Is addition or removal of seminal plasma able to compensate for the dilution effect of buffalo semen?

Andrologia ◽  
2021 ◽  
Author(s):  
V. Arjun ◽  
Pradeep Kumar ◽  
Ravi Dutt ◽  
Amit Kumar ◽  
Renu Bala ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Dilution Effect ◽  
Buffalo Semen

327 IDENTIFICATION OF OSTEOPONTIN IN WATER BUFFALO SEMEN

2007 ◽  
Vol 19 (1) ◽  
pp. 279 ◽  
Author(s):  
M. E. Pero ◽  
G. J. Killian ◽  
P. Lombardi ◽  
L. Zicarelli ◽  
L. Avallone ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Male Fertility ◽  
Water Buffalo ◽  
Polyclonal Antibodies ◽  
Biological Fluids ◽  
Bovine Milk ◽  
Genetic Progress ◽  
Standard Procedures ◽  
Buffalo Semen

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.

Seminal plasma addition attenuates the dilution effect in bovine sperm

2001 ◽  
Vol 56 (1) ◽  
pp. 31-40 ◽  
Author(s):  
D.L. Garner ◽  
C.A. Thomas ◽  
C.G. Gravance ◽  
C.E. Marshall ◽  
J.M. DeJarnette ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Dilution Effect ◽  
Bovine Sperm

The Physiology of Sea-Urchin Spermatozoa Action of Versene

1954 ◽  
Vol 31 (2) ◽  
pp. 252-259
Author(s):  
LORD ROTHSCHILD ◽  
A. TYLER
Keyword(s):  
Trace Metals ◽  
Sea Urchin ◽  
Seminal Plasma ◽  
Sea Water ◽  
Dilution Effect ◽  
Dense Suspension

1. The effect of adding sea water containing different concentrations of versene to suspensions of sea-urchin spermatozoa (Echinus esculentus) has been investigated, as regards their respiration, motility and fertilizing capacity. 2. The respiratory Dilution Effect is progressively reduced and finally abolished when, instead of sea water, sea water containing 10-6, 10-5, 10-4 or 10-3 M versene is added to sperm suspensions. 3. At the same time versene greatly delays the senescence of the spermatozoa, both as regards their motility and fertilizing capacity. For example, after seventeen hours, a 105 sperm/ml. suspension in sea water containing 10-3 M versene has 125 times the fertilizing capacity of a suspension containing 107 sperm/ml. without versene. 4. The change in the ratio of seminal plasma to sea water which occurs when a dense suspension is diluted does not explain the Dilution Effect. 5. These results are discussed in relation to the hypothesis which accounts for the Dilution Effect in terms of trace metals, particularly copper, normally occurring in sea water, and the amounts available per spermatozoon under various conditions of dilution.

The Respiration of Sea-Urchin Spermatozoa

1950 ◽  
Vol 27 (3) ◽  
pp. 420-436
Author(s):  
LORD ROTHSCHILD
Keyword(s):  
Oxygen Saturation ◽  
Sea Urchin ◽  
Seminal Plasma ◽  
Sea Water ◽  
Chelating Agent ◽  
Dilution Effect ◽  
O2 Uptake ◽  
Sperm Density ◽  
Dense Suspensions ◽  
Mammalian Spermatozoa

1. The O2 uptake of sea-urchin spermatozoa, Echinus esculentus, has been reexamined under varying conditions of sperm density, and in the presence of CuCl22H2O and sodium diethyldithiocarbamate (DDC). 2. The total O2 uptake of dilute sperm suspensions was previously thought to be higher than that of dense suspensions per unit quantity of spermatozoa (Dilution Effect I). This result is only obtained when the oxygen saturation of the dense suspension is inadequate, which may easily occur as the QOO2 of sea-urchin spermatozoa may reach 30 at 15.0° C. When oxygen saturation is satisfactory, the total O2 uptake of dense solutions is slightly greater than that of dilute ones. The experiment cannot be done in micro-respirometers of the normal Warburg type unless the density of spermatozoa per ml. suspension is less than about 109 corresponding to an initial semen dilution of 1:20 or 1:25. These figures apply to other manometric experiments on the O2 uptake of sea-urchin spermatozoa using normal amounts of material. 3. When movement ceases, there is a sharp increase in the O2 uptake of the suspension. 4. The addition of seminal plasma to dilute sperm suspensions does not inhibit the increased rate of O2 uptake, per unit quantity of spermatozoa, observed in these suspensions when compared with dense ones (Dilution Effect II). Dilution Effect II is therefore not caused by the dilution of an inhibitory substance in seminal plasma. 5. Sperm suspensions were prepared by diluting semen 1:50 with sea water and allowing them to respire for 45 mm. They were then centrifuged, the supernatant was discarded and the spermatozoa were re-suspended to different densities with sea water. This treatment has the following effects: (a) Centrifugation irreversibly damages the spermatozoa and reduces their O2 uptake. (b) Removal of the supernatant, which contains seminal plasma, and re-suspension in sea water also reduces O2 uptake. (c) The treatment markedly reduces Dilution Effect II. If the experiment is done in the same way but the suspensions are only allowed to respire for 10 min. before centrifugation, (a) and (b) are the same, but Dilution Effect II is normal. This shows that during metabolism, a regulatory substance is lost from dilute suspensions, as in mammalian spermatozoa; but this is not the cause of Dilution Effect II. 6. Dilution Effect II, considered as the reduced O2 uptake of dense suspensions, can be reversed by the addition of CuCl22H2O, 1 p.p.m., to the medium. 7. Dilution Effect II can be made to occur in sperm suspensions which do not normally exhibit it, by the addition of DDC, in concentrations as low as 3.64x10-5M (final concentration). The action of DDC is not greater when its concentration is increased to 10-3M, which suggests that in these conditions it acts as a chelating agent and not as a narcotic. For the same reasons its oxidation product, tetra ethyldithiocarbamyl disulphide, is unlikely to be responsible for DDC's inhibitory effect on sperm O2 uptake. 8. These results are consistent with the hypothesis that Dilution Effect II is due to the amounts of copper (or possibly zinc) in sea water being inadequate to satisfy the requirements of dense sea-urchin sperm suspensions. This situation is unlikely to arise during natural spawning as sperm densities are too low for the effect to occur in these conditions. Other interpretations of the stimulating action of copper and zinc are discussed. 9. The experiments remove several of the differences hitherto believed to exist between sea-urchin and mammalian spermatozoa.

Lactic dehydrogenase activity in buffalo (Bos-bubalis) bull semen and seminal plasma in relation to various physical characteristics

1980 ◽  
Vol 94 (1) ◽  
pp. 255-256 ◽  
Author(s):  
R. K. Tuli ◽  
Mehar Singh
Keyword(s):  
Dehydrogenase Activity ◽  
Seminal Plasma ◽  
Lactic Dehydrogenase ◽  
Physical Characteristics ◽  
Bull Semen ◽  
Ldh Activity ◽  
Buffalo Semen

In anaerobic glyoolysis lactic dehydrogenase (LDH) is the terminative enzyme in the sequence of reactions that promote the breakdown of sugars to lactate. The LDH activity of cattle semen and seminal plasma together with its relationship with various characteristics of semen have been reported (Stallcup & Hayden, 1960; Roussel & Stallcup, 1965). Such work in respect of buffalo semen is not available, although there are some reports on LDH activity of spermatozoa and seminal plasma (Singh & Sadhu, 1972; Chauhan & Srivastava, 1973). The present investigation was undertaken to determine the LDH activity of buffalo whole semen as well as of seminal plasma and to find correlations with some physical characteristics of semen.

Supplemented stallion seminal plasma can improve impaired motility due to the dilution effect in chilled Asian elephant sperm

2018 ◽  
Vol 53 (2) ◽  
pp. 525-533
Author(s):  
A Pinyopummin ◽  
S Mahasawangkul ◽  
G Nunklang ◽  
K Kornkaewrat ◽  
S Laopiem ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Dilution Effect ◽  
Asian Elephant

scholarly journals 329PREGNANCY RATE IN EWES INSEMINATED WITH SEXED SEMEN IN ARGENTINA

2004 ◽  
Vol 16 (2) ◽  
pp. 284 ◽  
Author(s):  
L. Cattaneo ◽  
P. Martín ◽  
J.O. Caballero ◽  
M. Calvi ◽  
B. Vietri ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Seminal Plasma ◽  
High Speed ◽  
Dilution Effect ◽  
Low Doses ◽  
Chi Square ◽  
Speed Flow ◽  
Frozen Semen ◽  
Significant Difference ◽  
Sexed Semen

Offspring of many species have been born from flow cytometrically sex-sorted sperm since its introduction in 1989 (LA Jonhson et al.; 1999, Theriogenology). Births of lambs after insemination of ewes with low doses of sexed semen were first reported by Hollinshead et al. (Rep. Fert. Dev. 2002). A field trial was carried out in Patagonia Argentina comparing the use of low doses of sexed and non-sexed frozen semen. This study is aimed at comparing pregnancy rates in ewes inseminated with sorted and nonsorted frozen-thawed sperm. Ejaculates from two Merino rams were diluted with modified tirode solution (Shenk et al., 1999, Theriogenology), stained with Hoescht 33342, and finally incubated at 35°C for 40 minutes. The addition of 10% ram seminal plasma to the sample was done to avoid sperm agglutination caused by dilution effect (Mann, 1964). Sorting was performed with a high speed flow cytometer (MOFLO®). Up to 8 million of sexed spermatozoa were collected in 15-mL conical tubes containing modified tirode solution plus 10% ram seminal plasma, centrifuged at 600g 12min and resuspended with test yolk media plus 11% skimmed milk and 5% glycerol. After being refrigerated at 4°C for at least 1.5h, sexed and nonsexed semen were packaged in 0.25-mL straws at a concentration of 4 to 5 total million sperm and then frozen in an automatic freezing machine (IMV®). One hundred and eighty-three ewes and hoggets were laparoscopically inseminated by the same technician with eight to ten million sorted and nonsorted frozen-thawed semen 14h after heat detection. Pregnancy diagnosis was performed by ultrasound at day 30 post-insemination. Data were analized by Chi-square. There was significant difference in the overall pregnancy rate between sorted and nonsorted sperm, although, for one of the rams that difference was not significant. There was, also, no significant difference in pregnancy rate between ram 1 and 2 in ewes inseminated with either sexed or nonsexed semen (Table 1). These results indicate that the different performance of sorted compared to non sorted sperm is mainly explained by the lower results achieved with semen from ram 2. According to this conclusion, further studies should be done in order to evaluate the degree to which the sorting process affects the sperm fertilizing capability in different rams. This research was supported by Fundación Margarita Perez Companc. Table 1

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