3 groupings according to their ages (young age (Y): ≤3 years, n: 20; middle age (M): 4-6 years, n: 20; old age (O): ≥7 years; n: 8). The sperm-rich portion of the ejaculate from each dog was divided into four aliquots and extended with either tris as a control (C) or tris loaded with 0.5, 1.0, and 1.5 mg/120 × 106 CLC as low (L), intermediate (I), and high (H) doses, respectively. Following equilibration for at least half an hour, the straws were frozen in nitrogen vapor and then stored in liquid nitrogen at least for 48 h. Later, the frozen straws were thawed in a water bath for spermatological evaluation. Significant differences were observed between different age groups in terms of the spermatological parameters (p < 0.05). The evidence suggests that increasing age is associated with poor in-vitro spermatological parameters and CLC was able to protect the acrosome integrity from cryo-damage during the freeze-thawing process. Better semen freezability characteristics were obtained at young ages, considering the overall parameters.
... From this perspective, the decrease in LIN, STR and WOB values was not more than 17% from 24 to 48 h of storage when ostrich semen was diluted with OS2 (adapted for the micro minerals, Se and Zn) compared to the 26% decrease when diluted with OS1 (not adapted for micro minerals). Sperm quality variables, such as LIN, STR and WOB, are important for improved in vivo fertility rates in mammals (Budworth et al., 1988;Farrell et al., 1998) and in high mobility phenotypic males such as turkeys (King et al., 2000) and the domestic fowl (Froman and Feltman, 1998;Froman et al., 1999). In mammals, ejaculates specifically suitable for cryopreservation have greater percentages of LIN, STR and WOB sperm (Perumal et al., 2014). ...
The dilution of semen is a very important initial process for semen processing and evaluation, storage and preservation in vitro and efficient artificial insemination. The aim of the study was to evaluate the effect of two synthetic diluents (OS1 and OS2) on ostrich sperm motility parameters during in vitro storage. Formulation of OS1 was based on macro minerals (Na, K, P, Ca, Mg) and OS2 on the further addition of micro minerals (Se and Zn), based on mineral concentration determined in the ostrich seminal plasma (SP). Sperm motility was evaluated at different processing stages (neat, after dilution, during storage and after storage) by measuring several sperm motility variables using the Sperm Class Analyzer® (SCA). Processing (dilution, cooling and storage) of semen for in vitro storage purposes decreased the values for all sperm motility variables measured. The percentage motile (MOT) and progressive motile (PMOT) sperm decreased 20% to 30% during 24 h of storage, independent of diluent type. Quality of sperm swim (LIN, STR and WOB), however, was sustained during the longer storage periods (48 h) with the OS2 diluent modified with Se and Zn additions. Quality of sperm swim with use of OS1 was 6% to 8% less for the LIN, STR, and WOB variables. Male fitted as a fixed effect accounted for >60% of the variation for certain sperm motility variables (PMOT, MOT, VCL, VSL, VAP and ALH) evaluated at different processing stages. Semen from specific males had sustained sperm motility characteristics to a greater extent than that of other males during the 24-h storage period.
... Spermatozoa motility is a critical factor in the maintenance of fertility. In birds, the vaginal portion of oviduct regulates spermatozoa entry and only motile spermatozoa are able to traverse the vagina and enter into the hen's spermatozoa storage tubules (King et al. 2000). As described by Froman and McLean (1996), spermatozoa motility is a primary determinant of fertility in the fowl. ...
The effect of taurine on the turkey spermatozoa motility and viability during the in vitro incubation was assessed. Experimental samples were prepared by diluting the raw semen in nine different concentrations of taurine – from 10 mg/ml to 0.078125 mg/ml. The motility parameters were evaluated by the CASA system (Computer Assisted Semen Analyser) using the program Sperm Vision® and for spermatozoa viability assessment the eosin-nigrosin staining was performed. Selected parameters were evaluated at six time periods: 0, 1, 2, 3, 4, and 5 h at 5°C and 41°C. At 5°C, a significantly lower percentage of motility and progressive motility was detected only in the samples with the highest concentration of taurine (10 mg/ml) at time 0 and 1. After 2 h of incubation a significant preventive effect of taurine on spermatozoa parameters was observed. The tendency of the taurine effect on motility parameters was different during the in vitro incubation at 41°C. Significantly lower values of motility parameters were detected in all experimental samples in comparison to the control after 5 h. The analysed concentrations of taurine did not significantly affect viability of turkey spermatozoa during all time periods. A higher percentage of dead spermatozoa were observed at 41°C (4.87–9.90%) if compared to 5°C (2.12–4.88%). The results indicated that the addition of taurine (from 2.5 to 7.5 mg/ml) to turkey spermatozoa positively affected the monitored spermatozoa parameters incubated at 5°C.
... Sperm mobility was assessed using the Sperm Motility Test (SMT) according to the Accudenz 1 procedure (Accurate Chemical & Scientific Corp., Westbury, NY 11590) described by Iaffaldano et al. [22] with the Accudenz 1 concentration suggested by King et al. [23] for turkey semen. The test is based on the ability of the spermatozoa with a forward progressive motility to penetrate a 4% Accudenz 1 layer. ...
Metabolic profile of fresh turkey spermatozoa at three different reproductive period ages, namely 32, 44 and 56 weeks, was monitored by Nuclear Magnetic Resonance (NMR) spectroscopy and correlated to sperm quality parameters. The age-related decrease in sperm quality as indicated by reduction of sperm concentration, sperm mobility and osmotic tolerance was associated to variation in the level of specific water-soluble and liposoluble metabolites. In particular, the highest levels of isoleucine, phenylalanine, leucine, tyrosine and valine were found at 32 weeks of age, whereas aspartate, lactate, creatine, carnitine, acetylcarnitine levels increased during the ageing. Lipid composition also changed during the ageing: diunsaturated fatty acids level increased from 32 to 56 weeks of age, whereas a reduction of polyunsaturated fatty acids content was observed at 56 weeks. The untargeted approach attempts to give a wider picture of metabolic changes occurring in ageing suggesting that the reduction of sperm quality could be due to a progressive deficiency in mitochondrial energy producing systems, as also prompted by the negative correlation found between sperm mobility and the increase in certain mitochondrial metabolites.
... Sperm motility is a critical factor in the maintenance of fertility. Historically, it has been difficult to use sperm motility assessment as a predictor of fertility potential in poultry, possibly because of the subjectiveness of the methods used (King et al., 2000). However, in the recent twenty years computer-aided sperm analysis (CASA) technology has been used to investigate sperm motility parameters from several species: rats (Moore and Akhondi, 1996), rabbits (Farrell et al., 1993), boars (Holt et al., 1997), bulls (Amann, 1989), turkeys (Bakst and Cecil, 1992), and humans (MacLeod and Irvine, 1995). ...
The objective of our study was to evaluate the influence of different storage temperature on rooster sperm motility. Semen was collected once a week from Lohmann Light breeder males into prepared sterile tube. The heterospermic pool was diluted at the ratio of 1:100 in a commercial avian extender, divided into two aliquots and incubated at 8°C or 37°C. The quality of semen samples were evaluated using CASA system (Sperm Vision™) after 30, 60 and 120 min of incubation. We observed significantly (P<0.05) the highest progressive motility of rooster sperm after 60 min of spermatozoa incubation at 8°C, that was also significantly (P<0.05) higher in comparison to spermatozoa incubated for 60 min at 37°C. Basing on the observed results, we hypothesized that law temperatures (about 8°C) would be better for long-term storage of rooster spermatozoa. Further experiments with different semen diluents and storage temperatures and intervals are required in order to prove our hypothesis.
... Several sperm velocity parameters: curvilinear velocity (VCL), average path velocity (VAP) and straight-line velocity (VSL), were evaluated. Sperm velocities contribute to overall sperm mobility and are important characteristics of sperm function [14,15]. Methods: The study is a single group pretest-posttest experimental prospective comparative self-control; clinical trial research, it has been conducted in Sudan -Khartoum state at andrology clinic in the Reproductive Health Care Centre (RHCC), in the period from September till December 2013. ...
Background: The use of minerals in treatment of different diseases is as old as man himself. zinc is the most famous trace mineral related to male sexual function. Oligoasthenozoospermic subfertile patients were treated with zinc sulphate for three months. Objectives: Aim of the research is to investigate the role of Zinc and if it affects the abnormalities of some semen parameters and to study the possible role of pharmaceutical preperations of zinc in amelioration of male subfertility as well as to assess the ability of Zinc to induce changes in the serum and semen zinc levels in addition to the levels of reproductive hormones (FSH and Testosterone). Type of the study: The study is a single group pretest-posttest experimental prospective comparative self-control; clinical trial research. Methods: The patients were tested before and after the treatment for semen analysis via Computerized Assisted Semen Analysis (CASA) dynamic analysis report I and II as well as for FSH and Testosterone hormonal levels, serum and semen zinc levels. Results: Zinc administration induced a significant increase(p≤0.001) in FSH, Testosterone, serum and semen zinc level as well as in the total and progressive sperm motility percentages. Conclusions: Zinc administration induced significant changes (p≤0.001) towards improvement in the total and progressive sperm motility percentages in oligoasthenozoospermic patients by CASA dynamic analysis report I and II.
... Generally, the effects were negative for sperm kinetics, compared to the control group. The reduced VAP, VSL, VCL and ALH imply that spermatozoa of supplemented boars swam slower and with less progressive movement, whereas the reduced DAP, DSL and DCL indicate that they travelled shorter distances (Cancel, Lobdell, Mendola, & Perreault, 2000;King, Holsberger, & Donoghue, 2000). Such effects may result from the reduced mitochondrial functionality, which would impair energy metabolism that is essential for spermatozoa movement. ...
... The low-density lipoproteins might have a reduced intensity of interactive losses along with better capacity to prevent efflux of membrane constituents that resulted in higher values. Average path velocity (VAP) indicates average path covered and is correlated with axial (X axis and Y axis) (King et al., 2000). The VCL and VSL are considered to be the most important characteristics to evaluate the kinetic characters that influence the fertilizing ability of spermatozoa (Jobling et al., 2002). ...
The present study was designed to evaluate kinematic response of sperm cell to low-density lipoproteins (LDL) in fresh diluted, short-term (4°C) or long-term (-196°C) stored semen. Four healthy bucks of similar age and weight were selected as semen donor. The semen was collected twice a week using artificial vagina. The semen after initial evaluation was pooled and divided into three aliquots, each diluted with TRIS based extender containing 8% LDL to reach final concentration of 200 million sperm/ ml. The first aliquot was evaluated after 15 to 20 minutes of its storage at 37 °C, second after it storage at 4°C for 48 hours and third was cryopreserved and evaluated after seven days of storage. Percent live sperm, sperm responsive to hypo osmotic swelling test and those exhibiting rapid progression were significantly (P ≤ 0.01) higher in fresh diluted followed by short and long term sored semen. A significant (P ≤ 0.01) decrease in the kinematic characters (average path velocity (VAP, µm/sec), straight line velocity (VSL, µm/sec), Linearty (Lin%), Straightness (Str %), Wobble (WOB%), beat cross frequency (BCF %) and maximum amplitude-lateral head displacement (ALH, µm) was observed in short term followed by long term store semen as compared to fresh diluted semen. Low-density lipoprotein was able to maintain the curvilinear velocity (VCL, µm/sec) of sperm subjected to 4°C during short term storage. In conclusion, decrease in temperature during semen storage alter the sperm path and its velocities, but LDL has a protective effect on sperm flagellar assembly and mitochondrial energy production system that sustained the sperm capacity to travel total distance per unit time upto 4°C during short term storage.
Eggshell breakage and broiler bone disorders are major problems for the breeder and broiler industries which are linked to mineral metabolism and animal genetics. The purpose of this work was to discover the link between individual animal phenotypic differences in mineral metabolism against concentrations of novel plasma biomarkers including tartrate resistant acid phosphatase (TRAP) and bone alkaline phosphatase (BAP). A subset of hens were selected from a flock of Cobb 500 breeders with the best or worst eggshell quality based upon dual energy x‐ray absorptiometry (DEXA) and specific gravity (SG). Breeders were defined as having good eggshell quality (SG ≥ 1.080), or poor eggshell quality (SG < 1.080). Progeny hatched from breeders with good or poor eggshell quality were reared to 2 week of age and blood and bone samples were obtained after euthanasia. In both breeders and progeny, plasma concentrations of BAP and TRAP were measured, and bone mineral density was evaluated by DEXA. Results showed that breeders selected for eggshell quality had significantly different plasma concentrations of BAP (Good = 326.5 pg/mL, Poor = 253.2 pg/mL), and TRAP activity (Good = 2203 U, Poor = 4985 U). Breeders selected for eggshell quality produced progeny with different bone breaking strength (Good = 1.61 kg/mm, Poor = 1.47 kg/mm), tibia ash (Good = 45.9%, Poor = 42.2%), plasma BAP (Good = 372.3 pg/mL, Poor = 312.4 pg/mL), and lower plasma TRAP activity (Good = 18010 U, Poor = 23590 U). These data suggest that there is a strong correlation between the eggshell quality of breeders, performance and bone strength of progeny, and plasma of concentrations of BAP and TRAP in both breeder hens and progeny.
Objective:
Numerous raptor species including some falcon species are facing continuous population decline in the wild and some are threatened by extinction. To support these species, captive breeding and reintroduction programs are attempted. Besides conservation, some large falcon species are commonly used in falconry and therefore bred commercially. Assisted reproduction is established in falcon breeding since the 1970s and semen analysis is an integral part to enable assessment of breeding males, inclusion or exclusion of semen donors and quality control of semen prior to artificial insemination. Conventional methods for semen analysis are widely used, but are time consuming and depend on the investigator's experience and ability. Computer-assisted semen analysis (CASA) would offer an objective, fast and reproducible alternative, but as they have not been established in large falcon species, this was the aim of this study.
Material and methods:
To this end, we examined in 3 breeding seasons 109 semen samples of gyr-saker hybrid falcons (n=2) and peregrine falcons (n=4) in 940 fields of view using the Minitube CASA SpermVision and compared these results to results of conventional methods of semen analysis. We used a preprogrammed setting and adapted two settings of CASA according to specific semen characteristics of falcons.
Results:
Sperm velocity, motility and viability parameters were recorded successfully using CASA. Correlation of conventional and computer-assisted motility analysis improved during the process of adaptation of CASA settings, but both methods differed significantly due to misinterpretation of round bodies and semen impurities by CASA. Viability values of conventional and computer assisted viability analysis using SYBR-PI correlated significantly while sperm concentration did not at all.
Conclusion:
CASA failed to replace conventional semen analysis regarding sperm motility and sperm concentration using 3 different settings, as a reliable differentiation between spermatozoa, spermatids and round bodies was not achieved.
Clinical relevance:
Using CASA, sperm velocity parameters were measured in spermatozoa of captive-bred large falcons for the first time and may be used as orientation values.
Objective: To summarize the currently available Phase I and II clinical trials of the effects of Nonoxynol-9 (N-9) on human sperm structure and functions.
Methods: A systematic review and meta-analysis aiming to evaluate the spermicidal activity of N-9 on motility, was conducted in PubMed, EMBASE, and Cochrane databases by 10 March 2021. Counted numbers of progressive motile (PR) sperm in cervical mucus and the vanguard sperm penetration distances were analyzed. Other effects on sperm structures and physiological activities were reviewed as well.
Results: In the pooled results, percentages or counted numbers of PR sperm decreased after the treatment of N-9. Vanguard sperm penetration distance was shortened in treated groups. N-9 has been confirmed to damage the structures of sperm, as well as other organelles like acrosome and mitochondria. The physiological activities such as generation of reactive oxygen species (ROS), superoxide dismutase (SOD) activity, acrosin activity, and hemizona binding were all inhibited in the reviewed studies.
Conclusions: N-9 has several impacts on sperm owing to its potency in reducing sperm motility and cervical mucus penetration, as well as other functional competencies.
1 ، به حاضر مطالعه مقایسه منظور اسپرم کیفیت بر خوراک در سلنیوم معدنی و آلی منابع منجمد های یخ خروس شده گشایی های تحقیق، این در شد. انجام اکسیداتیو تنش تحت گوشتی مادر گله 22 راس سویه خروس قطعه 303 سن با 23 قالب در هفتگی با تصادفی کامال طرح 2 و تیمار 6 تیم شدند. نگهداری تیمار هر در پرنده از: بودند عبارت آزمایشی ارهای 1 جیره شاهد: گروه) بدون پایه مکمل منابع با سازی (دگزامتازون تزریق یا و سلنیوم CON ،) 2 گروه) DEX : (به دگزامتازون تزریق با همراه پایه جیره مقدار 2 میلی طی بدن وزن کیلوگرم بر گرم 3 به و مرحله میان) در روز یک صورت 3 گروه) DEX(OSe) دریافت گروه : کننده دگزامتازون گروه همانند 2 مکمل جیره با همراه ، با شده سازی 3 / 0 میلی به سلنومتیونین از خوراک کیلوگرم بر سلنیوم گرم عنوان و آلی سلنیوم منبع 2 گروه) DEX (ISe) پیش تحرک و کل تحرک بهبود سبب ،) اسپرم رونده گروه سایر به نسبت شد آزمایشی های (00 / 0 > p) اگرچه DEX(OSe) معنی تفاوت پیش حرکت در شاهد گروه با داری اسپرم، رونده (نداشت 00 / 0 < p (معدنی سلنیوم مصرف همچنین .) DEX (ISe) تاثیر) معنی پیش تحرک و کل تحرک بهبود بر داری اسپرم، رونده گروه به نسبت DEX نداشت (00 / 0 < p) نه دیگر، طرف از. زنده تنها پارامترهای بلکه اسپرم، پالسمایی غشای یکپارچگی و مانی پراکسیداز (گلوتاتیون بیوشیمیایی گروه در نیز دیسموتاز) سوپراکسید و DEX(OSe) گروه سایر به نسبت بود باالتر آزمایشی های (00 / 0 > p ب .) ه طور به کلی می نظر مکمل رسد، خروس جیره سازی تنش شرایط تحت که آلی سلنیوم با مادر های تزریق با می باشند، گرفته قرار دگزامتازون معدنی سلنیوم به نسبت اسپرم کیفیت بهبود موجب تواند شود .
Turkey semen contains cysteine-rich secretory proteins (CRISPs) that belong to the dominant seminal plasma proteins. We aimed to isolate and characterize CRISP from turkey seminal plasma and evaluate its possible involvement in yellow semen syndrome (YSS). YSS, which is well characterized, causes reduced fertility and hatchability. The protein was purified using hydrophobic interaction, gel filtration, and reverse phase chromatography. It then was subjected to identification by mass spectrometry, analysis of physicochemical properties and specific antibody production. The biological function of the isolated protein was tested and included its effects on sperm motility and migration and sperm-egg interactions. Sperm motility was measured with the CASA system using Hobson Sperm Tracker. The reproductive tract of turkey toms was analyzed for gene expression; immunohistochemistry was used for protein localization in the male reproductive tract, spermatozoa, and inner perivitelline layer. The isolated protein was identified as cysteine-rich venom protein-like isoform X2 (CRVP X2; XP_010706464.1) and contained feature motifs of CRISP family proteins. Turkey CRVP X2 was present in both spermatozoa and seminal plasma. The extensive secretion of CRVP X2 by the epithelial cells of the epididymis and ductus deferens suggests its involvement in post-testicular sperm maturation. The internally localized CRVP X2 in the proximal part of the sperm tail might be responsible for stimulation of sperm motility. CRVP X2 on the sperm head might be involved in several events prior to fusion and may also participate in gamete fusion itself. Although the mechanisms by which CRPV X2 mediates fertilization are still unknown, the involvement of complementary sites cannot be excluded. The disturbance of CRVP X2 expression can serve as an etiologic factor of YSS in the turkey. This study expands the understanding of the detailed mechanism of fertilization in birds by clarifying the specific role of CRVP X2.
• PercollTM is one of the most widely used colloid for animal sperm preparation. The aim of this study was to evaluate whether PercollTM colloid centrifugation could be practical to improve cockerel sperm quality, and to compare the effects of PercollTM single layer centrifugation (SLC) and density gradient centrifugation (DGC) in order to obtain the most optimal protocol for cockerel semen.
• In the experiment with PercollTM SLC for fresh semen, an increase of motile sperm was seen after PercollTM 80% SLC and 90% SLC was conducted, at levels of 28.8% and 30.2% respectively (P<0.01). The increase of progressively motile sperm after PercollTM 80% SLC and 90% SLC was 177.2% and 202.4% respectively (P<0.01). Meanwhile, for semen stored at 4℃ for 24 hr, the increase of motile sperm after PercollTM 70% SLC and 80% SLC was 41.2% and 44.0% (P<0.01), and the increase of progressive sperm after PercollTM 70% SLC and 80% SLC was 71.3% and 83.1% respectively (P<0.01). Both the percentage of motile sperm and progressive sperm of the fresh and stored cockerel semen after appropriate PercollTM SLC was significantly enhanced.
• Sperm membrane integrity did not show any decrease after PercollTM centrifugation compared with non-centrifuged semen, which suggested that the PercollTM centrifugation treatment in this study did not cause damage to cockerel sperm membranes.
• In the experiment regarding the comparison of PercollTM SLC and DGC with fresh semen, the increase of motile sperm after PercollTM 80% SLC, 90% SLC and 40%/80% DGC was 29.5%, 36.4%, and 25.0% respectively; and the increase of progressive sperm was 44.7%, 58.5%, and 54.7%, respectively. For semen stored at 4℃ for 24 hr, the increase of motile sperm after PercollTM 70% SLC, 80% SLC and 35%/70% DGC were 41.2%, 44.0%, and 26.4%; and the increase of progressive sperm was 71.3%, 83.1%, and 43.7%, respectively. There were no significant differences between the increase of sperm motility after PercollTM 80%, 90% SLC or PercollTM 40%/80% DGC in fresh cockerel semen. There was no significant difference between PercollTM 70%, 80% SLC and PercollTM 35%/70% in stored cockerel semen. There was a tendency for sperm recovery rates with PercollTM SLC to be higher than PercollTM DGC, although this did not reach statistical significance in this study.
• It was concluded that PercollTM SLC was more suitable for cockerel sperm separation than PercollTM DGC. The results suggested that PercollTM 80% SLC was the most optimal procedure to separate fresh cockerel sperm and PercollTM 70% SLC was the most optimal procedure to separate stored cockerel sperm. PercollTM SLC is simpler, more user-friendlier, less time-consuming and more economical than DGC for cockerel semen processing.
Persistence of the semen quality and sperm function is one of the important factors in flock fertility preservation. The aim of this study was to assess the trends of semen and sperm parameters in selected cocks with superior and inferior sperm motility from Iranian native and Arian commercial line. Roosters were grouped into high (HML) and low motility (LML) at the early experiment (27 week). The trends for semen and sperm parameters were monitored during an eight-month period (30–60 weeks). Sperm motility in all groups showed an acceptable range (58–87%). The sperm motility trend of indigenous group was dissimilar to the commercial line. The trend line in the LML group from the indigenous roosters was ascending for the sperm motility, while the HML group showed a descending trend. Sperm concentration in both groups had a descending trend during the period of semen production. Sperm parameters, such as VAP, VSL, VCL, ALH and STR differed in the HML and LML groups in both strains (P < 0.05). In conclusion, selection of roosters based on the sperm motility at the early breeding is not sufficient for maintaining a desirable sire attributes and flock fertility. Therefore, increasing the rooster to hen ratio amid the production period may be a good managerial suggestion for maintaining the hen fertility and flock hatch rate.
Mounting an immunological response is energetically demanding and necessarily redirects allocation of resources toward immune system activation and away from other energetically expensive processes, such as reproduction. Lipopolysaccharide (LPS), a major component of the outer membrane of the cell wall of Gram-negative bacteria Escherichia coli, mimics a bacterial infection without producing the cost of replicating the pathogen and is one of the most commonly used agents to induce an acute phase immune response. Here, we ask if a trade-off can be induced between activation of the acute phase immune response and sperm function, a key indicator of sperm competitive ability. Further, we ask whether repeated exposure to this endotoxin in a social species such as the house sparrow (Passer domesticus), where repeated pathogen exposure may be common, may have a more pronounced effect. To address our questions, we exposed individuals to two rounds of LPS treatment or control, to mimic a repeated pathogen exposure in the wild. We predicted that repeated pathogen exposure would have detrimental effects on sperm quality, and therefore, reproductive success. We compared a measure of sperm quality (straight-line velocity) in captive male house sparrows between LPS-treated and control individuals. We found that although LPS treatment impaired circulating testosterone and induced a hypothermic state when compared with controls, it did not affect sperm quality within days or weeks following a single or repeated LPS exposure.
This study examined factors affecting the analysis of motility of chicken semen. The viability of spermatozoa was estimated using varying dilution ratios and supplementation with BSA or fatty acid free (FAF)-BSA as protein sources in semen diluent. Fresh semen was examined after preparing dilutions in beltsvile poultry semen extender (BPSE) of 1/8, 1/16 and 1/32 at 25^{\circ}C. The motility of incubated semen at each dilution was observed at 3 min (89.9%, 69.9% and 53.2%), 30 min (86.7%, 71.4% and 51.7%), 1 h (89.5%, 74.0% and 53.5%) and 3 h (78.5%, 66.5% and 45.7%), respectively. The addition of BSA or FAF-BSA to BPSE diluent significantly increased the viability of semen in 1/32 dilution with results of 53.2% (control), 84.8% (BSA) and 92.9% (FAF-BSA) (p
Males' fertility is one of the challenging problems in the poultry industry, and one of its biological indicators is sperm motility. Based on ,these biological properties were studied in a commercial (Arian) and a local (Urmia) strains. The samples consisted of 14 Arian birds and 36 local birds. Analysis of sperm motility were accomplished using computer and CASA software and statistical comparisons were performed by SAS. The results indicated that there are significant differences in Class A between the two strains, but in classes B, C and D did not differ significantly in terms of biological properties. This difference could be due to genetic mutations in genomic regions associated with the biological characteristics of sperm.
(1) The distributions of living and dead sperm in the oviducts of hens a t various time intervals after artificial insemination were determined by using sperm labelled with ³²P and assaying the radioactivity of serial sections of the oviduct. Appropriate. tests of the method showed it to be valid and reasonably accurate for short-term experiments. (2) The number of sperm reaching the site of fertilization at the upper end of the oviduct (the infundibulum) was dependent primarily on where in the lower genital tract the sperm were deposited. Following intravaginal insemination with 2 X 10⁸ sperm, from 7 X 10³ to 70 X 10³ sperm were detected in the infundibula of different hens up to 1 hr after insemination. After intra-uterine insemination with a like number, from 137 X 10³ to 2642 X 1010³ sperm were detected. (3) The junction of the vagina and uterus (or shell gland) proved to be a barrier to sperm progress, as was shown by the greater efficiency of sperm utilization above the junction than below it. (4) Dead sperm inseminated intravaginally did not pass into the uterus but those inseminated into the uterus reached the infundibulum in as great numbers as a similar sample of living sperm. This suggested that the mechanism of sperm transport differs on either side of the uterovaginal junction. (5) From the speed of transport of sperm and the passage up the oviduct of sperm-free fluid injected into the uterus, it is suggested that the spasmodic contraction of muscle investing the wall of the upper vagina and lower uterus induced as a response to tactile stimuli is mainly responsible for sperm movement from the uterovaginal junction to the infundibulum. (6) Motility of sperm is necessary only to traverse the vagina and perhaps to penetrate the vitelline membrane of the egg during the process of fertilization. At other stages of movement between the vagina and the egg, sperm play a passive role in their own transport.
Motility characteristics of turkey spermatozoa before and after storage for 24 h at 7 degrees C in diluent with and without bovine serum albumin (BSA; 1% final concentration) were measured by computer-assisted semen analysis. BSA significantly increased the percentage of motile spermatozoa and sperm velocity, linearity, lateral head displacement and beat frequency in each treatment, but BSA in fresh or stored semen in diluent did not augment hen fertility over 15 weeks of egg production. Fatty-acid-free BSA, globulin-free BSA and Fraction V BSA all significantly increased each sperm motility characteristic compared with semen in diluent alone. The lack of correlation between sperm motility and fecundity emphasizes the need to develop procedures for semen evaluation that accurately predict the fertilizing capacity of an aliquot of semen.
This study was conducted to determine the number of spermatozoa residing in the oviduct sperm-storage tubules (SST) and the relationship between these numbers and the number of spermatozoa embedded in the perivitelline layer of oviductal eggs after a single insemination of 200 x 10(6) spermatozoa. The SST of hens inseminated within one week before the expected onset of egg production were filled faster (4 h vs. 2 days) and possessed more spermatozoa (4.1 vs. 2.0 x 10(6)) than the SST of hens inseminated after the onset of egg production. Furthermore, for hens in egg production, significantly fewer spermatozoa were recovered from the SST if the hen was inseminated within 2 h before or after oviposition than if inseminated more than 2 h before or after the oviposition. There was a strong positive correlation between the number of spermatozoa in the SST and the number of spermatozoa embedded in the perivitelline layer of the oviductal eggs (r = 0.85, p less than 0.01). These data show that the population of spermatozoa actually accepted by the SST is quite small relative to the number of spermatozoa inseminated and that maximum sperm-storage is achieved when the hen is inseminated just prior to the onset of egg production. It is suggested that the sperm-storage capacity of the oviduct and the quality of the semen sample can be estimated on the basis of numbers of spermatozoa embedded in the egg perivitelline layer.
RATE of sperm transport in the female reproductive tract has been reported for many mammalian species (Austin and Bishop,
1957; Parkes, 1960; and Bishop, 1961). The time interval between coitus or artificial insemination and the arrival of spermatozoa
at the site of fertilization is surprisingly short: intervals of IS minutes or less have been recorded in the ewe (Mattner
and Braden, 1963), cow (Van Demark and Moeller, 1951), and sow (First et al., 1968). In the hen, spermatozoa labelled with inorganic 32P were recovered from the infundibulum within one hour after intravaginal insemination (Allen and Grigg, 1957). However, the
same authors also reported that 32P labelled spermatozoa deposited in the uterus reached the infundibulum in less than 15 minutes.
No literature was found concerning the rapidity of sperm transport in the turkey hen. Consequently, this investigation was
conducted to determine the distributions of living and dead spermatozoa in the reproductive . . .
Recent observations in turkey and chicken hens show that sperm storage in both species is a highly inefficient process. After artificial insemination (AI), less than 1% of spermatozoa inseminated are selected for transport to and enter the sperm storage tubules (SST). It has been shown that the sperm selection process is orchestrated within the vagina and not at the level of the SST. At least two mechanisms are involved in the selection of spermatozoa fit for sperm storage, one being mechanical (motility) and the other biochemical in nature (sperm-vaginal mucosa interactions). Furthermore, it was also observed that the sperm storage efficiency in the chicken is dependent upon the logarithm of the number of spermatozoa inseminated. From a practical standpoint, inseminations performed frequently with a moderate number of spermatozoa should be more efficient than inseminations performed with higher doses at longer intervals.
Maximal filling of the SST of hens in egg production requires only 1 day for the chicken and 2 days for the turkey. By contrast, the release of sperm from the SST is about seven times faster in the chicken than the turkey hen. The efficiency of oviducal sperm storage is related to a number of factors including age of the hen, stage of the ovulatory cycle when inseminated, and, in the turkey, if the hen was inseminated before or after the onset of egg production.
Two different categories should be considered among factors that affect sperm survival in vivo. 1) Factors affecting sperm storage. These factors, acting in the vaginal portion of the oviduct, regulate the migration of spermatozoa up to the SST by increasing (e.g., short intervals between oviposition and AI) or decreasing (e.g., sperm migration in prelaying hens) the barrier effect of the vagina. 2) Factors affecting sperm release. In chicken hens, the hen’s age does not impair the sperm storage efficiency but rather increases the rate of release of spermatozoa, thus contributing to a shorter duration of fertility.
When a suspension of rooster sperm was overlaid upon 6% (wt/vol) Accudenz, immotile sperm did not enter but motile sperm entered rapidly. The absorbance of the Accudenz layer increased as a result. These phenomena were used to measure sperm motility objectively at body temperature. The intra-assay coefficient of variation (CV) was 2.6% (n = 3). When roosters (n = 36) were ejaculated repeatedly and sperm motility data analyzed by two-way ANOVA, a male effect was observed (P < or = 0.001). When roosters were ranked by mean motility scores (n = 3 evaluations per male) and representative males selected as semen donors, a difference in fertility (P < or = 0.001) was observed between males characterized by minimal and maximal sperm motility. Frequency analysis with data from a second flock of roosters (n = 100) revealed a normal distribution. Roosters categorized by average sperm motility (n = 18) or sperm motility greater than one standard deviation above the mean (n = 17) were selected for further analysis by repeated measurements. A split-plot ANOVA revealed a difference between categories (P < or = 0.0001) and variation among males within a category (P < or = 0.0001). In contrast, sperm motility was independent of time and there was no interaction between category and time. Thereafter, five roosters from each group were ejaculated weekly and interassay CV estimated with semen pooled by category (n = 3 observations per category). During this interval, sperm motility of average roosters was 55 +/- 5.9% of that of roosters within the high motility category. Interassay CV were 18.1 and 9.2% for roosters originally categorized by average and high sperm motility, respectively. The assay described has potential for: 1) selecting males based on sperm motility, and 2) standardizing the measurement of poultry sperm motility.
Semen donors were selected from a population of 100 roosters based upon the extent to which sperm penetrated 6% (wt/vol) Accudenz from an overlay of extended semen. Semen donors categorized by average or high sperm motility (n = 5 per phenotype) were ejaculated weekly, their ejaculates pooled by phenotype, and pooled semen extended. A subsample of each sperm suspension was overlaid on 6% (wt/vol) Accudenz in a cuvette, the cuvette was placed in a 41 C water bath, and the absorbance of the Accudenz layer was measured after a 5-min incubation. The remainder of the sperm suspension was inseminated (n = 55 hens per phenotype). Each hen was inseminated weekly with 50 x 10(6) sperm for 14 wk. The hatchability of eggs laid by hens inseminated with sperm from the high motility phenotype was 10% greater (P < or = 0.001) than that of hens inseminated with sperm from the average phenotype. The difference in fecundity was explicable in terms of fertility (P < or = 0.001). A replicate experiment tested the effect of sperm motility as well as insemination dose on fertility. Roosters were treated as above, and hens (n = 41 to 45 per phenotype) were inseminated weekly with 25, 50, or 100 x 10(6) sperm per hen for 3 wk. Two-way ANOVA detected a sperm motility effect (P < or = 0.0001) but did not detect a dose effect (P > or = 0.05) or a motility by dose interaction (P > or = 0.05). A posteriori comparison among means revealed that the maximal fertility obtained with sperm from average roosters was 9% less (P < or = 0.05) than that obtained with only 25% as many sperm from the high motility phenotype. These experiments demonstrated that the fecundity of artificially inseminated hens can be increased when sperm penetration of Accudenz is used as a selection criterion for semen donors.
Given a pool of genetically superior male turkeys, the subsequent selection of toms as semen donors for artificial insemination should be based upon criteria that are predictive of the tom's fertility and fecundity over the course of a production cycle. Recently, sperm mobility phenotype has been shown to correlate highly with fertility. Therefore, the objectives of this study were to determine: 1) whether sperm mobility phenotypes of individual toms were independent of time, and 2) the extent to which traditional semen quality variables changed as a function of time during the study interval. Sperm mobility phenotype was determined by objectively measuring the ability of sperm to penetrate 2% Accudenz solution at body temperature. During the initial assessments of a flock (n = 94), sperm mobility indexes (SMI) were significantly higher for the High Mobility Phenotype toms (56.61 +/-1.03% SMI) compared to the Low Mobility Phenotype (30.46+/-1.27% SMI, P < or = 0.0001) toms. Over the 5 mo of this study, the High Mobility Phenotype toms consistently had higher (P < or = 0.05) SMI values than the Low Mobility Phenotype toms, with SMI values similar to those of the initial screen. Ejaculate volume, concentration, and plasma membrane integrity showed no significant differences between phenotypes (P > or = 0.05). Sperm viability remained significantly lower (P < or = 0.05) over the course of this study for the High Mobility Phenotype toms than for the Low Mobility Phenotype toms. Because sperm mobility phenotype remained consistent through time, the Sperm Mobility Test provides a potentially important tool for selecting semen donors in turkeys.
Study Objective To develop mathematical models using kinematic parameters from Computer-Aided Sperm Analysis (CASA) that predict the fertilization rate of sperm recovered from the caput epididymidis and to test the hypothesis that fertilization was enhanced by the presence of specific sperm subpopulations in the inseminate. Setting In vitro fertilization (IVF) program. Patients Thirteen patients with congenital absence of the vas deferens provided epididymal sperm for IVF as well as for CASA. Results The mathematical model that was most predictive of fertilization rates included kinematic parameters of the epididymal aspirate (percent motility), the inseminate used for IVF (curvilinear velocity [VCL]), and the change in sperm movement after in vitro processing by the mini-Percoll technique (difference in amplitude of lateral head displacement [ALH]). Multivariate cluster analysis revealed that inseminates that resulted in higher fertilization rates had subpopulations of sperm that were characterized by high VCL and high mean angular displacement, as well as a greater change in ALH after processing. Conclusion In vitro fertilization with epididymal sperm was more likely to succeed when the sperm population that was initially aspirated had a higher proportion of motile cells and when these sperm were capable of capacitation in vitro as indicated by the appearance of sperm subpopulations with motility that resembled hyperactivation.
Objective To determine the predictive value of quantitative motility characteristics produced by the Hamilton Thorn Motility (HTM) Analyzer (Hamilton Thorn Research, Beverley, MA) for in vivo conception. Design A prospective analysis of 222 couples attending a regional infertility clinic. The measurements were made on a semen sample, and the presence or absence of a treatment-independent conception up to 22 months later was determined. The semen variables were then correlated to conception. Setting University based center for reproductive medicine. Patients, Participants The presence or absence of an in vivo conception was recorded in 222 couples in whom the influence on fertility of the female partner was minimized, i.e., normal in terms of history and examination, a regular menstrual cycle, ovulatory (midluteal serum P > 18 nmol/L [5.6 ng/mL]), and the outcome of the hysterosalpingogram was normal. The median follow-up time was 13 months (range, 5 to 22 months). Interventions None. Main Outcome Measure Pregnancy. Results A number of variables were significantly related to time to conception. When a forward stepwise analysis was performed, the total number of spermatozoa was selected on the first step, and average path velocity was selected on the second step. No other variables were selected. The final variables consisted of the total number of spermatozoa and average path velocity. Conclusion The measurement of quantitative motility and sperm number using a HTM Analyser is of clinical value.
A semiautomatic computerized technique for the measurement of sperm swimming speed is presented. The equipment is easy to use and would be suitable for routine clinical laboratories. The value of the sperm speed measurements obtained from over 100 individuals in relation to fertility has been studied by the correlation of these results with human in vitro fertilization (IVF) data and sperm penetrating capability in the zona-free hamster egg assay. The results show that sperm speed measurements correlate very well with those of the IVF, both human and hamster, and can be used successfully, in conjunction with multivariate statistical methods, to predict the outcomes of such techniques with about 75% accuracy.
A REVIEW of the literature has revealed several methods of obtaining spermatozoa from the domestic fowl. None of these, however, seem quite adequate for obtaining supplies of undiluted spermatozoa in the amounts necessary for extensive work in the field of artificial insemination, fertility, and associated studies.
The method presented in this paper consists of a manual elicitation of an ejaculatory response which is undoubtedly reflex in nature. Repeated responses can be obtained in a short period of time. Relatively large amounts of semen can be collected easily and the bird can be held in the best position for obtaining clean samples.
The method is best handled by two operators. One operator holds the bird by its thighs in a head-downward position. The abdomen is toward the second operator, the legs are spread slightly and the abdomen otherwise well exposed. The second operator then massages the keel and the soft part . . .
Roosters homozygous for the rose comb allele (R/R) are subfertile. In previous research, these subfertile roosters were characterized by an in vitro sperm penetration assay as having limited sperm motility. The objectives in the present study were to characterize sperm motility by computer-assisted sperm motion analysis and to account for a mechanism underlying poor sperm motility. Percentages of motile sperm differed between subfertile males and fertile controls (r/r) by 29% (p < 0.001). The concentration of intracellular ATP in sperm form subfertile roosters was less than in that from fertile controls (p < 0.001). The genotypic difference is sperm motility, as measured with the sperm penetration assay, was maintained when ATP production was dependent on anaerobic glycolysis (p < 0.001). In this case, sperm were incubated with exogenous glucose and cyanide. Consequently, we could not attribute the genotypic difference in sperm mobility to mitochondrial respiration. In contrast, glucose transport, as measured by the uptake of [1,2-3H]-2-deoxy-D-glucose, was reduced in sperm from subfertile roosters (p < 0.001). Neither hexokinase nor glyceraldehyde-3-phosphate dehydrogenase activity differed between genotypes (p > 0.05). Likewise, lactate dehydrogenase activity did not differ between genotypes (p > 0.05). As evidenced by creatine kinase activity and dynein ATPase activity, neither the potential for energy transfer nor utilization within the axoneme differed between genotypes (p > 0.05). Therefore, we attribute the subfertility of roosters homozygous for the rose comb allele to decreased spermatozoal glucose transport.
A semiautomatic computerized technique for the measurement of sperm swimming speed is presented. The equipment is easy to use and would be suitable for routine clinical laboratories. The value of the sperm speed measurements obtained from over 100 individuals in relation to fertility has been studied by the correlation of these results with human in vitro fertilization (IVF) data and sperm penetrating capability in the zona-free hamster egg assay. The results show that sperm speed measurements correlate very well with those of the IVF, both human and hamster, and can be used successfully, in conjunction with multivariate statistical methods, to predict the outcomes of such techniques with about 75% accuracy.
BLDSC reference no.: DX173061. Thesis (doctoral)--University of Dundee, 1992.
To develop mathematical models using kinematic parameters from Computer-Aided Sperm Analysis (CASA) that predict the fertilization rate of sperm recovered from the caput epididymidis and to test the hypothesis that fertilization was enhanced by the presence of specific sperm subpopulations in the inseminate.
In vitro fertilization (IVF) program.
Thirteen patients with congenital absence of the vas deferens provided epididymal sperm for IVF as well as for CASA.
The mathematical model that was most predictive of fertilization rates included kinematic parameters of the epididymal aspirate (percent motility), the inseminate used for IVF (curvilinear velocity [VCL]), and the change in sperm movement after in vitro processing by the mini-Percoll technique (difference in amplitude of lateral head displacement [ALH]). Multivariate cluster analysis revealed that inseminates that resulted in higher fertilization rates had subpopulations of sperm that were characterized by high VCL and high mean angular displacement, as well as a greater change in ALH after processing.
In vitro fertilization with epididymal sperm was more likely to succeed when the sperm population that was initially aspirated had a higher proportion of motile cells and when these sperm were capable of capacitation in vitro as indicated by the appearance of sperm subpopulations with motility that resembled hyperactivation.
The fertilization rates observed in 122 attempts at in-vitro fertilization were examined in relation to sperm characteristics assessed by visual and automated screening. Using linear regression analysis, a significant correlation was found between the fertilization rate and (i) evaluations in fresh semen sperm concentration, percentages of sperm motility, vitality and normal morphology and velocity, (ii) measurements in swim-up preparations of percentages of sperm motility, vitality and morphology, velocity and amplitude of lateral head displacement. No significant correlation was found between the fertilization rate and any of the parameters studied in 24-h-old swim-up suspensions. Analysis by multiple variable stepwise linear regression showed an optimal correlation (R6 = 0.62) between the observed fertilization rate and theoretical calculation obtained from the following predictive function: fertilization rate = -0.3 + (0.008 x swim-up motility) + (0.004 x normal sperm morphology in fresh semen). Introduction of kinematic characteristics studied by automated screening improved the multiple correlation between the calculated and observed fertilization rate in cases of normal or mildly defective semen. Because of the limited availability of motile spermatozoa, automated analysis could not supersede classical sperm analysis in cases of more severe sperm defects.
To determine which sperm movement characteristics are related to in vitro fertilization rates, semen and swim-up preparations used for in vitro fertilization in 108 patients were assessed using the Hamilton-Thorn HTM-2030 Motility Analyzer (HTMA) and other sperm tests. There were highly significant correlations between manual and HTMA results for sperm concentration (Spearman r = 0.881; P less than 0.001) and the percentage of motile spermatozoa (Spearman r = 0.580; P less than 0.001). The percentage of motile spermatozoa with average path velocities greater than 10 microns/s and greater than 20 microns/s, straight line and curvilinear velocity, linearity (straight line velocity vs curvilinear velocity), amplitude of lateral head displacement, and beat-cross frequency were significantly higher in the insemination medium after selection of motile spermatozoa by the swim-up technique than in the semen. Linearity (P less than 0.01), the percentage of morphologically normal spermatozoa (P less than 0.05) and straight line velocity (P less than 0.05) in semen, and the percentage of motile spermatozoa with average path velocities greater than 10 microns/s in both semen (P less than 0.05) and insemination medium (P less than 0.05) were significantly correlated with in vitro fertilization rate when examined by a nonparametric (Spearman) test. With logistic regression analysis of all data, only the diagnoses of male infertility and tubal disease, linearity in semen, and the percentage of motile spermatozoa with average path velocities between 10 and 20 microns/s in insemination medium were significantly related to in vitro fertilization rates.(ABSTRACT TRUNCATED AT 250 WORDS)
The purpose of this study was to evaluate relationships between measured sperm velocity and in-vivo fertility, using donor semen samples from an artificial insemination (AID) programme. Seventy-one frozen semen samples were examined; measurements of sperm velocity were made immediately after thawing, upon a motile 'swim-up' fraction, and finally after 3.5 h incubation at 37 degrees C in the freezing mixture. Zona-free hamster egg penetration assays were performed upon all samples. Two groups of samples were identified; seven donors (11 samples) had failed to produce any pregnancies through AID from a range of 3 to 14 cycles tested, whilst the remaining samples (from 25 donors) had achieved at least one pregnancy each. The mean sperm velocity (+/- SEM) for the latter 'fertile' group was significantly higher than the corresponding value for the 'infertile' group; (i) after thawing, 65.9 +/- 1.8 versus 50.4 +/- 3.2 microns/s (P less than 0.001) and (ii) after 3.5 h incubation, 42.1 +/- 2.1 versus 24.7 +/- 5.7 microns/s (P less than 0.002). Using the maintenance of sperm velocity during incubation as an indicator of survival, life-table analyses were used to calculate monthly conception rates on various sub-groups of the semen samples. Poor survival (greater than 40% decline in velocity over 3.5 h) was associated with a monthly pregnancy rate of only 11.58% (362 cycles), whilst better survival (less than 40% decline) was associated with the significantly higher (P = 0.024) pregnancy rate of 16.87% (480 cycles).
This paper highlights the most critical aspects of the problem of predicting fertility. To determine if a laboratory test(s) is highly correlated with fertility it is essential to have: a) specific, precise and accurate laboratory tests, and b) precise and accurate fertility data. Acquisition of precise and accurate data for laboratory tests and fertility of spermatozoa in the same sample is not easy. Data derived from in vitro fertilization are not tests of fertility, because only a subset of the attributes important for fertilization in vivo are tested. Because of deficiencies in fertility data, there probably is no valid report for human spermatozoa correlating results of laboratory tests and fertility, and very few valid studies for laboratory or domesticated animals. There is little doubt that objective measures of sperm motion, acrosomal status, or other characteristics are significantly correlated with fertility. However, establishment of the correlations between a group of attributes and fertility is not the question of interest. The goal is prediction of fertility. There has been no recent effort to develop a prediction of fertility or fecundity based on sperm characteristics, and achievement of this goal may be elusive.
In this paper we examine the value of both conventional and computer-assisted semen analysis (CASA) using the Hamilton-Thorn HTM-S 2030 in predicting the in-vivo fertility of cryopreserved donor semen. Semen samples were examined prospectively and data on the conventional criteria of semen quality, sperm morphometry and movement were collected. Of 61 ejaculates identified, 33 achieved pregnancies ('successful') and 28 failed to do so ('unsuccessful'), despite insemination into at least four different normal female recipients. When the post-thaw semen profiles were compared, no differences were observed between the two groups in respect of the conventional criteria of semen quality determined by conventional laboratory techniques; however, there were differences in respect of both morphometry and movement characteristics determined by the HTM-S. When multiple logistic regression was used to examine the ability of the variables measured to predict the achievement of pregnancy, the conventional criteria of semen quality were of no value (chi 2 = 6.67, P = 0.353). However, the CASA assessment successfully predicted outcome in 86.9% of cases (chi 2 = 44.3, P = 0.0021). It was concluded that CASA assessment is of significant value in predicting the ability of an ejaculate to achieve pregnancy.
The relationship between the total number of sperm inseminated, semen quality, and fertility in rabbits was investigated, using fractionated or unfractionated semen and different diluting fluids. Semen was from Dutch-belted males collected twice weekly with an artificial vagina. All does were superovulated except in Experiment 3. In Experiment 1, sperm were fractionated on discontinuous 4% and 10% bovine serum albumin columns. Sperm from each portion of the gradient, along with unfractionated controls, were diluted to give 0.25 x 10(6), 0.5 x 10(6), 1.0 x 10(6), and 2.0 x 10(6) total sperm per insemination. In Experiment 2, sperm were diluted with Dulbecco's phosphate-buffered saline to provide 0.10 x 10(6), 0.50 x 10(6), and 1.0 x 10(6) total sperm per insemination, with minimal processing time. In Experiment 3, does were allowed to kindle after inseminating 0.1 x 10(6) or 1.0 x 10(6) sperm. In Experiment 4, sperm were diluted with TALP buffer: seminal plasma 1:1 to 0.025 x 10(6), 0.05 x 10(6), and 0.10 x 10(6) total sperm per insemination. Over 2,800 embryos or unfertilized oocytes were obtained either 24 or 48 hours after insemination to measure fertility. Sperm numbers required for normal fertility were 0.50 x 10(6) in Experiment 1 and only 0.05 x 10(6) in Experiment 4. This reduction presumably was due primarily to reduced processing time and diluent change. Litter size was normal with 0.1 x 10(6) sperm (Experiment 3). In Experiment 4, computer-assisted sperm analysis (HTM 2030 system; Beverly, Massachusetts) was adapted to successfully screen out some of the "interfering" granules in rabbit semen.(ABSTRACT TRUNCATED AT 250 WORDS)
To determine the predictive value of quantitative motility characteristics produced by the Hamilton Thorn Motility (HTM) Analyzer (Hamilton Thorn Research, Beverley, MA) for in vivo conception.
A prospective analysis of 222 couples attending a regional infertility clinic. The measurements were made on a semen sample, and the presence or absence of a treatment-independent conception up to 22 months later was determined. The semen variables were then correlated to conception.
University based center for reproductive medicine.
The presence or absence of an in vivo conception was recorded in 222 couples in whom the influence on fertility of the female partner was minimized, i.e., normal in terms of history and examination, a regular menstrual cycle, ovulatory (midluteal serum P > 18 nmol/L [5.6 ng/mL]), and the outcome of the hysterosalpingogram was normal. The median follow-up time was 13 months (range, 5 to 22 months).
None.
Pregnancy.
A number of variables were significantly related to time to conception. When a forward stepwise analysis was performed, the total number of spermatozoa was selected on the first step, and average path velocity was selected on the second step. No other variables were selected. The final variables consisted of the total number of spermatozoa and average path velocity.
The measurement of quantitative motility and sperm number using a HTM Analyser is of clinical value.
Rat spermatozoa recovered from different regions of the excurrent ducts of 10 adult males (proximal cauda epididymidis [PC], distal cauda epididymidis [DC], and vas deferens [VD]) were assessed by in vitro fertilization (LVF) using limited sperm numbers, and by continuous evaluation of motility parameters during 5 hours of incubation in vitro with automated computer-aided sperm analysis (CASA). Spermatozoa from the PC region fertilized (68 +/- 6%) a significantly greater (P < or = 0.005) number of oocytes than those from the DC (44 + 5%) or VD (47 +/- 7%). For pooled samples from all three regions, the mean fertilization rate (51 +/- 14%) was less tan for spermatozoa from the PC (P < 0.05) but was not significantly different from spermatozoa from the DC or VD. For each time point and sample, 1,592 +/- 428 sperm tracks were analyzed. CASA was verified by comparison with manual still-frame analysis of video recordings, by repeated analysis of the same or different samples of spermatozoa, and by examination of computer tracks. The coefficients of variation for various motion parameters suggested that the CASA obtained a high degree of precision. There were no significant differences in motility parameters for spermatozoa recovered from equivalent regions of the left or right tract or in motility parameters for spermatozoa from different regions of the tract immediately after recovery. However, during incubation in vitro, spermatozoa from the DC or VD regions exhibited a marked decline in straight-line velocity (VSL) compared with spermatozoa from the PC region. The reduction in VSL (combined values from right and left tract) for DC or VD spermatozoa compared with PC spermatozoa was significant at 2.5 hours of incubation (P < or = 0.05) and highly significant (P < or = 0.005) by the end of the incubation period. Differences in average path velocity (VAP) were also apparent after 4 hours (p < or = 0.05), but no significant differences were observed for measurements of curvilinear velocity (VCL) or lateral bead displacement (ALH). Overall, the decline in VSL over 5 hours was highly correlated (P < or = 0.001) with the outcome of fertilization in vitro. In contrast, initial VSL and changes in VCL of spermatozoa were not correlated with fertilization rate. These results indicate that the in vitro fertilizing capacity of rat spermatozoa is correlated with 1) the decline in straight-line velocity (VSL) as measured by repeated CASA during incubation in vitro and 2) with the site of recovery of mature rat spermatozoa from the distal excurrent duct. It is suggested that the deterioration of the quality of rat spermatozoa in the distal epididymidis and vas deferens during storage may occur sooner than previously realized, and therefore care must be taken when recovering samples for fertility assessment. In keeping with findings in other species, immediate "snapshot" analysis of rat motility was a poor predictor of sperm fertility. In contrast, continuous CASA provided significant information for determining sperm fertilizing capacity and will be a useful technique for reproductive toxicology.
We studied the prognostic value of sperm characteristics for the outcome of intra-uterine insemination with partner sperm (IUIPS). A total of 712 cycles of IUIPS following induction of ovulation with gonadotrophin (hMG/hCG) for 277 sterile couples attending the assisted reproductive technology centre of Poissy Hospital (78300-France) between January 1991 and December 1994 was studied retrospectively. Ninety-two clinical pregnancies were obtained giving an overall rate of 12.9% per cycle. None of the characteristics of the sperm as assessed initially correlated with outcome. In contrast, the number of motile spermatozoa given (n) affected outcome: for n < 1 x 10(6) the pregnancy rate was 2%; for n = 5 to 8 x 10(6) the rate was 19%. However, for +/- 8 x 10(6) the proportion of biochemical pregnancies and miscarriages was 40% which was significantly higher than for smaller concentration. The resort of IVF following 4 IUIPS failures leads to a pregnancy rate per cycle of only 6.7%.
Two fertility trials were undertaken to evaluate the relationship between boar semen quality and fertility (conception rate and litter size) after on-farm artificial insemination (AI). Trial 1 included 98 ejaculates from 27 boars, and trial 2 included 72 ejaculates from 26 boars. The semen quality was measured by computer-assisted semen analysis (CASA) using the Hobson Sperm Tracker. Boar semen was diluted in a standard extender (Beltsville Thawing Solution, BTS), dispensed into 75 ml allquots each containing 1.5 x 10(9) spermatozoa and dispatched to farms by overnight mail for use by their normal AI procedures. Randomly selected 75 ml aliquots of semen from each boar were also sent to the institute of Zoology for CASA measurement. Prior to CASA analysis, the spermatozoa were recovered from the BTS using Percoll gradients, resuspended in trisbuffered saline media containing 40 mM Ca++, and incubated at 39 degrees C. Parameters of sperm motion were measured after 0, 2, 4, and 6 hours of incubation. Various multiple regression models based on measured motion parameters could account for up to 24% of the variation in litter size. Using logistic regression, highly significant (P < 0.0001) models explaining conception rate in terms of sperm motion were derived for trial 2 only. The change in sperm velocity during the first 2 hours of incubation and the magnitude of the velocity parameters after 2 hours were identified as the most consistent indicators of fertility. Other attributes of sperm quality, i.e., frequency of spontaneous acrosome reactions (AR) and ARs induced by ionophore A23187 or solubilized pig zona pellucida, were also examined. When the "within-trial" median litter size was used as a way of allocating ejaculates to "high" or "low" litter-size groups, higher litter size was associated with lower frequency of both spontaneous and induced AR. These results demonstrate that fertility information can be derived from the CASA analysis of boar semen provided it is combined with a period of incubation in capacitating conditions.
Roosters homozygous for the rose comb allele (R/R) are subfertile. In previous research, these subfertile roosters were characterized by an in vitro sperm penetration assay as having limited sperm motility. The objectives in the present study were to characterize sperm motility by computer-assisted sperm motion analysis and to account for a mechanism underlying poor sperm motility. Percentages of motile sperm differed between subfertile males and fertile controls (r/r) by 29% (p < 0.001). The concentration of intracellular ATP in sperm form subfertile roosters was less than in that from fertile controls (p < 0.001). The genotypic difference is sperm motility, as measured with the sperm penetration assay, was maintained when ATP production was dependent on anaerobic glycolysis (p < 0.001). In this case, sperm were incubated with exogenous glucose and cyanide. Consequently, we could not attribute the genotypic difference in sperm mobility to mitochondrial respiration. In contrast, glucose transport, as measured by the uptake of [1,2-3H]-2-deoxy-D-glucose, was reduced in sperm from subfertile roosters (p < 0.001). Neither hexokinase nor glyceraldehyde-3-phosphate dehydrogenase activity differed between genotypes (p > 0.05). Likewise, lactate dehydrogenase activity did not differ between genotypes (p > 0.05). As evidenced by creatine kinase activity and dynein ATPase activity, neither the potential for energy transfer nor utilization within the axoneme differed between genotypes (p > 0.05). Therefore, we attribute the subfertility of roosters homozygous for the rose comb allele to decreased spermatozoal glucose transport.
Sperm from each rooster within a base population (n = 271) were evaluated with a mobility assay validated in previous work. Frequency analysis confirmed a normal distribution for the variable of sperm mobility. Repeated-measure analysis of males categorized by phenotype demonstrated that average and high sperm mobility phenotypes were distinct and independent of time. Sperm morphology, fertilizing ability, and ATP content were compared between phenotypes. Fertility and sperm ATP content differed (p < 0.001) between phenotypes, whereas sperm morphology did not (p > 0.05). Experimentation with washed sperm demonstrated that phenotype was fully expressed when mitochondrial respiration was the only source of ATP. Sperm mobility increased (p < 0.001) when sperm from average males were exposed to calyculin A, a protein phosphatase inhibitor. Correlation analyses were performed with data from a subpopulation (n = 46) whose range, mean, and variance were equivalent to those of the base population. Neither body weight nor the combined weight of the testes was correlated with sperm mobility (r = -0.02 and 0.01, respectively). In contrast, sperm ATP content was correlated with sperm mobility (r = 0.80). We attribute phenotypic differences in sperm mobility to differential rates of mitochondrial ATP synthesis.
Commercial turkey production relies on the artificial insemination (AI) of pooled semen. However, semen quality ultimately depends on the quality of individual ejaculates. The purpose of this study was to evaluate semen from individual toms by means of an objective sperm-mobility assay. Semen was then pooled by mobility phenotype and inseminated into hens, and the percentages of fertile and hatched eggs were determined after egg incubation. To indirectly evaluate hens' sperm storage, we determined the number of sperm holes in the perivitelline layer (PL) of freshly laid eggs. Semen from individual ejaculates (two trials, total of 169 toms) was evaluated by use of the sperm-mobility test (SMT). Semen was diluted to 1 x 10(9) sperm/ml, in prewarmed N-tris-[hydroxymethyl] methyl-2-amino-ethanesulfonic acid (TES)-buffered saline, and was placed over 3 ml of a 2% (w/v) Accudenz solution at 41degrees C. After a 5-minute incubation period, the cuvette was placed in a densimeter, and percentage transmission was recorded after 1 minute. Semen samples from toms ranked, according to sperm mobility, in the highest 10% and the lowest 10%, after three evaluations, were pooled by group and were used to inseminate hens weekly (trial 1: n = 20 hens/group, for 10 weeks, AI dose 150 x 10(6) spermatozoa inseminated fresh and after 24-hour in vitro storage at 5 degrees C; trial 2: n = 60 hens/group, for 16 weeks, AI dose = 75 x 10(6) spermatozoa inseminated fresh). Each week, eggs from day 6 post-AI were evaluated for holes in the PL, vestiges of acrosomal induced hydrolysis. Spermatozoa from toms of different mobility phenotypes were also evaluated individually, for sperm chromatin structure and motility variables, by use of the Hobson Sperm Tracker. Toms characterized by high and low in vitro sperm-mobility phenotype were categorized as "high mobility" and "low mobility," respectively. The percentage of fertile eggs from hens inseminated with semen from the high-mobility toms was higher than the percentage of fertile eggs from the low-mobility group, in each trial (95.8+/-1.3% vs. 90.4+/-2.2%, and 88.7+/-4.0% vs. 82.4+/-0.4%, trials 1 and 2, respectively; P < 0.05). More sperm holes were observed in the PL of eggs fertilized by the high-mobility toms than in the PL of eggs fertilized by the low-mobility toms (P < 0.05). No differences in susceptibility of sperm nuclear DNA to denature in situ, as measured by the flow-cytometric sperm chromatin-structure assay, were detected between toms of differing mobility phenotypes. Sperm-motility variables, straight-line velocity, and average-path velocity were significantly greater for high-mobility toms compared to low-mobility toms (P < 0.05). Sperm-mobility differences between toms (detected by means of the SMT) influenced sperm storage, as indicated by the number of sperm in the PL and by the percentage of fertile eggs produced.
We have devised a novel means of investigating competitive fertilization in turkeys, using microsatellite genotyping to identify male parentage. Our results demonstrate that sperm mobility is a mechanism responsible in part for paternity efficiency in turkeys. Sperm mobility is composed of several parameters in which sperm motility is a component. Differences between ejaculates in the number of sperm penetrating into a dense, insert, nontoxic solution were measured and used to classify males into high, average, or low sperm mobility phenotypes. Microsatellite genotyping was used to determine parentage of poults after equal numbers of sperm from 10 males (either high or average phenotype, n = 5, mixed with low phenotype, n = 5) were inseminated simultaneously. In a separate study, the numbers of sperm hydrolyzing the perivitelline layer of eggs were compared between hens inseminated with sperm from high-, average-, or low-phenotype males. Overall, heterospermic inseminations resulted in consistently fewer offspring produced by low-mobility phenotype males. This correlated with physiological data in which semen from the low-mobility males had reduced numbers of sperm at the fertilization site as determined by sperm hole counts in the perivitelline layer of eggs. This is the first illustration of a measurable sperm trait predictive of paternity success in a competitive fertilization trial in turkeys, a species that is predominately reproduced by artificial insemination of multiple-sire pools.
Previous research demonstrated that sperm mobility is a quantitative trait of the domestic fowl. The trait is quantified by measuring the absorbance of an Accudenz solution after overlay with a sperm suspension and brief incubation at body temperature. In the present work, average and high sperm mobility phenotypes (n = 30 males per phenotype) were selected from a base population. Differences were found between sperm oxygen consumption (p < 0.0001), acylcarnitine content (p < 0.05), linear velocity (p < 0.001), and straightness (p < 0.001), a trajectory variable measured with the Hobson SpermTracker. Oxygen consumption and stearoylcarnitine content of sperm from the high-mobility phenotype were twice those observed with sperm from average males, implying a pivotal role for mitochondria. On the basis of these results, a graded relationship was predicted between fertility and sperm mobility. Males (n = 48) were chosen at random from another base population, sperm mobility was measured per male, and each ejaculate was used to inseminate 8-12 hens (8 x 10(7) viable sperm per hen). When fertility was plotted as a function of sperm mobility, data points approximated a skewed logistic function. The hypothesis that vaginal immunoglobulins constitute an immunological barrier to sperm transport was tested and rejected. Therefore, we concluded that sperm mobility is a primary determinant of fertility in the fowl.
Characterization of a mechanism impeding sperm transport through the vagina of the chicken The Hague, The Netherlands
- Steele Mg
- Wishart
- Gj
Steele MG, Wishart GJ. Characterization of a mechanism impeding sperm transport through the vagina of the chicken. In: Proceedings of the 12th International Conference on Animal Reproduction. The Hague, The Netherlands; 1992:1593–1595.
Characterization of a mechanism impeding sperm transport through the vagina of the chicken
- Mg Steele
- Wishart
- Mg Steele
- Wishart
Steele MG, Wishart GJ. Characterization of a mechanism impeding sperm
transport through the vagina of the chicken. In: Proceedings of the
12th International Conference on Animal Reproduction. The Hague,
The Netherlands; 1992:1593-1595.
Characterization of a mechanism impeding sperm transport through the vagina of the chicken
- Steelemg Wishartgj