Journal articles on the topic 'Bulls Spermatozoa Storage'
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1
Zakharchuk, D. "SPERM PRODUCTIVITY AND FERTILIZATION CAPACITY OF SPERMATOZOA OF HOLSTEIN STUD BULLS IN CONDITIONS OF LLC “UKRAINIAN GENETIC COMPANY”." Animal Breeding and Genetics 62 (December 8, 2021): 136–44. http://dx.doi.org/10.31073/abg.62.18.
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Effective selection and breeding in dairy farming is impossible without artificial insemination, results of artificial insemination largely depend on the quality of semen of bulls-improvers. Objective. To ascertain parameters of sperm productivity and fertilization capacity of spermatozoa of Holstein stud bulls of foreign selection in conditions of LLC “Ukrainian Genetic Company”. Methods. We have analyzed ejaculates of 20 mature Holstein stud bulls elite-record class brought to animal breeding enterprise from Germany and the Netherlands. The duration of use of bulls at the breeding enterprise is on average 4–5 years.Quantitative indicators of sperm were taken per year to avoid the influence of seasonal factors. Semen was collected twice a week by artificial vagina on the false stud. Ejaculates obtained were analyzed according to the DSTU 35.35-97 incertified production laboratory of LLC “Ukrainian Genetic Company”, equipped with up-to-date equipment for sperm quality assessment, packing, freezing and storage. Quantity and quality characteristics of sperm were analyzed on computer sperm analyzer IVOS (HamiltonThorneResearch, USA). Sperm fit for cryopreservation is diluted at a temperature of +35°С in the environment AndroMed (Germany) at rate no less than 20 million spermatozoids for one sperm dose and packed into 0.25 ml straws. Frozen sperm doses after check are kept in the special bio-storage in liquid nitrogen at a temperature of -196°С.
Results. Fertilization capacity ofthe spermatozoaof experimental stud bulls was measured by the percentage of quantity of cows and female calves that were impregnated after the first insemination in Zhytomyr (PAF “Ierchyky”, SPDG “Nova Peremoga”, ALLC “Ptakhoplemzavod “Korobivskyi”) and Kyiv (LLC “Agrofirm “Kyivska”) regions. On the whole results of insemination of 12525 cows and 1230 female calves of puberty age were analyzed.
Sperm productivity index and average fertilization capacity of spermatozoa of stud bulls was assessed by the method of M. M. Maiboroda, S. H. Hermanchuk, Yu. P. Polupan, and D. M. Basovsʼkyy.
It was revealedsignificant variability of sperm productivity indicators of experimental stud bulls, which vary within: number of obtained high-quality ejaculates within one year –32–173 pcs., total volume of native sperm – 201–1016 ml, average value of spermatozoa concentration in 1 ml – 1.51–3.52 milliard, average sperm motility in ejaculate – 7.2–83 points. Bull studs of LLC “Ukrainian Genetic Company” are characterized by a rather high index of sperm productivity which equals from 5.19 to 15.29 billion of motile spermatozoids in ejaculate.
Average fertilization capacity of spermatozoa of experimental Holstein bulls in conditions of 4 farms of Zhytomyr and Kyiv region ranges from 37.1 to 61.4%, average figure for examined livestock is 50.9%. During the research we found that stud bulls whose sperm fertilization capacity was above 50% in most cases (6 out of 8) had index of sperm productivity above average – 9.86–15.25billion of motile spermatozoids in ejaculate.
We have detected positive and statistically significant correlation between fertilization capacity of spermatozoa and the average value of concentration, motility of spermatozoids in ejaculate (+0.474 and+0.417 respectively) and sperm productivity index (+0.639). Conclusion. Our research proves dependence of fertilization capacity of sperm on its quality parameters. The results we obtained suggest that it is expedient to use index of sperm productivity to assess quality of sperm and prove its importance for breedingbulls selection.
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Chrenek, P., E. Spaleková, L. Olexikova, A. Makarevich, and E. Kubovicova. "Quality of Pinzgau bull spermatozoa following different periods of cryostorage." Zygote 25, no. 2 (March 9, 2017): 215–21. http://dx.doi.org/10.1017/s0967199417000077.
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SummaryThe aim of this work was to examine the influence of cryostorage duration of Pinzgau bull's insemination doses (IDs) on some sperm traits. The IDs were frozen by a slow freezing method and stored in liquid nitrogen for different periods: less than 8 years (group 1), 8–13 years (group 2) and 14–18 years (group 3). Motility (CASA), pathological sperm rate (Giemsa staining), apoptotic (Yo-Pro-1-positive) and necrotic (propidium iodide-positive) cell occurrence and fertilizing ability (penetration/fertilization test) of spermatozoa were evaluated post-thaw. The average post-thaw sperm motility in all examined groups was over 40%. No significant influence of storage length either on the sperm total motility or progressive movement was revealed. In each tested group the average rate of malformed spermatozoa did not exceed 20%. No effect of cryostorage length on the occurrence of apoptotic or necrotic sperm was noted. Similarly, penetrating/fertilizing ability of sperm did not differ among the groups, excepting differences in the rate of pronuclei (PN) formation. In group 1, 72.9% of eggs showed two visible PN following 20 h incubation with sperm, whilst in groups 2 and 3 only 67 and 54.5% of zygotes, respectively, had both PN at this time. These results revealed no influence of storage time on the bull spermatozoa in all parameters excepting the rate of PN formation. As high inter-male variability was observed in the susceptibility of bull sperm to cryostorage, individual differences should be taken into account when semen from individual bulls is to be stored for a long time.
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Bochenek, M., and Z. Smorag. "258 THE EFFECT OF A PLANT PROTEIN COMPONENT OF MEDIA USED FOR BULL SPERM SEXING ON SPERM MEMBRANE STATUS." Reproduction, Fertility and Development 20, no. 1 (2008): 209. http://dx.doi.org/10.1071/rdv20n1ab258.
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The aim of the work was to examine the effect of modified TALP medium (TALP/Pp, Animal Pharma B.V., Hengelo, The Netherlands)—used in the sperm sexing procedure—on bull sperm membrane status. The TALP was modified by replacement of bovine serum albumin (BSA) with a mixture of several plant proteins and soya lecithin (Pp). The Pp component was prepared using a high pressure homogenization process. The TALP/Pp had the same pH and osmotic pressure as the original TALP medium (TALP/BSA). The work was divided into 2 parts: (1) Nine ejaculates collected from 2 bulls (Holstein and Polish Red) were used. Immediately after collection, each ejaculate was split into 2 parts and diluted (1:2) with TALP/BSA or TALP/Pp. The sperm membrane status was examined after 3 days of storage at 15�C. (2) Fifteen ejaculates collected from 5 bulls (Holstein, Polish Red, and Simmental) were used. Each ejaculate was split into 2 parts: the first part was diluted with TALP/BSA, stained, incubated, and sexed according to the XY Inc. bull semen sexing procedure; the second part was diluted, stained, incubated, and collected after sexing into TALP/Pp with no egg yolk addition. In both groups no red food due was used to identify and exclude the dead spermatozoa from the sorted fractions. The sperm sexing procedure was performed with an SX MoFlo high-speed sorter at a speed of 3000–4000 cells/s. After collecting about 10 million spermatozoa, both fractions, X andY, were mixed, centrifuged at 700g for 15 min to concentrate the spermatozoa (20 million mL–1), and the sperm membranes examined. For sperm membrane examination, 'live/dead' samples were stained with SYBR-14/propidium iodide fluorochromes and analyzed by flow cytometry. The data from 20 000 spermatozoa were collected for each sample. The percentage of membrane-intact ('live') spermatozoa was taken for statistical analysis. The mean percentage of live spermatozoa stored for 3 days in TALP/BSA v. TALP/Pp was 25.7% (SD = 7.48) v. 28.58% (SD = 7.04), respectively (P < 0.01). The mean percentage of live spermatozoa in samples of sexed semen was 33.57% (SD = 18.97) for TALP/BSA and 38.51% (SD = 20.22) for TALP/Pp (P < 0.01). It can be concluded that Pp should be considered as a replacement for BSA in the TALP medium used for bull sperm sexing because (1) it results in significantly higher numbers of live spermatozoa after storage and/or sexing; (2) it eliminates a possible source of transmissible diseases (such as bovine spongiform encephalopathy); and (3) it decreases the total cost of the basic media used for the bull sperm sexing procedure.
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Brillianti, Febby Fairy, Pudji Srianto, Trilas Sardjito, Tri Wahyu Suprayogi, Indah Norma Triana, and Dadik Rahardjo. "Kualitas semen sapi pejantan berdasarkan umur, suhu, dan kelembaban di Taman Ternak Pendidikan Universitas Airlangga." Ovozoa : Journal of Animal Reproduction 10, no. 3 (December 1, 2021): 81. http://dx.doi.org/10.20473/ovz.v10i3.2021.81-89.
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This research was conducted at the Teaching Farm of the Faculty of Veterinary Medicine, Airlangga University, to know the effect of age, temperature, and humidity on the quality of semen of bulls which are turned into frozen semen. This research uses fresh semen samples of bulls of various ages grouped into 3 groups, young, middle age, and old groups and measures temperature and humidity during the storage process. Fresh semen is collected using an artificial vagina and then examined macroscopic and microscopic tests which is then checked for post thawing motility. Data analysis uses Analysis of Variants (ANOVA) followed by Duncan. The results showed that the percentage of spermatozoa motility of fresh semen and post thawing differed and age affected the quality of semen produced. There was no effect of temperature and humidity when processing of collecting semen. Bull’s semen at Teaching Farm Faculty of Veterinary Medicine Airlangga University has good quality and is suitable for Artificial Insemination.
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Pero, M. E., G. J. Killian, P. Lombardi, L. Zicarelli, L. Avallone, and B. Gasparrini. "327 IDENTIFICATION OF OSTEOPONTIN IN WATER BUFFALO SEMEN." Reproduction, Fertility and Development 19, no. 1 (2007): 279. http://dx.doi.org/10.1071/rdv19n1ab327.
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The competitiveness of buffalo breeding will depend on the utilization of reproductive biotechnologies that allows acceleration of genetic progress. A major factor hampering the efficiency of both artificial insemination and in vitro embryo production programs in this species is male hypofertility. Reports for several species suggest that seminal plasma contains factors that influence male fertility. Osteopontin is a glycoprotein found in several biological fluids including seminal plasma, and its presence is associated with spermatozoa concentration. In cattle, expression of osteopontin was highly correlated with bull fertility, and it was proposed to be a marker to predict male fertility (Cancel et al. 1999 Biol. Reprod. 60, 454–460). No data are available about the presence or activity of osteopontin in water buffalo. The aim of this preliminary study was to determine if osteopontin is present in buffalo semen and to evaluate whether freezing procedures cause the loss of osteopontin from spermatozoa. Semen was collected in authorized semen collection centers from 6 buffalo bulls by using an artificial vagina. A collection of bovine semen was used as a positive control. An aliquot from each sample was frozen using standard procedures for semen storage. Each ejaculate was centrifuged at 600g for 10 min at room temperature, and the supernatant was recovered and centrifuged at 10 000g for 1 h at 4�C. The total protein concentration in seminal plasma and spermatozoa was determined by the Bradford method, using ovoalbumin as the standard. Proteins (50 �g) were separated by electrophoresis and analyzed by western blotting (Cancel et al. 1999). Polyclonal antibodies against bovine milk osteopontin were prepared as previously described (Cancel et al. 1997 Biol. Reprod. 57, 1293–1301). The intensities of bands indicated by western blot were quantified by densitometer. Osteopontin was detected in all samples of buffalo semen. Most of the osteopontin detected was in the seminal plasma. Relative amounts of osteopontin detected in spermatozoa were 50% or less of that detected in seminal plasma; furthermore, the protein was not found in sperm from all bulls. These results suggest that most osteopontin is produced by the ampullae and seminal vesicles, similar to what was reported for cattle (Cancel et al. 1999). Semen frozen by standard procedures showed a reduction in amount of osteopontin by up to 50%. These studies suggest that the fertility-associated protein osteopontin may be useful as a sensitive tool to evaluate whether sperm storage procedures are detrimental and result in excess loss of osteopontin from sperm. In conclusion, the results have demonstrated that osteopontin is present in buffalo seminal plasma and sperm. Further studies will examine whether the expression of osteopontin is correlated with the fertility of buffalo bulls, as has been demonstrated in bovine bulls.
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Khaled Taïbi, Mohamed Achir,, Leila Ait Abderrahim, Mohamed Boussaid, Kada Souana, Abdelkader Tadj, Toufik Benaissa, and Tayeb Gouchich. "Dissecting the relationship between artificial insemination success and bull semen quality in the arid region of Tiaret (Algeria)." Bionatura 7, no. 1 (February 15, 2022): 1–5. http://dx.doi.org/10.21931/rb/2022.07.01.18.
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Despite being subject to prior assortment, frozen bull sperms commercialized for artificial insemination may present certain morphological defects. The present study aims (i) to assess the artificial insemination success of the most common cattle breeds in Algeria and (ii) to evaluate the possible effects of commercialized bull’s semen quality on this operation. Artificial insemination was assessed through four years of field monitoring by inseminating different cattle breeds of normal fertility. However, semen quality was evaluated using light microscopy by measuring viability, motility, and morphological abnormalities of spermatozoa. Field study revealed a high percentage of normal calving in red and white Holstein breed (44.83 %) against the high percentage of embryonic mortality (46.43 %) and calving with a malformation (10.71 %) in Montbéliarde breed. Semen quality assessment revealed that sperm viability and motility were higher in Holstein breeds than in Montbéliarde. Furthermore, significant differences between semen bulls were found in the proportion of abnormal spermatozoa; a higher rate of sperms with the abnormal head was observed in the black and white Holstein breed (69.3±10.98 %). However, the percentage of abnormal sperms with tail defects was higher in the Montbéliarde breed (67.5±10.74 %). The lousy quality of the selected semen and/or the poor handling and storage of frozen semen constitute a determinant factor that hinders the success of artificial insemination in the arid region of Tiaret (Algeria).
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Susandani, Oky, Tri Wahyu Suprayogi, Ratna Damayanti, and Anwar Ma'ruf. "Factors Affecting Fresh Semen Quality in Pasundan Cattle at UPTD BPPIBTSP Ciamis." Journal of Applied Veterinary Science And Technology 2, no. 2 (October 30, 2021): 37. http://dx.doi.org/10.20473/javest.v2.i2.2021.37-42.
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Background: Pasundan cattle are local cattle native to Indonesia. One way to conserve beef cattle genetics is to use Artificial Insemination technology. The success of Artificial Insemination can be influenced by the quality of semen. Purpose: To determine factors affecting fresh semen quality in Pasundan cattle at UPTD BPPIBTSP Ciamis. Methods: The data were obtained through observations on seven Pasundan bulls in March 2021 towards fresh semen quality and some factors influencing it. The Pasundan bulls observed were seven productive males. Results: The fresh semen quality of Pasundan cattle, such as volume, color, and pH, showed good result,s but the average consistency and concentration of spermatozoa were still below the standard. The factors that can affect the fresh semen quality are the breed of beef cattle, age, body weight, feed, season, exercise, and frequency of semen storage. Conclusion: The determining factors that can cause the consistency and concentration of Pasundan cattle’s spermatozoa at UPTD BPPIBTSP Ciamis are feed and season.
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Malcotti, V., V. Pelufo, N. Bergamo, and E. Aisen. "Recovery of epididymal spermatozoa from bull and red deer, stored at different times and temperatures before freezing - thawing." Animal Production Science 52, no. 8 (2012): 741. http://dx.doi.org/10.1071/an11366.
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In order to preserve male germoplasm, the recovery and cryopreservation of spermatozoa from the epididymides of hunted animals represents an accessible source of gametes. As a first experimental model, epididymal spermatozoa from slaughtered bulls were recovered at 30, 54, 78 and 102 h after death. The scrotal contents were stored at either 5 or 20°C. The sperm cells of each treatment (time + temperature combinations) were frozen with Triladyl (T) or Triladyl + Trehalose (TT) diluents. In order to assess sperm viability and integrity, post-thawing evaluation included individual motility, supravital stain, hyperosmotic swelling test (E+), acrosome status and sperm chromatin structure assay. Both at raw and post-refrigerated states, the sperm motility rate was higher in sperm obtained from epidydmes stored at 5°C, compared with those stored at 20°C for all collection times. Sperm collected at 102 h after death from epididymides stored at 5°C maintained a motility of 20% (120 h, raw state). When comparisons were carried out after thawing, motility was higher in the 5°C group, achieving the best results with TT diluent (7.5%) at 102 h. However, when supravital stain and E+ tests were observed, viability and membrane integrity were well preserved even at 102 h post mortem (30 and 36%, respectively, with TT diluent at 5°C). These results suggest that frozen-thawed epididymal spermatozoa could have a low motility rate while most of them remain alive. Acrosome status was not greatly affected by storage time. In a second experiment, epididymal spermatozoa from hunted red deer stags (Cervus elaphus) were recovered at 4 and 30 h after death. The scrotal contents were stored at 20°C, because that temperature is closer to field and shipment conditions. The sperm cells were frozen with TT diluent. Post-thawing evaluation included the same parameters indicated for bull spermatozoa. The assessment of spermatozoa collected at 30 hours post mortem and then subsequently frozen and thawed indicated that at this time an acceptable motility rate (35%) and viability (39.7%) were achieved. Frozen and subsequently thawed epididymal spermatozoa showed 47.9% of membrane integrity, 59.3% of acrosome integrity and 26.5% of chromatin damage, using TT diluent. A preliminary in vivo trial demonstrated that the pregnancy rate in artificially inseminated deer decreased when sperm were obtained at 30 h post mortem. According to these results, it may be concluded that storage at 5°C is better than 20°C to obtain well preserved epididymal spermatozoa from bulls, and that TT could be a useful cryoprotectant to preserve viable and fertile sperm cells after the freezing–thawing process. Before these results can be applied to assisted reproduction programs in endangered deer species, some adaptations must be developed.
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Amorim, E. A. M., J. K. Graham, M. Meyers, and B. Spizziri. "67 DELIVERING CHOLESTANOL OR DESMOSTEROL TO BULL SPERM MEMBRANES IMPROVES CRYOSURVIVAL." Reproduction, Fertility and Development 20, no. 1 (2008): 114. http://dx.doi.org/10.1071/rdv20n1ab67.
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Altering the lipid composition of sperm plasma membranes affects sperm cryosurvival. Cryopreservation induces many stresses on the spermatozoa, including destabilization of the plasma membrane, which results in the loss of sperm motility and function. Treating bull spermatozoa with cholesterolloaded cyclodextrin (CLC) prior to cryopreservation increases sperm cryosurvival rates. This study compared the effect of adding other sterols, which should incorporate into the membrane and increase membrane fluidity at low temperatures, thereby increasing cryosurvival. Ejaculates from four bulls were divided into two experiments (E). In E1, ejaculates were extended with Tris, and then subdivided into four treatments: No additive (control), 1.5 mg CLC/120 million sperm (positive control), and 1.5 mg/120 million sperm in cyclodextrin pre-loaded with either cholestanol or desmosterol. Spermatozoa were incubated for 15 min at 22�C after which both the ability of fresh spermatozoa to bind to the zona pellucida (ZP) and chicken egg perivitelline membrane (EPM) and their osmotic tolerance were evaluated. In E2, sperm were diluted to 120 million cells mL–1 in a Tris diluent and treated as described for E1. Then, samples were diluted 1:1 (v:v) in Tris with 20% Egg Yolk (EY) and cooled to 5�C. After dilution 1:1 (v:v) with Tris containing 10% EY and 16% glycerol, samples were allowed to equilibrate for 15 min, and then were packaged into 0.5-mL straws, frozen in static liquid nitrogen vapor for 20 min, and plunged into liquid nitrogen for storage. Straws were thawed and the motility and zona-binding ability were determined using a Hamilton Thorne Motility Analyzer (Hamilton Thorne Biosciences, Beverly, MA, USA) and epifluorescence microscopy, respectively. Treatment differences for sperm motility, osmotic tolerance, and zona binding were determined using analysis of variance. Treating spermatozoa with CLC resulted in more fresh bull spermatozoa binding to the EPM and ZP compared to cholestanolor desmosterol-loaded cyclodextrin-treated spermatozoa or control cells (P < 0.05). No differences were observed between EPM and ZP binding (P > 0.05). The percentages of total and progressively motile spermatozoa were higher for fresh samples treated with cholesterol-, cholestanol-, or desmosterol-loaded cyclodextrin than for control cells (P < 0.05) when spermatozoa were exposed to anismotic conditions, and then returned to isosmolality. After cryopreservation, the percentages of motile spermatozoa and number of spermatozoa binding to ZP were similar for spermatozoa treated with CLC (56% and 115 sperm/ZP) and cholestanol (53% and 108 sperm/ZP) compared to spermatozoa treated with desmosterol (42% and 86 sperm/ZP; P < 0.05). All treatments provided higher motility and binding efficiency than control spermatozoa (32% and 62 sperm/ZP; P < 0.05). Therefore, adding cholesterol or cholestanol to bull sperm membranes improved cell cryosurvival. Studies to determine if cholestanol affects sperm capacitation need to be conducted.
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Durfey, C. L., T. Rowlison, C. U. Lagu, C. Sente, M. L. Khaitsa, H. J. Clemente, P. L. Ryan, S. T. Willard, and J. M. Feugang. "111 Improvement of cat and bull sperm quality using nanotechnology as a model for wild species." Reproduction, Fertility and Development 31, no. 1 (2019): 181. http://dx.doi.org/10.1071/rdv31n1ab111.
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Assisted reproductive techniques (ART) are widely used in domestic species, with increasing applications in wildlife conservation. However, the currently available techniques for semen preparations are not fully reliable or applicable in all species to ensure successful ART outcomes. Recent developments in nanotechnology offer new horizons for further optimization of sperm preparations. The use of conjugated magnetic nanoparticles allows for selective targeting and removal of moribund spermatozoa through the technique known as nanopurification, which has shown to be beneficial in domestic boars and bulls. Nanopurification is a rapid, straightforward, effortless, and noninvasive technique that, when applied in wildlife or endangered species, is expected to have a tremendous impact on ART outcomes. In this study, we evaluated the effectiveness of magnetic (iron oxide) nanoparticles designed to target moribund spermatozoa (e.g. acrosome reacted) on domestic felid and threatened bovid species. Fresh epididymal spermatozoa of domestic cats and ejaculated spermatozoa from genetically valuable threatened bulls (Ankole and Sahiwal) were collected and mixed with iron-oxide magnetic nanoparticles (MNP). After 30min of incubation at 38°C, semen mixtures were placed on an electromagnetic field (10-15min) to trap MNP-bound spermatozoa (moribund), followed by the elution of viable nontrapped spermatozoa (nanopurified). All samples were analysed for motility characteristics using a computer-assisted sperm analyzer. Aliquots of cat spermatozoa were subjected to (1) morphology and acrosome analyses (n=4-8 replicates), (2) cryotolerance to liquid nitrogen storage (n=4 replicates), and (3) IVF (n=1 replicate). Data were analysed with Student’s t-test and P<0.05 indicated significant differences. Regardless of the species, both control and nanopurified spermatozoa revealed comparable motility characteristics (P>0.05). Sperm MNP-bound samples exhibited higher proportions of moribund spermatozoa as compared to their nanopurified and control nonpurified counterparts (57.5v. 47.5 and 49.5%, respectively; P<0.05). Cryopreservation revealed a trend for higher post-thaw motility of nanopurified cat spermatozoa (18%) v. the controls (12%). The use of frozen-thawed control v. nanopurified cat spermatozoa to in vitro fertilize oocytes (n=13 per group) resulted in comparable embryo cleavage (15v. 31%, respectively) and blastocyst (15v. 8%, respectively) rates. This preliminary study indicates the successful interactions of MNP nanoparticles with feline and bovine spermatozoa. The nanopurification seems to improve the cryotolerance of cat spermatozoa without affecting their fertilization potential. Additional research is being conducted to confirm the current findings and optimize this novel technique for future implementation in conservation breeding programs. Work supported by USDA-ARS 58-6402-3-018 and Morris Animal Veterinary Student Scholarship - D18ZO-608.
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Szczesniak-Fabianczyk, B., M. Bochenek, A. T. Palasz, J. De la Fuente, and Z. Smorag. "95 EFFICACY OF FIVE DIFFERENT SEMEN EXTENDERS FOR THE CRYOPRESERVATION OF BULL SEMEN." Reproduction, Fertility and Development 20, no. 1 (2008): 128. http://dx.doi.org/10.1071/rdv20n1ab95.
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Replacement of animal-origin components in extenders used for bull semen freezing is of high importance for individuals involved in cattle breeding. The experiment was designed to compare efficacy of 5 different semen extenders in cryopreservation of bull semen: sodium citrate-based extender containing egg yolk (CT), commercially available Bioxcell� (IMV Technologies, L'Aigle, France), and 3 custom-made homogenized plant lipidsbased, egg yolk-free extenders (Y-1, Y-2, and Lipo) . The objective was to determine whether homogenization procedures of lipids improve the quality of the extender. Lipid homogenates of custom-made extenders were prepared in Tris buffer using a high pressure homogenizer (Nira Saovi, Parma, Italy). Ten (Y-1) or 5 (Y-2) homogenization cycles were applied and then 8% glycerol was added. Lipid liposomes were produced by simultanous high pressure homogenization of lipids and glycerol supplementation (Lipo). Semen was collected from young bulls of 3 different breeds (Simmental, Polish Red, and Holstein; 1 ejaculate/bull). Each ejaculate with at least 70% motility was split into 5 parts and processed further by a standard freezing protocol: semen was diluted at 35�C with each of the 5 extenders to a concentration of 100 � 106 spermatozoa per mL, cooled to 4�C over 5 h, aspirated into 0.25-mL plastic straws, frozen in liquid nitrogen vapor to –140�C, and then plunged into LN2. Straws were thawed in a water bath at 37�C for 20 s. Sperm motility was estimated microscopically immediately after thawing and after 5 h of storage at 22�C. Immediately after thawing, flow cytometry and SYBR-14/PI staining were used for examination of sperm membrane integrity (live/dead assay). A total of 20 000 spermatozoa of each sample were counted. Student's t-test was used to estimate statistical differences between experimental groups. The mean sperm motility after thawing ranged from 45.6% (SD = 13.7) for CT (egg yolk extender) to 57.8% (SD = 7.1) for Lipo. A significant difference (P < 0.05) was observed betweenY-1 (50.0%, SD = 9.7) and Lipo and Bioxcell (56.1%, SD = 8.6). After 5 h of storage at 22�C, the mean motility for all tested bulls ranged from 25.0% (SD = 7.1) for CT to 42.2% (SD = 7.5) for Lipo. Significant differences were observed between Lipo (P < 0.01), Y-2 (P < 0.05) and CT, and between Y-1 and Lipo (P < 0.01). Mean percentage of 'live' spermatozoa with intact membrane after freezing/thawing was 51.85% (SD = 11.49) for Y-1, 45.72% (SD = 9.36) for Y-2, 47.57% (SD = 7.93) for Lipo, 45.47% (SD = 8.35) for Bioxcell, and 49.06 (SD = 11.59) for CT. No significant differences were observed except forY-1 and Bioxcell extenders (P < 0.05). It can be concluded that both methods of lipid/glycerol homogenization can be successfully applied in the preparation of bull semen extender. In addition, extensive lipid homogenization (10 cycles) produced more transparent extender that in turn improved visualization of sperm. Custom-made plant origin lipids homogenization may provide a valuable alternative for the preparation of extenders that more closely match the membrane composition of bull sperm cells and thus contribute to development of an efficient extender free of animal-origin components for bull semen freezing.
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Bochenek, M., T. Herjan, and Z. Smorag. "361 INFLUENCE OF SEXING PROCEDURE ON BULL SPERM CHROMATIN STRUCTURE." Reproduction, Fertility and Development 19, no. 1 (2007): 296. http://dx.doi.org/10.1071/rdv19n1ab361.
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Flow cytometry is the only reliable and relatively fast method allowing separation of live X and Y spermatozoa for sex regulation. Many thousands of animals of different mammalian species have been born after insemination with sexed semen during the past 20 years. Nevertheless, the question is still open: does the bull sperm sexing technology affect chromatin structure? A case of serious chromatin damage after sexing stallion semen was reported previously (Bochenek et al. 2006 Havemeyer Foundation Monograph Series No. 18, 13 –14). The aim of this work was to examine the effect of the sexing procedure and different UV laser powers on bull sperm chromatin structure. The ejaculates of 28 bulls (one ejaculate/bull) were used in the study. Each ejaculate was divided into 5 groups: (1) control, unprocessed; (2) sorted strictly according to XY Inc. protocol (Schenk et al. 1999 Theriogenology 52, 1375 –1391); (3) as group 2, but without the Red Food dye staining used for dead spermatozoa discrimination; (4) as group 2, but with double UV laser power (300 mW); and (5) as group 3, but with double UV laser power (300 mW). Sperm sorting was performed with a MoFLoSX flow cytometer at speeds of 3000 –5000 cells/s. Sorted fractions of X and Y spermatozoa were mixed again and stored for 24 h at 15 °C. A sperm chromatin structure assay (SCSA) was performed twice on each sorted sample, immediately after sorting and after 24 h. The chromatin of control samples was examined according to the same time schedule. The percentage of spermatozoa with damaged chromatin was calculated (COMP α-t) as well as standard deviation of the α-t parameter (SD α-t). The latter parameter, although less intuitive, is considered as even more precise than COMP α-t in chromatin investigations. The mean percentage of spermatozoa with abnormal chromatin was 1.12% (SD = 0.47) for control samples. The highest level of chromatin abnormality was noted for the 300 mW group with no dead cell discrimination (Red Food staining): 1.29% (SD = 1.05). After 24 h of storage, the mean level of chromatin abnormality increased to 1.97% (SD = 0.96) in control samples whereas that in all sorted samples was lower: from 1.06% (SD = 0.4) to 1.16% (SD = 0.62) in the 150 mW/non-Red Food-stained and the 300 mW/Red Food-stained groups, respectively. This difference appeared to be statistically significant (t; P ≤ 0.05). Interestingly, the percentage of abnormal spermatozoa decreased slightly after 24 h of storage in the 300 mW/Red Food-stained and the 300 mW/non-Red Food-stained groups ( –0.13% and –0.08%, respectively). Calculation of the SD α-t parameter showed statistically significant differences in chromatin abnormality between the control group vs. the 300 mW/non-Red Food-stained group immediately after sorting and the control group vs. the 150 mW/Red Food-stained group after 24 h of storage. In conclusion, although the statistically significant increase of chromatin damage was found after sexing in some investigated groups, it seems that the level of this abnormality is far too low to affect sexed semen fertility.
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Carini, M. E., R. Cavia, G. Larraburu, and G. M. Brogliatti. "86 MOTILITY AND VIABILITY OF BULL SEMEN DURING 24 HOURS OF STORAGE AT 4°C." Reproduction, Fertility and Development 18, no. 2 (2006): 151. http://dx.doi.org/10.1071/rdv18n2ab86.
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Currently, cryopreservation process of fresh bull semen is carried out between 3 and 6 hours of refrigeration at 4°C post-collection (Hafez, 1989). However, it is sometimes difficult when the cryopreservation process is not available at the site of collection. The objective of this study was to determine seminal motility and viability in samples maintained at 4°C during 24 hours. A total of 98 ejaculates from 23 adult bulls (Angus, Brangus, Braford and Hereford) were collected and diluted in a semi-defined semen extender (Andromed, Minitüb, Tiefenbach, Germany) and stored at 4°C. Parameters of velocity average path (VAP, µm/s), velocity straight line (VSL, μm/s), amplitude lateral head (ALH, μm), linearity (LIN, %), percentage of rapid cells (RAPID, %), percentage of slow and static cells (SL/ST, %), and viability (VIA, %) were determined by Computer Assisted Semen Analysis (CASA, HTM-ceros 12.1, Berkeley, CA, USA). Measurements were done at 6, 9, 12, and 24 h. The obtained results were analyzed statistically with one-way ANOVA and Dunnet Multiple Comparison Test and are summarized in Table 1. There were no significant differences (P > 0.05) in the VAP, RAPID, or SL/ST during 24 h of storage at 4°C. Measurements were significantly different (P < 0.01) for VSL and VIA at 24 h. Measurements of ALH were increased from 12 h (P < 0.01) and consequently, LIN decreased at the same time (P < 0.01). These results suggest that there are no differences in velocity, except in VSL at the end of the storage time. The type of movement of the spermatozoa change, because ALH increases and the trajectory loses linearity. A decrease in viability suggests that from 24 h of storage, the membrane of the spermatozoa becomes more susceptible. More research needs to be done to evaluate the competence of this time-storage semen in the artificial insemination trial. Table 1. Parameters of motility and viability of semen maintained at 4°C during 24 h This research was supported by Centro Genético Bovino de EOLIA S.A.
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Imrat, P., S. Mahasawangkul, J. Gosálvez, P. Suthanmapinanth, P. Sombutputorn, S. Jansittiwate, N. Thongtip, et al. "Effect of cooled storage on quality and DNA integrity of Asian elephant (Elephas maximus) spermatozoa." Reproduction, Fertility and Development 24, no. 8 (2012): 1105. http://dx.doi.org/10.1071/rd11309.
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Artificial insemination (AI) is a potentially useful tool for breeding captive elephants because it facilitates efforts to minimise inbreeding. However, cooled storage of elephant semen markedly reduces fertility. This study compared the effects on semen-quality parameters, including sperm DNA fragmentation, of storing elephant semen at 4°C or 15°C in a commonly-used diluent (TEST) or a diluent developed to protect against sperm DNA damage (BullMax). Storing elephant semen for >24 h in either extender at either temperature resulted in decreases in sperm motility, viability, acrosome integrity and DNA integrity (P < 0.05); the decrease in motility was especially rapid. A subjective impression of circular sperm movement in TEST was confirmed by a higher curvilinear velocity and amplitude of lateral head displacement, but lower straight-line velocity and linearity than in BullMax. Initial percentages of spermatozoa with fragmented DNA (%SDF) did not differ between extenders or temperatures, but the rate of increase in %SDF during a 48-h incubation at 37°C was higher in TEST than in BullMax (P < 0.05). In conclusion, BullMax allows more linear movement and better preserves DNA stability of stored elephant spermatozoa than TEST. Sperm DNA stability during incubation at 37°C is a promising, discriminative parameter for selecting semen storage conditions of bulls for elephant AI.
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Solomina, O. N., M. I. Sergushkina, А. А. Shirokikh, T. V. Polezhaeva, I. G. Shirokikh, O. O. Zaitseva, and A. N. Khudyakov. "Hericium erinaceus BP16 as a source of polysaccharides stabilizing the functions of bulls spermatozoa during hypothermic storage." Theoretical and Applied Ecology, no. 3 (2021): 212–18. http://dx.doi.org/10.25750/1995-4301-2021-3-212-218.
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Tamargo Miguel, C., S. S. Pérez-Garnelo, P. Beltrán Breña, A. T. Palasz, J. De la Fuente, A. Rodriguez, and C. O. Hidalgo. "162 FREEZING OF SEMEN FROM ENDANGERED ASTURIANA DE LA MONTAÑA BULLS IN ZWITTERONIC LIPIDS-BASED EXTENDERS." Reproduction, Fertility and Development 20, no. 1 (2008): 161. http://dx.doi.org/10.1071/rdv20n1ab162.
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This experiment was designed to test the efficacy of 2 different preparation protocols of zwitteronic soyabean-origin lipids for the production of lipidsglycerol liposomes for use in bull semen cryopreservation. Lipids liposomes were prepared at 10% concentration in Tris buffer by 1. highpressure homogenization (Panda 2K, Parma, Italy) and then 8% glycerol were added, extender-1 (E-1); Lipids were homogenized together with glycerol, extender-2 (E-2). Bioxcell extender (E-3) was used as control. Semen was collected 3 times from 3 endangered Asturiana de la Monta�a bulls by the means of an artificial vagina. Ejaculates with at least 70% motility were processed further by a standard freezing protocol used in our AI station. Semen was diluted at 37�C with each of the 4 extenders to a concentration of 92 � 106 spermatozoa per mL, cooled to 4�C over 4 h, aspirated into 0.25-mL plastic straws (IMV Technologies, Aigle, France), frozen in a bio-freezer (IMV Technologies) in 3 steps from 4 to –140�C, and then plunged into liquid N2. Straws were thawed in a water bath at 37�C for 30 s. Sperm motility was analyzed microscopically immediately after collection, after dilution, and after 4, 24, 48, and 72 h of storage at 4�C. Post-thaw semen progressive motility was assessed microscopically, and sperm movement characteristics were analyzed by computer-assisted semen analysis (CASA) (SCA�, Microptic, Barcelona, Spain). Data were compared between extenders and bulls by 2-way ANOVA; percentages were transformed by arcsine transformation before ANOVA. Total and progressive sperm motility at 0 h after dilution ranged from 90 to 70% and was not different between extenders and bulls. There was no difference between bulls in total and progressive motility after 24 h of cold storage; however, both were significantly greater (P < 0.05) for Control (62.4 � 14.7 and 41.4 � 14.9) and E-1 (70.1 � 12 and 33.8 � 7.0) extenders than for the E-2 extender (22.5 � 17 and 1.2 � 1.3). Average post-thaw sperm motility was not different between bulls for either extender, but motility for Bioxcell (Control, 48.1 � 14.6%) and E-1 extenders (43.2 � 13.0%) were significantly greater (P < 0.05) than for E-2 extender (18.7 � 8.8%). There were no differences between bulls for all kinematic semen parameters: curvilinear (VCL), straight line (VSL), average path (VAP) velocities, linearity (LIN) and straightness (STR), evaluated by CASA before and after freezing; however, all were lower (P < 0.05) for the E-2 extender and not different between Control and E-1 extenders. Sperm movements derived from heads (VCL) and linearity of sperm(LIN), both closely related to field fertility, were in the range of 90.9 � 2.1 and 63.0 � 5.5 for E-3 (Control) extender; 99.1 � 3.4 and 49.4 � 3.5 for E-1; and 21.8 � 2.2 and 29.9 � 4.0 for E-2. In summary, zwitteronic soyabean lipid liposomes are an effective egg yolk substitute for the cryopreservation of Asturiana de la Monta�a bull semen; however, the homogenization protocol of the lipids-glycerol mixture must be improved.
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Nain, Dipti, Tushar Kumar Mohanty, Raju Kr Dewry, Mukesh Bhakat, Sapna Nath, Vinod Kumar Gupta, and Mohsin Ahmad Parray. "Butylated Hydroxytoluene (BHT) Improves the Post-Thaw Semen Quality in Low-Dose Sperm Cryopreservation in Murrah Buffalo Bull." Cryoletters 44, no. 1 (January 1, 2023): 57–65. http://dx.doi.org/10.54680/fr23110110612.
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BACKGROUND: Cryopreservation is an important technique for the long-term storage of semen for artificial insemination (AI). Buffalo spermatozoa are sensitive to cryopreservation procedures because of the presence of a high amount of polyunsaturated fatty acids in the plasma membrane. OBJECTIVE: To study the effect of different concentrations of BHT on the quality of Murrah buffalo bull semen for low-dose cryopreservation. MATERIAL AND METHODS: Semen was collected from four high fertile Murrah buffalo bulls (6 ejaculates each) using an artificial vagina. A total of 24 ejaculates were collected from each bull twice a week using an artificial vagina. Every sample was split into four parts: Control without additives; and three treatments with BHT at 0.5 mM, 1 mM or 2 mM. Semen was cryopreserved at low-dose sperm cryopreservation of 20, 15, 10 and 5 million sperm per aliquot after supplementation of BHT. Semen samples were evaluated for fresh, pre-freeze and post-thaw stages. RESULTS:There was a significant increase (p<0.05) in sperm quality parameters, such as progressive motility (%), viability (%), HOST response (%), acrosome integrity (%) and post-thaw motility, with the addition of 0.5-1 mM BHT. CONCLUSION: The addition of BHT in Murrah buffalo semen improves the low dose cryopreservation quality in a dose-dependent manner.
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Olivier, F., T. Spies, F. Martinez-Pastor, D. M. Barry, and P. Bartels. "211QUALITY CHANGES IN BLUE WILDEBEEST (CONNOCHAETES TAURINUS) EPIDIDYMAL SPERMATOZOA MAINTAINED AT 4°C." Reproduction, Fertility and Development 16, no. 2 (2004): 227. http://dx.doi.org/10.1071/rdv16n1ab211.
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Wildlife management in southern Africa often involves the harvesting of animals on ranches and reserves, providing unique opportunities to collect and assess the quality of epididymal spermatozoa for possible future conservation actions. The black wildebeest (Connochaetes gnu) is facing renewed threats to its survival, including the production of fertile hybrids from crossing with the more common blue wildebeest (Connochaetes taurinus). The close relationship between the two wildebeest species allows for the blue wildebeest to be used as a model to assess epididymal sperm quality over time while maintained at 4°C. Field conditions often preclude the immediate availability of liquid nitrogen, necessitating the development of alternative short-term storage methods. All chemicals were provided by Sigma (South Africa) unless otherwise stated. Testes were harvested from 6 blue wildebeest bulls at a local game farm, Savannah, and kept at 5°C during transportation to the lab. Epididymides were dissected out and spermatozoa were flushed out of the cauda epididymis using 1mL of Tris-citrate egg yolk extender (Fraction A, Biladyl, Minitub, Germany), followed by storage at 4°C and assessment at 12h intervals. At each interval, an aliquot was removed, washed with a modified buffered HEPES solution (20mM HEPES, 355mM sucrose, 10mM glucose, 2.5mM KOH;; 400mOsm/kg, pH 7; Sigma) and visually assessed with a phase contrast microscope (×200, at 37°C) to determine the percentage of motile (MS) and progressive motile (PS) spermatozoa. In addition, plasma membrane integrity (PMI) was assessed with eosin-nigrosin staining and active mitochondrial status (MIT) assessed with an epifluorescent microscope (×400) using the fluorescent probe JC-1 (Molecular Probes, The Netherlands;; 7.5μM; 30min at 37°C). Resilience to hypo-osmotic shock was also evaluated by incubating the sample in a modified hypo-osmotic medium (100mOsmkg−1; 15min RT), and staining with PI to assess plasma membrane integrity (HOSPMI). A summary of results is presented in the table 1. The MS, MIT and HOSPMI did not decrease significantly during the 48h storage period. The only parameters that showed a significant decrease were PS and PMI (P<0.01, Kruskall-Wallis test). However, PMI showed a slow but steady decrease (13%), whereas PS underwent a significant drop (52%). In conclusion, epididymal spermatozoa from the blue wildebeest, kept at 4°C for 48h, may still be useful for some assisted-reproduction techniques. The use of spermatozoa from a common but closely related wildebeest species allows for the development of assisted-reproduction techniques that may one day aid the conservation of threatened wildebeest species. Additional research is needed to confirm these findings and to test the effect of longer storage times on spermatozoa of this species as well as closely related endangered species. Table 1 Parameters measured during the 12-h time periods (mean±SD)
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Perumal, P., Kezhavituo Vupru, and K. Khate. "Effect of Addition of Melatonin on the Liquid Storage (5°C) of Mithun (Bos frontalis) Semen." International Journal of Zoology 2013 (2013): 1–10. http://dx.doi.org/10.1155/2013/642632.
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The present study was undertaken to assess the effect of melatonin (MT) on sperm motility, viability, total sperm abnormality, acrosomal and plasma membrane integrity, DNA abnormality, antioxidant profiles such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and total antioxidant capacity (TAC), enzymatic profiles such as aspartate amino transaminase (AST), alanine amino transaminase (ALT), and biochemical profiles such as malonaldehyde (MDA) production and cholesterol efflux. Total numbers of 30 ejaculates were collected twice a week from eight mithun bulls and semen was split into five equal aliquots, diluted with the TEYC extender. Group 1 has semen without additives (control) and group 2 to group 5 have semen that was diluted with 1 mM, 2 mM, 3 mM, and 4 mM of melatonin, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 0, 6, 12, 24, and 30 h of incubation. Inclusion of melatonin into diluent resulted in significant (P<0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, melatonin at 3 mM has significant improvement in quality of mithun semen than melatonin at 1 mM, 2 mM or 4 mM stored inin vitrofor up to 30 h. It was concluded that the possible protective effects of melatonin on sperm parameters are it prevents MDA production and preserve the antioxidants and intracellular enzymes during preservation.
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Wirtu, G., C. E. Pope, A. Cole, R. A. Godke, D. L. Paccamonti, and B. L. Dresser. "115 SPERM CRYOPRESERVATION IN TRAGELAPHINE ANTELOPES." Reproduction, Fertility and Development 18, no. 2 (2006): 166. http://dx.doi.org/10.1071/rdv18n2ab115.
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As an integral aspect of our program to develop assisted reproductive technologies in tragelaphine antelopes, we have been evaluating the effects of various extenders on cryosurvival of spermatozoa. Semen was collected by electroejaculation (EEJ) from an eland (Taurotragus oryx, n = 14 EEJs) and a bongo (Tragelaphus euryceros, n = 7 EEJs) male during sedation and standing restraint in a hydraulic chute. Epididymal spermatozoa were collected from the proximal vas deferens, and distal epididymides recovered from four common eland bulls after elective castration at the Hattiesburg Zoo, Mississippi (age = 46 months) and the San Diego Wild Animal Park, California (ages = 18, 20, and 23 months). Testes from a bongo (age = 6.5 yr) and a greater kudu bull (Tragelaphus strepsiceros, age = 7.5 yr) at the Audubon Nature Institute were recovered and processed immediately postmortem. Kudu, bongo, and the Hattiesburg eland testes were transported at ambient temperature whereas the testes from San Diego elands were shipped overnight in a cold-pack container. Sperm processing was done at room temperature (RT = 23°C) in HEPES-buffered Tyrode's solution. Four extenders containing 7% glycerol were evaluated: Biladyl®, TEST Yolk buffer (TYB), Beltsville extender (BF5F), and skim milk. For freezing, sperm samples were initially extended and gradually cooled to 4°C before sequential addition of extender containing glycerol at 4°C; the procedure was then modified by addition of the glycerol fraction at RT before the sample was cooled to 4°C. Spermatozoa were vapor-frozen in 0.5 mL straws placed 5 cm above liquid nitrogen before storage at −196°C. Straws were thawed in a water bath (38°C, 30 s) to evaluate cryosurvival. All eland and bongo electroejaculation procedures produced spermatozoa, although sperm quality varied. Ejaculate volume averaged 5.4 ± 1.2 mL and 3.7 ± 1.1 mL in the bongo and the eland, respectively. Sperm motility at collection, after cooling, and after freezing in the different extenders is presented in Table 1. No immediate decline in sperm motility was observed after adding glycerol to samples, whether at 23°C or 4°C. Bongo spermatozoa lost more motility during freezing than during cooling (15.0 ± 4.4 vs. 5.8 ± 2.0%; P = 0.006); whereas, in eland, motility loss during cooling and freezing was similar (25.3 ± 16.2 vs. 21.1 ± 7.7; P = 0.44). The present results indicate that cooling and freezing tolerance of spermatozoa in tragelaphine antelopes is influenced by species, extender type, and temperature at which a cryoprotectant is added. Table 1. Cryosurvival of ejaculated or epididymal spermatozoa of three antelope species in different extenders
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SRIVASTAVA, N., MEGHA PANDE, T. V. RAJA, S. TYAGI, SURESH KUMAR, SUSHIL KUMAR, RAVINDER KUMAR, et al. "Prognostic value of post thaw semen quality parameters, mitochondrial integrity and cholesterol content of sperm membrane vis-à-vis conception rate in Frieswal bulls." Indian Journal of Animal Sciences 88, no. 8 (September 6, 2018): 892–98. http://dx.doi.org/10.56093/ijans.v88i8.82911.
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The present study aimed to analyze the quantitative relationship of membrane cholesterol content (Chol content), mitochondrial integrity (Δψm), and quality parameters of spermatozoa (SQP) at post-thaw stage with conception rate (CR) and estimated relative conception rate (ERCR) in Frieswal bulls. For the experiment, frozen semen straws (32) were collected from the SF laboratory and CR (Total insemination 3482) was obtained from Field Progeny Testing units. Based on the CR, bulls were grouped into low, medium or high groups (G I, G II and G III, respectively). SQP, viz. viability (Eosin Nigrosin), post thaw motility, biochemical integrity of the membrane (HOS res), acrosome integrity (Giemsa, and fluorochromes fluorescein isothiocyanate Pisum sativum agglutinin and propidium iodide, respectively), chol-content, and Δψm using fluorescent probe JC-1 (5,5’,6,6’-tetrachloro- 1,1’,3,3’-tetraethylbenzimi-dazolylcarbocyanine iodide) were determined. The values thus obtained were subjected to the stepwise regression analysis using the least squares principles for each group, and the CR and ERCR were regressed on the various SQP. Pearson’s correlation coefficients were estimated between the CR and ERCR values and the various SQPs. The coefficient of determination (R2) moderately higher for all the models and ranged from 63.70–93.40% (high and medium group, respectively). High R2 value of the prediction equation for the herd and bulls with medium CR (75.9 and 93.4%, respectively) reveal their suitability to predict the CR and ERCR potential of the cryopreserved semen. Study results point to inclusion of cholesterol content and Δψm estimation in routine semen analysis before long-term storage or usage for insemination purposes.
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Kulkarni, Nitish A., A. K. Roy, Sujata Pandita, C. G. Shashank, and H. S. Chethan. "Time Dependent Impact of Reactive Oxidants on Seminal Attributes, Mitochondrial Membrane Potential, Lipid Peroxidation and Capacitation-Like Changes of Karan-Fries (KF) Bulls During Cryopreservation." Cryoletters 43, no. 4 (July 1, 2022): 227–36. http://dx.doi.org/10.54680/fr22410110212.
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BACKGROUND: Cryopreservation of semen is a valuable technique; however, it is also known to be detrimental to the structure of spermatozoa and fertility due to cryo-injury and subsequent generation of reactive oxidants. OBJECTIVE:To determine the time-dependent impact of reactive oxidants on seminal attributes, mitochondrial membrane potential (MMP), lipid peroxidation status (LPO) and early capacitation like changes. MATERIALS AND METHODS:Semen samples were collected by artificial vagina technique from six Karan-Fries (KF) bulls and subsequently examined at 0 h (before cryopreservation) and at 24 hours, 15 days and 2-months of storage for various seminal attributes, MMP (Δψm) , and early capacitation-like changes. Simultaneously, LPO (TBARS) was determined in fresh and post-thaw seminal plasma. RESULTS: A sharp decrease (P<0.01) in semen quality was observed only after 24 h of cryopreservation except for viability and acrosomal integrity. Sperm viability and acrosome integrity reduced significantly up to 2 months of cryopreservation. The lipid peroxidation status was found to be lower in fresh seminal plasma (2.63±0.22 vs. 3.51±0.34 units/mL) as compared to post-thaw. Furthermore, the active Δψm of fresh semen showed a significant (P <0.01) decrease after 24 hours (77.92±0.387 vs. 54.52±0.28%) of cryopreservation, while there was a non- significant decrease in active MMP after 15 d and 2-months (53.68±0.138 and 52.76±0.16%). Moreover, significant (P <0.01) early capacitation-like changes were found in post-thaw spermatozoa (25.72±0.12 vs. 9.1±0.19%) as compared to fresh ejaculate. CONCLUSION: Spermatozoa incur the majority of damages during the early phase of cryopreservation, however the damage associated by different stressors cannot be neglected.
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Perumal, P. "Effect of Superoxide Dismutase on Semen Parameters and Antioxidant Enzyme Activities of Liquid Stored (5°C) Mithun (Bos frontalis) Semen." Journal of Animals 2014 (March 13, 2014): 1–9. http://dx.doi.org/10.1155/2014/821954.
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The present study was undertaken to assess the effect of superoxide dismutase (SOD) on sperm motility, and viability; total sperm abnormality; acrosomal and plasma membrane integrity; DNA abnormality; antioxidant profiles such as catalase (CAT), reduced glutathione (GSH), and total antioxidant capacity (TAC); enzymatic profiles such as aspartate amino transaminase (AST), and alanine amino transaminase (ALT); and biochemical profiles such as malondialdehyde (MDA) production and cholesterol efflux. Total numbers of 50 ejaculates were collected twice a week from eight mithun bulls and semen was split into four equal aliquots, diluted with the TEYC extender. Group 1: semen without additives (control), and group 2 to group 4: semen was diluted with 50 U/mL, 100 U/mL, and 150 U/mL of SOD, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 1, 6, 12, 24, and 30 h of incubation. Inclusion of SOD into diluent resulted in significant (P<0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, SOD at 100 U/mL has significant improvement in quality of mithun semen than SOD at 50 or 150 U/mL stored in in-vitro for up to 30 h. It was concluded that the possible protective effects of SOD on sperm parameters are that it prevents MDA production and preserves the antioxidants and intracellular enzymes during preservation.
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AD, Omur. "Antioxidant Potential of Ferulic Acid on the Freezability of Bull Semen." Open Access Journal of Veterinary Science & Research 4, no. 3 (2019): 1–4. http://dx.doi.org/10.23880/oajvsr-16000187.
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Ejaculates were collected twice a week from the bulls, via an artificial vagina, during two weeks. The suitable ejaculates obtained for sperm density (≥ 1.4 × 10 9 spermatoz oa / ml) and for motility (≥ 75%) were used for dilution and freezing of semen. A Tris - based extender (Tris 297.58mM, citric acid 96.32mM, fructose 82.66mM, egg yolk 15% (v/v), glycerol 5% (v/v), gentamicin 0.1 ml / 100ml, pH 6.8 - 7.0) was used as the base extender (cryopreservation diluent). Pooled ejaculate was split into 2 equal aliquots and diluted at 32 °C with base extender containing ferulic acid (100 μM) and no antioxidant (control), respectively. Each aliquot was diluted to a final semen concentrati on of approximately 1.2 × 10 8 sperm/ml (single step dilution), in 15 - ml polypropylene centrifuge tubes. After dilution, semen samples were kept at room temperature for 10 minutes then, the diluted semen samples were aspirated into 0.25 ml French straws, seal ed with polyvinyl alcohol powder and equilibrated at 5 °C for 3 h. After equilibration, the straws were frozen in liquid nitrogen vapour (4 cm above the liquid nitrogen, ~ - 100 o C ) for 10 min and then plunged into liquid nitrogen for storage, - 196 o C . In the study, sperm samples containing antioxidant and non - antioxidant were evaluated for spermatozoa motility and membrane integrity after freezing / thawing. In the present study, no statistically significant difference was found between the control and experim ental groups for motility and membrane integrity after freeze - thawing. The application consisted of 4 replications.
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Yekti, Aulia Puspita Anugra, Jois Harsah, Muchamad Luthfi, Muhammad Dikman, Asri Nurul Huda, Kuswati Kuswati, and Trinil Susilawati. "Kualitas Semen dengan Berbagai Formulasi Pengencer Dasar Air Kelapa Hijau Selama Simpan Dingin pada Sapi Madura." Jurnal Ilmu dan Teknologi Peternakan Tropis 5, no. 2 (December 16, 2018): 37. http://dx.doi.org/10.33772/jitro.v5i3.4738.
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ABSTRAKInseminasi Buatan dengan menggunakan semen cair digunakan untuk daerah yang sulit nitrogen cair dan mempunyai kualitas yang lebih baik dari pada semen beku. Tujuan penelitian ini adalah untuk mengetahui kualitas berbagai bahan pengencer dasar air kelapa penyimpanan dingin 2-5°C.Penelitian ini dilaksanakan di Loka Penelitian Sapi Potong Grati, Pasuruan. Semen yang digunakan berasal dari sapi madura sebanyak dua ekor, berumur 5 dan 3 tahun dan berat badan yaitu 397 kg dan 360,5 kg. Sapi madura ditampung seminggu 2 kali dengan motilitas > 70% , sedangkan air kelapa yang digunakan adalah air kelapa hijau yang masih muda. Pengenceran semen cair dibagi menjadi 4 yaitu P0 (CEP-3 + 20% kuning telur) sebagai kontrol, P1 (air kelapa hijau +20% kuning telur), P2 (P1 + 0,4% putih telur + 1% fruktosa) dan P3 (P1 + 0,4% putih telur kuning telur +2% fruktosa). Data dianalisis menggunakan uji Pearson’s Chi Square dan Uji Deskriptif. Hasil penelitian menunjukkan motilitas spermatozoa sesuai standar SNI yaitu motilitas> 40% pada pengencer CEP-3 dapat disimpan selama hari ke-8 (40,50±6,43%) sedangkan pada pengencer dasar air kelapa hijau pada P1, P2 dan P3 tidak menunjukkan perbedaan yang nyata (P>0,05) selama disimpan 6 hari yaitu 40,50±10,12%, 38,00±4,22%, 40,00±8,50%. Abnormalitas dari semua perlakuan menunjukan nilai <20%. Viabilitas didapatkan nilai tertinggi pada perlakuan P0(89,58±2,16%) kemudian P1(89,39±3,79%), P2(88,62±4,59%) dan P3(87,93±4,41%).Kata kunci: CEP-3, semen cair, sapi madura, simpan dingin, air kelapa hijau ABSTRACTArtificial Insemination using liquid semen is performed for areas that where liquid nitrogen is difficult to find and havng better quality than frozen semen. Purpose of this research was to investigate the quality on various coconut water base diluents on liquid semen of madura bull during cold storage of 2-5°C. Research was conducted at Laboratory of Reproduction of Grati Beef Cattle Research Station, Pasuruan.Semen that is used comes from two madura bulls aged 5 and 3 years with body weight is 397 kg and 360.5 kg. The semen was collected twice a week with motility> 70%, and the coconut water used is unripe green coconut water. The research treatments were P0 (CEP-3 + 20% egg yolk) as control, P1 (unripe green coconut water + 20% egg yolk), P2 (P1+ 1% fructose + 0.4% egg white) and P3 (P1+ 0.4% egg white + 2% fructose). Data were analyzed using Pearson's Chi Square test and Descriptive Test. The results showed that the motility of spermatozoa was within Indonesian National Standard (SNI) with more than 40% motility in the CEP-3 diluent and it can be stored until the 8th day (40.50 ± 6.43%). The basic diluents of green coconut water at P1, P2 and P3 was not significantly affected (P> 0.05) until 6 days storing with the motility number average are 40.50 ± 10.12%, 38.00 ± 4.22%, 40, 00 ± 8.50%. The abnormality of all treatments was under 20%. The highest viability was showed by treatment P2 (89.58±2.16%), followed by P4 (89.39 ± 3.79%), P3 (88.62 ± 4.59%) and the lowest was P4 (87.93 ± 4.41%). Keywords:CEP-3, liquid semen, madura bull, cool storage, green coconut water ABSTRACT Artificial Insemination using liquid semen is performed for areas that where liquid nitrogen is difficult to find and havng better quality than frozen semen. Purpose of this research was to investigate the quality on various coconut water base diluents on liquid semen of madura bull during cold storage of 2-5°C. Research was conducted at Laboratory of Reproduction of Grati Beef Cattle Research Station, Pasuruan.Semen that is used comes from two madura bulls aged 5 and 3 years with body weight is 397 kg and 360.5 kg. The semen was collected twice a week with motility> 70%, and the coconut water used is unripe green coconut water. The research treatments were P0 (CEP-3 + 20% egg yolk) as control, P1 (unripe green coconut water + 20% egg yolk), P2 (P1+ 1% fructose + 0.4% egg white) and P3 (P1+ 0.4% egg white + 2% fructose). Data were analyzed using Pearson's Chi Square test and Descriptive Test. The results showed that the motility of spermatozoa was within Indonesian National Standard (SNI) with more than 40% motility in the CEP-3 diluent and it can be stored until the 8th day (40.50 ± 6.43%). The basic diluents of green coconut water at P1, P2 and P3 was not significantly affected (P> 0.05) until 6 days storing with the motility number average are 40.50 ± 10.12%, 38.00 ± 4.22%, 40, 00 ± 8.50%. The abnormality of all treatments was under 20%. The highest viability was showed by treatment P2 (89.58±2.16%), followed by P4 (89.39 ± 3.79%), P3 (88.62 ± 4.59%) and the lowest was P4 (87.93 ± 4.41%).
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Susilowati, Suherni, Trilas Sardjito, Imam Mustofa, Oky Setio Widodo, and Rochmah Kurnijasanti. "Effect of green tea extract in extender of Simmental bull semen on pregnancy rate of recipients." Animal Bioscience 34, no. 2 (February 1, 2021): 198–204. http://dx.doi.org/10.5713/ajas.20.0025.
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Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows.Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination.Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12).Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.
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Magalhães, L. C. O., C. M. Melo, M. J. Sudano, D. M. Paschoal, L. F. Crocomo, F. O. Papa, F. C. Landim-Alvarenga, and M. D. Lopes. "100 A COMPARISON OF GLYCEROL AND EGTA FOR ULTRARAPID FREEZING OF BULL EPIDIDYMAL SPERM." Reproduction, Fertility and Development 22, no. 1 (2010): 209. http://dx.doi.org/10.1071/rdv22n1ab100.
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The ultrarapid freezing technique was developed as an alternative to slow conventional freezing to avoid formation of ice crystals (Tucker MJ and Liebermann J 2007, Vitrification in Assisted Reproduction, 87-92). The recent use of ethylene glycol tetraacetic acid (EGTA) instead of glycerol as a cryoprotectant for sperm is a result of efforts to reduce osmotic and cytotoxic effects. Accordingly, the objective of the present study was to compare the cryoprotective effects of EGTA v. glycerol on bovine sperm frozen using an ultrarapid method. A pool of epididymal sperm collected from 5 bulls was maintained in Botusui extender (Biotech, Botucatu, Brazil) containing 5% BSA, either EGTA (5%) or glycerol (5%), and 0 or 2 mg mL of polyvinyl alcohol (PVA). All samples were loaded into 0.25-mL French straws and either placed into a freezing machine (TK 4000 compacta, TK equipamentos para reproducao, Uberaba, Brazil) and cooled at a rate of 50°C/min, or exposed to liquid nitrogen (LN2) vapor for 5 to 15 min before plunging into LN2. After overnight storage in LN2, straws were thawed and examined using CASA (HTM IVOS 12, Hamilton Thorne Research, Beverly, MA, USA) for total motility (TM), progressive motility (PM), path velocity (VAP), progressive velocity (VSL), and track speed (VCL). Acridine Orange (AO) was used to check DNA integrity of sperm cells (Chirinea VH et al. 2006 Ciencia Animal Brasileira 7, 407-415). Our results indicate that, despite the potentially damaging effects of the extreme temperature changes, epididymal sperm showed 95% of DNA integrity and, therefore, should be able to participate in the fertilization process. Addition of PVA had a negative effect on sperm motility characteristics. CASA revealed that glycerol was a more suitable cryoprotectant for ultrarapid freezing of bull epididymal spermatozoa than was EGTA (see Table 1). Table 1. CAPES.
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Nichi, M., T. Rijsselaere, A. Van Soom, J. B. P. De Clercq, I. G. F. Goovaerts, V. H. Barnabe, and P. E. J. Bols. "15 EFFECT OF BULL EPIDIDYMIS STORAGE CONDITIONS ON CRYOPRESERVED EPIDIDYMAL SPERM IN VITRO FERTILITY AND LIPID PEROXIDATION STATUS." Reproduction, Fertility and Development 19, no. 1 (2007): 126. http://dx.doi.org/10.1071/rdv19n1ab15.
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Although cryopreservation of epididymal sperm has been studied extensively in several species, some factors that could negatively influence its quality are still unknown, such as the storage conditions of the epididymides prior to sperm collection. Studies indicate that the lower the storage temperature, the better the sperm quality after collection (Kaabi et al. 2003 Theriogenology 60, 1249–1259). An additional factor is lipid peroxidation in which sperm membrane resistance against reactive oxygen species (ROS) attacks is an important component. The objective of this experiment was to study whether the epididymis storage temperature following slaughter, as well as the intrinsic resistance against oxidative stress, affects the fertilizing capacity of cryopreserved epididymal bull sperm in vitro. Twelve epididymides (6 bulls) were collected after slaughter and divided into 2 groups, (stored at either 4 or 37�C for 2), after which semen was collected from the caudae epididymides and cryopreserved. Subsequently, one aliquot of the frozen–thawed semen samples was subjected to induced lipid peroxidation with ferrous sulfate and ascorbate (37�C; 2 h), after which tiobarbituric acid-reactive substances (TBARS), as an index of lipid peroxidation, were measured according to a method previously described (Beorlegui et al. 1997 Andrologia 29, 37–42). A second aliquot was used for in vitro fertilization in a routine IVF–IVC setup in duplicate (24-h maturation, SOF culture medium in 5% CO2, 5% O2, and 90% N2). In vitro embryo production results at Day 7 and TBARS levels were statistically analyzed using SAS (SAS Institute, Inc., Cary, NC, USA). No influence of storage temperature was observed at either TBARS level (4�C: 943.6 � 173.4; 37�C: 751.4 � 136.2 ng of TBARS/108 spermatozoa; P = 0.3) or on blastocyst rates (4�C: 23.0 � 2.8; 37�C: 18.7 � 3.6% of blastocysts; P = 0.2). However, the percentage of hatched blastocysts tend to be higher for epididymides stored at 4�C when compared to those stored at 37�C (6.4 � 1.8 and 2.3 � 0.9, respectively; P = 0.06). In addition, a negative correlation was found between TBARS concentrations and blastocyst rates (R = –0.57; P < 0.05). Compared to fresh samples collected from epididymides under the same conditions (unpublished data), levels of TBARS were two- to threefold higher for the cryopreserved sperm, indicating that lipid peroxidation appears to play a role in the decrease in quality of cryopreserved epididymal sperm. In conclusion, temperature during the epididymides short-term storage prior to sperm cryopreservation did not seem to influence the sperms' in vitro fertilizing capacity. On the other hand, an alternative to improve cryopreserved epididymal sperm in vitro fertility (or fertilizing capacity) could be the addition of antioxidants to semen extenders. Further studies are necessary to confirm this hypothesis.
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Niasari-Naslaji, A., S. Mosaferi, A. A. Gharahdaghi, A. Abarghani, A. Ghanbari, and A. Gerami. "96 A NOVEL EXTENDER FOR PRESERVATION OF BACTRIAN CAMEL (CAMELUS BACTRIANUS) SEMEN." Reproduction, Fertility and Development 17, no. 2 (2005): 198. http://dx.doi.org/10.1071/rdv17n2ab96.
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Lactose and Green buffer (IMV, France) are the most commonly used extenders for camel semen. The viability of Bactrian camel spermatozoa in lactose extender is reduced after 4 h of incubation at 4°C (unpublished data). Although Green buffer is used for dromedary camel semen, there are no data indicating its effectiveness for Bactrian camel semen. More recently, we reported that the osmolality and pH of Bactrian camel semen are 316.1 ± 1.48 mOsm/kg and 7.4 ± 0.03, respectively. The objective of this study was to compare three different semen extenders, to determine if a TRIS-based diluent (SHOTOR* Diluent), a completely defined diluent, can maintain cooled camel sperm as effectively as established diluents. SHOTOR Diluent consists of 2.6 g TRIS, 1.35 g citric acid, 1.2 g glucose, and 0.9 g fructose in 100 mL of deionized water, with an osmolality of 330 mOsm/kg and pH of 6.9. SHOTOR Diluent, lactose, 10% (w/v), with an osmolality of 330 mOsm/kg and pH of 6.9, and Green buffer were compared in this study. All extenders contained 20% egg yolk. Semen was collected from bulls with a sound history of semen quality and fertility (n = 3), using a modified artificial vagina, and divided equally into the different extenders (Mosaferi S et al. 2004 15th Int. Cong. Anim. Reprod. 2, 520; Mosaferi S et al. 2004 Theriogenology, in press). Progressive forward motility and percentage of live spermatozoa were examined at the time of semen collection (time 0) and after 4, 12, and 24 h of incubation at 4°C. Data were analyzed using the GLM procedure in SAS/STAT after arcsine transformation. The forward progressive motilities of spermatozoa at 0, 4, 12, and 24 h after semen collection were 65.5, 54, 44.5, and 36.5% in SHOTOR Diluent; 31, 18.5, 8.5, and 0% in 10% lactose; and 60.5, 54.5, 33, and 32.5 % in Green buffer, respectively (Table 1). The percentage of live spermatozoa at 0, 4, 12, and 24 h were 84.5, 84, 81 and 74.5% in SHOTOR Diluent; 80, 79.5, 72.5, and 56.5% in 10% lactose; 89, 82.5, 82.5, and 77.5% in Green buffer, respectively (P > 0.05). The progressive forward motility of spermatozoa did not significantly decrease by 12 h at 4°C in SHOTOR Diluent (P > 0.05; Table 1), whereas it significantly decreased after 4 h and 12 h of incubation at 4°C in Green buffer and 10% lactose, respectively (P < 0.05; Table 1). Further decrease in the progressive forward motility occurred in all extenders after 24 h at 4°C (P < 0.05; Table 1). In conclusion, SHOTOR Diluent is better than Green buffer and 10% lactose as an extender for chilled storage of Bactrian camel semen for 12 hat4°C. Table 1. The progressive forward motility of Bactrian camel spermatozoa extended in SHOTOR Diluent (1), 10% lactose (2) and Green buffer (3) at the time of semen collection (time 0) and after 4, 12, and 24 h of incubation at 4°C *Shotor means camel in the Persian language. The authors wish to thank the director and station staff of Bactrian Camel Reseach Center at Meshkinshahr, Ardabil, for providing facilities and kind assistance throughout the experiment.
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Harayama, Hiroshi, Kazuhiro Nishijima, Tetsuma Murase, Mitsuhiro Sakase, and Moriyuki Fukushima. "Relationship of protein tyrosine phosphorylation state with tolerance to frozen storage and the potential to undergo cyclic AMP-dependent hyperactivation in the spermatozoa of Japanese Black bulls." Molecular Reproduction and Development 77, no. 10 (September 15, 2010): 910–21. http://dx.doi.org/10.1002/mrd.21233.
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Lamy, Julie, Perrine Nogues, Lucie Combes-Soia, Guillaume Tsikis, Valérie Labas, Pascal Mermillod, Xavier Druart, and Marie Saint-Dizier. "Identification by proteomics of oviductal sperm-interacting proteins." Reproduction 155, no. 5 (May 2018): 457–66. http://dx.doi.org/10.1530/rep-17-0712.
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The interactions between oviductal fluid (OF) proteins and spermatozoa play major roles in sperm selection, storage and capacitation before fertilization. However, only a few sperm-interacting proteins in the OF has been identified and very little is known about the regulation of sperm-oviduct interactions across the estrous cycle. Samples of bovine frozen-thawed sperm from three bulls were incubated with OF at pre-, post-ovulatory stages (Pre-/Post-ov) or luteal phase (LP) of the estrous cycle (7 mg/mL proteins, treated groups) or with a protein-free media (control). The proteomes of sperm cells were assessed by nanoLC–MS/MS and quantified by label-free methods. A total of 27 sperm-interacting proteins originating in the OF were identified. Among those, 14 were detected at all stages, eight at Post-ov and LP and five only at LP. The sperm-interacting proteins detected at all stages or at LP and Post-ov were on average more abundant at LP than at other stages (P < 0.05). At Pre-ov, OVGP1 was the most abundant sperm-interacting protein while at Post-ov, ACTB, HSP27, MYH9, MYH14 and OVGP1 were predominant. Different patterns of abundance of sperm-interacting proteins related to the stage were evidenced, which greatly differed from those previously reported in the bovine OF. In conclusion, this study highlights the important regulations of sperm-oviduct interactions across the estrous cycle and provides new protein candidates that may modulate sperm functions.
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Kowalczyk, Alicja, Elżbieta Gałęska, Anna Szul, Katarzyna Łącka, Anna Bubel, Jose P. Araujo, Riaz Ullah, and Marcjanna Wrzecińska. "Fertility Rate and Assessment of the Cytoprotective Capacity of Various Types of Holothuroidea Extracts on Spermatozoa." Veterinary Sciences 9, no. 4 (April 15, 2022): 189. http://dx.doi.org/10.3390/vetsci9040189.
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For years, compounds of natural origin have been the subject of extensive biomedical research due to very interesting, new ingredients potentially useful for various pharmaceutical, medical and industrial applications. The therapeutic properties and healing benefits of sea cucumbers may result from the presence of numerous, biologically active ingredients. Sperm subjected to processing and subsequent storage at low temperatures experience a number of damage, including the loss of the integrity of the cytoplasmic membrane, DNA and acrosome defragmentation. Therefore, the aim of this experiment was to investigate the cytoprotective potential of sea cucumber extract against cryopreserved sperm and semen fertility rate. Commercially available sea cucumber extract was taken from the cellulose shell, then 790 mg of powder was weighed out and placed in 3 glass tubes containing, respectively: 10 mL of water-glycerin solution (WG), water-ethanol (EC), glycerin-ethanol (GE), glycerin-DMSO (DG). Tubes were mixed with vortex for 3 min, then placed in a water bath and incubated for 16 h at 40 °C. Six simmental bulls, 3 years old, of known health status were used for the experiment. Semen was collected from each male once a week (for 18 weeks) using an artificial vagina. After an initial assessment of semen quality, the ejaculates were pooled to eliminate individual differences between males, then diluted to a final concentration of 80 × 106 sperm/mL with a commercial extender (Optixcell, IMV, L’Aigle, France) and divided into 16 equal samples. Control (C) without additive, the test samples contained 2, 4, 6, 8 and 10 µL WG, 2, 4, 6, 8 and 10 µL WE, 2, 4, 6, 8 and 10 µL GE, 2, 4, 6, 8 and 10 µL DG. Semen was frozen/thawed and assessed for motility, viability, DNA defragmentation, mitochondrial membrane potential and acrosome integrity. It was shown a positive effect of water-glycerin (WG) and glycerine-ethanol (GE) extracts on the efficiency of sperm preservation at low temperatures. Established that, depending on the type of prepared extract, the sea cucumber can have both cytoprotective (WG, GE, WE) and cytotoxic (DG) effects. Moreover, too high concentrations of the extract can adversely affect the sperm in terms of parameters such as viability, motility, mitochondrial potential, and the integrity of the acrosome or DNA of cells. The present study, thanks to the use of model animals to study the cytoprotective potential of the sea cucumber extract, proves that it can be a potential candidate for use in semen cryopreservation technology to improve the efficiency of storage at low temperatures. Further research is needed to optimize the composition of individual types of extracts and their effect on sperm. The highest effectiveness of female fertilization was observed when doses from GE groups (2 and 4) were used for insemination. The results of this analysis prove that the addition of the tested extract may improve the fertilization efficiency.
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Nichi, M., J. B. P. De Clercq, I. G. F. Goovaerts, V. H. Barnabe, and P. E. J. Bols. "307 EFFECT OF BULL EPIDIDYMIS STORAGE CONDITIONS ON SPERM RESISTANCE AGAINST LIPID PEROXIDATION AND SUBSEQUENT IN VITRO EMBRYO PRODUCTION." Reproduction, Fertility and Development 18, no. 2 (2006): 261. http://dx.doi.org/10.1071/rdv18n2ab307.
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Sperm recovery from the cauda epididymis can be a usefull tool in case of unexpected death of genetic high-value animals or endangered species or when the collection of sperm by other means becomes impossible. Studies indicate that the lower the temperature of epididymis storage, the better the sperm quality after collection (Kaabi et al. 2003 Theriogeneology 60, 1249-1259). One of the main factors that can negatively affect sperm viability during storage is lipid peroxidation, where sperm membrane resistance against reactive oxygen species (ROS) attacks is an important factor. The objective of this experiment was to study whether the temperature of epididymis storage following slaughter would have an influence on the membrane's resistance against lipid peroxidation and on the sperm cell's fertilizing capacity. Sixteen epididymides (from eight bulls) were collected after slaughter and divided into two groups, one stored at 4�C and the other at 37�C for 2 h, after which semen was collected from the caudae epididymides. Sperm concentration was measured and an aliquot containing 108 sperm cells was submitted to induced lipid peroxidation with ferrous sulfate and ascorbate (37�C; 2 h). Subsequently, thiobarbituric acid reactive substances (TBARS), as an index of lipid peroxidation, were measured according to a method previously described (Beorlegui et al. 1997 Andrologia 29, 37-42). A second aliquot of the sample was used for fertilization in a routine IVF-IVC set up in duplicate (24-h maturation, SOF culture medium in 5% CO2, 5% O2, and 90% N2). In vitro embryo production and level of TBARS were statistically analyzed using SAS (SAS Institute, Inc., Cary, NC, USA). TBARS levels were transformed to logarithm form in order to obey the residue normality being analyzed using PROC GLM. The percentage of blastocysts was analyzed using the Wilcoxon test. When compared to the samples stored at 4�C, semen of caudae epididymides stored at 37�C showed higher levels of TBARS and lower mean blastocyst rates (324.7 � 59.6 and 36.6 � 1.6 vs. 466.9 � 67.9 ng of TBARS/108 spermatozoa and 28.8 � 2.9%, respectively; P < 0.05). A negative correlation was found between TBARS and blastocyst rates (R = -0.43). The lower quality of sperm collected from epididymides maintained at higher temperatures may be related to a decrease in sperm resistance against lipid peroxidation which would further impair sperm fertilizing capacity. However, further studies are necessary in order to study the effect of temperature on the sperm membrane lipid profile, because the content of polyunsaturated fatty acids may be affected by temperature; this is an important factor relative to sperm membrane lipid peroxidation susceptibility (Ollero et al. 2000 Mol. Reprod. Dev. 55, 326-334). Another important factor is the epididymal environment because interactions between the sperm membrane and its surroundings can play an important role on the membrane's antioxidant protection.
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Widjaya, Nilawati. "Pengaruh Pemberian Susu Skim dengan Pengencer Tris Kuning Telur terhadap Daya Tahan Hidup Spermatozoa Sapi pada Suhu Penyimpanan 5ºC." Sains Peternakan 9, no. 2 (February 6, 2017): 72. http://dx.doi.org/10.20961/sainspet.9.2.72-76.
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<p>The objective of the study was to determine the effect of skim milk and Tris yolk<br />extender on the viability bovine spermatozoa at 5 ºC. This study used Simmental bull semen. The design used was Randomized Design Group (RAK) with 5 treatments and 5 groups of ejaculate. The treatments were: P0 = skim milk 100% without Tris yolk extender; P1 = 95% skim milk + 5% Tris yolk extender, P2 = 90% skim milk + Tris yolk extender 10%, P3 = skim milk 85% + Tris yolk extender 15%, P4 = skim milk 80% + Tris yolk extender 20%. The observed variables were the percentage of live spermatozoa and the motility of spermatozoa after storage for 2 days at 5ºC. Skim milk with Tris yolk extender affect the viability and motility of spermatozoa stored at 5ºC (P<0.01). Provision of extender up to 15% gave the best results and suppressed the decline in the viability of spermatozoa storaged for 2 days at 5ºC.</p><p>Key words: skim milk, spermatozoa, Tris yolk extender, viability</p>
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Widjaya, Nilawati. "Pengaruh Pemberian Susu Skim dengan Pengencer Tris Kuning Telur terhadap Daya Tahan Hidup Spermatozoa Sapi pada Suhu Penyimpanan 5ºC." Sains Peternakan 9, no. 2 (February 6, 2017): 72. http://dx.doi.org/10.20961/sainspet.v9i2.4796.
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<p>The objective of the study was to determine the effect of skim milk and Tris yolk<br />extender on the viability bovine spermatozoa at 5 ºC. This study used Simmental bull semen. The design used was Randomized Design Group (RAK) with 5 treatments and 5 groups of ejaculate. The treatments were: P0 = skim milk 100% without Tris yolk extender; P1 = 95% skim milk + 5% Tris yolk extender, P2 = 90% skim milk + Tris yolk extender 10%, P3 = skim milk 85% + Tris yolk extender 15%, P4 = skim milk 80% + Tris yolk extender 20%. The observed variables were the percentage of live spermatozoa and the motility of spermatozoa after storage for 2 days at 5ºC. Skim milk with Tris yolk extender affect the viability and motility of spermatozoa stored at 5ºC (P<0.01). Provision of extender up to 15% gave the best results and suppressed the decline in the viability of spermatozoa storaged for 2 days at 5ºC.</p><p>Key words: skim milk, spermatozoa, Tris yolk extender, viability</p>
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Ardhani, Fikri, Hayatul Mufidah, Rahmah Samsuriati, and Hilman Pratama Putra. "Efek Lama Penyimpanan Semen Beku Sapi Bali pada Pos Inseminasi Buatan terhadap Membran Plasma, Tudung Akrosom Utuh, dan DNA Spermatozoa." Jurnal Ilmu Peternakan Terapan 3, no. 2 (July 10, 2020): 58–66. http://dx.doi.org/10.25047/jipt.v3i2.1950.
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The purpose of this study was to determine the effect of frozen storage time for Bali Bull in artificial insemination station in Samarinda City, East Kalimantan on the quality of motility, viability, velocity, abnormality, plasma membrane integrity (MIn), acrosome integrity (AIn), and DNA damage of spermatozoa. The study design used a completely randomized design (CRD) with 5 treatments (storage time) and 5 replications. Frozen semen of Bali Bull used in 2009 (10 years of storage), 2011 (7 years of storage), 2013 (5 years of storage), 2015 (3 years of storage), and 2017 (1 year of storage). The storage time of frozen semen stored for one to ten years in liquid nitrogen at the artificial insemination station in Kota Samarinda, East Kalimantan was still suitable for use in artificial insemination based on motility quality (44.99±2.40%), viability (55.33±2,60%), velocity (0.050±0.002 mm/sec), abnormality (12.87±1.09%), plasma membrane integrity (58.83 ± 1.86%), acrosome integrity (75.48 ± 1 , 61%), and DNA damage of spermatozoa (1.60 ± 0.21%).
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Murphy, Craig, Shauna A. Holden, Edel M. Murphy, Andrew R. Cromie, Patrick Lonergan, and Sean Fair. "The impact of storage temperature and sperm number on the fertility of liquid-stored bull semen." Reproduction, Fertility and Development 28, no. 9 (2016): 1349. http://dx.doi.org/10.1071/rd14369.
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In Ireland, liquid bull semen is stored at unregulated ambient temperatures, typically at 5 × 106 spermatozoa per dose, and inseminated within 2.5 days of collection. In Experiment 1, the effect of storage temperature (5, 15, 22, 32°C and fluctuations (Flux) between these temperatures) on progressive motility, viability, acrosomal status, DNA fragmentation and osmotic resistance was assessed. In Experiment 2, the field fertility of liquid semen at 5, 4 and 3 × 106 spermatozoa per dose, up to Day 2 after collection, was assessed in comparison to frozen–thawed semen at 20 × 106 spermatozoa per dose (n = 35 328 inseminations). In Experiment 1, storage at 15°C resulted in the highest progressive motility (P < 0.01). The osmotic resistance of spermatozoa declined with duration of storage; however, after Day 3 this decline was reduced in the 5°C and Flux 15°C treatments (P < 0.01). In Experiment 2, the non-return rate of liquid semen stored at 4 and 3 × 106 spermatozoa per dose on Day 2 of storage was reduced in comparison to frozen–thawed semen (P < 0.01). In conclusion, liquid semen is versatile between storage temperatures of 5 and 22°C, but demonstrates reduced fertility on Day 2 of storage at lower sperm numbers in comparison to frozen–thawed semen.
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Shedova, E. N., G. N. Singina, V. A. Bagirov, and N. A. Zinovieva. "147 THE DEVELOPMENTAL CHARACTERISTICS OF IN VITRO-PRODUCED CATTLE-WISENT (BOS TAURUS-BISON BONASUS) HYBRID EMBRYOS." Reproduction, Fertility and Development 29, no. 1 (2017): 182. http://dx.doi.org/10.1071/rdv29n1ab147.
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Interspecies hybrids are important resources for research and agriculture. Therefore, the aim of this study was to evaluate development, quality, and viability of embryos produced in vitro using cattle (Bos taurus) oocytes and European bison (Bison bonasus) epididymal sperm. The epididymes were obtained following a forced slaughter of one bull aged 7 years. The sperm was collected by scraping the inner surface of the epididymes, diluted with the cryopreservation medium, and equilibrated for 4 h at 4°C. Thereafter, sperm aliquots (0.2 mL) were frozen in liquid nitrogen vapor for 5 min and then plunged into liquid nitrogen for storage. Prior to fertilization, frozen semen was thawed in pre-warmed medium for 1 min at 37°C and prepared by the swim-up method. The frozen-thawed ejaculated sperm from the Russian Black Pied bulls was used as a positive control. Slaughterhouse-derived cumulus-oocyte complexes were matured for 24 h in TCM-199 supplemented with 10% FCS, 0.2 mM sodium pyruvate, 10 μg mL−1 porcine FSH, and 10 μg mL−1 ovine LH. Matured oocytes (35–40 oocytes per group) were co-incubated for 18 h with homologous (n = 266 oocytes) or heterologous (n = 292 oocytes) sperm (spermatozoa/mL) in 500 µL of TALP containing 10 μg mL−1 heparin, 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine, and 0.1% minimal essential medium nonessential amino acids. After IVF, the oocytes were cultured in CR1aa medium (Rosenkrans 1994 J. Anim. Sci. 72, 434–437) to the blastocyst stage. All the cultures were performed at 38.5°C and 5% CO2 in humidified air. At Days 2 and 7 after insemination, the cleavage and blastocyst rates were determined. In addition, a part of obtained blastocysts was fixed with 4% paraformaldehyde, and the total cell number and apoptotic cell ratio were determined by 4’,6-diamidino-2-phenylindole and TUNEL staining. The remaining blastocysts were cultured up to Day 10, and the hatching rates were assessed. The data (3–5 replicates) were analysed by ANOVA. The cleavage rates did not differ among both male species (72.4 and 77.1%). Furthermore, no significant effects of interspecies fertilization on the blastocyst rate or total cell number per blastocyst were found (27.4 ± 1.6% and 77.0 ± 5.7 for cattle embryos and 26.2 ± 1.9% and 83.1 ± 8.9 for cattle-wisent hybrid embryos). On the other hand, the significant differences between homologous and heterologous fertilization were detected in the rate of hatched blastocysts (60.3 ± 5.1 v. 38 ± 2.9, P < 0.05) and apoptotic cell ratio 7.3 ± 0.8 v. 11.6 ± 1.04, P < 0.05). Our findings demonstrate that hybrid embryos produced by IVF of bovine oocytes with the epididymal sperm of European bison can be developed up to advanced blastocyst stages. However, the hybrid embryos have a lower quality and viability than cattle embryos. Research was supported by the Program of Presidium of the Russian Academy of Science, project no. IV.13.3.
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Suarez, S. S. "Interactions of spermatozoa with the female reproductive tract: inspiration for assisted reproduction." Reproduction, Fertility and Development 19, no. 1 (2007): 103. http://dx.doi.org/10.1071/rd06101.
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Artificial insemination with sexed semen, in vitro fertilisation and intracytoplasmic sperm injection have been used to reproduce animals, but often not as successfully as natural mating. Learning more about how spermatozoa normally interact with the female tract can provide inspiration for developing improvements in assisted reproduction. The present review focuses on Bos taurus, because more is known about this species than others. At coitus, bull spermatozoa are deposited into the anterior vagina, where they rapidly enter the cervix. Cervical mucus quickly filters out seminal plasma from spermatozoa, unlike most assisted reproduction protocols. Spermatozoa that reach the uterus may require certain cell surface proteins to swim through the uterotubal junction. Shortly after passing through the junction, most spermatozoa are trapped in a storage reservoir by binding to oviducal epithelium, in the case of cattle via bovine seminal plasma (BSP) proteins coating the sperm head. As ovulation approaches, spermatozoa capacitate and shed BSP proteins. This reduces sperm binding to the epithelium and releases them from storage. Motility hyperactivation assists spermatozoa in leaving the storage reservoir, swimming through oviducal mucus and the cumulus oophorus, and penetrating the oocyte zona pellucida. Chemotactically regulated switching between asymmetrical (i.e. hyperactivated) and symmetrical flagellar beating may also guide spermatozoa to the oocyte.
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Klein, Erin Kathleen, Allan John Gunn, Cyril Perumamthadathil Stephen, Aleona Swegen, Robert John Aitken, and Zamira Gibb. "Maintaining the fertility of bull spermatozoa during room temperature storage." Animal Reproduction Science 220 (September 2020): 106369. http://dx.doi.org/10.1016/j.anireprosci.2020.106369.
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Ratnawati, D., M. Luthfi, D. Pamungkas, and L. Affandhy. "Motility characterization of albumin sexed spermatozoa in two different diluents and additional antioxidant." Journal of the Indonesian Tropical Animal Agriculture 45, no. 4 (November 12, 2020): 277–86. http://dx.doi.org/10.14710/jitaa.45.4.277-286.
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The purpose of this study was to characterize spermatozoa motility of sexed semen (X and Y) using albumin by diluents and antioxidant treatments. The material used in this study was fresh semen of Ongole Crossbreed (OC) bull with progressive motility >70%. The sperm sexing methodology used albumin gradient of 5%, 10%, and 15%. The diluents used were CEP-2 and andromed with or without the addition of 1mM Glutathione as an antioxidant. The sexed semen was made into liquid semen and stored at 3-5°C. The motility was observed at day 0 (H0) and day 5 (H5). Motility was analyzed using SCA v.2.1. The parameters measured were total motility, progressive motility, VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF and sperm hyperactive. The experimental applied was the factorial pattern 2 x 2. The first factor was the type of diluents (CEP-2 and Andromed), and the second factor was additional glutathione (with or without). Data was analyzed by general linear model of SPSS-IBM 24 program. Sexed semen with CEP-2 diluents showed better motility of spermatozoa than andromed in the upper layers (spermatozoa X) and lower layers (spermatozoa Y). The CEP-2 diluent had a big role to maintain motility, progressive motility and velocity spermatozoa during cold storage. The addition of Glutathione 1mM could support the motility of sexed spermatozoa at cold storage, especially for LIN, STR, and WOB values.
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Azura, Sarah, Hermin Ratnani, Suherni Susilowati, Mas'ud Hariadi, Abdul Samik, and Koesnoto Soepranianondo. "Effect of α-tocopherol supplementation in diluents on the motility, viability and plasma membrane integrity of Simmental bull spermatozoa after cooling." Ovozoa : Journal of Animal Reproduction 9, no. 1 (May 11, 2020): 1. http://dx.doi.org/10.20473/ovz.v9i1.2020.1-6.
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Semen storage in cold temperatures might cause an increase in reactive oxygen species (ROS) production. This condition resulted in spermatozoa damage and quality decrease. This study was conducted to investigate the effects of α-tocopherol supplementation in diluents on the motility, viability, and plasma membrane integrity of Simmental bull spermatozoa after cooling. Semen samples were diluted in skim milk egg yolk supplemented with 0, 0.5, 1.0, and 1.5 mM α-tocopherol respectively for control, Tl, T2, and T3. Spermatozoa were evaluated for their motility, viability, and membrane integrity in cooling temperature (5°C). The daily evaluation showed that 1.5 mM α-tocopherol was the best in maintaining motility, viability, and plasma membrane integrity, while 1.0 mM α-tocopherol was only good for maintaining viability. Therefore, it can be concluded that α-tocopherol at the concentration of 1.5 mM was an efficient antioxidant supplement for Simmental cattle semen in skim milk egg yolk diluent.
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Arat, S., S. Pabuccuoglu, H. Sagirkaya, K. Demir, R. Arici, B. Ustuner, S. Alcay, et al. "22 SEMEN AND REPRODUCTIVE PROFILES OF CLONED ANATOLIAN GREY CATTLE." Reproduction, Fertility and Development 27, no. 1 (2015): 103. http://dx.doi.org/10.1071/rdv27n1ab22.
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Anatolian grey cattle (endangered native Anatolian cattle) as 1 male (clone 1) and 4 females (clones 2–5) were produced from cells of 1 male and 1 female cattle by somatic cell nuclear transfer (SCNT) in a previous study. In this study, we examined the reproductive potential of these cloned animals, which are now 4 and 5 years old. The parameters evaluated by phase contrast microscopy for motility, TUNEL for DNA fragmentation, eosin staining for viability, Hoechst 33258 staining and hypo-osmotic swelling test (HOST) for membrane integrity, and fluorescein isothiocyanate-Pisum sativum agglutinin (FITC-PSA) for acrosome integrity of frozen-thawed spermatozoa, as well as birth and survival of calves following insemination with frozen-thawed semen of cloned and nuclear donor bull and normal bull. Six ejaculates and 3 samples per ejaculate from each bull were tested, and the Mann-Whitney U test was used to analyse the data. The spermatological parameters of cloned bull semen – volume, concentration, and motility of fresh – were within accepted limits for artificial insemination (4.60 ± 0.47 mL, 1.55 ± 0.21 × 109 spermatozoa mL–1, 80.00 ± 1.07%, respectively). Frozen-thawed sperm motility and viability rate were higher in the cloned bull (56.6%, 56.7%) than in its nuclear donor (47%, 43%; P < 0.05). Intact membrane and DNA fragmentation rate of cloned bull and its nuclear donor bull sperm were similar (P > 0.05) but the intact acrosome rate of cloned bull was higher than that of its nuclear donor (P < 0.05). Low rates in frozen-thawed sperm of nuclear donor can be related to storage time of sperm which were frozen 5 years before. One (clone 4) of the cloned grey heifers was artificially inseminated with frozen semen from nuclear donor bull and the other (clone 5) was naturally mated with a Holstein bull. Two healthy calves were delivered naturally. When same cloned cows (clones 4–5) and 2 other cloned heifers (clones 2–3) were artificially inseminated with frozen semen of the cloned grey bull, clones 2 and 4 gave birth to 2 healthy female calves. One cloned cow (clone 3) aborted in the third month of gestation and other one (clone 5) is currently 8 months pregnant. Two calves of clone 4 and 5 are 17 months old and 2 other calves of clone 2 and 4 are now 6 and 1 months old. Except for clone 3, our results show that cloned Anatolian grey bull and cows produced from frozen cells in gene bank have normal fertility.
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Vera-Munoz, Oscar, Lamia Amirat-Briand, Djemil Bencharif, Marc Anton, Serge Desherces, Eric Shmitt, Chantal Thorin, and Daniel Tainturier. "Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C." Asian Journal of Andrology 13, no. 2 (November 29, 2010): 281–86. http://dx.doi.org/10.1038/aja.2010.84.
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Amirat, Lamia, Marc Anton, Daniel Tainturier, Gérard Chatagnon, Isabelle Battut, and Jean Luc Courtens. "Modifications of bull spermatozoa induced by three extenders: Biociphos, low density lipoprotein and Triladyl, before, during and after freezing and thawing." Reproduction 129, no. 4 (April 2005): 535–43. http://dx.doi.org/10.1530/rep.1.00011.
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The success of artificial insemination with frozen semen implies the reduction of the deleterious effects on the cells induced by this technique. These effects can occur as early as during the first dilution in an extender, as well as at any step, during or after the freezing process.In this work, we have compared the modifications induced by Triladyl, low density lipoproteins (LDL) and Biociphos extenders, after dilution and cooling to 4 °C for 1, 4 and 24 h. Alterations in the cell structures were visualized by electron microscopy (EM). More than 80% of spermatozoa were injured after incubation for 4 h in Triladyl, while 3% and 47% were counted in LDL and Biociphos respectively. This latter extender was deleterious to cell membrane integrity after incubation for 4 h or longer.The ultrastructure of frozen spermatozoa was studied by EM of cryofixed-cryosubstituted samples obtained from regular 0.5 ml French straws frozen using our usual protocol. The main differences between samples concerned the size and appearance of the frozen extender veins, while very few cell defects were found to be added by the freezing process at any depth in the straws.After thawing, semen motility was twofold higher (P< 0.05) in Biociphos (64%) and LDL (61%) than in Triladyl (32%) and the cells were less altered in LDL. We concluded that the LDL extender offers a better protection for storage of frozen spermatozoa, and can probably also be used for the preservation of fresh semen for short periods.
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shahba, mohamed, Reda El-Sheshtawy, Abdel-Salam El-Azab, Alaa Abdel-Ghaffar, Maha Ziada, and Adel Zaky. "The Effect of Freeze Drying Using Different Media and Storage Temperatures on Some Parameters of Buffalo Bull Spermatozoa." Egyptian Journal of Veterinary Sciences 47, no. 1 (July 1, 2016): 51–61. http://dx.doi.org/10.21608/ejvs.2016.1162.
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Shahba, Mohamed I., Reda I. El-Sheshtawy, Abdel-Salam I. El-Azab, Alaa E. Abdel-Ghaffar, Maha S. Ziada, and Adel A. Zaky. "The effect of freeze-drying media and storage temperature on ultrastructure and DNA of freeze-dried buffalo bull spermatozoa." Asian Pacific Journal of Reproduction 5, no. 6 (November 2016): 524–35. http://dx.doi.org/10.1016/j.apjr.2016.11.002.
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Lloyd, R. E., A. Fazeli, P. F. Watson, and W. V. Holt. "The oviducal protein, heat-shock 70-kDa protein 8, improves the long-term survival of ram spermatozoa during storage at 17°C in a commercial extender." Reproduction, Fertility and Development 24, no. 4 (2012): 543. http://dx.doi.org/10.1071/rd11173.
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Poor fertility rates are often observed when fresh ram semen stored in conventional extenders is used for cervical artificial insemination (AI). Heat-shock 70-kDa protein 8 (HSPA8), found within the oviduct, prolongs boar, ram and bull sperm survival at body temperatures in vitro. Here, we aimed to determine whether supplementing extenders (INRA-96 and RSD-1) with HSPA8 (4 µg mL–1) would improve their performance in maintaining freshly collected ram sperm viability and sperm nuclear DNA integrity during storage over 48 h at 17°C. Sperm function was assessed at 1, 6, 24 and 48 h and this experiment was repeated using 25 × 106 and 800 × 106 spermatozoa mL–1. INRA96 supplemented with HSPA8 maintained sperm viability significantly better than INRA96 alone at both sperm concentrations. However, sperm nuclear DNA fragmentation (DF) increased significantly during storage using the higher sperm concentration, irrespective of the extender and the protein treatment used. Increasing levels of sperm nuclear DF over time could explain why poor fertility rates are often observed following cervical AI using stored ram semen. However, further research is required to ascertain whether supplementing the commercially available INRA96 extender with HSPA8 will improve fertility rates following cervical AI using stored ram semen.
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Ducha, Nur. "The test about blood serum capabilities in maintaining the quality of bull spermatozoa during storage in cep diluent at refrigerator temperature." IOP Conference Series: Earth and Environmental Science 130 (March 2018): 012043. http://dx.doi.org/10.1088/1755-1315/130/1/012043.
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Mardian, Bayu Anke, Zumarni Zumarni, and Anwar Effendi Harahap. "KUALITAS SEMEN CAIR SAPI SIMENTAL MENGGUNAKAN LARUTAN ISOTONIS KOMERSIAL PADA KONSENTRASI DAN LAMA PENYIMPANAN BERBEDA." JURNAL PETERNAKAN 14, no. 2 (November 17, 2017): 70. http://dx.doi.org/10.24014/jupet.v14i2.3676.
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Artificial insemination (AI) is a technique to insert the sperm or semen of bull thawed and processed first into the genital tract of female methods and a special tool called insemination gun, efforts to optimize the management of the cement in order to obtain the quality of cement is optimal to do with the selection of diluent cement. One alternative materials cement diluent is an isotonic solution Commercial (LIK). This study aimed to examine the interaction between the concentration and duration of storage of diluent solution isotonic different to the quality of spermatozoa cow simental. This research was conducted at the Regional Technical Implementation Unit of Artificial Insemination Centres (UPTD BIB) Tuah Sakato, Payakumbuh. In April 2016. This study used a randomized block design (RAK) factorial design to Factor A (Concentration LIK) ie A1 = 100 ml Tris Egg Yolk, A2 = (60 ml) Tris Egg Yolk + (40 ml) LIK, A3 = (55 ml) Tris Egg Yolk + (45 ml) LIK, A4 = (50 ml) Tris Egg Yolk + (50 ml) LIK, and factor B (retention) is B1 = 0 day, B2 = 1, B3 = 2 day, days, B4 = 3 days. Each treatment consists of three replicates. Parameters measured were motility, percentage of survival, abrnormalitas, and MPU. These results indicate that an isotonic solution can be used as an alternative Commercial diluent with the best results on the percentage LIK 40 ml, but the effectiveness of the efficiency of storage use was only on the first day (24 hours).