Abstract: The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P
Abstract: The present study was carried out on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to assess the effect of cooling using mist cooling and fanning on Sahiwal bulls in the dry hot summer season. Fourteen Sahiwal bulls were divided into two groups of seven each. Sexual behavior and semen quality traits considered were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-1 was the control, whereas group-2 (treatment group) bulls were exposed to mist cooling and fanning (thrice a day 15 min each) in the dry hot summer season. Group-2 showed significantly (p < 0.01) higher value in DMT (sec), ES, PS, ITS, LS, semen volume (ml), semen color density, mass activity, initial motility, progressive motility and live sperm.
Abstract: The study was conducted on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to determine the effect of exercise on the sexual behavior and semen quality. Fourteen Sahiwal bulls were classified into two groups of seven each. Group-1, bulls were exercised by walking in a bull exerciser once a week one hour before semen collection, whereas bulls in group-2 were exercised daily. Sexual behavior and semen quality traits studied were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-2 showed significantly (p < 0.01) higher value in RT (sec), DMT (sec), TTTM (sec), ES, PS, ITS, LS, semen volume, semen color density and mass activity.
Abstract: This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P
Abstract: In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P
Abstract: The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P
Abstract: Oxidative stress associated with semen cryopreservation may result in lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Curcumin (CUR), a natural phenol isolated from Curcuma longa Linn. has been presented as a possible supplement for a more effective semen cryopreservation because of its antioxidant properties. This study focused to evaluate the effects of CUR on selected oxidative stress parameters in cryopreserved bovine semen. 20 bovine ejaculates were split into two aliquots and diluted with a commercial semen extender containing CUR (50 μmol/L) or no supplement (control), cooled to 4 °C, frozen and kept in liquid nitrogen. Frozen straws were thawed in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, and reactive oxygen species (ROS) generation was quantified by using luminometry. Superoxide generation was evaluated with the NBT test, and LPO was assessed via the TBARS assay. CUR supplementation significantly (P
Abstract: Target of this study was the analysis of the impact of
crude glycerol on canine spermatozoa motility, morphology,
viability, and membrane integrity. Experiments were realized in vitro.
In the study, semen from 5 large dog breeds was used. They were
typical representatives of large breeds, coming from healthy rearing,
regularly vaccinated and integrated to the further breeding. Semen
collections were realized at the owners of animals and in the
veterinary clinic. Subsequently the experiments were realized at the
Department of Animal Physiology of the SUA in Nitra. The
spermatozoa motility was evaluated using CASA analyzer
(SpermVisionTM, Minitub, Germany) at the temperature 5 and 37°C
for 5 hours. In the study, 13 motility parameters were evaluated.
Generally, crude glycerol has generally negative effect on
spermatozoa motility. Morphological analysis was realized using
Hancock staining and the preparations were evaluated at
magnification 1000x using classification tables of morphologically
changed spermatozoa. Data clearly detected the highest number of
morphologically changed spermatozoa in the experimental groups
(know twisted tails, tail torso and tail coiling). For acrosome
alterations swelled acrosomes, removed acrosomes and acrosomes
with undulated membrane were detected. In this study also the effect
of crude glycerol on spermatozoa membrane integrity were analyzed.
The highest crude glycerol concentration significantly affects
spermatozoa integrity. Results of this study show that crude glycerol
has effect of spermatozoa motility, viability, and membrane integrity.
Detected changes are related to crude glycerol concentration,
temperature, as well as time of incubation.
Abstract: More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.
Abstract: Rainbow trout homogametic males, (XX or YY sex genotype), can be obtained, respectively, through masculinisation of genetic females or induced androgenesis. Aim of this study was to compare reproductive potential of neo-males (XX) and super-males (YY) with heterogametic males (XY). We measured spermatozoa motility parameters, sperm concentration, osmolality and characterized protein profiles in samples of stripped and testicular sperm obtained from XY and YY males, and testicular sperm of XX males. The motile spermatozoa, as measured by both subjective method and CASA, showed no differences between testicular sperm of XX males and stripped sperm of XY and YY males whereas testicular sperm of XY and YY males had significantly lower sperm motility. Result of protein densitometry showed similarities in protein profile between seminal plasma of XY and YY males and testicular fluids of XX males. Testis of XX males showed specific histological structures of cysts consists hypertrophied Sertoli cells.
Abstract: The objective of the present study was to determine
the effect of different concentration of spermatozoa and length of
storage in 5 0C on sperm motility. Semen was collected using
artificial vagina from goat aged 2 to 2.5 years. Fresh goat semen
with sperm motility ≥ 70% was used as material. Semen was
divided into 4 treatments of concentration (40 x 10 6 / ml, 50 x
106/ml, 60x106/ml, 70x106/ml) with length of storage 0,12,24,36 h. in
5 0C. There were interactions (P
Abstract: The aim of this study was to investigate the effects of
supplementing the diluent of roosters' semen with different levels of
olive oil on motility, viability, morphology and acrosome integrity of
chicken spermatozoa after in vitro storage for up to 72 h. Semen was
collected from 60 White Layer males (62 wk of age) kept in
separated floor pens and randomly divided into six treatment groups
(10 males in each group). Experimental groups were as follows: T1
:fresh semen, T2 : semen extended 1:1 with Al – Daraji 2 diluent
(AD2D) alone, T3 – T6 :semen samples extended 1:1 with AD2D
supplemented with 2 ml, 4 ml, 6 ml or 8 ml of olive oil / 100 ml of
diluent, respectively. Semen samples were then stored at 5 °C for 24
h, 48 h or 72 h. There was a clear influence of diluent
supplementation with olive oil on the spermatozoa motility profile;
olive oil groups (T3, T4, T5 and T6) recorded the highest scores of
mass activity and individual motility during all storage periods
compared to T1 and T2 groups. In addition, the inclusion of olive oil
into semen diluent (T3, T4, T5 and T6) gave significantly higher
percentages of viable spermatozoa, normal morphologically
spermatozoa and intact acrosomes irrespective of storage period.
These results clearly show that supplementation the diluent of
roosters' semen with olive oil can improve semen quality when
semen samples in vitro stored at 5 °C for up to 72 h.
Abstract: The aim of current study was to investigate the
changes in the quality parameters of Holstein bull semen during the
heat stress and the effect of feeding a source of omega-3 fatty acids
in this period. Samples were obtained from 19 Holstein bulls during
the expected time of heat stress in Iran (June to September 2009).
Control group (n=10) were fed a standard concentrate feed while
treatment group (n=9) had this feed top dressed with 100 g of an
omega-3 enriched nutriceutical. Semen quality was assessed on
ejaculates collected after 1, 5, 9 and 12 weeks of supplementation.
Computer-assisted assessment of sperm motility, viability (eosinnigrosin)
and hypo-osmotic swelling test (HOST) were conducted.
Heat stress affected sperm quality parameters by week 5 and 9
(p
Abstract: Uterine and oviducal fluids are necessary for
capacitation of the spermatozoa and early embryonic development.
The aim of the present study was to determine the effects of estrous
cycle phases (follicular and luteal) on some biological parameters
(enzymes, electrolytes and total proteins) in uterine and oviducal
secretions of ewes. Oviducal and uterine fluids were collected,
diluted and centrifuged. According to our results, concentrations of
GPT, G6PDH, total proteins, K and Na were significantly (P