3 Animal Protein-Free Semen Extender for Fixed-Time Insemination of Beef Cows

2018 ◽  
Vol 30 (1) ◽  
pp. 141
Author(s):  
S. X. Yang ◽  
E. M. Zwiefelhofer ◽  
G. P. Adams ◽  
M. Anzar
Keyword(s):  
Sperm Motility ◽  
Egg Yolk ◽  
Fixed Time ◽  
Beef Cows ◽  
Pregnancy Rates ◽  
Computer Assisted ◽  
Plant Origin ◽  
Development Fund ◽  
Semen Cryopreservation ◽  
Semen Extender

Current semen cryopreservation protocols include the use of animal products, such as egg yolk, in semen extenders, which raises important biosecurity concerns related to the transmission of infectious agents. Therefore, there is a need for alternatives to animal proteins in semen extenders. Cholesterol-cyclodextrin (CC) has been used as an adjunct in semen extenders to facilitate the delivery of exogenous cholesterol into sperm plasma membranes. The purpose of this study was to determine the fertility potential of semen extended with cholesterol of plant origin (PhytoChol, Wilshire Technologies, NJ, USA) in a fixed-time insemination program for beef cows. Semen was collected by electroejaculation from mature Simmentals bulls (n = 4 bulls, 4 replicates/bull) and analysed using computer-assisted semen analysis (CASA). Ejaculates from different bulls with ≥200 × 106 sperm mL−1 and ≥60% total motility were pooled. The pooled ejaculates were distributed into 3 treatments and diluted to 50 × 106 mL−1 in Tris-citric-acid base extenders containing either egg yolk (20% vol/vol egg yolk, control), 0.5 mg of CC per mL of semen, or 1.0 mg of CC per mL of semen. Glycerol (7% vol/vol) was added to each extender. Extended semen was loaded into 0.5-mL straws and frozen to –196°C. Post-thaw sperm motility was analysed using CASA. Ovulation was synchronized among multiparous beef cows using 1 of 3 protocols: (1) 5-day intravaginal progesterone-releasing device (PRID) and prostaglandin F2α PGF2α) treatment on the day of PRID removal (n = 37 on random days of cycle); (2) oestradiol + progesterone treatment and 7-day PRID followed by PGF2α on the day of PRID removal (n = 37 on random days of cycle); or (3) PGF2α alone (n = 19 on Day 7 to 9 from ovulation). Cows were treated with gonadotropin-releasing hormone (GnRH) and assigned randomly to be inseminated with 1 of the 3 semen treatments 72 h after PGF2α treatment. Ovulations were confirmed and pregnancies were diagnosed 28 to 35 days after insemination by ultrasonography. Post-thaw sperm motility was compared among extenders by analysis of variance. Pregnancy rates were compared among groups by generalized linear mixed model. Total motility (52 ± 3%, 59 ± 3%, and 62 ± 7%) and progressive motility (47 ± 3%, 54 ± 3%, and 58 ± 6%) did not differ among the egg yolk, 0.5 mg CC, and 1.0 mg CC extenders, respectively. There were no differences in pregnancy rates among synchronization groups; therefore, data were combined to compare the effects of semen extender. Four cows were excluded due to improper synchronization treatment. Pregnancy rates of cows inseminated with egg yolk (n = 31), 0.5 mg CC (n = 31), and 1.0 mg CC (n = 27) semen were 32, 74, and 52%, respectively (P < 0.05). Fertility in cattle inseminated using semen extended with cholesterol of plant origin has not been previously reported. We conclude that plant cholesterol may be used to replace animal proteins (egg yolk or milk origin) in bovine semen cryopreservation. Research supported by grants from the Saskatchewan Agricultural Development Fund, Agriculture and Agri-Food Canada, and Alberta Livestock and Meat Agency.

Cooled semen for fixed-time artificial insemination in beef cattle

2016 ◽  
Vol 28 (7) ◽  
pp. 1004 ◽  
Author(s):  
Juliana C. Borges-Silva ◽  
Márcio R. Silva ◽  
Daniel B. Marinho ◽  
Eriklis Nogueira ◽  
Deiler C. Sampaio ◽  
...  
Keyword(s):  
Beef Cattle ◽  
Sperm Motility ◽  
Artificial Insemination ◽  
Egg Yolk ◽  
Fixed Time ◽  
Pregnancy Rates ◽  
Reproductive Efficiency ◽  
Sperm Abnormalities ◽  
Hypoosmotic Swelling Test ◽  
Interesting Approach

This study evaluated the use of cooled semen in a fixed-time artificial insemination (FTAI) program compared with frozen–thawed semen to improve pregnancy rates in beef cattle. Ejaculates of three bulls were collected and divided into two treatments: (1) frozen–thawed semen and (2) cooled semen. Egg-yolk extender without glycerol was used for the cooled semen treatment. Straws (25 × 106 spermatozoa) were submitted to cooling for preservation at 5°C for 24 h, after which FTAI was performed. Nelore cows (n = 838) submitted to FTAI were randomly inseminated using frozen–thawed semen or cooled semen. There was a 20% increase in the pregnancy per AI (P AI–1) using cooled semen compared with frozen–thawed semen (59.9 ± 4.7 vs 49.4 ± 5.0%; P < 0.005). There was no difference in P AI–1 among the bulls (P = 0.40). The frozen–thawed semen had fewer functional spermatozoa than did the cooled semen when evaluated by sperm motility (61.7 vs 81.0%), slow thermoresistance test (41.7 vs 66.7%) and hypoosmotic swelling test (38.3 vs 53.7%; P < 0.05). The percentage of sperm abnormalities did not differ between the freeze–thawing and cooling processes (18.6 vs 22.1%; P > 0.05). Because there was less damage to spermatozoa and improvement in P AI–1, the use of cooled semen instead of frozen–thawed semen is an interesting approach to increase reproductive efficiency in cattle submitted to a FTAI protocol.

scholarly journals Efficiency of Commercial Egg Yolk-Free and Egg Yolk-Supplemented Tris-Based Extenders for Dromedary Camel Semen Cryopreservation

Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 999 ◽  
Author(s):  
Ayman Abdel-Aziz Swelum ◽  
Islam M. Saadeldin ◽  
Hani Ba-Awadh ◽  
Mohsen G. Al-Mutary ◽  
Abdullah F. Moumen ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Dna Integrity ◽  
Computer Assisted ◽  
Dromedary Camel ◽  
Plasma Membrane Integrity ◽  
Semen Cryopreservation

This study compared the efficiency of commercial egg yolk-free (AndroMed, OPTIXcell) and egg yolk-supplemented (Triladyl, Steridyl) Tris-based extenders for semen cryopreservation in seven adult dromedary camels. The camel-specific extender SHOTOR was used as control. The collected semen samples were evaluated and diluted with SHOTOR, Triladyl, Steridyl, AndroMed, or OPTIXcell. The diluted semen was gradually cooled and equilibrated for two hours before liquid nitrogen freezing. Semen was evaluated prior to freezing and after freeze-thawing cycles for motility, kinetics, vitality, abnormality, plasma membrane integrity, and DNA fragmentation using computer-assisted sperm analysis. In pre-freezing evaluation, progressive sperm motility was higher in SHOTOR-diluted samples (21.54 ± 1.83) than in samples diluted with Steridyl, OPTIXcell, or AndroMed (15.76 ± 1.80, 17.43 ± 1.10, and 13.27 ± 1.07, respectively). Moreover, Triladyl and SHOTOR resulted in significantly (p < 0.05) better sperm vitality and DNA integrity than all other diluents, but Triladyl resulted in a significantly (p < 0.05) better plasma membrane integrity (87.77 ± 0.31) than SHOTOR (85.48 ± 0.58). In the post-thawing evaluation, Triladyl led to significantly (p < 0.05) higher sperm motility (38.63 ± 0.81%; p < 0.05) when compared to SHOTOR, Steridyl or AndroMed (35.09 ± 1.341%, 34.4 ± 0.84%, and 31.99 ± 1.48%, respectively), with OPTIXcell being the least efficient (28.39 ± 0.86%). Progressive sperm motility was the highest when using Triladyl. Post-thawing curvilinear, straight line and average path sperm velocities were highest with Triladyl and lowest with AndroMed. Triladyl led to the highest linearity coefficient and straightness sperm coefficient, while SHOTOR to the highest DNA and plasma membrane integrity. OPTIXcell and AndroMed resulted in poor post-thawing sperm vitality, while Steridyl was less efficient than Triladyl. The highest rate of sperm abnormalities was recorded with OPTIXcell and the lowest with SHOTOR or Triladyl. In conclusion, SHOTOR, Triladyl, Steridyl, AndroMed, and OPTIXcell can all be used for camel semen cryopreservation; however, SHOTOR and Triladyl provided the best post-thawing sperm quality. Based on our findings, Triladyl is the best commercially available extender for dromedary camel semen cryopreservation to date.

scholarly journals Effect of quercetin or butylated hydroxytoluene on cooled or frozen-thawed ram sperm quality

2022 ◽  
Vol 43 (2) ◽  
pp. 841-854
Author(s):  
Lucas Emanuel Ferreira Canuto ◽  
◽  
Lorenzo Garrido Teixeira Martini Segabinazzi ◽  
Endrigo Adonis Braga de Araújo ◽  
Luis Fernando Mercês Chaves Silva ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Butylated Hydroxytoluene ◽  
Epifluorescence Microscopy ◽  
Computer Assisted ◽  
Semen Extender ◽  
Ram Sperm ◽  
Motility Parameters

Cooling and freezing processes cause physical and chemical damage to sperm by cold shock and oxidative stress. This study aimed to evaluate the effect of two antioxidants on sperm parameters of cooled and frozen-thawed ram semen diluted in an egg yolk-based extender. Semen was collected from 30 rams and processed in two consecutive experiments to test the inclusion of different concentrations of quercetin and butylated hydroxytoluene (BHT) in an egg yolk-based semen extender. Dimethyl sulfoxide (DMSO) was added as a solvent to the semen extender in a ratio of 1 mL DMSO for 90 mg of quercetin and 1 mL DMSO for 880 mg of BHT. After collection, semen was diluted at 200 × 106 motile sperm/mL (control) and split into different groups in each experiment. In experiment 1, semen was diluted with the extender containing quercetin (Q5, 5 μg/mL; Q10, 10 μg/mL; Q15, 15 μg/mL) or DMSO alone (DMSO1, 0.055 μL DMSO per mL; DMSO2, 0.165 μL DMSO per mL). In experiment 2, semen was diluted with the extender with BHT (BHT1, 0.5 μg/mL; BHT2, 1 μg/mL; BHT3, 1.5 μg/mL) or DMSO alone (DMSO3, 0.375 μL DMSO per mL; DMSO4, 1.125 μL DMSO per mL). After dilution, the semen was divided into two aliquots. Treated ram sperm samples were also subjected to different storage methods. The first set of samples was cooled at 5 °C for 24 h, whereas the second set of samples was frozen-thawed. Sperm motility parameters and plasma membrane integrity (PMI) were evaluated immediately after dilution (0h) and 24 h after cooling and in the frozen-thawed samples via computer-assisted sperm analysis and epifluorescence microscopy, respectively. The inclusion of quercetin or BHT did not affect sperm motility parameters or PMI of fresh, cooled, or frozen-thawed sperm in this study (P < 0.05). However, further studies are needed to test the effects of these antioxidants on the fertility of cryopreserved ram semen.

scholarly journals Ram Semen Cryopreservation Using Egg Yolk or Egg Yolk-free Extenders: Preliminary Results

2019 ◽  
Vol 50 (2) ◽  
pp. 96-103
Author(s):  
M. Ptáček ◽  
M. Stádníková ◽  
F. Savvulidi ◽  
L. Stádník
Keyword(s):  
Semen Analysis ◽  
Egg Yolk ◽  
Field Conditions ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Preliminary Results ◽  
Straight Line ◽  
Custom Made ◽  
Semen Processing ◽  
Semen Extender

Abstract Kinematic parameters of thawed ram semen frozen under field conditions were analysed with the use of two commercial (egg yolk vs egg yolk free) semen extenders in different year-seasonal periods. The semen was collected from Suffolk (n = 2) and Charollais (n = 1) rams kept in private breeding farm on 3 test days (pre-mating, at mating, post mating) during year 2016. Two commercial semen extenders (egg yolk-based BullXcell® or egg yolk-free AndroMed®) were used for semen processing. Processed semen was frozen in 0.25 ml plastic cryostraws using the custom-made portable freezing box for ram semen cryopreservation under field conditions. Thawed semen characteristics were evaluated using computer-assisted semen analysis (CASA) system immediately after thawing and after 2 h of heat incubation (±38°C). Significantly higher total motility (+12.3%), straight line velocity (+5.6 μm s−1), and average-path velocity (+6.9 μm s−1) were detected for the semen processed and cryopreserved using egg yolk-based semen extender. Year-seasonal variation and introducing the ram to service had shown to have a significant effect on the cryopreserved ram spermatozoa. These preliminary results confirmed the feasibility of ram semen processing and cryopreservation under field conditions.

Response of beef heifers and non-suckling beef cows to different oestrous synchronization protocols

2002 ◽  
Vol 74 (3) ◽  
pp. 547-552 ◽  
Author(s):  
D. Cavestany ◽  
N. Negrin ◽  
R. Negrin ◽  
J. F. Groth
Keyword(s):  
Beef Cattle ◽  
Medroxyprogesterone Acetate ◽  
Artificial Insemination ◽  
Experimental Period ◽  
Fixed Time ◽  
Beef Cows ◽  
Pregnancy Rates ◽  
Gnrh Analogue ◽  
Beef Heifers

AbstractThe objective was to evaluate different oestrous synchronization schemes in beef cattle under range conditions. In experiment 1, 202 heifers averaging 26 months of age were assigned randomly to three treatments: (a) GnRH-PG. (no. = 44) day 0 (D0): injection of GnRH; D7: injection of PGF2α; D0 to D25 oestrous detection (OD) and artificial insemination (AI); (b) OD-PG. (no. = 45) D0 to D4; OD + AI; D5: injection of PGF2α; D5 to D25 OD + AI; and (c)Ovsynch. (no. = 113) D0: injection of GnRH; D7: injection of PGF2α; D9: injection of GnRH and 16 h later AI at fixed time. In experiment 2, 318 non-suckling cows were assigned randomly to three treatments: (a) GnRH-PG. (no. = 106) D0: injection of GnRH; D7: injection of PGF2α; D0 to D25 OD + AI; (b) GnRH-MAP-PG. (no. = 106) D0: injection of a GnRH analogue and insertion of an intravaginal sponge impregnated with 250 mg of medroxyprogesterone acetate; D7: injection of PGF2α and sponge withdrawal; D0 to D25 OD + AI; (c) controls. (no. = 106) OD and AI. The experimental period lasted 25 days and, with exception of the Ovsynch treatment, oestrous detection was carried out twice a day (a.m./p.m.) and inseminations performed 12 h later. The pregnancy rates assessed by ultrasonography 30 days after AI were: experiment 1: (a) 30·0%; (b) 28·6% and (c) 62·3% (P < 0·05); experiment 2: (a) 60·2%; (b) 57·8% and (c) 45·5% (P < 0·05). It is concluded that in heifers and non-suckling cows, oestrous synchronization treatments result in higher pregnancy rates. In non-suckling cows, the addition of a progestagen did not improve the response.

scholarly journals Changes in Quality of Native and Frozenthawed Semen in Relation to Two Collections Performed in a 24-hour Interval and Adition of Clarified Egg Yolk to Extender

2016 ◽  
Vol 47 (2) ◽  
pp. 60-67
Author(s):  
P. Folková ◽  
J. Šichtař ◽  
O. Šimoník ◽  
A. Dokoupilová ◽  
R. Rajmon
Keyword(s):  
Sperm Motility ◽  
Egg Yolk ◽  
Semen Collection ◽  
Semen Extender

Abstract The aim of the study was to evaluate the effect of repeated semen collection and the substitution of normal egg yolk with clarified egg yolk to commercially produced semen extender on qualitative parameters of frozen-thawed canine semen. Two semen collections were scheduled in a 24-hour interval and in each of six dogs, three 1st and three 2nd collections were performed. The frozen-thawed sperm samples were prepared either with clarified or normal egg yolk and motility and viability were evaluated. The effect of the sequence of semen collection was demonstrated by significant differences in motility and also in viability of sperms both in native and frozen-thawed ejaculate. The percentage of viable sperms was significantly higher in samples from the 2nd compared to the 1st collection. This trend was the same also in motility except in native ejaculate. The addition of clarified egg yolk was beneficial for higher survival of sperms immediately after thawing and also after 30 min of incubation, compared to samples with normal egg yolk. Sperm motility evaluated after thawing was higher in samples with clarified egg yolk, without an apparent connection with semen collection sequence. The decrease of values of the qualitative parameters of sperms observed in the period of 30 min of incubation was significantly slowed down when clarified egg yolk was used. This was especially obvious in samples from the 2nd collection.

Embryo recovery and pregnancy rates after the delay of ovulation and fixed time insemination in superstimulated beef cows

2002 ◽  
Vol 57 (6) ◽  
pp. 1625-1634 ◽  
Author(s):  
Marcelo F. Gouveia Nogueira ◽  
Breno J.P. Barros ◽  
Andrey B. Teixeira ◽  
Luzia A. Trinca ◽  
Michael J. D’Occhio ◽  
...  
Keyword(s):  
Fixed Time ◽  
Beef Cows ◽  
Pregnancy Rates ◽  
Embryo Recovery

10 INFLUENCE OF ADMINISTRATION OF HUMAN CHORIONIC GONADOTROPIN ON DAY 4 AFTER FIXED-TIME AI ON CORPUS LUTEUM SIZE, FUNCTION, AND PREGNANCY RATE IN BEEF COWS

2014 ◽  
Vol 26 (1) ◽  
pp. 119
Author(s):  
J. B. S. Borges ◽  
D. X. Thedy
Keyword(s):  
Corpus Luteum ◽  
Pregnancy Rate ◽  
Fixed Time ◽  
Beef Cows ◽  
Pregnancy Rates ◽  
Control Group ◽  
Chi Square ◽  
Prostaglandin F2a ◽  
Successful Establishment ◽  
Early Rise

Successful establishment of pregnancy in the cow depends on early rise of progesterone (P4) to improve embryo development. The present study was carried out to determine the corpus luteum (CL) function and pregnancy rate following the administration of 1.500 IU of hCG in suckled beef cows on Day 4 after fixed-time AI (FTAI). Crossbred Angus cows (n = 429), 40 to 70 days postpartum, were synchronized using 2 mg of oestradiol benzoate (EB) i.m. and an intravaginal progesterone-releasing device containing 1 g of P4 for 7 days. At device removal, prostaglandin F2a (150 mg of D-cloprostenol) was injected i.m.; 24 h later, 1 mg of EB was given and the FTAI were done 30 to 36 h after EB using semen of just 1 bull. Cows were assigned alternatively to receive 1.500 IU of hCG i.m. (n = 200) or 1 mL of saline i.m. (n = 229) on Day 4 after FTAI. Ovaries were examined by transrectal ultrasonography (n = 71) on Days 4 and 7 to detect the presence and to measure the area of CL. Blood samples were collected on Days 4, 7, 10, 14, and 30 post-oestrus to determine P4 concentration by RIA. Pregnancy diagnoses were done 30 days after FTAI. The effects of treatments on CL area and serum P4 levels were analysed by ANOVA (Proc GLM, SAS Institute Inc., Cary, NC, USA) and pregnancy rates compared by chi-square (Proc CATMOT, SAS). The induction of accessory CL determined on Day 7 was 44% in cows treated with hCG and not observed in the control group. The area of the CL on Day 7 was significantly higher (P < 0.01; 3.37 ± 2.56 v. 2.56 ± 0.22 cm2) in the hCG group. Despite the increase in CL size, the mean levels of P4 on Days 4, 7, 10, 14, and 30 post-AI indicated no difference (P > 0.05) in CL function between control and hCG-treated cows. On Day 30, pregnancy rates were not different (P = 0.08), at 46.7 and 55% for control and hCG-treated cows, respectively. In conclusion, administration of 1.500 IU of hCG on Day 4 post-FTAI did not improve pregnancy rate in suckled beef cows.

242 EFFECTS OF TWO COMMERCIAL BOVINE SEMEN EXTENDERS FOR SHORT-TERM STORAGE ON MOTILITY PATTERN OF CHILLED BISON SEMEN

2011 ◽  
Vol 23 (1) ◽  
pp. 219
Author(s):  
B. M. Toosi ◽  
G. Gratton ◽  
C. Lessard ◽  
G. P. Adams
Keyword(s):  
Sperm Motility ◽  
Repeated Measures ◽  
Test Tube ◽  
Computer Assisted ◽  
San Jose ◽  
Short Term ◽  
Development Fund ◽  
Straight Line ◽  
Wood Bison

Difficulties of adequate cryopreservation of bison semen has limited the success of artificial insemination and in-vitro embryo production in bison. We evaluated the effects of short-term cooling on motility of bison sperm using two commercial semen extenders (Triladyl® and Andromed®; Minitube, Ingersoll, ON, Canada). Semen was collected by electroejaculation of mature wood bison (n = 3) and plains bison (n = 3) twice a week for 2 wk. Upon collection, the ejaculate was divided into 3 equal aliquots, which were then diluted 1:2 (vol/vol) in Triladyl or Andromed, or were not extended (n = 24 samples per treatment). Samples were maintained at 37°C until transfer to the laboratory (≤2 h). One millilitre of each sample was then placed into a test tube (15 mL, BD Falcon, BD Biosciences, San Jose, CA, USA) and were kept in water bath set at 5°C inside a walk-in refrigerator (4°C). Characteristics of sperm motility were evaluated before cooling (Day 0) and every 24 h after cooling for 5 days using a computer-assisted semen analyzer. Total motility (TM), progressive motility (PM), velocity curved line (VCL), velocity average path (VAP), and velocity straight line (VSL) were compared among treatments by ANOVA for repeated-measures. Values are expressed as mean ± SEM. After collection, the PM of the raw semen and semen extended in Triladyl or Andromed were not significantly different (63.1 ± 4.4%, 63.3 ± 3.1%, and 56.9 ± 4.5%, respectively). Cooling semen for a period of 24 h resulted in a decrease (P < 0.05) in PM in all 3 groups (4.4 ± 2.0%, 22.7 ± 2.9%, and 28.7 ± 4.3%, respectively). The PM of semen extended in Tryladyl or Andromed was greater than that of raw semen on Day 1 (P < 0.05). Semen extended in Triladyl and Andromed maintained PM on Day 2 (24.7 ± 3.3% and 21.8 ± 3.8%, respectively), but PM declined progressively to 1.1 ± 0.6% and 6.3 ± 2.1% by Day 5. A similar pattern was observed for the TM. The VCL, VAP, and VSL parameters for semen extended with Triladyl and Andromed decreased gradually between Day 0 and Day 5 (P < 0.05). From Day 1 to 4 after cooling, these velocity parameters were not significantly different between semen extended with Triladyl or Andromed; however, they were greater than those of raw semen (P < 0.05). All sperm velocity parameters for the raw semen declined by more than 60% between Days 0 and 2 (P < 0.05). On Day 0, VCL for semen extended with Andromed (152.2 ± 4.3) was greater than that of semen extended with Triladyl and raw semen (P < 0.05; 122.5 ± 7.0 and 122.4 ± 6.6, respectively). The VCL then decreased to 98.9 ± 12.9, 100.5 ± 10.8, and 18.6 ± 6.8 in Andromed, Triladyl and raw groups respectively on Day 2 (P < 0.05), followed by a further decline to 51.8 ± 14.3, 19.9 ± 10.0, and 9.0 ± 5.0, respectively, observed on Day 5. In conclusion, both Triladyl and Andromed improved characteristics of sperm motility of chilled bison semen. Despite an initial decrease within the first 24 h, bison sperm extended with Triladyl or Andromed maintained an acceptable degree of motility for up to 2 days after chilling to 5°C. Supported by Agriculture and Agri-Food Canada, Agriculture and Development Fund, and Canadian Animal Genetic Resources program.

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