Meta data analysis of conception rate in relation to sperm motility in Madura superior bulls

Abstract Madura bulls are Indonesian germplasm with a very high capacity to adapt to dry environments. Madura bulls come from a crossbreed between Zebu (Bos indicus) and banteng (Bos javanicus). One of the breeding strategies of Madura cattle is the use of artificial insemination (AI) with frozen semen. Regarding sperm motility as one of the standard parameters of good semen quality, it is good to know the reliability of sperm motility with the bull fertility rate. This study aimed to determine the conception rate percentage (%CR) relation to sperm motility in Superior Madura bulls. The frozen semen from eight Madura bulls belonging to the National Singosari and Lembang AI centre were used. They were classified based on the selected field reproductive efficiency data from the year 2018 until 2020. Sperm motility was evaluated using Computer Assisted Sperm Analysis (CASA). The data were analyzed using oneway ANOVA and Pearson correlation. The data showed that %CR was significantly higher (P<0.05) and positively correlated with sperm motility. It is proved that sperm motility represents good quality sperm as one of the fertility parameters in Madura bulls.

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Kualitas Semen Segar Sapi Bali Umur Produktif dan Non-produktif serta Penentuan Konsentrasi Krioprotektan dalam Pengencer Tris Kuning Telur

jurnal veteriner ◽

10.19087/jveteriner.2018.19.2.242 ◽

2018 ◽

Vol 19 (2) ◽

pp. 242 ◽

Cited By ~ 2

Author(s):

Anna Nabilla ◽

Raden Iis Arifiantini ◽

Bambang Purwantara

Keyword(s):

Semen Quality ◽

Egg Yolk ◽

Dimethyl Formamide ◽

Computer Assisted ◽

Sperm Analysis ◽

Bull Semen ◽

Frozen Semen ◽

Semen Volume ◽

Microscopic Evaluation

The aim of this study was to compare the quality of Bali bull fresh semen in productive group (below 10 years old) with over productive group and to evaluate the frozen semen quality of bali bull after cryopreservation in Tris egg yolk extender using Dimethyl formamide (DMF) and glycerol in three different concentrations (5%, 6%, and 7%). Semen was collected using artificial vagina. Semen then evaluated macro and microscopically. Macroscopic evaluation conducted visually and microscopic evaluation with the aid of Computer Assisted Sperm Analysis CASA. Immediatelly after evaluation, semen were divided into six aliquots and diluted with Tris egg yolk (TEY) glycerol 5% (TEYG5), TEY-glycerol 6% (TEYG6), TEY glycerol 7% (TEYG7), TEY-DMF 5% (TKTD5), TEY-DMF 6% (TEYD6) and TEY-DMF 7% (TKTD7). Results of experiment showed that there was no difference (P>0.05) among the all parameters, except for semen volume. Semen volume of productive group was higher than over productive age group. No difference was found in post thawing motility of the spermatozoa in TEY extender with 5% glycerol, and TEY with 5%, 6% and 7% DMF. The higher recovery rate of bali bull spermatozoa demonstrated by spermatozoa in TEY extender with glycerol 5% and DMF 6% (63.33±2.40 dan 68.67±2.33). It was concluded that bali bull semen quality did not differ between productive and over productive ages and post thawing semen quality of bali bull in TEY with 5-7% DMF or 5% glycerol was not also different.

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17 Increased inbreeding levels negatively affect sperm kinetics and motility in Purebred Spanish horses

Reproduction Fertility and Development ◽

10.1071/rdv33n2ab17 ◽

2021 ◽

Vol 33 (2) ◽

pp. 116

Author(s):

Y. Pirosanto ◽

A. Molina ◽

M. Valera ◽

J. Dorado ◽

E. Terán ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Reproductive Traits ◽

Study Groups ◽

Inbreeding Coefficient ◽

Computer Assisted ◽

Effective Population ◽

Mean Velocity ◽

Sperm Analysis ◽

Head Displacement

Reproductive performance is one of the key factors in livestock production. It is well known that reproductive traits are influenced by several genetic factors, such as the increase of individual inbreeding levels, which are associated with changes in sperm motility and shape in several species. In horses, the increase in inbreeding is a common problem because of the reduction in effective population size and the increase in selection intensity observed in several breeds. However, studies assessing the effect of high levels of inbreeding on the sperm quality of stallions are scarce. In the present study, we aimed to determine the effect of increased inbreeding levels and age on the sperm motility patterns of Purebred Spanish horses (PRE). We performed kinetic characterisation of 557 sperm samples of 82 PRE stallions aged between 3 and16 years, using computer-assisted sperm analysis (Androvision™, Minitube). We evaluated 5 parameters in 6 different fields per sample: curved line velocity (VCL, µm/s), velocity average path (VAP, µm/s), velocity straight line (VSL, µm/s), amplitude of lateral head displacement (ALH, µm), and beat-cross frequency (BCF, Hz). We determined the pedigree-based inbreeding coefficient (Fped) based on ∼300,000 PRE pedigree records to evaluate the inbreeding effect. Individuals were separated into 2 groups: highly inbred (n=339) and lowly inbred (n=218) according to an F value of 12.5%. Differences between groups were analysed using a generalized linear model. The analysis did not show significant differences (P>0.05) in the variables analysed with respect to the age of stallions. However, VAP, VCL, and AHL were lower in highly inbred than in lowly inbred animals (P<0.05), suggesting less velocity and amplitude of head displacement. In the case of BCF, no significant differences (P>0.05) were observed between the two study groups. In conclusion, age did not affect sperm quality parameters in the age group of stallions analysed. In addition, we demonstrated that high inbreeding coefficient reduced the mean velocity and trajectory pattern of spermatozoa in PRE.

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149 The second isoform of gonadotrophin-releasing hormone and its receptor affect boar semen quality

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab149 ◽

2020 ◽

Vol 32 (2) ◽

pp. 201

Author(s):

C. E. Ross ◽

F. H. Choat ◽

K. N. Plager ◽

A. T. Desaulniers ◽

R. A. Cederberg ◽

...

Keyword(s):

Semen Quality ◽

Computer Assisted ◽

Functional Protein ◽

Littermate Control ◽

Normal Morphology ◽

Releasing Hormone ◽

Sperm Analysis ◽

Sperm Characteristics ◽

Straight Line ◽

Gonadotrophin Releasing Hormone

Pigs are the only livestock species encoding a functional protein for both the second isoform of gonadotrophin-releasing hormone (GnRH-II) and its cognate receptor (GnRHR-II). Unlike the classical GnRH system (GnRH-I and GnRHR-I), GnRH-II and GnRHR-II are abundantly produced in porcine testes. Moreover, GnRH-II binding its receptor on Leydig cells stimulates luteinizing hormone-independent testosterone secretion. Interestingly, GnRHR-II is also localised to the connecting piece of mature, ejaculated spermatozoa, whereas GnRH-II is detected in seminal plasma, an interaction possibly influencing the function of sperm. To examine the role of GnRH-II and its receptor in the testis, we produced a swine line with reduced endogenous GnRHR-II levels (GnRHR-II KD). The objectives of this study were to (1) compare sperm characteristics between mature GnRHR-II KD and littermate control boars on the day of collection and following semen extension and (2) determine whether a GnRHR-I and GnRHR-II antagonist alters sperm characteristics after storage of extended semen. In Experiment 1, GnRHR-II KD (n=3) and littermate control (n=3) ejaculates were collected (Day 1) and computer-assisted sperm analysis (CASA) was performed (IVOS II Animal; Hamilton Thorne) to determine measures of sperm motion (motility, progressive motility, slow, and static), morphology (normal morphology, bent tail, coiled tail, distal droplet, proximal droplet (PD), distal midpiece reflex, elongation, and area), and kinematics (length of average path (DAP), length of straight line path (DSL), length of curvilinear path (DCL), average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), straightness (STR), linearity (LIN), amplitude of lateral head displacement (ALH), beat-cross frequency, and wobble (WOB)). Next, 3 billion sperm were extended with Androstar Plus (80-mL doses; Minitube) and stored at 17°C until Day 7 CASA. Data were analysed with the MIXED procedure of SAS (SAS Institute Inc.). On Day 1, semen doses from GnRHR-II KD boars had reduced DSL, VSL, STR, LIN, and WOB (P<0.05), whereas sperm from control boars possessed more PD (P<0.01). Day 7 CASA revealed that transgenic sperm had reduced DAP, DCL, VAP, and VCL, although sperm from control boars were slower (P<0.05). In Experiment 2, control ejaculates (n=3) were extended as above, treated with increasing concentrations (0, 0.0001, 0.001, 0.01, 0.1, 1, and 10μM) of a GnRH antagonist inhibiting both GnRHR-I and GnRHR-II (SB-75, cetrorelix), and stored at 17°C until Day 7 and 9 CASA. On Day 7, only sperm characteristics in doses treated with 10μM SB-75 were significantly lower (normal morphology, DAP, DCL, VAP, VCL, and ALH) or higher (PD, WOB, and area) than controls. Similar differences (except ALH; P<0.10) for the 10μM SB-75 treatment were detected on Day 9; however, motility, slow, static, STR, and LIN were also reduced (P<0.05). Thus, these data suggest that GnRH-II and its receptor are important to sperm function, representing a potential avenue to improve semen preservation. This research was funded by USDA/NIFA AFRI (2017-67015-26508; BRW).

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

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab23 ◽

2020 ◽

Vol 32 (2) ◽

pp. 137

Author(s):

Y. Pirosanto ◽

M. Valera ◽

A. Molina ◽

J. Dorado ◽

S. Demyda-Peyrás

Keyword(s):

Sperm Motility ◽

Inbreeding Depression ◽

Negative Correlation ◽

Semen Quality ◽

Sperm Quality ◽

Sperm Concentration ◽

Inbreeding Coefficient ◽

Semen Parameters ◽

Computer Assisted

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

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Effects of chilled storage and pH of activating solution on different motility parameters in burbot (Lota lota) sperm

Czech Journal of Animal Science ◽

10.17221/122/2017-cjas ◽

2018 ◽

Vol 63 (No. 11) ◽

pp. 429-434

Author(s):

Zoltán Bokor ◽

Balázs Csorbai ◽

Levente Várkonyi ◽

Zsolt Szári ◽

Ferenc Fodor ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Quality ◽

Computer Assisted ◽

Sperm Analysis ◽

Chilled Storage ◽

Straight Line ◽

H 26 ◽

Computer Assisted Sperm Analysis ◽

Motility Parameters ◽

Activating Solution

The effects of a simple saline solution prepared using two different pH (4.4 and 8.5) on sperm motility in burbot were investigated. Results were recorded during a 96-hour chilled storage (4°C) in 24-hour intervals. Measurements were focused on the detailed characteristics of motility using 12 parameters obtained from the Computer-assisted Sperm Analysis (CASA). Significantly higher progressive motility (pMOT), distance average path (DAP), distance curved line, distance straight line (DSL), average path velocity (VAP), curvilinear velocity, straight line velocity, and beat cross frequency (BCF) were observed with the activating solution buffered at pH 8.5 in comparison with pH 4.4. Already after 24 h a significant reduction was measured in pMOT (0 h: 49 ± 24%, 24 h: 12 ± 7%). Similar decreasing tendency was recorded only after 72 h in DAP (0 h: 26 ± 4 µm/s, 72 h: 19 ± 9 µm/s), DSL (0 h: 21 ± 5 µm/s, 72 h: 17 ± 8 µm/s), VAP (0 h: 59 ± 9 µm/s, 72 h: 43 ± 21 µm/s), and BCF (0 h: 28 ± 2 Hz, 72 h: 18 ± 10 Hz). The response of different investigated CASA parameters to different treatments varied in our experiments. According to our studies, numerous burbot sperm motility parameters are sensitive to chilled storage and to low pH of the activating solution. Our results could support the effective sperm quality assessment and successful artificial propagation process in burbot.

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Effect of the addition of sodium caseinate on the viability of cryopreserved buffalo semen

Semina Ciências Agrárias ◽

10.5433/1679-0359.2020v41n5supl1p2209 ◽

2020 ◽

pp. 2209-2218

Author(s):

Fernando Evaristo da Silva ◽

Jaqueline Candido Carvalho ◽

Camila de Paula Freitas Dell'Aqua ◽

Frederico Ozanam Papa ◽

Marc Roger Jean Marie Henry ◽

...

Keyword(s):

Cell Damage ◽

Membrane Integrity ◽

Sodium Caseinate ◽

Egg Yolk ◽

Freezing Process ◽

Computer Assisted ◽

Semen Cryopreservation ◽

Sperm Analysis ◽

Frozen Semen ◽

Buffalo Semen

The use of cooled semen in artificial insemination operations results in higher pregnancy rates than the use of frozen semen. This result seems to be related to the more severe damage triggered by the freezing process than that observed during refrigeration. Due to its ability to bind to sperm-binding proteins and calcium ions, sodium caseinate has been studied as a substance capable of preventing early sperm capacitation, a significant cause of the decreased pregnancy rate resulting from the use of frozen semen. The first objective of this study was to evaluate whether a commercial egg yolk diluent developed for frozen bovine semen could be used for buffalo semen cryopreservation; the second objective was to investigate the effect of this diluent in combination with sodium caseinate during the procedures of buffalo sperm cryopreservation using flow cytometry and computer-assisted sperm analysis. In the first part of the study, comparing the results of spermatic kinetics and plasma and acrosomal membrane integrity, it was observed that the freezing process resulted in more cell damage than the cooling process. In the second part of the study, no effects of the addition of sodium caseinate to the egg yolk diluent were observed. From the results of the present study, it was possible to conclude that the egg yolk-based diluent was suitable for buffalo semen cryopreservation and that the addition of sodium caseinate did not decrease the harmful effects related to seminal cryopreservation.

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12PREDICTION OF BULL FERTILITY BY COMPUTER ASSISTED SEMEN ANALYSIS

Reproduction Fertility and Development ◽

10.1071/rdv16n1ab12 ◽

2004 ◽

Vol 16 (2) ◽

pp. 128 ◽

Cited By ~ 3

Author(s):

S. Cseh ◽

T. Polichronopoulos ◽

L. Solti

Keyword(s):

Sperm Motility ◽

Semen Analysis ◽

Computer Assisted ◽

Reproductive Process ◽

Return Rates ◽

Frozen Semen ◽

Chi Square Analysis ◽

Fertilizing Capability ◽

Weighted Values

Sperm motility is clearly essential for fertilization both in vivo and in vitro. Motility is necessary for successful sperm transport, a step that is bypassed with in vitro fertilization. Recently, increasing attention has been paid to the objective evaluation and characterization of sperm motility more than simply determining the total proportion of motile spermatozoa. The purpose of computerassisted semen analysis (CASA) is to provide values for sperm concentration and sperm motility more rapidly and accurately than those obtained with traditional semen analyses methods. The objective of our experiment was to investigate the effect of specific aspects of sperm movement, such as the velocity of progression and the actual pattern of movement, to the fertilizing capability of sperm. Frozen semen samples of 10 HF breeding bulls were used in the study. For the motility analyses, Medealab CASA system (Medealab, Germany, Ver. 4.1) was used, and the velocity parameter of VCL (curvalinear velocity, μms−1), VSL (straight line velocity, μms−1), and VAP (average path velocity, μms−1) were evaluated and compared with the Day 30 and 75 non−return rates (NR30 and NR75). For every sample, a total of 10 fields were examined for 8s using a disposable 20 micron capillary chamber (CellVision, USA) giving a total of 1165 to 2831 cells evaluated. Chi square analysis, analyses of variance and linear correlation coefficient was applied to the statistical evaluation and comparison of the results. Data are based on weighted values. From the same batch of the analyzed frozen semen, a total of 8099 females were inseminated in more than 100 farms with a total of 6590 animals being positive for pregnancy at Day 30 and 4525 animals at Day 75. Within the bulls, differences were found in the values of NR30 and NR75 (P<0.05). Our data indicate very strong differences between the males’ NR30 and NR75 values (NR30: 65.6%±13.04 to 79.6%±11.17; P<0.001 and NR75: 37.8%±10.38 to 58.3%±15.53; P<0.001) reflecting the individual differences in the fertilizing capability of the males. All velocity parameters show very high correlation with strong significance both non−return rates but the best values belong to VAP (NR30 and NR75; P<0.02). Our data indicate that the bulls with lower VCL (25.51±33.04 to 79.54±58.03), VSL (11.35±19.45 to 36.36±35.71), and VAP (12.67±19.06 to 41.75±34.45) values showed lower fertilization rates both at NR30 and NR75. Computer and video technologies have advanced rapidly in recent years; thus the capability and accuracy of the latest versions of CASA systems are considerably better and they give more information about the different motion characteristics of spermatozoa. Because of the vital role of sperm motility in the reproductive process, such systems will enable us to move into a new era of diagnostic andrology and predict the fertilizing capability of semen. Supported by NKFP-Grants 4/040/2001 and 4/031/2001.

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Effects of Astaxanthin on Miniature Pig Sperm Cryopreservation

BioMed Research International ◽

10.1155/2018/6784591 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

Eunjoo Lee ◽

Daeyoung Kim

Keyword(s):

Sperm Motility ◽

Semen Parameters ◽

Control Group ◽

Computer Assisted ◽

Miniature Pig ◽

Oxidation Stress ◽

Sperm Analysis ◽

Semen Parameter ◽

Boar Sperm ◽

Sperm Plasma Membrane

The purpose of this study is to evaluate the effects of astaxanthin added to freezing buffer on semen parameters, total sperm oxidation stress after postthawing of boar sperm, and lipid peroxidation (LPO) which is caused by reactive oxygen species (ROS) in sperm membrane. Varying concentrations of astaxanthin (0, 10, 50, 100, and 500 μM) were used in the freezing buffer during cryopreservation to protect the DNA of thawed miniature pig sperm. Semen parameter was measured using computer-assisted sperm analysis (CASA) for sperm motility, and then ROS rate and oxidative stress of boar sperm were determined using fluorescence-activated cell sorting (FACS). Sperm motility was higher (p<0.05) in the astaxanthin group than in the control group. Sperm motility and the number of progressive motile sperm were higher (p<0.05) in the astaxanthin 500 μM group than in the control group. In ROS evaluation, the astaxanthin group had lower intracellular O2 and H2O2 in viable sperm. Yo-Pro-I/HE and PI/H2DCFDA staining as revealed using flow cytometry was lower in astaxanthin groups than in the other groups. As a result, we found that astaxanthin could protect the sperm plasma membrane from free radicals and LPO during boar sperm postthawing.

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Chlamydia trachomatis-induced death of human spermatozoa is caused primarily by lipopolysaccharide

Journal of Medical Microbiology ◽

10.1099/jmm.0.04836-0 ◽

2003 ◽

Vol 52 (3) ◽

pp. 193-200 ◽

Cited By ~ 57

Author(s):

S. Hosseinzadeh ◽

A.A. Pacey ◽

A. Eley

Keyword(s):

Chlamydia Trachomatis ◽

Sperm Motility ◽

Polymyxin B ◽

Probit Analysis ◽

Human Spermatozoa ◽

Similar Proportion ◽

Computer Assisted ◽

Osmotic Swelling ◽

Potent Effect ◽

Sperm Analysis

Elementary bodies (EBs) of Chlamydia trachomatis serovar E are more toxic to sperm than those from serovar LGV. In this study, lipopolysaccharide (LPS) was prepared from the EBs of both serovars and incubated with human spermatozoa at concentrations that matched the LPS concentration of EBs. The effects of EBs and LPS on sperm motility, viability and acrosomal status were then determined. Sperm motility was measured by computer-assisted sperm analysis and the hypo-osmotic swelling test was used to determine the proportion of dead cells. Acrosomal status was examined using a standard mAb assay. Over a 6 h incubation, LPS from both serovars resulted in a marked reduction in sperm motility (and a concomitant increase in the proportion of dead spermatozoa) in a manner similar to that seen in response to EBs of serovar E. In addition, when sperm were incubated with a range of doses of EBs and LPS, probit analysis revealed that the greater spermicidal effects of EBs from serovar E (when compared with serovar LGV) were not observed when sperm were incubated with LPS from the two serovars. This suggests that the more potent effect of EBs of serovar E cannot be explained entirely by differences in the composition of LPS. Interestingly, Escherichia coli LPS was required in doses 500 times more concentrated than chlamydial LPS in order to kill a similar proportion of sperm, suggesting that bacterial LPSs may differ in their spermicidal properties. However, that chlamydial LPS was spermicidal was demonstrated by the use of polymyxin B (a polycationic antibiotic known to neutralize LPS effects), confirming that the effects observed were primarily a result of LPS activity.

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Swim-up of tammar wallaby (Macropus eugenii) spermatozoa in Biggers, Whitter and Whittingham (BWW) medium: maximisation of sperm motility, minimisation of impairment of sperm metabolism and induction of sperm hyperactivation

Reproduction Fertility and Development ◽

10.1071/rd15152 ◽

2017 ◽

Vol 29 (2) ◽

pp. 345 ◽

Cited By ~ 1

Author(s):

Minjie Lin ◽

Xiyi Zhang ◽

Ray N. Murdoch ◽

R. John Aitken

Keyword(s):

Sperm Motility ◽

First Record ◽

Tammar Wallaby ◽

The Other ◽

Computer Assisted ◽

Sperm Analysis ◽

Rapid Induction ◽

Marsupial Species ◽

Accurate Quantification ◽

Better Than

A variety of media were compared for their ability to sustain the motility of tammar wallaby spermatozoa over an 8-h period following swim–up from coagulated semen. The study demonstrated that a modified Tyrode’s solution, Biggers, Whitter and Whittingham medium (BWW) was significantly better than any of the other assessed media in supporting wallaby sperm motility. After 8 h of incubation in BWW, motility was maintained at 79.3 ± 9.3%, with 77.0 ± 10.4% rapid and 65.7 ± 8.7% progressively motile spermatozoa. By contrast, motility was <10% at the same 8-h time point in all of the other media assessed. After 2 h of incubation in BWW, tammar spermatozoa consumed more oxygen than their counterparts in PBS (52.0 ± 2.7 vs 75.0 ± 6.6 μL per 108 spermatozoa per 2 h; P < 0.001). Motility was not enhanced in any of these media by the addition of 5 mM N-acetyl-D-glucosamine, the major energy substrate in wallaby semen. However, addition of dibutyryl cAMP and pentoxifylline in BWW resulted in the extremely rapid induction of hyperactivated motility in the entire sperm population. This burst of hyperactivated motility was entirely dependent on calcium in BWW and significantly inhibited by calmidazolium, a calmodulin inhibitor. A set of computer-assisted sperm analysis parameters were identified that permitted the accurate quantification of hyperactivation rates in this species. This is the first comparative analysis of media for harvesting and incubating marsupial spermatozoa and the first record of hyperactivated motility in any marsupial species.

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