Yet regardless of the reason, it appears that concentrations are highly variable, and that some studs have quite different average outputs when compared to others. PIC Boar Stud Management Guidelines . In general terms, the different methodologies of dilution and fractions of the ejaculate used in the study present optimal results after 5 days of conservation . Sperm concentration (thousands//*].) The testosterone concentration, libido, percentage of motile spermatozoa, volume of ejaculate, sperm concentration in the ejaculate and the total amount of the abnormal spermatozoa of the experimental boars during periods are presented in Table Table1 1. The Pig 4MID® Kit is the 1st functional test to evaluate objectively sperm quality and boar fertility. Sperm concentration affects the amount of seminal plasma surrounding each spermato-zoa, both in raw and extended semen. Based on sperm concentration measurements, two groups of boars were identified: Group 1 - boars providing ejaculates with a sperm concentration of at least 500 × 10 ³ /mm ³ and Group 2 - boars. The preferential binding of viable, low-Ca (2+) cells with suppressed tyrosine phosphorylation and slower functional modulation of boar spermatozoa attached to oviductal epithelial cells might represent a mechanism for selecting functionally competent spermatozoa and prolonging their lifespan by delaying capacitation in the oviductal reservoir. Six ejaculates from each boar and a total of 30 . These results indicate that the FACSCount AF System is a valuable tool for precise and accurate assessment of sperm concentration in boar semen and that use of this system may lead to production of more uniform insemination doses containing a specific number of sperm per dose. In exp. Assessment of semen concentration. Download. 1990). increases sperm concentration and motility in men with idiopathic asthenozoospermia (Vitali et al. For example, selenium and Vitamin E tend to produce better results in improving boar sperm motility, concentration and/or morphology when given together . concentration or total numbers of spermatozoa is not a component of semen quality evaluation, but more so, as a tool to monitor the health and productive output of the boar and as the primary feature in processing boar ejaculates for optimizing the genetic potential of a single individual. Both the viability and motility of sperm post-thaw were highest when preserved with 3% glycerol vs. other glycerol concentrations and a control sample without CPA. A minimal concentration of 30 x 10 6 and no more than 100 x 10 6 sperm/ml is recommended by European and American boar studs when extending boar semen. Figure 2. Epididymis is an elongated organ on the posterior surface of a testis that stores sperm while they mature. A final concentration of 25 to 50 million sperm/mL in extended semen generally maximizes sperm survivability in vitro. This number depends upon age, breed, frequency of collection, and individual boar. The software settings for the HTR Ceros 12.1 were those recommended by the manufacturer for analysis of boar sperm, namely: frames per second (Hz) 60, number of frames 45, minimum contrast 18, minimum cell size (pix) 7, cell size (pix) 9, cell inten- Capacitation, as a complex biological process, is influenced by many molecular factors, among which steroidal hormone estrogens play their role. If selection is based on semen quality, it must be taken into account that the sperm quality of boars younger than 8 months is lower than in older boars [].Therefore, selecting boars for AI at young ages based on semen quality can be misleading. Its concentration in epidid- Preliminary research tested whether 50, 100 or 150 mLs of semen extender should be included in the collection bag at the time of collection. The optimum progressive motility of boar spermatozoa was observed in extender with 0.5&# . 1.5-2 years. It is well noted that semen samples evaluated over the last four decades have ejaculates in the range of 40-60 billion/ejaculate and a concentration of 150-350 million/mL. 4VDX-18K2) allows to quantify proAKAP4 concentraion in boar semen as a marker of sperm quality and fertility. each boar ejaculate. September 13, 2010. Before and after freeze-thawing, sperm quality was evaluated through analysis of viability, motility, integrity of outer acrosome membrane, ROS levels . It is the concentration of a solute in a liquid or gas expressed as a unit of solute dissolved in one million units of solution. Individual estrogens have relatively same effect on capacitation progress. The nonfortified diets contained .06 ppm Se and 4.4 IU vitamin E/kg. Concentration-dependent Short-term Effects of AMPs on Sperm Motility (Experiment 1) To determine concentrations applicable for sperm preservation, AMPs were screened for their short-term effects on sperm motility. The Pig 4MID® Kit (Ref. Estrogens, present in a high concentration in the female . total number of sperm cells were not different among treatments. We evaluated the effect of polyphenol-rich aqueous extract of murtilla ( Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. However, because of some unique features the calibration and operation techniques are quite different. To ensure high sperm cell concentration and semen volume, do not collect boars more than three times a week. Satisfactory staining can be achieved within 2-3 min and the . To maximize sperm survival, a minimum of a 1:4 (i.e., 1 part semen to 4 parts extender) dilution should be obtained to ensure that the final seminal plasma concentration in extended semen is less than 20%. This measure is important for determining the number of doses and how many sperm (motile and normal) will be in each dose. The effects of Percoll density gradient centrifugation on sperm quality, in vitro fertilizability and developmental capacity of frozen-thawed boar sperm were evaluated. Appropriate boar selection is crucial and only boars with the best traits for sperm production should be retained. . On the contrary, porcine in vitro sperm capacitation and fertilisation media usually contains a standard concentration of 25 mmol/L, and one of the main problems presented is the . ded for the boars included in the study. It measures the sample turbidity in a manner similar to the standard units previously discussed. The authors conclude that 2-3% glycerol is the best concentration range for boar sperm with respect to motility post-thaw and gene expression patterns. To establish the best sperm concentration range that is suitable for the WST-8 assay, the above-mentioned semen mixtures were subsequently diluted with pre-warmed (37 °C) phosphate buffered saline (PBS) (Lonza, Basel, Switzerland) to obtain a sperm concentration ranging from 300, 150, 75, to 37.5 × 106sperm cells/mL. Application : in vitro determination of proAKAP4 concentrations in boar semen (all breeds) Semen Motility should be evaluated both in the ejaculate as well as in a diluted semen analysed after 2-3 days storage at 16-18 degree Celsius. A total of 10 mature boars were fed from weaning to 18 mo of age diets fortified with two levels of supplemental Se (0 or .5 ppm) or vitamin E (0 or 220 IU/kg diet). Introduction. Semen should have a milky white colour and no noticeable odour. Evaluation of the Karras Spermdensimeter in Relation to the Neubauer Counting Chamber for Sperm Concentration Measurement in Boar Semen. Based on sperm concentration measurements, two groups of boars were identified: Group 1 - boars providing ejaculates with a sperm concentration of at least 500 × 10 3/mm and Group 2 - boars providing ejaculates with a sperm concentration of less than 500 × 10 3/mm . Similarly, Vitamin B12 and folic acids tend to produce better results on folate and homocysteine metabolism in pigs during early pregnancy . The quality of raw, post-holding, and post-thaw semen was evaluated at the farm. c. Sperm concentration - The most common way of determining sperm concentration in gel-free boar semen is by measuring the degree of sample opacity. The importance of sperm quality to fertility when used for artificial insemination (AI) or natural breeding requires evaluation efforts. Junpen Suwimonteerabutr ; Swine Reproduction Research Unit, Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, Thailand . The motility and mobility of the sperm can be influenced by many sources. 7-6 5-2 7-2 12-6 5-4 6-8 5-6 5-7 5-4 4-6 6-5 3-8 4-5 2-5 . lution was made and the sperm concentration of the . 1995; Matalliotakis and Koumantakis 2000), in adult male chickens (Neuman et al. In the ejaculates of Landrace boars, these relationships are more pronounced than in Large White boars. In addition, the head size of boar sperm is larger than that of the giant panda and is more sensitive to freezing ( 31 ). After 5 min of incubation at 38°C, c-WFW and MK5E exerted contrary effects on boar sperm motility. In the Landrace boars group III were ejaculates with sperm concentration of more than 490 × 10 6 /mL. Due to the high concentration of sperm cells in a semen dose, it is necessary to dilute the semen sample to a known dilution factor to decrease the sperm concentration so that individual cells can be counted manually. Comparison of the calculated sperm concentration between a colorimeter and the Bürker's chamber for the same semen samples. In our experiments Excessive collection in a short time reduces the quality and quantity of semen and the boar's sex drive. Citing Literature Volume 37, Issue 6 December 2002 Pages 330-334 High-resolution two-dimensional polyacrylamide gel electrophoresis analyses have indicated that several major boar sperm plasma membrane polypeptides (PMPs) increased in concentration during maturation in the epididymis. The three primary way of measuring concentration include: hemacytometer, photometer, and computer assisted semen analysis (CASA). Sperm concentration was greater for DI than SI (P = 0.011), and tended to be greater for DI compared to CNT (P = 0.102). The plasma membrane of boar sperm contains a higher concentration of phospholipids and a lower concentration of cholesterol . SQA-V FUNCTIONAL SPERM CONCENTRATION: FSC (Functional Sperm Concentration, M/ml) is the concentration of progressively motile spermatozoa with normal morphology in a semen sample. 2, we examined the effects of 1 × MIC and 2 × MIC of two selected lipopeptides with the least negative impact on sperm quality, C16-KKK-NH 2 and C16-KKKK-NH 2, in extended boar semen . The fluid portion of this suspension is known as seminal plasma and it helps to carry and protect the sperm cells. The aim of the present study was to investigate the efficacy of the Improvac on testosterone concentration in blood serum, sexual behavior and sperm quality in matured AI boars. sperm levels from all three tests, it could be seen that the latter were considerably higher than the results of the first testing and almost equal for the other two tests. Validation of measurement systems can be obtained by performing measurements against a reference method. Though these results are promising for potential use of immunological castration in boar stud facilities, semen characteristics were evaluated for a limited number of animals. 125 82 177 171 158 81 150 120 77 95 159 73 116 60 79 73 153 141 95 50 Fructose (mg./lOO ml.) L-carnitine also has a positive effect on boar reproduction. In the Large White boars, group III were ejaculates with sperm concentration above 480 × 10 6 /mL. Boar sperm is sensitive to dilution during semen processing [8,23]. Concentration of sperm cells >150 mill./ml. Each falcon tube was subsequently brought to the final volume of 20 mL with semen preservation reagent. Boar semen is a suspension of sperm cells and secretions from the boar reproductive tract, including the accessory glands. Sperm cell concentration and motility were analysed using computer-assisted sperm analysis (CASA) system (HT CASA - Ceros II, Hamiltan Thorne, Beverly, MA) as per guidelines of the supplier. If you do not have a sperm counting device, you will have to follow some guidelines. The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. It's color can be somewhat yellowish, but is normally has an similar appearance to that of skim milk. Because sperm cells are necessary for fertilization, sperm quality is normally evaluated by assessing measurements such as semen volume, sperm concentration, motility, morphology, and other characteristics that are important to test sperm functionality. Make a a proper quality evaluation. Fifty ejaculates were collected from 37 different boars at an artificial . The dependence of the ejaculate characteristics and the size and shape of sperm on the sperm concentration is influenced by the breed. Sperm concentration is linked to age of the boar and differences in tens of billions have been observed (Clark et al., 2003). Mammalian sperm must undergo a series of controlled molecular processes in the female reproductive tract called capacitation before they are capable of penetrating and fertilizing the egg. Standard tests used to evaluate boar semen include sperm motility, morphology, concentration, total numbers, and ejaculate volume. Related Papers. Reprod Domest Anim 2002; 37:330- 334. To ensure uniformity of the semen doses, the accurate and precise determination of sperm concentration is a prerequisite for AI-stations, especially as AI-stations are trying to reduce the number of sperm per semen dose1. Using 150 mLs of semen extender consistently resulted in a concentration of sperm that was too dilute to extend to a concentration of 37.5 × 10 6 sperm/mL, which is the swine industry standard concentration used in artificial insemination (AI) doses. Capacitation is a physiological process that spermatozoa must experience in the female reproductive tract or in vitro to obtain the ability to bind, penetrate and fertilize the egg [1,2,3].The capacitation is based on many molecular processes including changes in the intracellular calcium concentration [], rearrangement of the acrosomal matrix [], rearrangement of the sperm cytoskeleton [6,7,8 . Sample opacity is estimated most commonly using a photometer, an instrument that measures the percentage of transmittance or absorbency of light through a sample. Method agreement was evaluated on the basis of scatter plots with Deming regression line, absolute bias plots with limits of agreement, and relative bias. midpiece. Accurate assessment of sperm Frozen-thawed boar spermatozoa are less fertile than fresh or cooled semen [], for reasons not yet completely understood.Boar spermatozoa are susceptible to cold shock [], possibly relating to the lipid composition of the membrane, which contains a high concentration of polyunsaturated fatty acids . Genetics and boar selection. Due to the high concentration of sperm cells in a semen dose, it is necessary to dilute the semen sample to a known dilution factor to decrease the sperm concentration so that individual cells In conclusion, it could be presumed that the trace element selenium influenced considerably the concentration of lysozyme in boar sperm. Immediately after collection and filtration, a 1:1 di-. The trial period lasted for 15 weeks and was divided into four periods: Control period: three weeks before vaccination; Period I - four weeks . Single immunized boars had semen volume, gel weight, sperm concentration, total sperm and sperm motility that did not differ from CNT boars. Table 3. A total of nine Danish Landrace AI boars were included in the analysis. Easy steps for boar evaluation. Poster presentations 502 sperm concentration and total and progressive motilities in Duroc boars. 700 x g for 5 minutes for sperm recovery. In boars, the semen also contains large quantities of gel. The most common way of estimating sperm concentration in gel-free boar semen is by measuring the degree of sample opacity. The equation that follows would be used to convert the counts in 5 squares to concentration/ml: Concentration/ml = (Dilution Factor)(Count in 5 squares)(0.05 X 106) By convention, sperm concentration is usually expressed in terms of sperm X 106/ml. For this reason, thirty boar ejaculates were divided into three equal volume fractions and cryopreserved with or without GSH at a final concentration of either 2 or 5. mM in freezing media. Concentration Concentration is an essential measure of total sperm cells in the ejaculate. The pellet was resuspended to 107 sperm/ml in different media according to the experiment, as stated in each case. Since the method which is used may influence the results considerably, the aim of the present study was to compare 5 frequently used techniques to determine boar semen concentration. Sperm cryopreservation is not common in swine artificial insemination programs. The effect of motility, ejaculate volume, ejaculate sperm concentration and diluted sperm concentration, morphology, acrosomal integrity, cell membrane functionality, boar age, rest days between ejaculations, days of semen storage, farrowing, and wean-to-estrus interval were assessed on pregnancy rate and number of piglets born. The in vivo concentration of bicarbonate (HCO3−), one of the essential sperm capacitating effectors, varies greatly in the different environments sperm go through from cauda epididymis to the fertilisation site. In the present study, we hypothesized that buckwheat honey (BH) should be regarded as a potential alternative to antibacterial and antioxidant agent in liquid storage of boar semen. However, normal boar semen as ejaculated differs in several ways from the semen of a ram or bull. Raw semen was evaluated for volume, sperm concentration (by means of a Neubauer chamber), and an aliquot was fixed in 4% formaldehyde for sperm morphological analysis (using differential interference contrast microscopy). The agreement between determining sperm concentration using Makler chamber and a photometer has been assessed. As sperm concentration increases, the amount of seminal plasma per sperm cell decreases. Both for research and practical purposes, accurate and repeatable methods are required to assess the concentration of boar semen samples. As sperm concentration increases, the amount of seminal plasma per sperm cell decreases. In fact, concentration of sperm inseminated, may actually compensate for many deficiencies involving the technology when using AI, when conditions are less than optimal. After analyzing the sperm concentration, semen was added to each of the above-mentioned falcon tubes to achieve a final sperm concentration of 1 × 10 7 spz/mL in a total volume of 20 mL. The normal whole boar ejaculated must be of considerable volume (>150 ml) and depending of the concentration, should be a milky white color. Linear and . The sperm concentration was determined with the aid of spectrophotometer using the following formula (76) × [21.39 × (Absorbance) − 1.09] for boar sperm concentration determinations. Overall, our work and the above studies suggest a . Ejaculate quality, extender type and length of time sperm are to be stored should be used to dictate optimal dilution rate. This is a very difficult parameter to measure manually since you have to kill the progressive cells to assess the morphology. The SpermaCue is a specialized photometic that is widely used in boar semen laboratories. Semen quality and reproductive performance after artificial insemination with boar sperm stored in a gelatin-supplemented extender. However, perhaps another concern should address the expected or actual levels of boar sperm concentration observed in the filed today compared to past decades. Figure 2 compares the sperm concentration (number of sperm x 10 6 /ml) of semen samples from different ejaculates (1-14) obtained with a colorimeter and the Bürker chamber. This is expressed in a larger head area and perimeter and in greater total sperm length. The results show that there are no permanent effects of immunocastration on First o allf, its volume is extraordinarily large, . The stimulatory effect depends on capacitation time and is highly influenced by differences in the response to estrogens such as E2 by individual animals. sperm. To this end, boar semen was firstly studied for in vitro dose tolerability to BH by measuring sperm progressive motility. Collect twice a day or on two successive days to re-inseminate females bred 8 to 12 hours before. 2002), and in rats (Palmero et al. MATERIALS AND METHODS Repeated observations of volume, concentration and total and progressive motilities of in natura ejaculates of 113 Duroc boars housed at the same boar stud were collected from February 2015 until May 2016. Sperm motility tended to be greater for DI boars compared to CNT boars (P = 0.066). Evaluation of the effect of heparin concentration on the physiology of boar spermatozoa Spermatozoa (107 sperm/ml) were suspended in CM and separated in three 1 ml aliquots. Four ejaculates were collected manually from each boar. Briefly, 3 mL of a 2.9% sodium citrate was placed in a cuvette and placed in a spectrophotometer set at 650 nm wavelength before calibration. Two-step density gradient centrifugation by Percoll enhanced significantly the motility parameters of sperm compared with a simple centrifugation procedure. Comments Minelli A, Moroni M, Castellini C. Isolation of the IGF-I protein complex from rabbit seminal plasma: effects on sperm mobility and viability. Determination of boar sperm concentration using a photometer is used routinely by many artificial insemination (AI) laboratories. 2.3. Concentration and Total Number of Spermatozoa Present per Sample The first step is to calculate the concentration of spermatozoa per mL of extended semen. Estrogens stimulate sperm capacitation of boar sperm collected from different individuals. Hansen C, Christensen P, Stryhn H, Hedeboe AM, Rode M, Boe-Hansen G. Validation of the FACSCount AF system for determination of sperm concentration in boar semen. Evaluating semen Volume Colorimeter. Therefore, properly estimating the concentration of the boar ejaculate is critical for efficient boar use and for influencing the effectiveness of the AI technology to optimize . Contents A flow cytometric method has been developed for rapid determination of sperm concentration in semen from various mammalian species. The ejaculates, which met the minimum spermio-gram standards for fresh semen, were used for the study. However, the impact of this factor when different ejaculate fractions are used has not been yet elucidated. The aim of the present investigation was to study the influence of boar, breed of the boar, sperm concentration and weight of the ejaculate The boar ejaculate may contain from a low of 10 to 15 billion up to over 100 billion sperm or enough sperm for 2 or 3 bottles up to 25 or more bottles. Determination of the optimal concentration of Minitube Equex paste for boar semen cryopreservation based on sperm motility characteristics. * All cells containing DNA are stained with SYBR‐14 or propidium iodide (PI) and sperm concentration is determined in relation to an internal standard of fluorescent microspheres (beads). Evaluation of semen Evaluation of semen and sperm morphology was performed immediately after semen collection. Kinetic characterization of the changes in protein tyrosine phosphorylation of membranes, cytosolic Ca2+ concentration and viability in boar sperm populations selected by binding to oviductal epithelial cells. The aim of the present investigation was to study the influence of boar, breed of the boar, sperm concentration and weight of the ejaculate on motility and acrosome integrity of liquid boar semen stored in BTS for 5 days. The experiment was a 2 x 2 arrangement of treatments in a randomized complete block design. Sample opacity is usually estimated using a photometer, an instrument that measures the percentage transmittance/absorbency of light through a sample. Reproduction 122, 469-480. The first step is to calculate the concentration of spermatozoa per mL of extended semen. was used to evaluate the sperm concentration (Vyt et al., 2004). 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