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Sample records for del semen tras

  1. TRIHALOMETHANE LEVELS AND SEMEN QUALITY

    EPA Science Inventory

    Trihalomethanes (THMs) are common byproducts of chlorinating drinking water. The effects of disinfection byproducts on semen quality have not yet been studied in humans, despite animal studies linking exposure to sperm abnormalities. We are currently analyzing the relationship of...

  2. Avian artificial insemination and semen preservation

    USGS Publications Warehouse

    Gee, G.F.; Risser, Arthur C.; Todd, Frank S.

    1983-01-01

    Summary: Artificial insemination is a practical propagation tool that has been successful with a variety of birds. Cooperative, massage, and electroejaculation and modifications of these three basic methods of semen collection are described for a variety of birds. Semen color and consistency and sperm number, moti!ity, and morphology, as discussed, are useful indicators of semen quality, but the most reliable test of semen quality is the production of fertile eggs. Successful cryogenic preservation of avian semen with DMSO or glycerol as the cryoprotectant has been possible. Although the methods for preservation require special equipment, use of frozen semen requires only simple insemination supplies

  3. Cryopreservation of American kestrel semen with dimethylsulfoxide

    USGS Publications Warehouse

    Gee, G.F.; Morrell, C.A.; Franson, J. Christian; Pattee, Oliver H.

    1993-01-01

    Semen samples from 15 male American Kestrels (Falco sparverius) were frozen in dimethyl sulfoxide (DMSO). The semen was thawed 1-14 mo later and used to inseminate six females during three breeding seasons. Kestrels inseminated with thawed semen containing 4% DMSO produced only infertile eggs (N = 14). Kestrels inseminated with thawed semen containing 6%, 8%, or 10% DMSO produced fertile eggs (N = 14) and live chicks (N = 6). Progressive motility of spermatozoa in thawed semen containing 10% DMSO was less (44 ? 6%) than in thawed semen containing 6% (62 ? 10%) or 8% (61 ? 1%) DMSO.

  4. Recent advances in cooled-semen technology.

    PubMed

    Aurich, Christine

    2008-09-01

    The majority of horse registries approve the use of artificial insemination, and horse breeding has widely taken benefit from the use of cooled-stored semen. New insights into cooled-semen technology open possibilities to reduce problems such as impaired semen quality after cooled-storage in individual stallions. The stallion itself has major impacts on quality and fertility of cooled-stored semen. Dietary supplementation of antioxidants and polyunsaturated fatty acids improves semen quality in a variety of species, but only few studies on this topic exist in the horse. Proper semen collection and handling is the main key to the maintenance of semen quality during cooled-storage. Semen collection should be achieved by minimal sexual stimulation with a single mount; this results in high sperm concentration, low content of seminal plasma and minimal contamination with bacteria. Milk-based semen extenders are most popular for semen processing and storage. The development of more defined extenders containing only the beneficial milk ingredients has made extender quality more constant and reliable. Semen is often centrifuged to decrease the seminal plasma content. Centrifugation results in a recovery rate of only 75% of spermatozoa in the semen pellet. Recovery rates after centrifugation may be improved with use of a "cushion technique" allowing higher centrifugation force and duration. However, this is not routinely used in cooled-semen technology. After slow-cooling, semen-storage and shipping is best performed at 5 degrees C, maintaining semen motility, membrane integrity and DNA integrity for up to 40 h after collection. Shipping containers created from Styrofoam boxes provide maintenance of semen quality at low cost.

  5. Seasonal and genera-specific variations in semen availability and semen characteristics in large parrots.

    PubMed

    Bublat, A; Fischer, D; Bruslund, S; Schneider, H; Meinecke-Tillmann, S; Wehrend, A; Lierz, M

    2017-03-15

    In large parrots electro-stimulation is suitable for collecting semen, and therefore, to facilitate semen examination and artificial insemination. Previous studies have detected differences in the semen collection success rate and semen parameters between psittacine genera. It remained unclear whether these differences were genera-related, seasonal variations or depend on the males' relationship status. To answer these questions, semen collection and spermatological analysis were performed for four psittacine groups (macaws, amazons, eclectus parrots and cockatoos) over 13 months. In one breeding facility, semen collection was attempted in 82 males using electro-stimulation twice monthly. A complete spermatological evaluation was performed on 435 semen samples. Volume, color, consistency, contamination and pH of semen, as well as motility, progressive motility, sperm concentration, total sperm count, viability, and morphology of spermatozoa were evaluated. Seasonality affected the collection success rate in macaws and amazons. Thereby, in amazons a distinct peak was observed several days before and around oviposition, whereas eclectus parrots and cockatoos produced semen all year round. The average sperm concentration was highest in eclectus parrots (2.7 × 10(6) sperm/μl) and lowest in macaws (35.6 × 10(3) sperm/μl). The differences in the semen collection success rate and semen parameters seem to coincide with the bird's breeding biology. The collected data allows a prognostic estimation when semen collection seems favorable, and may be taken as orientation values for semen analysis in these species.

  6. Bulls grazing Kentucky 31 tall fescue exhibit impaired growth, semen quality, and decreased semen freezing potential

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Serum prolactin (PRL) and testosterone concentrations, body weight, body composition, semen quality, and semen freezing potential for bulls grazing the toxic tall fescue (Lolium arundinaceum [Schreb.] Darbysh. ¼ Schedonorous arundinaceum [Schreb.] Dumort.) cultivar Kentucky 31 (E+) compared with a n...

  7. Evaluation of semen from nondomestic birds

    USGS Publications Warehouse

    Gee, G.F.; Bakst, M.R.; Cecil, H.C.

    1997-01-01

    Aspects of poultry Al technology are applicable to nondomestic birds. However, modifications in the methods of semen collection, evaluation, and insemination are often necessary to accomodate either the bird's size, sperm numbers, or. female anatomy. This section provides a brief overview of procedures used to evaluate semen from nondomestic birds. Unless specified, materials, reagents, etc., are identical to those used in evaluating poultry semen (see appropriate chapters).

  8. Ebola Virus Persistence in Semen Ex Vivo.

    PubMed

    Fischer, Robert J; Judson, Seth; Miazgowicz, Kerri; Bushmaker, Trent; Munster, Vincent J

    2016-02-01

    On March 20, 2015, a case of Ebola virus disease was identified in Liberia that most likely was transmitted through sexual contact. We assessed the efficiency of detecting Ebola virus in semen samples by molecular diagnostics and the stability of Ebola virus in ex vivo semen under simulated tropical conditions.

  9. Ebola Virus Persistence in Semen Ex Vivo

    PubMed Central

    Fischer, Robert J.; Judson, Seth; Miazgowicz, Kerri; Bushmaker, Trent

    2016-01-01

    On March 20, 2015, a case of Ebola virus disease was identified in Liberia that most likely was transmitted through sexual contact. We assessed the efficiency of detecting Ebola virus in semen samples by molecular diagnostics and the stability of Ebola virus in ex vivo semen under simulated tropical conditions. PMID:26811984

  10. Semen characteristics in pubertal boys. I. Semen quality after first ejaculation.

    PubMed

    Janczewski, Z; Bablok, L

    1985-01-01

    Semen specimens from 134 pubertal boys were examined, and some 274 assays were made. An analysis of the biological quality of semen in relation to the period of time after first ejaculation brings high values of statistical dependence of the volume of semen, its liquefaction, spermatozoal concentration, percentage of morphologically normal forms of spermatozoa, and normal spermatozoal motility on the period of time after first ejaculation. Normal figures for semen volume, semen liquefaction, spermatozoal concentration, and morphology are observed 12-14 months after first ejaculation. The percentage of normally motile spermatozoa becomes standard 21-23 months after first ejaculation. There were changes in semen characteristics from azoospermia through cryptozoospermia, oligozoospermia, and asthenozoospermia to normospermia. Azoospermia dominates until the fifth month after the first ejaculation, oligozoospermia from the sixth to the eleventh month, asthenozoospermia from the twelfth to the twentieth month, and normospermia from the twenty-first month.

  11. [Identification of semen Cuscutae by HPCE].

    PubMed

    Hu, P; Luo, G A; Wang, R J; Guo, H Z; Li, J S

    1997-07-01

    A method for identification of semen Cuscutae, a traditional Chinese herb, was developed. The proteins in semen Cuscutae were extracted under acid or basic conditions and separated by high performance capillary electrophoresis. The electrophoretograms of acidic or basic extractants from Cuscuta chinensis Lam., Cuscuta australis R. Br. and Cuscuta japonica Choisy showed significant differences, which can be used to identify the three different semen Cuscutae. The results of the identification for 13 pharmacognosical samples agreed well with those of scanning electronic microscopy and tissue microanalysis.

  12. Quality parameters for alpaca (Vicugna pacos) semen are affected by semen collection procedure.

    PubMed

    Morton, K M; Thomson, P C; Bailey, K; Evans, G; Maxwell, W M C

    2010-08-01

    Artificial insemination (AI) is poorly developed in camelids owing to the difficulty in collecting high quality semen and the highly viscous nature of the semen. Semen collected by artificial vagina (AV) is often of low quality and must be improved before any further development of AI technology can occur. The present study investigated the effects of adding a cervix-like stricture to the AV, presence of females, collecting semen into Androhep, skim-milk or Tris diluents, and catalase supplementation (0, 100, 200 or 600 units/ml) of Tris diluent on alpaca semen quality parameters. The addition of a cervix-like stricture increased mating length (p < 0.05), whilst the presence of females during semen collection did not improve semen quality parameters (p > 0.05). Collection of semen into Tris diluent improved sperm motility (58.0 +/- 11.9%) compared with the control (34.0 +/- 10.8%; p < 0.05), Androhep (33.5 +/- 10.7%) and skim-milk diluents (28.2 +/- 10.4%). Semen viscosity was reduced by collection into Androhep (4.6 +/- 1.7 mm) and skim-milk diluents (3.6 +/- 1.3 mm) compared with Tris diluent (5.7 +/- 2.1 mm) and no collection medium (9.3 +/- 3.5 mm; p < 0.05). Tris diluent supplemented with 100, 200 or 600 units/ml catalase increased semen viscosity (5.0 +/- 3.2 and 4.9 +/- 3.2 mm). Collection of alpaca semen by AV into Tris diluent increased semen quality facilitating further development of AI technology in alpacas.

  13. Alternatives to Antibiotics in Semen Extenders: A Review

    PubMed Central

    Morrell, Jane M.; Wallgren, Margareta

    2014-01-01

    Antibiotics are added to semen extenders to be used for artificial insemination (AI) in livestock breeding to control bacterial contamination in semen arising during collection and processing. The antibiotics to be added and their concentrations for semen for international trade are specified by government directives. Since the animal production industry uses large quantities of semen for artificial insemination, large amounts of antibiotics are currently used in semen extenders. Possible alternatives to antibiotics are discussed, including physical removal of the bacteria during semen processing, as well as the development of novel antimicrobials. Colloid centrifugation, particularly Single Layer Centrifugation, when carried out with a strict aseptic technique, offers a feasible method for reducing bacterial contamination in semen and is a practical method for semen processing laboratories to adopt. However, none of these alternatives to antibiotics should replace strict attention to hygiene during semen collection and handling. PMID:25517429

  14. Artificial insemination of cranes with frozen semen

    USGS Publications Warehouse

    Gee, G.F.; Sexton, T.J.; Lewis, J.C.

    1979-01-01

    For the first time (1978) artificial insemination (AI) with frozen greater sandhill crane (Grus canadensis tabida) semen resulted in fertile eggs and chicks. During the 2 year (1977-78) study, 6 of 27 eggs produced were fertile. Three chicks hatched. Semen samples used for insemination were frozen and stored in liquid nitrogen for two months or less. Recent improvements in the laboratory indicated that a more effective sample can be prepared and greater fertility rates should be expected.

  15. [Morphological semen changes in Chlamydia trachomatis infection].

    PubMed

    Maciejewski, Z; Dziecielski, H; Swierczyński, W; Semmler, G

    1989-06-01

    Semen was examined in 150 men patients of the Andrology Clinic for demonstration of Chlamydia trachomatis and for analysis of the effect of this infection on semen quality depression. A correlation was noted between the degree of infection (large number of organisms per field of vision) and such changes as cryptozoospermia, azoospermia, asthenozoospermia, teratozoospermia, oligoasthenozoospermia, asthenoteratozoospermia. Of interest was a high proportion of infection (56%) with Ch. trachomatis in this group.

  16. Applications of sexed semen in cattle production.

    PubMed

    Hohenboken, W D

    1999-12-01

    Sexed semen will contribute to increased profitability of dairy and beef cattle production in a variety of ways. It could be used to produce offspring of the desired sex from a particular mating to take advantage of differences in value of males and females for specific marketing purposes. Commercial dairy farmers, those who produce and market milk, could use sexed semen to produce replacement daughters from genetically superior cows and beef crossbred sons from the remainder of their cow population. To increase the rate of response to selection, seedstock dairy cattle breeders could produce bulls for progeny testing from a smaller number of elite dams by using sexed semen to ensure that all of them produced a son. Using sexed semen could then reduce the cost of progeny testing those bulls, because fewer matings would be necessary to produce any required number of daughters. Commercial beef cattle farmers, producing animals for eventual slaughter, could use sexed semen to capitalize on the higher value of male than female offspring for meat production. They could also use sexed semen to produce specialized, genetically superior replacement heifers from as small a proportion of the herd as possible. This would allow the remainder of the herd to produce male calves from bulls or breeds with superior genetic merit for growth, feed conversion efficiency, and carcass merit. Single-sex, bred-heifer systems, in which each female is sold for slaughter soon after weaning her replacement daughter, would be possible with the use of X-chromosome-sorted semen. Use of sexed semen would make terminal crossbreeding systems more efficient and sustainable in beef cattle. Fewer females would be required to produce specialized maternal crossbred daughters, and more could be devoted to producing highly efficient, terminal crossbred sons.

  17. Update on sexed semen technology in cattle.

    PubMed

    Seidel, G E

    2014-05-01

    The technology in current use for sexing sperm represents remarkable feats of engineering. These flow cytometer/cell sorters can make over 30 000 consecutive evaluations of individual sperm each second for each nozzle and sort the sperm into three containers: X-sperm, Y-sperm and unsexable plus dead sperm. Even at these speeds it is not economical to package sperm at standard numbers per inseminate. However, with excellent management, pregnancy rates in cattle with 2 million sexed sperm per insemination dose are about 80% of those with conventional semen at normal sperm doses. This lowered fertility, in part due to damage to sperm during sorting, plus the extra cost of sexed semen limits the applications that are economically feasible. Even so, on the order of 2 million doses of bovine semen are sexed annually in the United States. The main application is for dairy heifers to have heifer calves, either for herd expansion or for sale as replacements, often for eventual export. Breeders of purebred cattle often use sexed semen for specific matings; thawing and then sexing frozen semen and immediately using the few resulting sexed sperm for in vitro fertilization is done with increasing frequency. Beef cattle producers are starting to use sexed semen to produce crossbred female replacements. Proprietary improvements in sperm sexing procedures, implemented in 2013, are claimed to improve fertility between 4 and 6 percentage points, or about 10%.

  18. Semen collection using phantom in dromedary camel.

    PubMed

    Ziapour, S; Niasari-Naslaji, A; Mirtavousi, M; Keshavarz, M; Kalantari, A; Adel, H

    2014-12-10

    Semen collection is relatively long, unsafe, and tedious procedure in dromedary camel. The innovation of safe, hygienic, and simple approach to collect semen could make great progress in development of AI program in this species. This study investigated two methods of semen collection using phantom and artificial vagina in dromedary camel. Semen was collected using phantom (n = 4 bulls; 26 collections) and artificial vagina (n = 6 bulls; 11 collections) and diluted with INRA96 at the ratio of 1:10. The duration of semen collection, semen parameters, and morphometric features of sperm were evaluated. For specimen collected through phantom and AV, the respected duration of semen collection (411.2 ± 48.19 vs 326 ± 37.05 sec), volume (6.6 ± 0.87 vs 6 ± 1.57 ml), osmolarity (328 ± 1.6 vs 319.4 ± 3.21 mOsm/kg H2O), pH (7.7 ± 0.06 vs 7.9 ± 0.16) of semen, concentration (161.4 ± 44.05 × 10(6)/mL vs 160.2 ± 58.42 × 10(6)/mL), total motility (84.1 ± 1.89 vs 78.3 ± 3.97%), progressive forward motility (45.5 ± 3.69 vs 44.3 ± 6.41%), live percentage (72.2 ± 3.11 vs 76 ± 2.53%), and plasma membrane integrity (61.5 ± 2.49 vs 58.9 ± 4.19%) of sperm were similar (P > 0.05). The number of specimens contaminated with visible particles was greater using AV (72.7%) compared to phantom (0%; P < 0.05). Total length, head, middle-piece, and tail length of sperm were 45.9 ± 0.1, 5.6 ± 0.01, 7 ± 0.02, and 34.2 ± 0.16 μm, respectively. The frequency of abnormal sperm was 13.28% among which coiled tail, detached head, and proximal protoplasmic droplets had greater incidence. In conclusion, phantom could be considered as a suitable approach to collect semen due to simplicity, safety, and lack of specimen contamination in dromedary camel.

  19. Semen quality in Peruvian pesticide applicators: association between urinary organophosphate metabolites and semen parameters

    PubMed Central

    Yucra, Sandra; Gasco, Manuel; Rubio, Julio; Gonzales, Gustavo F

    2008-01-01

    Background Organophosphates are broad class of chemicals widely used as pesticides throughout the world. We performed a cross-sectional study of associations between dialkylphosphate metabolites of organophosphates and semen quality among pesticide applicators in Majes (Arequipa), Peru. Methods Thirty-one men exposed to organophosphate (OP) pesticides and 31 non-exposed were recruited (age, 20–60 years). In exposed subjects, semen and a blood sample were obtained one day after the last pesticide application. Subjects were grouped according to levels of OP metabolites in urine. Semen samples were analyzed for sperm concentration, percentage of sperm motility, percentage of normal morphology, semen leucocytes and concentrations of fructose and zinc. Exposure to OP was assessed by measuring six urinary OP metabolites (dimethyl and diethyl phosphates and thiophosphates) by gas chromatography using a single flame photometric detector. Results Diethyldithiophosphate (p = 0.04) and diethylthiophosphate (p = 0.02) better reflected occupational pesticide exposure than other OP metabolites. Semen analysis revealed a significant reduction of semen volume and an increase in semen pH in men with OP metabolites. Multiple regression analysis showed that both occupational exposure to pesticides and the time of exposure to pesticides were more closely related to alterations in semen quality parameters than the single measurement of OP metabolites in urine. Conclusion The study demonstrated that occupational exposure to OP pesticides was more closely related to alterations in semen quality than a single measurement of urine OP metabolites. Current measurement of OP metabolites in urine may not reflect the full risk. PMID:19014632

  20. Teacher Reporting Attitudes Scale (TRAS): Confirmatory and Exploratory Factor Analyses with a Malaysian Sample

    ERIC Educational Resources Information Center

    Choo, Wan Yuen; Walsh, Kerryann; Chinna, Karuthan; Tey, Nai Peng

    2013-01-01

    The Teacher Reporting Attitude Scale (TRAS) is a newly developed tool to assess teachers' attitudes toward reporting child abuse and neglect. This article reports on an investigation of the factor structure and psychometric properties of the short form Malay version of the TRAS. A self-report cross-sectional survey was conducted with 667 teachers…

  1. Urethral anatomy and semen flow during ejaculation

    NASA Astrophysics Data System (ADS)

    Kelly, Diane

    2016-11-01

    Ejaculation is critical for reproductive success in many animals, but little is known about its hydrodynamics. In mammals, ejaculation pushes semen along the length of the penis through the urethra. Although the urethra also carries urine during micturition, the flow dynamics of micturition and ejaculation differ: semen is more viscous than urine, and the pressure that drives its flow is derived primarily from the rhythmic contractions of muscles at the base of the penis, which produce pulsatile rather than steady flow. In contrast, Johnston et al. (2014) describe a steady flow of semen through the crocodilian urethral groove during ejaculation. Anatomical differences of tissues associated with mammalian and crocodilian urethral structures may underlie these differences in flow behavior.

  2. Ejaculatory process and related semen characteristics.

    PubMed

    Bravo, P W; Moscoso, R; Alarcon, V; Ordoñez, C

    2002-01-01

    South American camelids are dribble ejaculators, and urethral contractions occur throughout copulation, which may last 25 min. The urethral contractions and their association with semen characteristics during copulation were determined in llamas and alpacas. A transrectal probe was held in the rectum of the male while copulating an artificial vagina, which was accessed underneath the dummy through a hole. The semen-collecting tube was changed every 5 min. Semen characteristics, color, volume, consistency, motility, concentration, and percentage of live sperm were determined at 5-min intervals. Urethral contractions were evenly distributed during copulation: 40 in alpacas and 63 in llamas (p < .05), with a general range of 11 to 132. Semen color was milky in 63%, and translucent in 36.5% for alpacas; and creamy (9.9%), milky (47%), and translucent (42%) for llamas. The mean volume of ejaculate was 0.3, 0.4, 0.6, 0.7, 0.6, 0.8, 0.3, and 3.0 mL for 5, 10, 15, 20, 25, and 30 min, respectively. Semen consistency was variable: viscous (65%) and semiviscous (34%) in alpacas; and viscous (57%) and semiviscous (42%) in llamas. Spermatic motility varied between 60 and 80% for the llama, and 40 and 80% for the alpaca. Spermatic concentration varied between 60 and 188 x 10(3)/mm3 in llamas, and 30 and 170 x 10(3)/mm3 in alpacas. Percentage of live sperm varied the least: 81 to 90% in llamas and 65 to 90% in alpacas. The ejaculate of llamas and alpacas is not fractionated, urethral contractions are evenly distributed, during copulation, and semen characteristics are present throughout the copulatory period.

  3. Strategies for Processing Semen from Subfertile Stallions for Cooled Transport.

    PubMed

    Varner, Dickson D

    2016-12-01

    Subfertility can be a confusing term because some semen of good quality can have reduced fertility following cooled transport if the semen is processed in an improper manner. General procedures aimed at processing stallion semen for cooled transport are well described. An array of factors could exist in reduced fertility of cool-transported semen. This article focuses on centrifugation techniques that can be used to maximize sperm quality of stallions whose semen is intended for cooled transport. Clinical cases are also provided for practical application of techniques.

  4. Effects of storage temperature and semen extender on stored canine semen.

    PubMed

    Hori, Tatsuya; Yoshikuni, Ryuta; Kobayashi, Masanori; Kawakami, Eiichi

    2014-03-01

    The objective of the present study was to determine an optimum temperature and extender for short-term transport of canine ejaculated semen. There was no significant difference in the qualities of semen diluted with two kinds of extender, egg yolk Tris-citrate fructose (EYT-FC) or glucose (EYT-GC) extender, between the 2, 8 or 12 and the 4°C control groups during storage for up to 48 hr, while the 16-24°C groups showed decreased sperm motility during storage for 48 hr. However, the 2°C group showed slightly lower sperm motility and slightly higher sperm abnormality than the 4°C group. Therefore, we concluded that semen qualities can be maintained for up to 48 hr when canine semen samples are extended with EYT-FC or EYT-GC and stored at a temperature in the range of 4-12°C.

  5. Effects of seasons on some semen parameters and bacterial contamination of Awassi ram semen.

    PubMed

    Azawi, O I; Ismaeel, M A

    2012-06-01

    The objectives of the present study were to determine the effects of season on some semen parameters and bacterial contamination of Awassi ram semen. Semen samples from six mature Awassi rams were used in this study. Semen collection was performed with artificial vagina every week, from September 2009 to October 2010. Volume, sperm concentration, mass motility, individual motility, percentage live sperm and sperm abnormalities were evaluated. Moreover, determination of viable bacterial count of the rams was also recorded weekly. Higher (p < 0.05) semen volume in the hot summer and spring months was observed of August (1.55 ± 0.08 ml) and March (1.27 ± 0.15 ml). Sperm concentration was highest (p < 0.05) in the breeding season (late summer to early autumn) of September (4.21 ± 0.86 × 10(9) sperm/ml). Sperm individual motility and percent of live sperm observed in August (summer) and May (end of spring) when the environmental temperature started to increase were recorded highest values and differed significantly (p < 0.05) from December and January (winter). The highest value of the mean sperm acrosomal defects (13.33 ± 0.63%) was recorded in December. The highest value of the mean viable bacterial count (138.3 ± 21.6) was recorded in July (summer). A significant decrease (p < 0.01) in the mean viable bacterial count was observed from the middle of winter towards the end of spring. The lowest bacterial count was noted in January (60.5 ± 2.98). It could be concluded from the results of the present study that there is an effect of season on ram semen quality, and summer high temperature in northern Iraq has no effect on Awassi ram semen. There is a significant effect of season on bacterial count on Awassi ram semen.

  6. Correlation of phthalate exposures with semen quality

    SciTech Connect

    Pant, Niraj Shukla, Manju; Kumar Patel, Devendra; Shukla, Yogeshwar; Mathur, Neeraj; Kumar Gupta, Yogendra; Saxena, Daya Krishna

    2008-08-15

    Phthalates are widely used man-made chemical released in the environment and human exposure is mainly through diet. As the phthalate plasticizers are not covalently bound to PVC, they can leach, migrate or evaporate into the environment and as a result have become ubiquitously contaminants. The present study investigates the correlation, if any, between the phthalate esters (DEP, DEHP, DBP, DMP, DOP) and sperm mitochondrial status, ROS, LPO, SCSA, and sperm quality. The study was conducted in the urban/rural population of Lucknow visiting Obstetrics and Gynecology Department, CSMMU, Lucknow. Semen analysis was performed according to the WHO guidelines while phthalate analysis by HPLC and LPO by spectrophotometer and the sperm mitochondrial status, ROS, SCSA using flow cytometry. The questionnaire data showed no significant difference in the demographic characteristics among the groups. In general, urban population was found to have statistically significant higher levels of phthalate esters than the rural. Further, infertile men showed statistically significant (p < 0.05) higher levels of pollutants in the semen than fertile men. A negative correlation between semen phthalate level viz DEHP and sperm quality and positive association with depolarized mitochondria, elevation in ROS production and LPO, DNA fragmentation was established. The findings are suggestive that phthalates might be one among the contributing factors associated with the deterioration in semen quality and these adverse effects might be ROS, LPO and mitochondrial dysfunction mediated.

  7. AIR POLLUTION EFFECTS ON SEMEN QUALITY

    EPA Science Inventory

    The potential impact of exposure to periods of high air pollution on male reproductive health was examined within the framework of an international project conducted in the Czech Republic. Semen quality was evaluated in young men (age 18) living in the Teplice District who are ex...

  8. A comparison of ABAcard(®) p30 and RSID™-Semen test kits for forensic semen identification.

    PubMed

    Boward, Emily S; Wilson, Stacey L

    2013-11-01

    The screening and confirmatory tests available to a forensic laboratory allow evidence to be examined for the presence of bodily fluids. With the majority of evidence being submitted involving sexual assaults, it is important to have confirmatory tests for the identification of semen that are straightforward, quick, and reliable. The purpose of this study was to compare two commonly used semen identification kits utilized by forensic laboratories: ABAcard(®) p30 and Rapid Stain Identification of Human Semen (RSID™-Semen). These kits were assessed with aged semen stains, fresh and frozen post-vasectomy semen, post-coital samples collected on different substrates, post-vasectomy semen mixed with blood, saliva, and urine, a series of swabs collected at increasing time intervals after sexual intercourse, and multiple non-semen samples. The test kits were compared on the basis of sensitivity, specificity, and the cost and time effectiveness of each protocol. Overall, both semen identification tests performed well in the studies. Both kits proved specificity for identifying semen, however the ABAcard(®) p30 test surpassed the RSID™-Semen test in sensitivity, cost per test, and simplified test protocol.

  9. Cryopreservation of Sambar deer semen in Thailand.

    PubMed

    Vongpralub, Thevin; Chinchiyanond, Wittaya; Hongkuntod, Pornchai; Sanchaisuriya, Pitcharat; Liangpaiboon, Sanan; Thongprayoon, Areeya; Somphol, Noppadon

    2015-01-01

    Little is known of the different freezing and thawing techniques for post-thaw survival of spermatozoa in Sambar deer. So, this study determined the effect of seminal plasma, egg yolk and glycerol extenders and their concentrations, plus cooling, freezing, and thawing protocols on the post-thaw quality of their semen. Semen samples were collected by electro-ejaculation from four Thai Sambar deer stags (Cervus unicolor equinus). As evaluated by post-thaw progressive motility and acrosome integrity removal of seminal plasma was beneficial; Tris-egg yolk was the most efficient extender; a 20% egg yolk concentration was better than the 0%, 10%, or 30%; and a 3% glycerol concentration was better than 5%, 7%, or 9%. Using the optimum dilution techniques, semen was loaded in 0.5 ml plastic straws. Cooling times from ambient temperature to 5°C in 3 hr resulted in higher post-thaw progressive motility and acrosome integrity than 1, 2, or 4 hr. Suspending the straws 4 cm above the surface for 15 min before plunging into liquid nitrogen was better than suspending at 2 or 6 cm. For thawing frozen semen, an intermediate thawing (50°C, 8 sec) protocol was more effective than the slower (37°C, 10 sec) or faster (70°C, 5 sec) thawing rates. Timed insemination following estrus synchronization of 10 hinds resulted in six confirmed pregnancies at 60 days. Five hinds delivered live fawns. This study provides an effective approach for semen cryopreservation and artificial insemination (AI), which should be valuable to scientists for genetics and reproductive management of Sambar deer in developing countries.

  10. Semen collection and fertility in naturally fertile sandhill cranes

    USGS Publications Warehouse

    Chen, G.; Gee, G.F.; Nicolich, Jane M.; Taylor, J.A.; Urbanek, R.P.; Stahlecker, D.W.

    1997-01-01

    Aviculturists often ask if semen collection will interfere with fertility in naturally fertile pairs of cranes. We used 12 naturally fertile Florida sandhill crane (Grus canadensis pratensis) pairs for this study, 6 control and 6 experimental. All pairs had produced fertile eggs in previous years and were in out-of-doors pens scattered throughout different pen complexes, within auditory range but physically isolated. Semen was collected on Tuesday mornings and Friday afternoons from 26 February 1993 to 4 June 1993. We used standard artificial insemination methods to collect and to evaluate the semen and spermatozoa. Semen collection did not affect semen quality or quantity. Semen volume, sperm density, sperm motility, sperm morphology, sperm live, sperm number per collection, and male response to semen collection exhibited significant daily variation (P < 0.05). Although semen collection began 13 days before the first egg in the experimental group, we observed no differences in the date of first egg laid or in fertility between experimental and control groups. Also, we observed no differences in the interval between clutches or in the percentage of broken eggs between experimental and control groups. Sires consistently producing better semen samples produced fewer fertile eggs than sires producing poorer semen samples (r = 0.60).

  11. Recognition of FT-IR Data Cuscutae Semen, Japanese Dodder, and Sinapis Semen Using Discrete Wavelet Transformation and RBF Networks

    PubMed Central

    Hu, Tao; Weng, Xuexiang; Xu, Lishan; Cheng, Cungui; Yu, Peng

    2013-01-01

    Horizontal attenuation total reflection Fourier transformation infrared spectroscopy (HATR-FT-IR) studies on cuscutae semen and its confusable varieties Japanese dodder and sinapis semen combined with discrete wavelet transformation (DWT) and radial basis function (RBF) neural networks have been conducted in order to classify them. DWT is used to decompose the FT-IRs of cuscutae semen, Japanese dodder, and sinapis semen. Two main scales are selected as the feature extracting space in the DWT domain. According to the distribution of cuscutae semen, Japanese dodder, and sinapis semen's FT-IRs, three feature regions are determined at detail 3, and two feature regions are determined at detail 4 by selecting two scales in the DWT domain. Thus five feature parameters form the feature vector. The feature vector is input to the RBF neural networks to train so as to accurately classify the cuscutae semen, Japanese dodder, and sinapis semen. 120 sets of FT-IR data are used to train and test the proposed method, where 60 sets of data are used to train samples, and another 60 sets of FT-IR data are used to test samples. Experimental results show that the accurate recognition rate of cuscutae semen, Japanese dodder, and sinapis semen is average of 100.00%, 98.33%, and 100.00%, respectively, following the proposed method. PMID:24282653

  12. Recognition of FT-IR Data Cuscutae Semen, Japanese Dodder, and Sinapis Semen Using Discrete Wavelet Transformation and RBF Networks.

    PubMed

    Hu, Tao; Weng, Xuexiang; Xu, Lishan; Cheng, Cungui; Yu, Peng

    2013-01-01

    Horizontal attenuation total reflection Fourier transformation infrared spectroscopy (HATR-FT-IR) studies on cuscutae semen and its confusable varieties Japanese dodder and sinapis semen combined with discrete wavelet transformation (DWT) and radial basis function (RBF) neural networks have been conducted in order to classify them. DWT is used to decompose the FT-IRs of cuscutae semen, Japanese dodder, and sinapis semen. Two main scales are selected as the feature extracting space in the DWT domain. According to the distribution of cuscutae semen, Japanese dodder, and sinapis semen's FT-IRs, three feature regions are determined at detail 3, and two feature regions are determined at detail 4 by selecting two scales in the DWT domain. Thus five feature parameters form the feature vector. The feature vector is input to the RBF neural networks to train so as to accurately classify the cuscutae semen, Japanese dodder, and sinapis semen. 120 sets of FT-IR data are used to train and test the proposed method, where 60 sets of data are used to train samples, and another 60 sets of FT-IR data are used to test samples. Experimental results show that the accurate recognition rate of cuscutae semen, Japanese dodder, and sinapis semen is average of 100.00%, 98.33%, and 100.00%, respectively, following the proposed method.

  13. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... POULTRY) AND ANIMAL PRODUCTS IMPORTATION OF CERTAIN ANIMAL EMBRYOS AND ANIMAL SEMEN Certain Animal Semen... clinical signs of scrapie at the time of semen collection nor developed scrapie between the time of...

  14. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... POULTRY) AND ANIMAL PRODUCTS IMPORTATION OF CERTAIN ANIMAL EMBRYOS AND ANIMAL SEMEN Certain Animal Semen... clinical signs of scrapie at the time of semen collection nor developed scrapie between the time of...

  15. Involuntary reduction in vigour of calves born from sexed semen.

    PubMed

    Djedović, Radica; Bogdanović, Vladan; Stanojević, Dragan; Nemes, Zsolt; Gáspárdy, András; Cseh, Sándor

    2016-06-01

    The objective of this study was to compare the reproductive traits of heifers and the development characteristics of their calves following artificial insemination (AI) with sexed and non-sexed semen. The analysed characteristics included conception rate, gestation length, calf birth weight, calf vigour, stillbirth rate, and twinning rate. Data of 530 calves produced with sexed and 1,163 calves produced with non-sexed semen were analysed. The General Linear Model (GLM) was applied to assess the influence of semen type, farm, season of insemination, the calf's sex and the inseminating sire on gestation length and calf birth weight. With the exception of gestation length (P > 0.05), all other traits studied were significantly (P < 0.01) influenced by the type of semen. The conception rate was 55% for conventional and 44% for sexed semen, and the average gestation length was 274.6 and 274.9 days, respectively. The mean calf birth weight was 37.47 kg for non-sexed and 36.75 kg for sexed semen. The stillbirth rate was 6.19% for conventional and 7.54% for sexed semen, while the twinning rate was 3.78% for conventional and 1.13% for sexed semen. The calves produced with non-sexed and sexed semen differed significantly in viability (P < 0.001), the latter having a lower calf vigour score. The use of conventional semen did not affect the ratio of female and male calves (52.7:47.3%; P > 0.05); however, artificial insemination with X-sorted sexed semen significantly altered the sex ratio of calves (85.1:14.9%, P < 0.01). The results obtained in this investigation are in agreement with the majority of studies which compared the fertility traits, sex ratio and calf characteristics depending on the application of artificial insemination with sexed or conventional semen.

  16. Sperm ubiquitination in epididymal feline semen.

    PubMed

    Vernocchi, Valentina; Morselli, Maria Giorgia; Varesi, Sara; Nonnis, Simona; Maffioli, Elisa; Negri, Armando; Tedeschi, Gabriella; Luvoni, Gaia Cecilia

    2014-09-01

    Ubiquitin is a 8.5-kDa peptide that tags other proteins for proteasomal degradation. It has been proposed that ubiquitination might be responsible for the elimination of defective spermatozoa during transit through the epididymis in humans and cattle, but its exact biological function in seminal plasma has not yet been clarified. In the domestic cat (Felis catus), the percentage of immature, unviable, and abnormal spermatozoa decreases during the epididymal transit, indicating the existence of a mechanism that removes defective spermatozoa. Magnetic cell separation techniques, based on the use of magnetic beads coated with anti-ubiquitin antibodies, may allow the selective capture of ubiquitinated spermatozoa from semen, thus contributing to the identification of a potential correlation between semen quality and ubiquitination process. Moreover, the selective identification of all the ubiquitinated proteins in different epididymal regions could give a better understanding of the ubiquitin role in feline sperm maturation. The aims of this study were as follows: (1) to verify the possibility of separating ubiquitinated spermatozoa with magnetic ubiquitin beads and identify the morphological and acrosomal differences between whole sample and unbound gametes, (2) to characterize all the ubiquitinated proteins in spermatozoa retrieved in the three epididymal regions by a proteomic approach. The data indicated the presence of ubiquitinated proteins in cat epididymal semen. However, a correlation between abnormal and ubiquitinated spermatozoa has not been found, and ubiquitin cannot be considered as a biomarker of quality of epididymal feline spermatozoa. To the author's knowledge, this is the first identification of all the ubiquitinated proteins of cat spermatozoa collected from different epididymal regions. The proteomic pattern allows a further characterization of cat epididymal semen and represents a contribute to a better understanding of the ubiquitin role in

  17. Zooplankton Feeding on the Nuisance Flagellate Gonyostomum semen

    PubMed Central

    Johansson, Karin S. L.; Vrede, Tobias; Lebret, Karen; Johnson, Richard K.

    2013-01-01

    The large bloom-forming flagellate Gonyostomum semen has been hypothesized to be inedible to naturally occurring zooplankton due to its large cell size and ejection of long slimy threads (trichocysts) induced by physical stimulation. In a grazing experiment using radiolabelled algae and zooplankton collected from lakes with recurring blooms of G. semen and lakes that rarely experience blooms, we found that Eudiaptomus gracilis and Holopedium gibberum fed on G. semen at high rates, whereas Daphnia cristata and Ceriodaphnia spp. did not. Grazing rates of E. gracilis were similar between bloom-lakes and lakes with low biomass of G. semen, indicating that the ability to feed on G. semen was not a result of local adaptation. The high grazing rates of two of the taxa in our experiment imply that some of the nutrients and energy taken up by G. semen can be transferred directly to higher trophic levels, although the predominance of small cladocerans during blooms may limit the importance of G. semen as a food resource. Based on grazing rates and previous observations on abundances of E. gracilis and H. gibberum, we conclude that there is a potential for grazer control of G. semen and discuss why blooms of G. semen still occur. PMID:23667489

  18. Chronic boron exposure and human semen parameters.

    PubMed

    Robbins, Wendie A; Xun, Lin; Jia, Juan; Kennedy, Nola; Elashoff, David A; Ping, Liu

    2010-04-01

    Boron found as borates in soil, food, and water has important industrial and medical applications. A panel reviewing NTP reproductive toxicants identified boric acid as high priority for occupational studies to determine safe versus adverse reproductive effects. To address this, we collected boron exposure/dose measures in workplace inhalable dust, dietary food/fluids, blood, semen, and urine from boron workers and two comparison worker groups (n=192) over three months and determined correlations between boron and semen parameters (total sperm count, sperm concentration, motility, morphology, DNA breakage, apoptosis and aneuploidy). Blood boron averaged 499.2 ppb for boron workers, 96.1 and 47.9 ppb for workers from high and low environmental boron areas (p<0.0001). Boron concentrated in seminal fluid. No significant correlations were found between blood or urine boron and adverse semen parameters. Exposures did not reach those causing adverse effects published in animal toxicology work but exceeded those previously published for boron occupational groups.

  19. [Analysis and identification of Semen Glycines Nigrae and Semen Pharbitidis by infrared spectroscopy].

    PubMed

    Du, Juan; Peng, Xi-Yuan; Ma, Fang; Chen, Jian-Bo; Zhou, Qun; Jin, Zhe-Xiong; Sun, Su-Qin

    2014-09-01

    Semen Glycines Nigrae and Semen Pharbitidis containing a large amount of fats and proteins are commonly used in Chinese herbal medicine. Tri-step infrared spectroscopy was applied to fast analyze and identify the two samples. In the conventional infrared spectroscopy, the samples both have obvious characteristic absorption peaks at 1,745 cm(-1) assigned to the stretching mode of C==O in esters. Furthermore, the two kinds of herbs have the peaks at 1,656 and 1,547 cm(-1) assigned to the amide I and II bands of protein. Obviously, the infrared spectra of herbs demonstrate that protein and fat is the major component in two kinds of herbs, and the relative intensity of the peaks assigned to fat and protein indicate their relative content is different. And the result is consistent with the reported. In the second derivative spectra, Semen Pharbitidis has a peak at 1,712 cm(-1) assigned to the organic acid, however, Semen Glycines Nigrae has not this absorption peak. In addition, in the second derivative spectra, appeared more differences between the two samples in shape and intensity of the peaks. In two-dimensional correlation infrared spectra, the two samples were visually distinguished due to their significant differences in auto-peak position and intensity. In the region of 1,500-1,700 cm(-1), Semen Glycines Nigrae has two autopeaks and Semen Pharbitidis has three autopeaks. In the region of 2,800-3,000 cm(-1), the samples both have two autopeaks, but the position of the strongest autopeak is different. It was demonstrated that the Tri-step infrared spectroscopy were successfully applied to fast analyze and identify the two kinds of samples containing the same major component, and made sure the foundation for future researches.

  20. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis

    PubMed Central

    Anand, Mukul; Yadav, Sarvajeet

    2016-01-01

    Aim: The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. Materials and Methods: A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck). Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Results: Significantly (p<0.01) higher value of live percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (p<0.01) higher followed by slow and rapid progression. Conclusion: This study showed that the best results for motion, vitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats. PMID:27051209

  1. Semen phthalate metabolites, semen quality parameters and serum reproductive hormones: A cross-sectional study in China.

    PubMed

    Wang, Yi-Xin; Zeng, Qiang; Sun, Yang; Yang, Pan; Wang, Peng; Li, Jin; Huang, Zhen; You, Ling; Huang, Yue-Hui; Wang, Cheng; Li, Yu-Feng; Lu, Wen-Qing

    2016-04-01

    Exposure to phthalates has been found to have adverse effects on male reproductive function in animals. However, the findings from human studies are inconsistent. Here we examined the associations of phthalate exposure with semen quality and reproductive hormones in a Chinese population using phthalate metabolite concentrations measured in semen as biomarkers. Semen (n = 687) and blood samples (n = 342) were collected from the male partners of sub-fertile couples who presented to the Reproductive Center of Tongji Hospital in Wuhan, China. Semen quality parameters and serum reproductive hormone levels were determined. Semen concentrations of 8 phthalate metabolites were assessed using high-performance liquid chromatography and tandem mass spectrometry. Associations of the semen phthalate metabolites with semen quality parameters and serum reproductive hormones were assessed using confounder-adjusted linear and logistic regression models. Semen phthalate metabolites were significantly associated with decreases in semen volume [mono-n-butyl phthalate (MBP), mono-(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-oxohexyl) phthalate (MEOHP)], sperm curvilinear velocity [monobenzyl phthalate (MBzP), MEHP, the percentage of di-(2-ethylhexyl)-phthalate metabolites excreted as MEHP (%MEHP)], and straight-line velocity (MBzP, MEHP, %MEHP), and also associated with an increased percentage of abnormal heads and tails (MBzP) (all p for trend <0.05). These associations remained suggestive or significant after adjustment for multiple testing. There were no significant associations between semen phthalate metabolites and serum reproductive hormones. Our findings suggest that environmental exposure to phthalates may impair human semen quality.

  2. Teacher Reporting Attitudes Scale (TRAS): confirmatory and exploratory factor analyses with a Malaysian sample.

    PubMed

    Choo, Wan Yuen; Walsh, Kerryann; Chinna, Karuthan; Tey, Nai Peng

    2013-01-01

    The Teacher Reporting Attitude Scale (TRAS) is a newly developed tool to assess teachers' attitudes toward reporting child abuse and neglect. This article reports on an investigation of the factor structure and psychometric properties of the short form Malay version of the TRAS. A self-report cross-sectional survey was conducted with 667 teachers in 14 randomly selected schools in Selangor state, Malaysia. Analyses were conducted in a 3-stage process using both confirmatory (stages 1 and 3) and exploratory factor analyses (stage 2) to test, modify, and confirm the underlying factor structure of the TRAS in a non-Western teacher sample. Confirmatory factor analysis did not support a 3-factor model previously reported in the original TRAS study. Exploratory factor analysis revealed an 8-item, 4-factor structure. Further confirmatory factor analysis demonstrated appropriateness of the 4-factor structure. Reliability estimates for the four factors-commitment, value, concern, and confidence-were moderate. The modified short form TRAS (Malay version) has potential to be used as a simple tool for relatively quick assessment of teachers' attitudes toward reporting child abuse and neglect. Cross-cultural differences in attitudes toward reporting may exist and the transferability of newly developed instruments to other populations should be evaluated.

  3. Seasonal and cryopreservation impacts on semen quality in boars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seasonal boar infertility occurs worldwide and contributes to economic loss to the pork industry. The current study evaluated cooled vs cryopreserved semen quality of 11 Duroc boars collected in June (cool season) and August 2014 (warm season). Semen was cooled to 16°C (cooled) or frozen over liquid...

  4. Association of oxidative status and semen characteristics with seminal plasma proteins of buffalo semen

    PubMed Central

    Sharma, L.; Pandey, V.; Nigam, R.; Saxena, A.; Swain, D. K.; Yadav, B.

    2016-01-01

    To study the influence of season on oxidative status of buffalo semen and their association with semen characteristics and seminal plasma proteins, ejaculates were collected twice a week in winter, summer and rainy seasons from six adult Bhadawari buffalo bulls. The neat semen was analyzed for semen characteristics immediately after collection and oxidative status viz. lipid peroxidation (LPO), catalase (CAT), super oxide dismutase (SOD) activity, and total protein (TP) were estimated in seminal plasma. The protein profiling was carried out by one-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The significant effect of season was observed on TP, SOD activity and % protein fractions of seminal plasma proteins of buffalo bulls. The TP values showed positive correlation with ejaculate volume (EV), sperm concentration (SC), and % live-dead (LD) and negative correlation with progressive motility (PM), and hypo-osmotic swelling test (HOST). The SOD activity showed positive correlation with PM, LD, HOST and % acrosoamal integrity (AI). Besides that, results showed correlation of TP with 6.5, 38 and 66 kDa proteins, LPO with 70, 72, 84 and 86 kDa proteins, CAT with 70 kDa and 86 kDa proteins, and SOD with 6.5, 24.5, 44.5, 70 and 72 kDa proteins. In conclusion, this study indicated that TP and SOD activity of seminal plasma of buffalo bulls were influenced by season and were found to be associated with some of the semen characteristics and expression of seminal plasma proteins. PMID:28224004

  5. Perfluorochemicals and Human Semen Quality: The LIFE Study

    PubMed Central

    Chen, Zhen; Schisterman, Enrique F.; Kim, Sungduk; Sweeney, Anne M.; Sundaram, Rajeshwari; Lynch, Courtney D.; Gore-Langton, Robert E.; Barr, Dana Boyd

    2014-01-01

    Background: The relation between persistent environmental chemicals and semen quality is evolving, although limited data exist for men recruited from general populations. Objectives: We examined the relation between perfluorinated chemicals (PFCs) and semen quality among 501 male partners of couples planning pregnancy. Methods: Using population-based sampling strategies, we recruited 501 couples discontinuing contraception from two U.S. geographic regions from 2005 through 2009. Baseline interviews and anthropometric assessments were conducted, followed by blood collection for the quantification of seven serum PFCs (perfluorosulfonates, perfluorocarboxylates, and perfluorosulfonamides) using tandem mass spectrometry. Men collected a baseline semen sample and another approximately 1 month later. Semen samples were shipped with freezer packs, and analyses were performed on the day after collection. We used linear regression to estimate the difference in each semen parameter associated with a one unit increase in the natural log–transformed PFC concentration after adjusting for confounders and modeling repeated semen samples. Sensitivity analyses included optimal Box-Cox transformation of semen quality end points. Results: Six PFCs [2-(N-methyl-perfluorooctane sulfonamido) acetate (Me-PFOSA-AcOH), perfluorodecanoate (PFDeA), perfluorononanoate (PFNA), perfluorooctane sulfonamide (PFOSA), perfluorooctane sulfonate (PFOS), and perfluorooctanoic acid (PFOA)] were associated with 17 semen quality end points before Box-Cox transformation. PFOSA was associated with smaller sperm head area and perimeter, a lower percentage of DNA stainability, and a higher percentage of bicephalic and immature sperm. PFDeA, PFNA, PFOA, and PFOS were associated with a lower percentage of sperm with coiled tails. Conclusions: Select PFCs were associated with certain semen end points, with the most significant associations observed for PFOSA but with results in varying directions. Citation

  6. Risk of Salmonella transmission via cryopreserved semen in turkey flocks.

    PubMed

    Iaffaldano, N; Reale, A; Sorrentino, E; Coppola, R; Di Iorio, M; Rosato, M P

    2010-09-01

    To investigate the possibility to carry pathogen bacteria in turkey flocks via cryopreserved semen, research was carried out 1) to investigate the microbial contamination of fresh and frozen thawed turkey semen and 2) to evaluate the effect of the freezing-thawing process on the survival of 3 serovars of Salmonella spp. experimentally inoculated in turkey semen. Five pools of semen diluted 4-fold were cooled, added with 8% of dimethylacetamide as a cryoprotectant, and aliquots of 80 muL were directly plunged into liquid nitrogen to form frozen pellets. Mesophilic viable counts, total and fecal coliforms, Enterobacteriaceae, enterococci, Campylobacter spp., and Salmonella spp. were investigated on fresh and thawed samples. Further, 5 pools of diluted semen were each divided into 3 subsamples, inoculated with 7.8 +/- 0.2 log cfu.mL(-1) of Salmonella Liverpool, Salmonella Montevideo, and Salmonella Braenderup, respectively, and cryopreserved before to assess the postthaw viability of Salmonella spp. strains. Fresh semen was highly contaminated by all of the saprophytic bacteria investigated and the cryopreservation process reduced the amount of mesophilic viable count and total coliforms (P < 0.05) and fecal coliforms, Enterobacteriaceae, and enterococci (P < 0.01) by about 1 log cfu.mL(-1). Conversely, neither Campylobacter spp. nor Salmonella spp. were found as endogenous bacteria in semen. In the inoculated semen, both Salmonella Liverpool, Salmonella Montevideo, and Salmonella Braenderup colonies were recovered postthaw, showing a significant reduction of 2.03 +/- 0.28, 3.08 +/- 0.22, and 2.72 +/- 0.23 log cfu.mL(-1), respectively, compared with the fresh semen (P < 0.001). In conclusion, the cryopreservation process allowed us to obtain a low reduction of microbial count both in endogenous saprophytic bacteria and artificially inoculated Salmonella spp. strains; therefore, the possibility of Samonella spp. transmission to flocks through the use of infected

  7. A successful new approach to honeybee semen cryopreservation.

    PubMed

    Wegener, Jakob; May, Tanja; Kamp, Günter; Bienefeld, Kaspar

    2014-10-01

    Honeybee biodiversity is under massive threat, and improved methods for gamete cryopreservation could be a precious tool for both the in situ- and ex situ-conservation of subspecies and ecotypes. Recent cryoprotocols for drone semen have improved the viability and fertility of frozen-thawed semen by using increased diluent:semen-ratios, but there is still much room for progress. As semen cryopreserved after dilution often appeared hyperactive, we speculated that the disruption of sperm-sperm interactions during dilution and cryopreservation could reduce the fertile lifespan of the cells. We therefore developed protocols to reduce admixture, or abolish it altogether by dialyzing semen against a hypertonic solution of cryoprotectant. Additionally, we tested methods to reduce the cryoprotectant concentration after thawing. Insemination of queens with semen cryopreserved after dialysis yielded 49%, 59% and 79% female (= stemming from fertilized eggs) pupae in three separate experiments, and the numbers of sperm found in the spermathecae of the queens were significantly higher than those previously reported. Post-thaw dilution and reconcentration of semen for cryoprotectant removal reduced fertility, but sizeable proportions of female brood were still produced. Workers stemming from cryopreserved semen did not differ from bees stemming from untreated semen with regard to indicators of fluctuating asymmetry, but were slightly heavier. Cryopreservation after dialysis tended to increase the proportion of cells with DNA-nicks, as measured by the TUNEL-assay, but this increase appears small when compared to the baseline variations of this indicator. Overall, we conclude that cryoprotectant-addition through dialysis can improve the quality of cryopreserved drone semen. Testing of offspring for vitality and genetic integrity should continue.

  8. Effect of semen extender and storage temperature on ram sperm motility over time

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Storage of ram semen for long period of time depends on a number of factors, including type of extender and storage temperature. A study compared the effect of semen extender and storage temperature on motility of ram semen stored for 72 h. Semen collected via electroejaculator from 5 mature Katahd...

  9. Application of liquid semen technology improves conception rate of sex-sorted semen in lactating dairy cows.

    PubMed

    Xu, Z Z

    2014-11-01

    The objective was to compare reproductive performance of liquid sex-sorted (SS) semen with that of conventional (CON) semen in lactating dairy cows. Between 2011 and 2013, commercial dairy herds (n = 101, 203, and 253 for 2011, 2012, and 2013, respectively) with predominantly Holstein-Friesian cows were enrolled in a contract mating program to produce surplus heifers for export using liquid SS semen. During the spring mating period, each herd was allocated with liquid SS semen at 50% of its daily requirement and the remaining daily requirement was allocated with CON liquid semen. Sperm for producing SS semen was sorted by Sexing Technologies NZ Ltd. (Hamilton, New Zealand) and then packaged using the liquid semen technology of LIC (Hamilton, New Zealand) at a dose of 1 × 10(6) sperm. Artificial insemination (AI) with liquid SS semen was carried out between 43 and 46 h after collection. Conventional semen straws contained 1.25 × 10(6), 1.75 × 10(6), or 2 × 10(6) sperm for semen to be used on d 1, 2, or 3 after collection, respectively. Only CON inseminations on the same days as when SS semen was used were included in the comparison. Herd managers biased usage of SS semen toward cows with a longer postpartum interval before the mating start date (64.0 vs. 62.8 d), cows of higher genetic merit (NZ$107.0 vs. NZ$98.4), younger cows (5.1 vs. 5.2 yr), and cows in which they had more confidence of being genuinely in estrus as measured by a lower percentage of short returns between 1 and 17 d (5.3 vs. 7.5%). After adjusting for these factors, the estimated difference in nonreturn rate between AI with SS and CON semen over the 3 seasons was -3.8 percentage points (SS = 70.2% vs. CON = 74.0%; SS/CON = 94.9%). The estimated maximum difference in calving rate per AI between SS and CON semen was -3.1 percentage points for 2011 (SS = 51.2% vs. CON = 54.3%; SS/CON = 94.3%) and -3.0 percentage points for 2012 (SS = 49.7% vs. CON = 52.6%; SS/CON = 94.5%). Calving data for 2013

  10. Environmental Exposure to Triclosan and Semen Quality.

    PubMed

    Zhu, Wenting; Zhang, Hao; Tong, Chuanliang; Xie, Chong; Fan, Guohua; Zhao, Shasha; Yu, Xiaogang; Tian, Ying; Zhang, Jun

    2016-02-17

    Triclosan (2,4,4'-trichloro-2'-hydroxy-diphenyl ether, TCS) is widely used in personal care, household, veterinary and industrial products. It was considered as a potential male reproductive toxicant in previous in vitro and in vivo studies. However, evidence from human studies is scarce. Our study aims to investigate the relationship between TCS exposure and semen quality. We measured urinary TCS concentrations in 471 men recruited from a male reproductive health clinic. TCS was detected in 96.7% of urine samples, with a median concentration of 0.97 ng (mg·creatinine)(-1) (interquartile range, 0.41-2.95 ng (mg·creatinine)(-1)). A multiple linear regression analysis showed a negative association between natural logarithm (Ln) transformed TCS concentration (Ln-TCS) and Ln transformed number of forward moving sperms (adjusted coefficient β = -0.17; 95% confidence interval (CI) (-0.32, -0.02). Furthermore, among those with the lowest tertile of TCS level, Ln-TCS was negatively associated with the number of forward moving sperms (β = -0.35; 95% CI (-0.68, -0.03)), percentage of sperms with normal morphology (β = -1.64; 95% CI (-3.05, -0.23)), as well as number of normal morphological sperms, sperm concentration and count. Our findings suggest that the adverse effect of TCS on semen quality is modest at the environment-relevant dose in humans. Further studies are needed to confirm our findings.

  11. Persistent Organic Pollutants and Semen Quality: The LIFE Study

    PubMed Central

    Mumford, Sunni L.; Kim, Sungduk; Chen, Zhen; Gore-Langton, Robert E.; Barr, Dana Boyd; Buck Louis, Germaine M.

    2014-01-01

    Growing evidence suggests that persistent environmental chemicals such as polychlorinated biphenyls may adversely affect human fecundity. The purpose of this study was to evaluate associations between persistent environmental chemicals and semen quality among 501 male partners of couples discontinuing contraception for purposes of becoming pregnant. Men provided a blood specimen and two fresh semen samples collected approximately a month apart that underwent next day analysis for 35 semen quality endpoints. Serum samples were analyzed for 36 polychlorinated biphenyls (congeners #18, 28, 44, 49, 52, 66, 74, 87, 99, 101, 114, 118, 128, 138, 146, 149, 151, 153, 156, 157, 167, 170, 172, 177, 178, 180, 183, 187, 189, 194, 195, 196, 201, 206, 209); 1 polybrominated biphenyl (#153); 9 organochlorine pesticides; and 10 polybrominated diphenyl ethers (congeners #17, 28, 47, 66, 85, 99, 100, 153, 154, 183) using high resolution mass spectrometry. To estimate the effect of chemicals on semen quality, we regressed each semen marker on each chemical while adjusting for research site, age, body mass index, serum lipids, and cotinine levels. Males with chemical concentrations in the fourth quartile, as compared to the first quartile, showed significant associations for several individual chemicals in each chemical class and type of semen quality parameter indicating negative and positive associations with semen quality. Polybrominated diphenyl ethers in particular were associated with several measures of increased abnormal morphology. These exploratory results highlight the role of environmental influences on male fecundity, and are of particular interest given the ubiquitous exposures to these compounds. PMID:25441930

  12. Effects of sperm concentration at semen collection and storage period of frozen semen on dairy cow conception.

    PubMed

    Haugan, T; Gröhn, Y T; Kommisrud, E; Ropstad, E; Reksen, O

    2007-01-01

    The present study was based on data obtained from artificial inseminations (AIs) performed with cryopreserved semen from elite bulls used in the Norwegian breeding program. Semen was diluted to standardize the number of spermatozoa to 18 million per AI dose. The aim of the study was to investigate whether the net sperm concentration at semen collection and the storage period in liquid nitrogen have any effect on probability of conception in dairy cattle. We demonstrated that the natural range of sperm concentration at semen collection within some of the bulls was associated with the probability of conception. However, no primary trend among bulls was found on the effect of sperm concentration at semen collection. This appears to be due to differences among bulls in their response to the dilution ratio of seminal plasma to extender. The effect of storage time was investigated in semen that had been stored between 1000 days and 2400 days in AI straws in liquid nitrogen at the AI center. Our findings showed that use of semen with the longest storage period, i.e. 1951-2400 days, resulted in a more than one percentage point lower probability of conception than semen with a shorter storage period. In conclusion, the net sperm concentration at semen collection, which affects the dilution ratio of seminal plasma to extender, should be considered individually among bulls to achieve optimal reproductive performance. Furthermore, this study gives support to the idea that a measurable degree of damage to the spermatozoa could occur during the preservation time in liquid nitrogen.

  13. Structural analysis of TRAS1, a novel family of telomeric repeat-associated retrotransposons in the silkworm, Bombyx mori.

    PubMed Central

    Okazaki, S; Ishikawa, H; Fujiwara, H

    1995-01-01

    We characterized TRAS1, a retrotransposable element which was inserted into the telomeric repetitive sequence (CCTAA)n of the silkworm, Bombyx mori. The complete sequence of TRAS1, a stretch of 7.8 kb with a poly(A) tract at the 3' end, was determined. No long terminal repeat (LTR) was found at the termini of the element. TRAS1 contains gag- and pol-like open reading frames (ORFs) which are similar to those of non-LTR retrotransposons. The two ORFs overlap but are one nucleotide out of frame (+1 frameshift). Most of the approximately 250 copies of TRAS1 elements in the genome were highly conserved in the structure. Chromosomal in situ hybridization showed that TRAS1 elements are clustered at the telomeres of Bombyx chromosomes. A phylogenetic analysis using the amino acid sequence of the reverse transcriptase domain within the pol-like ORF revealed that TRAS1 falls into one lineage with R1, which is a family of non-LTR retrotransposons inserted into the same site within the 28S ribosomal DNA unit in most insects. TRAS1 may have been derived from R1 and changed the target specificity so that TRAS1 inserts into the telomeric repetitive sequence (CCTAA)n. Southern hybridization and Bal 31 exonuclease analyses showed that TRAS1 elements are clustered proximal to the terminal long tract of (CCTAA)n. TRAS1 is a novel family of non-LTR retrotransposons which are inserted into the telomeric repetitive sequences as target sites. PMID:7623845

  14. History of commercializing sexed semen for cattle.

    PubMed

    Garner, D L; Seidel, G E

    2008-04-15

    Although the basic principles controlling the sex of mammalian offspring have been known for a relatively long time, recent application of certain modern cellular methodologies has led to development of a flow cytometric system capable of differentiating and separating living X- and Y-chromosome-bearing sperm in amounts suitable for AI and therefore, commercialization of this sexing technology. After a very long history of unsuccessful attempts to differentiate between mammalian sperm that produce males from those that produce females, a breakthrough came in 1981 when it was demonstrated that precise DNA content could be measured. Although these initial measurements of DNA content killed the sperm in the process, they led to the ultimate development of a sperm sorting system that was capable, not only of differentiating between live X- and Y-sperm, but of sorting them into relatively pure X- and Y-sperm populations without obvious cellular damage. Initial efforts to predetermine the sex of mammalian offspring in 1989 required surgical insemination, but later enhancements provided sex-sorted sperm in quantities suitable for use with IVF. Subsequent advances in flow sorting provided minimal numbers of sperm sufficient for use in AI. It was not until the flow cytometric sorting system was improved greatly and successful cryopreservation of sex-sorted bull sperm was developed that efficacious approaches to commercialization of sexed semen could be implemented worldwide in cattle. A number of companies now offer sex-sorted bovine sperm. Innovative approaches by a diverse group of scientists along with advances in computer science, biophysics, cell biology, instrumentation, and applied reproductive physiology provided the basis for commercializing sexed semen in cattle.

  15. Environmental Exposure to Triclosan and Semen Quality

    PubMed Central

    Zhu, Wenting; Zhang, Hao; Tong, Chuanliang; Xie, Chong; Fan, Guohua; Zhao, Shasha; Yu, Xiaogang; Tian, Ying; Zhang, Jun

    2016-01-01

    Triclosan (2,4,4′-trichloro-2′-hydroxy-diphenyl ether, TCS) is widely used in personal care, household, veterinary and industrial products. It was considered as a potential male reproductive toxicant in previous in vitro and in vivo studies. However, evidence from human studies is scarce. Our study aims to investigate the relationship between TCS exposure and semen quality. We measured urinary TCS concentrations in 471 men recruited from a male reproductive health clinic. TCS was detected in 96.7% of urine samples, with a median concentration of 0.97 ng (mg·creatinine)−1 (interquartile range, 0.41–2.95 ng (mg·creatinine)−1). A multiple linear regression analysis showed a negative association between natural logarithm (Ln) transformed TCS concentration (Ln-TCS) and Ln transformed number of forward moving sperms (adjusted coefficient β = −0.17; 95% confidence interval (CI) (−0.32, −0.02). Furthermore, among those with the lowest tertile of TCS level, Ln-TCS was negatively associated with the number of forward moving sperms (β = −0.35; 95% CI (−0.68, −0.03)), percentage of sperms with normal morphology (β = −1.64; 95% CI (−3.05, −0.23)), as well as number of normal morphological sperms, sperm concentration and count. Our findings suggest that the adverse effect of TCS on semen quality is modest at the environment-relevant dose in humans. Further studies are needed to confirm our findings. PMID:26901211

  16. Evidence of excretion of Schmallenberg virus in bull semen

    PubMed Central

    2014-01-01

    Schmallenberg virus (SBV) is a novel orthobunyavirus, discovered in Germany in late 2011. It mainly infects cattle, sheep and goats and could lead to congenital infection, causing abortion and fetal abnormalities. SBV is transmitted by biting midges from the Culicoides genus and there is no evidence that natural infection occurs directly between ruminants. Here, we could detect SBV RNA in infected bull semen using qRT-PCR (three bulls out of seven tested positive; 29 positive semen batches out of 136). We also found that highly positive semen batches from SBV infected bulls can provoke an acute infection in IFNAR-/- mice, suggesting the potential presence of infectious virus in the semen of SBV infected bulls. PMID:24708245

  17. Indian story on semen loss and related Dhat syndrome

    PubMed Central

    Prakash, Om; Kar, Sujit Kumar; Sathyanarayana Rao, T. S.

    2014-01-01

    India is a country of many religions and ancient cultures. Indian culture is largely directed by the Vedic culture since time immemorial. Later Indian culture is influenced by Buddhism, Islam, and Christianity. Indian belief system carries the footprints of these cultures. Every culture describes human behaviors and an interpretation of each human behavior is largely influenced by the core cultural belief system. Sexuality is an important domain which is colored by different cultural colors. Like other cultures, Indian culture believes “semen” as the precious body fluid which needs to be preserved. Most Indian beliefs consider loss of semen as a threat to the individual. Ancient Indian literature present semen loss as a negative health related event. Dhat syndrome (related to semen loss) is a culture-bound syndrome seen in the natives of Indian subcontinent. This article gathers the Indian concepts related to semen loss. It also outlines belief systems behind problems of Dhat syndrome. PMID:25568479

  18. Risk factors for bacterial contamination during boar semen collection.

    PubMed

    Goldberg, Ana Maria G; Argenti, Laura E; Faccin, Jamil E; Linck, Lídia; Santi, Mônica; Bernardi, Mari Lourdes; Cardoso, Marisa R I; Wentz, Ivo; Bortolozzo, Fernando P

    2013-10-01

    The aim of this study was to evaluate the influence of multiple factors on bacterial contamination in 213 ejaculates from four boar studs. Semen contamination by aerobic mesophiles increased in ejaculates where the preputial fluid flowed into the collection container, collection glove was dirty, preputial hair was long (>1.0 cm), the collection lasted >7 min and boars were older than 18 months. An increase in coliforms occurred when preputial fluid dripped into the collection container, collections lasted >7 min or when penis escaped during collection. Semen contamination increased when two or more factors related to hygiene (poor hygiene of the boar, dirty preputial ostium, large preputial diverticulum, long preputial hair, dirty gloves, preputial liquid trickling from the hand of the technician into the semen container and penis escaping) were present. A vigilant protocol of collection must be followed to minimize bacterial contamination, especially avoiding dripping of preputial liquid into the semen container.

  19. Cryogenic preservation of semen from the Aleutian Canada goose (Branta canadensis leucopareia)

    USGS Publications Warehouse

    Gee, G.F.; Sexton, T.J.

    1990-01-01

    Aleutian Canada geese (Branta canadensis leucopareia) were inseminated with frozen-thawed semen containing 6% or 7% dimethylsulfoxide (DMSO) resulting in 32 fertile eggs and 17 goslings; with 7% DMSO, 19 of 31 eggs were fertile. Beltsville Poultry Semen Extender (BPSE), adjusted to 270 ? 30 mOs and 7.5 ? 0.4 pH, was used to dilute semen samples and the DMSO before cryopreservation. About half of the live spermatozoa in the fresh semen (92.9 ? 2.5% live cells, laboratory studies; 87.3 ? 7.3%, insemination trials) survived the freeze-thaw process (46.7 ? 7.8%, laboratory; 33.3 ? 17.8%, insemination trials). Samples of frozen-thawed semen contained a greater percentage of bent spermatozoa (27.1 ? 8.4% of live cells) than fresh semen (14.4 ? 3.0% of live cells). Fecal- and urate-contaminated semen (a common problem when collecting goose semen) reduced the sperm motility score from 3.2 ? 0.6 to 2.7? 0.7 and number of live spermatozoa in frozen-thawed semen from 49 ? 9% to 24 ?18%. Other variables examined that had less of an effect on semen quality included semen extenders, semen holding temperature, dilution and equilibration, relationship between hour of semen collection and level of semen contamination, and the relationship between season and sperm concentration.

  20. Semen Quality and Time-to-Pregnancy, the LIFE Study

    PubMed Central

    Buck Louis, Germaine M.; Sundaram, Rajeshwari; Schisterman, Enrique F.; Sweeney, Anne; Lynch, Courtney D.; Kim, Sungduk; Maisog, José M.; Gore-Langton, Robert; Eisenberg, Michael L.; Chen, Zhen

    2013-01-01

    Objective To assess semen parameters and couple fecundity as measured by time-to-pregnancy (TTP). Design Observational prospective cohort with longitudinal measurement of TTP. Setting 16 Michigan/Texas counties Participants 501 couples discontinuing contraception were followed for one year while trying to conceive; 473 (94%) men provided one semen sample and 80% provided two samples. Interventions None Main Outcome Measures Using prospectively measured TTP, fecundability odds ratios (FORs) and 95% confidence intervals (CIs) were estimated for 36 individual semen quality parameters accounting for repeated semen samples, time off contraception, abstinence, enrollment site, and couples’ ages, body mass indices (BMI), and serum cotinine concentrations. Results In adjusted models, semen quality parameters were associated with significantly shorter TTP as measured by FORs >1: percent motility, strict and traditional morphology, sperm head width, elongation factor, and acrosome area. Significantly longer TTPs or FORs <1 were observed for morphologic categories amorphous and round sperm heads neck/midpiece abnormalities. No semen quality parameters achieved significance when simultaneously modeling all other significant semen parameters and covariates, except for percent coiled tail when adjusting for sperm concentration (FOR=0.99; 95% CI 0.99, 1.00). Male age was consistently associated with reduced couple fecundity (FOR=0.96; 95% CI 0.93–0.99) reflecting a longer TTP across all combined models. Female but not male BMI also conferred a longer TTP (FOR=0.98; 95% CI 0.96–0.99). Conclusions Several semen measures were significantly associated with TTP when modeled individually, but not jointly and in the context of relevant couple based covariates. PMID:24239161

  1. [Quality of semen Hydnocarpi anthelminiticae introduced to various places].

    PubMed

    Lin, L; Xu, H; Chen, J; Chen, Y

    1996-02-01

    The result shows that the Semen Hydnocarpi Anthelminticae introduced in Guangdong, Hainan and Yunnan Provinces is similar to the control medicinal materials in fatty oil content, index of refraction, effective component content and trace element content. But the saponification value of the introduced products is higher. The quality of the Semen Hydnocarpi Anthelminticae introduced to the above places is up to the level of the control medicinal materials.

  2. Artificial insemination and cryopreservation of semen from nondomestic birds

    USGS Publications Warehouse

    Gee, G.F.; Bakst, M.R.; Wishart, G.J.

    1995-01-01

    Studies of Al and cryopreservation of semen from nondomestic birds began because of the increased emphasis on conservation of avian species threatened with extinction. Over the years, aviculturists have developed techniques for Al and cryopreservation of semen obtained from a variety of birds ranging from passerines to Andean condors. Generally, for each new species, we develop a practical semen collection technique and then evaluate the semen. A commercial semen extender (Beltsville Poultry Semen Extender) is modified and used to dilute the semen and provide support for the sperm during the freezing process (the pH and osmolality of the extender is adjusted to reflect the pH and osmolality of the semen being frozen). We find that the freezing schedule developed by Sexton (1977), which utilizes dimethylsulfoxide (DMS0) as cryoprotectant, works well for many species. We cool the sample sequentially in an ethanol bath, in liquid nitrogen vapor, and lastly in liquid nitrogen. Although we have experimented with a variety of freezing protocols, we prefer a 15-min equilibration period in DMSO at 5 C. We begin the freezing process by cooling at -1 C/min from 5 to -20 C in the ethanol bath. The samples are transferred into a vapor tank at a location just above liquid nitrogen and frozen at -50 C/min to -80 C. To complete the freezing process, the samples are plunged into the liquid nitrogen in the bottom of the vapor tank. The samples remain in liquid nitrogen until they are thawed just before insemination. If necessary, the freezing equipment can be transported in a van to remote locations.

  3. Enumeration of semen leucocytes by fluorescence in situ hybridisation technique

    PubMed Central

    Conte, R A; Luke, S; Verma, R S

    1995-01-01

    Aim—To determine whether the fluorescent in situ hybridisation technique (FISH) using a total human DNA genomic probe can be used to enumerate semen leucocytes. Methods—Semen samples from five donors were subjected to a mild KC1 solution. These samples were then biotin labelled under FISH conditions using a total human DNA genomic probe and the leucocyte counts were determined. To check the accuracy of the technique a monoclonal antibody against the common leucocyte antigen CD45 [KC56(T-200)] served as a control. An isotypic control for [KC56(T-200)], the immunoglobulin [MsIgG1], served as a secondary control. Results—Semen leucocytes stained by the FISH technique were easily detected because of their distinct bright yellow colour, while the sperm cells were red. The leucocyte count ranged from 0·5 to 4·9 × 106 per ml of semen. KC56(T-200) and its isotypic control MsIgG1, which served as control for the FISH technique, accurately identified 94% and 97% of the semen leucocytes of a control donor, respectively. Conclusions—The FISH technique using a total human DNA probe can accurately and effectively enumerate the overall leucocyte population in semen. Images PMID:16696031

  4. Semen and reproductive profiles of genetically identical cloned bulls.

    PubMed

    Tecirlioglu, R Tayfur; Cooney, Melissa A; Korfiatis, Natasha A; Hodgson, Renee; Williamson, Mark; Downie, Shara; Galloway, David B; French, Andrew J

    2006-06-01

    In this comparative study, reproductive parameters and semen characteristics of cloned bulls (n = 3) derived from somatic cell nuclear transfer (SCNT) were compared to their original cell donor Holstein-Friesian (n = 2) bulls from the same enterprise to assess the differences in reproductive potential between a donor bull and its clones. The parameters evaluated included motility of fresh, frozen-thawed and Percoll-treated frozen-thawed spermatozoa, as well as in vitro fertilization (IVF) ability, embryo quality, birth and survival of calves following IVF and embryo transfer with frozen-thawed semen. With fresh semen, spermatozoa from one cloned bull had lower motility than its donor. Cloned bulls had higher velocity parameters in fresh semen, but those effects were not obvious in frozen-thawed or frozen-thawed semen selected with a Percoll gradient. Semen collected from cloned bulls had significantly higher IVF rates compared to donors; however, embryo development per cleaved embryo or quality of blastocysts did not differ between donors and cloned bulls. Pregnancy and live offspring rates from one donor and its cloned bull did not differ between fresh (40%, 16/40 versus 46%, 17/37) and vitrified/thawed (13%, 2/16 versus 25%, 4/16) embryo transfer following IVF. A total of 26 calves were obtained from genotypically identical donor and cloned bulls with no signs of phenotypical abnormalities. These preliminary results suggested that the physiology of surviving postpubertal cloned bulls and quality of collected semen had equivalent reproductive potential to their original cell donor, with no evidence of any deleterious effects in their progeny.

  5. Lifestyle and semen quality: role of modifiable risk factors.

    PubMed

    Jurewicz, Joanna; Radwan, Michał; Sobala, Wojciech; Ligocka, Danuta; Radwan, Paweł; Bochenek, Michał; Hanke, Wojciech

    2014-02-01

    The relationship between exposure to lifestyle factors and adverse effects on human reproductive health is debated in the scientific literature and these controversies have increased public and regulatory attention. The aim of the study was to examine the association between modifiable lifestyle factors and main semen parameters, sperm morphology, and sperm chromatin structure. The study population consisted of 344 men who were attending an infertility clinic for diagnostic purposes with normal semen concentration of 20-300 M/ml or with slight oligozoospermia (semen total concentration of 15-20 M/ml) [WHO 1999]. Participants were interviewed and provided semen samples. The interview included questions about demographics, socio-economic status, medical history, lifestyle factors (consumption of alcohol, tobacco, coffee intake, cell phone and sauna usage), and physical activity. The results of the study suggest that lifestyle factors may affect semen quality. A negative association was found between increased body mass index (BMI) and semen volume (p = 0.03). Leisure time activity was positively associated with sperm concentration (p = 0.04) and coffee drinking with the percentage of motile sperm cells, and the percentage of sperm head and neck abnormalities (p = 0.01, p = 0.05, and p = 0.03, respectively). Drinking red wine 1-3 times per week was negatively related to sperm neck abnormalities (p = 0.01). Additionally, using a cell phone more than 10 years decreased the percentage of motile sperm cells (p = 0.02). Men who wore boxer shorts had a lower percentage of sperm neck abnormalities (p = 0.002) and percentage of sperm with DNA damage (p = 0.02). These findings may have important implications for semen quality and lifestyle.

  6. A comparison of semen diluents on the in vitro and in vivo fertility of liquid bull semen.

    PubMed

    Murphy, Edel M; Murphy, Craig; O'Meara, Ciara; Dunne, Gemma; Eivers, Bernard; Lonergan, Patrick; Fair, Sean

    2017-02-01

    The aim of this study was to assess the effect of semen diluent on calving rate (CR) following artificial insemination with liquid bull semen stored for up to 3 d postcollection. In experiment 1, the effect of storing liquid semen maintained at a constant ambient temperature in 1 of 7 different diluents [Caprogen (homemade), OptiXcell, BioXcell, BullXcell, INRA96, NutriXcell, or AndroMed (all commercially available)] on total and progressive motility was assessed on d 0, 1, 2, and 3 postcollection. In experiment 2, the field fertility of liquid semen diluted in Caprogen, BioXcell, or INRA96 and inseminated on d 1, 2, or 3 postcollection was assessed in comparison to frozen-thawed semen (total of n = 19,126 inseminations). In experiment 3, the effect of storage temperature fluctuations (4 and 18°C) on total and progressive motility following dilution in Caprogen, BioXcell, and INRA96 was assessed on d 0, 1, 2, and 3 postcollection. In experiment 1, semen stored in Caprogen, BioXcell, and INRA96 resulted in the highest total and progressive motility on d 1, 2, and 3 of storage compared with OptiXcell, BullXcell, NutriXcell, and AndroMed. In experiment 2, an effect of diluent on CR was found as semen diluted in BioXcell had a lower CR on d 1, 2, and 3 of storage (46.3, 35.4, and 34.0%, respectively) in comparison with Caprogen (55.8, 52.0, and 51.9%, respectively), INRA96 (55.0, 55.1, and 52.2%, respectively), and frozen-thawed semen (59.7%). Effects were found of parity, cow fertility sub-index, as well as the number of days in milk on CR. In experiment 3, when the storage temperature of diluted semen fluctuated between 4 and 18°C, to mimic what occurs in the field (nighttime vs. daytime), BioXcell had the lowest total and progressive motility in comparison to Caprogen and INRA96. In conclusion, diluent significantly affected sperm motility when stored for up to 3 d. Semen diluted in INRA96 resulted in a similar CR to semen diluted in Caprogen and to frozen

  7. EVALUATION OF CHROMOSOME BREAKAGE AND DNA INTEGRITY IN SPERM: AN INVESTIGATION OF REMOTE SEMEN COLLECTION CONDITIONS

    EPA Science Inventory

    Home collection of ejaculated semen would facilitate participation rates and geographic diversity in reproductive epidemiology studies. Our study addressed concerns that home collection and overnight mail return might induce chromosome/DNA damage. We collected semen from 10 hea...

  8. Expected net present value of pure and mixed sexed semen artificial insemination strategies in dairy heifers.

    PubMed

    Olynk, N J; Wolf, C A

    2007-05-01

    Sexed semen has been a long-anticipated tool for dairy farmers to obtain more heifer calves, but challenges exist for integrating sexed semen into commercial dairy farm reproduction programs. The decreased conception rates (CR) experienced with sexed semen make virgin heifers better suited for insemination with sexed semen than lactating dairy cows. This research sought to identify when various sexed semen breeding strategies provided higher expected net present value (NPV) than conventional artificial insemination (AI) breeding schemes, indicating which breeding scheme is advisable under various scenarios. Budgets were developed to calculate the expected NPV of various AI breeding strategies incorporating conventional (non-sexed) and sexed semen. In the base budgets, heifer and bull calf values were held constant at $500 and $110, respectively. The percentage of heifers expected to be born after breeding with conventional and sexed semen used was 49.2 and 90%, respectively. Breeding costs per AI were held constant at $15.00 per AI for conventional semen and $45.00 per AI for sexed semen of approximately the same genetic value. Conventional semen CR of 58 and 65% were used, and an AI submission rate was set at 100%. Breeding strategies with sexed semen were assessed for breakeven heifer calf values and sexed semen costs to obtain a NPV equal to that achieved with conventional semen. Breakeven heifer calf values for pure sexed semen strategies with a constant 58 and 65% base CR in which sexed semen achieved 53% of the base CR are $732.11 and $664.26, respectively. Breakeven sexed semen costs per AI of $17.16 and $22.39, compared with $45.00 per AI, were obtained to obtain a NPV equal to that obtained with pure conventional semen for base CR of 58 and 65%, respectively. The strategy employing purely sexed semen, with base CR of both 58 and 65%, yielded a lower NPV than purely conventional semen in all but the best-case scenario in which sexed semen provides 90% of

  9. The utility of nanowater for ram semen cryopreservation.

    PubMed

    Murawski, Maciej; Schwarz, Tomasz; Grygier, Joanna; Patkowski, Krzysztof; Oszczęda, Zdzisław; Jelkin, Igor; Kosiek, Anna; Gruszecki, Tomasz M; Szymanowska, Anna; Skrzypek, Tomasz; Zieba, Dorota A; Bartlewski, Pawel M

    2015-05-01

    Nanowater (NW; water declusterized in the low-temperature plasma reactor) has specific physicochemical properties that could increase semen viability after freezing and hence fertility after artificial insemination (AI) procedures. The main goal of this study was to evaluate ram semen quality after freezing in the media containing NW. Ejaculates from 10 rams were divided into two equal parts, diluted in a commercially available semen extender (Triladyl®; MiniTüb GmbH, Tiefenbach, Germany) prepared with deionized water (DW) or NW, and then frozen in liquid nitrogen. Semen samples were examined for sperm motility and morphology using the sperm class analyzer system and light microscopy. Cryo-scanning electron microscopy (cryo-SEM) was employed to determine the size of extracellular water crystals in frozen semen samples. Survival time at room temperature, aspartate aminotransferase (AspAT) and alkaline phosphatase (ALP) concentrations post-thawing as well as conception/lambing rates after laparoscopic intrauterine AI of 120 ewes were also determined. There were no significant differences between DW and NW groups in sperm progressive motility (26.4 ± 12.2 and 30.8 ± 12.4%) or survival time (266.6 ± 61.3 and 270.9 ± 76.7 min) after thawing and no differences in the percentages of spermatozoa with various morphological defects before or after freezing. There were, however, differences (P < 0.05) in AspAT (DW: 187.1 ± 160.4 vs. NW: 152.7 ± 118.3 U/l) and ALP concentrations (DW: 2198.3 ± 1810.5 vs. NW: 1612.1 ± 1144.8 U/l) in semen samples post-thawing. Extracellular water crystals were larger (P < 0.05) in ejaculates frozen in NW-containing media. Ultrasonographic examinations on day 40 post-AI revealed higher (P < 0.05) conception rates in ewes inseminated with NW (78.3%) compared with DW semen (58.3%), and the percentages of ewes that carried lambs to term were 73.3% and 45.0% in NW and DW groups, respectively (P < 0.01). In summary, the use of a semen

  10. Semen characteristics in pubertal boys. IV. Semen quality and hormone profile.

    PubMed

    Janczewski, Z; Bablok, L

    1985-01-01

    The biological quality of semen was contrasted with levels of FSH, LH, and testosterone in plasma. The level of FSH rises significantly from azoospermia/cryptozoospermia (3.87 +/- 1.1 mU/ml) to asthenozoospermia (5.73 +/- 2.11 mU/ml). In normospermia, however (4.63 +/- 1.88 mU/ml), the level of FSH decreases in a statistically significant manner and remains at the standard level. Comparing the level of LH to the quality of semen, it rises in a statistically significant manner from azoospermia/cryptozoospermia (6.46 +/- 1.35 mU/ml) to oligozoospermia (9.03 +/- 3.35 mU/ml). The level decreases in a statistically significant manner in normospermia (7.15 +/- 1.69 mU/ml). The level of testosterone shows a progressive linear growth from azoospermia/cryptozoospermia (6.03 +/- 2.09 micrograms/ml) to normospermia (6.55 +/- 2.12 micrograms/ml). The growth is statistically insignificant.

  11. Quality and fertility of cooled-shipped stallion semen at the time of insemination.

    PubMed

    Heckenbichler, Sabine; Deichsel, Katharina; Peters, Pamela; Aurich, Christine

    2011-03-15

    Stallion semen processing is far from standardized and differs substantially between AI centers. Suboptimal pregnancy rates in equine AI may primarily result from breeding with low quality semen not adequately processed for shipment. It was the aim of the study to evaluate quality and fertility of cooled-shipped equine semen provided for breeding of client mares by commercial semen collection centers in Europe. Cooled shipped semen (n = 201 doses) from 67 stallions and 36 different EU-approved semen collection centers was evaluated. At arrival, semen temperature was 9.8 ± 0.2 °C, mean sperm concentration of AI doses was 68 ± 3 x 10(6)/ml), mean total sperm count was 1.0 ± 0.1 x 10(9), total motility averaged 83 ± 1% and morphological defects 45 ± 2%. A total of 86 mares were inseminated, overall per season-pregnancy rate in these mares was 67%. Sperm concentration significantly influenced semen motility and morphology at arrival of the shipped semen. Significant effects of month of the year on volume, sperm concentration and total sperm count of the insemination dose were found. The collection center significantly influenced all semen parameters evaluated. Semen doses used to inseminate mares that became pregnant had significantly higher total and progressive motility of spermatozoa and a significantly lower percentage of morphological semen defects than insemination doses used for mares failing to get pregnant. Results demonstrate that insemination with semen of better quality provides a higher chance to achieve pregnancy. Besides the use of stallions with good semen quality, appropriate semen processing is an important factor for satisfying results in artificial insemination with cooled-shipped horse semen.

  12. Semen and the diagnosis of infertility in Aristotle.

    PubMed

    Trompoukis, C; Kalaitzis, C; Giannakopoulos, S; Sofikitis, N; Touloupidis, S

    2007-02-01

    Aristotle (384-322bc) was one of the leading intellectual figures of all time. In his work he systematised a massive amount of knowledge on a diverse range of subjects, including medicine. This article discusses the observations and hypotheses of this great philosopher on semen and infertility, as they are presented in his work Generation of Animals. This is combined with an evaluation of his positions in relation to those of the Hippocratic Corpus on the same subject. An extensive review of Aristotle's work Generation of Animals was performed with particular focus on his perspectives about semen and infertility. Publications referring to this work were also reviewed. According to Aristotle, semen is that which contains the principles that come from both parents when they unite. He believed that semen was formed by the secretion of nutriments by the body, developing his theories of sterility on this basic principle. A lack of fertility is attributed to genetic or acquired causes. He proposed methods for diagnosing sterility, primarily the 'water test' for men and the 'pessary' method for women. Even if his observations contain clear mistakes, such as attributing only secondary functions to male testicles and the identification of menses as women's 'seed', Aristotle's views also contain keen observations and exceptional thinking, both on the characteristics of semen and the causes of sterility (infertility).

  13. Lipid Concentrations and Semen Quality: The LIFE Study

    PubMed Central

    Schisterman, Enrique F.; Mumford, Sunni L.; Chen, Zhen; Browne, Richard W.; Barr, Dana Boyd; Kim, Sungduk; Louis, Germaine M. Buck

    2014-01-01

    The decline in sperm count rates over the last 50 years appears to parallel the rising prevalence of obesity. As lipids levels are strongly associated with obesity, high lipids levels or hyperlipidemia may thus play an important role in the decline in fertility in addition to other environmental or lifestyle factors. The objective of this population based cohort study was to evaluate the association between men’s serum lipid concentrations and semen quality parameters among 501 male partners of couples desiring pregnancy and discontinuing contraception. Each participant provided prospectively up to two semen samples (94% of men provided one or more semen samples, and 77% of men provided a second sample approximately one month later). Linear mixed effects models were used to estimate the associations between baseline lipid concentrations and semen quality parameters, adjusted for age, body mass index, and race. We found that higher levels of serum total cholesterol, free cholesterol and phospholipids were associated with a significantly lower percentage of sperm with intact acrosome and smaller sperm head area and perimeter. Our results suggest that lipid concentrations may affect semen parameters, specifically sperm head morphology, highlighting the importance of cholesterol and lipid homeostasis for male fecundity. PMID:24596332

  14. HIV-1 Populations in Semen Arise through Multiple Mechanisms

    PubMed Central

    Dibben, Oliver; Jabara, Cassandra B.; Arney, Leslie; Kincer, Laura; Tang, Yuyang; Hobbs, Marcia; Hoffman, Irving; Kazembe, Peter; Jones, Corbin D.; Borrow, Persephone; Fiscus, Susan; Cohen, Myron S.; Swanstrom, Ronald

    2010-01-01

    HIV-1 is present in anatomical compartments and bodily fluids. Most transmissions occur through sexual acts, making virus in semen the proximal source in male donors. We find three distinct relationships in comparing viral RNA populations between blood and semen in men with chronic HIV-1 infection, and we propose that the viral populations in semen arise by multiple mechanisms including: direct import of virus, oligoclonal amplification within the seminal tract, or compartmentalization. In addition, we find significant enrichment of six out of nineteen cytokines and chemokines in semen of both HIV-infected and uninfected men, and another seven further enriched in infected individuals. The enrichment of cytokines involved in innate immunity in the seminal tract, complemented with chemokines in infected men, creates an environment conducive to T cell activation and viral replication. These studies define different relationships between virus in blood and semen that can significantly alter the composition of the viral population at the source that is most proximal to the transmitted virus. PMID:20808902

  15. HIV-1 Populations in Semen Arise through Multiple Mechanisms.

    PubMed

    Anderson, Jeffrey A; Ping, Li-Hua; Dibben, Oliver; Jabara, Cassandra B; Arney, Leslie; Kincer, Laura; Tang, Yuyang; Hobbs, Marcia; Hoffman, Irving; Kazembe, Peter; Jones, Corbin D; Borrow, Persephone; Fiscus, Susan; Cohen, Myron S; Swanstrom, Ronald

    2010-08-19

    HIV-1 is present in anatomical compartments and bodily fluids. Most transmissions occur through sexual acts, making virus in semen the proximal source in male donors. We find three distinct relationships in comparing viral RNA populations between blood and semen in men with chronic HIV-1 infection, and we propose that the viral populations in semen arise by multiple mechanisms including: direct import of virus, oligoclonal amplification within the seminal tract, or compartmentalization. In addition, we find significant enrichment of six out of nineteen cytokines and chemokines in semen of both HIV-infected and uninfected men, and another seven further enriched in infected individuals. The enrichment of cytokines involved in innate immunity in the seminal tract, complemented with chemokines in infected men, creates an environment conducive to T cell activation and viral replication. These studies define different relationships between virus in blood and semen that can significantly alter the composition of the viral population at the source that is most proximal to the transmitted virus.

  16. The effects of semen collection on fertility in captive, naturally fertile, sandhill cranes

    USGS Publications Warehouse

    Chen, G.; Gee, G.F.; Nicolich, Jane M.; Taylor, J.A.

    2001-01-01

    We tested to see if semen collection interferes with fertility in naturally fertile pairs of cranes. We used 12 naturally fertile, Florida sandhill crane (Grus canadensis pratensis) pairs for this study, 6 control and 6 experimental. All pairs had previously produced fertile eggs. Semen was collected on Tuesday mornings and Friday afternoons from 26 February 1993 to 4 June 1993. We used standard artificial insemination methods to collect and to evaluate the semen and spermatozoa. Semen collection had minimal effect on semen quality and semen quantity. Semen volume, sperm density, sperm motility, sperm morphology, sperm viability, sperm number per collection, and male response to semen collection exhibited significant daily variation. Although semen collection began 13 days before the first egg in the experimental group, we did not observe differences in the date of first egg laid or in fertility between experimental and control groups. Also, we observed no statistically significant differences in the interval between clutches or in the percentage of broken eggs between experimental and control groups. However, 4 eggs were broken by adults during the disturbance associated with capturing birds for semen collection. We found that females with mates from which we consistently gathered better semen samples produced fewer fertile eggs than females with sires producing poorer semen samples (r = 0.60). We interpret these results to mean that males that were successfully breeding with their mates had little left at the time of our collection.

  17. Rooster Semen Cryopreservation: Effect of Pedigree Line and Male Age on Post-Thaw Sperm Function

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fertility rates of cryopreserved poultry semen are highly variable and not reliable for use in preservation of commercial genetic stocks. Our objective was to evaluate the cryosurvival of semen from 8 pedigreed layer lines at the onset and end of production. Semen from 160 roosters (20/line) was...

  18. Seasonal variations in semen characteristics, semen cryopreservation, estrus synchronization, and successful artificial insemination in the spotted deer (Axis axis).

    PubMed

    Umapathy, Govindhaswamy; Sontakke, Sadanand D; Reddy, Anuradha; Shivaji, S

    2007-05-01

    Ten adult male spotted deer were monitored over a 2-year interval to determine seasonal variations in testicular size, semen characteristics and serum testosterone concentrations, and to determine if there was an association between season and type of antler. Mean (+/-S.E.M.) testicular volume (118.8+/-4.6 cm(3)), serum testosterone concentration (1.2+/-0.1 ng/mL), semen volume (4.1+/-0.6 mL), sperm concentration (338.3+/-24.9 x 10(6) mL(-1)), percentage of morphologically normal sperm (79.1+/-2.8%), and percentage of motile sperm (66.5+/-1.5%) were higher (P<0.05) in hard antler deer (peaked from March to May) than in deer with velvet antlers or in deer in which the antler has been shed. Thus, March-May was considered the physiologic breeding season for these deer; at this time, all stags had hard antlers. Furthermore, a Tris-citrate-based semen extender containing 4% glycerol and 20% egg-yolk was adequate for cryopreservation of semen. Estrus was induced with an implant containing norgestomet, timed transcervical AI was done with fresh semen, and 3 of 10 females were pregnant at 60 days, with fawns born 120 (premature), 240 and 243 days after AI. These results were considered a model for the use of assisted reproductive techniques to conserve other critically endangered deer species of India.

  19. Rotation of Boar Semen Doses During Storage Affects Sperm Quality.

    PubMed

    Schulze, M; Rüdiger, K; Waberski, D

    2015-08-01

    It is common practice to rotate boar semen doses during storage for prevention of sperm sedimentation. In this study, the effect of rotation of boar semen doses during storage on sperm quality was investigated. Manual turning twice daily and automatic rotation five times per hour resulted in the following effects: alkalinization of the BTS-extender, loss of membrane integrity at day 3, and loss of motility and changes in sperm kinematics during a thermoresistance test at day 5. Using a pH-stabilized variant of BTS extender, sperm motility and velocity decreased in continuously rotated samples, whereas membrane integrity and mitochondrial activity remain unaffected. It is concluded that rotation of semen samples adversely affects sperm quality and, therefore, should no longer be recommended for AI practice.

  20. [Semen analysis: spermiogram according to WHO 2010 criteria].

    PubMed

    Gottardo, F; Kliesch, S

    2011-01-01

    Semen analysis plays a key role in the diagnostics of male infertility. Semen analysis has to be performed according to World Health Organisation (WHO) criteria. The updated version of the WHO manual was completed at the end of 2009 and published in 2010. Standard procedures in semen analysis include evaluation of sperm concentration, motility, morphology and vitality. In this new version particular attention has been paid to internal and external quality control, helping to identify and correct incidental and systematic errors both in routine analysis as well as in the field of research. The new manual describes all laboratory solutions, procedures and calculation formulas, and focuses on the definition of cryptozoospermia or azoospermia. A chapter concerning cryopreservation of spermatozoa has been newly integrated. The following overview presents the most important aspects of the updated WHO manual.

  1. Are serum zinc and copper levels related to semen quality?

    PubMed

    Yuyan, Li; Junqing, Wu; Wei, Yuan; Weijin, Zhou; Ersheng, Gao

    2008-04-01

    Low serum zinc levels are harmful to semen quality in Chinese men. In this study, eligible men aged 20-59 years old-excluding those who had ever had urinary or genital disease, tuberculosis, or occupational heavy metal contact-were examined for semen quality and serum zinc and copper concentrations. Progressive motility showed differences among the five copper groups, but multiple logistic analyses did not show that higher or lower serum copper levels had a significant effect on sperm quality. When serum zinc concentration was low, the risk of asthenozoospermia was higher. The ratio of Cu/Zn was higher in the progressive motility abnormal group than in the normal group.

  2. A trial of semen collection by transrectal electroejaculation method from Amur leopard cat (Prionailurus bengalensis euptilurus)

    PubMed Central

    TAJIMA, Hideo; YOSHIZAWA, Madoka; SASAKI, Shinichi; YAMAMOTO, Fujio; NARUSHIMA, Etsuo; OGAWA, Yuka; ORIMA, Hiromitsu; TSUTSUI, Toshihiko; TOYONAGA, Mari; KOBAYASHI, Masanori; KAWAKAMI, Eiichi; HORI, Tatsuya

    2016-01-01

    We collected semen from a male Amur leopard cat using the transrectal electroejaculation method and investigated the semen qualities for about four years. In addition, the influence of the season on the spermatogenic function of the Amur leopard cat was investigated with regard to the semen qualities, testicular volume and serum testosterone level. As a result, we could collect semen with good sperm qualities that would be useable for artificial insemination. Some seasonality was noted in the testicular volume and serum testosterone level. We clarified that the semen qualities were favorable before and during the female breeding season compared with those after the breeding season. PMID:26935841

  3. Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics.

    PubMed

    Guimarães, T; Lopes, G; Pinto, M; Silva, E; Miranda, C; Correia, M J; Damásio, L; Thompson, G; Rocha, A

    2015-01-15

    Freezability of equine semen may be influenced by microorganism population of semen. The objective of this study was to verify the effect of single-layer density gradient centrifugation (SLC) of fresh semen before cryopreservation on semen's microbial load (ML) and sperm cells kinetics after freezing-thawing. For that, one ejaculate was collected from 20 healthy stallions and split into control (C) samples (cryopreserved without previous SLC) and SLC samples (subjected to SLC). Semen cryopreservation was performed according to the same protocol in both groups. Microbial load of each microorganism species and total microbial load (TML) expressed in colony-forming units (CFU/mL) as well as frozen-thawed sperm kinetics were assessed in both groups. Additional analysis of the TML was performed, subdividing the frozen-thawed samples in "suitable" (total motility ≥ 30%) and "unsuitable" (total motility < 30%) semen for freezing programs, and comparing the C and SLC groups within these subpopulations. After thawing, SLC samples had less (P < 0.05) TML (88.65 × 10(2) ± 83.8 × 10(2) CFU/mL) than C samples (155.69 × 10(2) ± 48.85 × 10(2) CFU/mL), mainly due to a reduction of Enterococcus spp. and Bacillus spp. A relationship between post-thaw motility and SLC effect on ML was noted, as only in samples with more than 30% total motility was ML reduced (P < 0.05) by SLC (from 51.33 × 10(2) ± 33.26 × 10(2) CFU/mL to 26.68 × 10(2) ± 12.39 × 10(2) CFU/mL in "suitable" frozen-thawed semen vs. 240.90 × 10(2) ± 498.20 × 10(2) to 139.30 × 10(2) ± 290.30 × 10(2) CFU/mL in "unsuitable" frozen-thawed semen). The effect of SLC on kinetics of frozen-thawed sperm cells was negligible.

  4. New methods and media for the centrifugation of honey bee (Hymenoptera: Apidae) drone semen.

    PubMed

    Wegener, Jakob; May, Tanja; Kamp, Günter; Bienefeld, Kaspar

    2014-02-01

    Centrifugation of Apis mellifera L. drone semen is a necessary step in the homogenization of semen pools for the enlargement of the effective breeding population, as well as in the collection of semen by the so-called washing technique. It is also of interest for the removal of cryoprotectants after cryopreservation. The adoption of methods involving semen centrifugation has been hampered by their damaging effect to sperm. Here, we tested four new diluents as well as three additives (catalase, hen egg yolk, and a protease inhibitor), using sperm motility and dual fluorescent staining as indicators of semen quality. Three of the new diluents significantly reduced motility losses after centrifugation, as compared with the literature standard. Values of motility and propidium iodide negativity obtained with two of these diluents were not different from those measured with untreated semen. The least damaging diluent, a citrate-HEPES buffer containing trehalose, was then tested in an insemination experiment with centrifuged semen. Most queens receiving this semen produced normal brood, and the number of sperm reaching the storage organ of the queen was not significantly different from that in queens receiving untreated semen. These results could improve the acceptance of techniques involving the centrifugation of drone semen. The diluent used in the insemination experiment could also serve as semen extender for applications not involving centrifugation.

  5. Reproduction in nondomestic birds: Physiology, semen collection, artificial insemination and cryopreservation

    USGS Publications Warehouse

    Gee, G.F.; Bertschinger, H.; Donoghue, A.M.; Blanco, J.; Soley, J.

    2004-01-01

    Pioneering work by Quinn and Burrows in the late 1930s led to successful artificial insemination (AI) programs in the domestic poultry industry. A variety of species specific modifications to the Quinn and Burrows massage technique made AI possible in nondomestic birds. Massage semen collection and insemination techniques span the entire range of species from sparrows to ostriches. Also, cooperative semen collection and electroejaculation have found limited use in some nondomestic species. Artificial insemination produces good fertility, often exceeding fertility levels in naturally copulating populations. However, aviculturists should explore other ways to improve fertility before resorting to AI. Artificial insemination is labor intensive and may pose risks to nondomestic birds as well as handlers associated with capture and insemination. Semen collection and AI makes semen cryopreservation and germ plasma preservation possible. Yet, semen cryopreservation techniques need improvement before fertility with frozen-thawed semen will equal fertility from AI with fresh semen.

  6. Maintaining semen quality by improving cold chain equipment used in cattle artificial insemination

    PubMed Central

    Lieberman, Daniel; McClure, Elizabeth; Harston, Stephen; Madan, Damian

    2016-01-01

    Artificial insemination of dairy cattle is a common practice in the developing world that can improve farmer incomes and food security. Maintaining the fertilizing potential of frozen semen as it is manipulated, transported and stored is crucial to the success of this process. Here we describe simple technological improvements to protect semen from inadvertent thermal fluctuations that occur when users mishandle semen using standard equipment. We show that when frozen semen is mishandled, characteristics of semen biology associated with fertility are negatively affected. We describe several design modifications and results from thermal performance tests of several improved prototypes. Finally, we compare semen that has been mishandled in standard and improved equipment. The data suggest that our canister improvements can better maintain characteristics of semen biology that correlate with fertility when it is mishandled. PMID:27313137

  7. Maintaining semen quality by improving cold chain equipment used in cattle artificial insemination

    NASA Astrophysics Data System (ADS)

    Lieberman, Daniel; McClure, Elizabeth; Harston, Stephen; Madan, Damian

    2016-06-01

    Artificial insemination of dairy cattle is a common practice in the developing world that can improve farmer incomes and food security. Maintaining the fertilizing potential of frozen semen as it is manipulated, transported and stored is crucial to the success of this process. Here we describe simple technological improvements to protect semen from inadvertent thermal fluctuations that occur when users mishandle semen using standard equipment. We show that when frozen semen is mishandled, characteristics of semen biology associated with fertility are negatively affected. We describe several design modifications and results from thermal performance tests of several improved prototypes. Finally, we compare semen that has been mishandled in standard and improved equipment. The data suggest that our canister improvements can better maintain characteristics of semen biology that correlate with fertility when it is mishandled.

  8. Short communication: Progressive motility of frozen-thawed canine semen is highest five minutes after thawing.

    PubMed

    Karger, S; Geiser, B; Grau, M; Heuwieser, W; Arlt, S P

    2017-04-01

    Progressive motility is usually estimated by visual inspection using a light contrast microscope at X 100 immediately after semen collection or immediately after thawing frozen semen. Standard operating procedures have never been established for this test. The objective of this experiment was to examine time-dependent changes of motility after thawing cryopreserved canine semen. Semen of 35 dogs was collected, and volume, concentration, progressive motility, morphology, membrane integrity and HOS test were evaluated. For cryopreservation, CaniPRO(®) Freeze A&B was used. Semen was thawed and diluted using CaniPRO(®) culture medium. After thawing, semen was evaluated as before. In addition, every sample was evaluated for progressively motile sperm cells 0, 5, 20 and 60 min after thawing. Progressive semen motility was significantly highest five minutes after thawing.

  9. 19 CFR 12.32 - Honeybees and honeybee semen.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Honeybees and honeybee semen. 12.32 Section 12.32 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY SPECIAL CLASSES OF MERCHANDISE Wild Animals, Birds, and Insects § 12.32 Honeybees and...

  10. Magnesium in human semen: possible role in premature ejaculation.

    PubMed

    Omu, A E; Al-Bader, A A; Dashti, H; Oriowo, M A

    2001-01-01

    Although magnesium is involved in many biological process and it is found higher levels in semen than serum, its role in human semen has not been elucidated. This investigation was conducted to evaluate the relationship between premature ejaculation and the levels of seminal magnesium. The levels of magnesium, zinc, copper, and selenium were evaluated with an atomic absorption spectrophotometer in serum and seminal plasma in 3 groups of men: (a) normal sperm parameters (15) (b) oligoasthenozoospermia (15), and genuine premature ejaculation (9). There were normal serum and semen levels of all the elements in the three groups, but significantly lower seminal plasma magnesium levels in men with premature ejaculation. The hormonal profile, body mass index (BMI) had no association with premature ejaculation. Decreased levels of magnesium gives rise to vasoconstriction from increased thromboxane level, increased endothelial intracellular Ca2+, and decreased nitric oxide. This may lead to premature emission and ejaculation processes. Magnesium is probably involved in semen transport. More research into the role of magnesium in the male physiology of reproductive tract, especially its association with premature ejaculation, is advocated.

  11. Effect of Dursban 44 on semen output of Holstein bulls.

    PubMed

    Everett, R W

    1982-09-01

    Dursban 44, an insecticide for lice control, was applied to 185 Holstein bulls 9 to 52 mo of age. These sires were in various stages of progeny testing at an artificial insemination center. Application of this product killed 7 bulls, and the remaining bulls exhibited varying severity of illness with 6 classified as very sick. This study evaluated the effect of this illness on semen production. Semen output on 40,950 ejaculates from 583 Holstein bulls collected from July 1, 1975, through March 31, 1981, was analyzed to establish normal semen production and to estimate the effect of illness caused by Dursban 44 treatment. Ejaculate number, days between collections by previous number of ejaculates, calendar months, years, and ages of bulls affected the semen output characteristics, original volume, sperm concentration, percent motile sperm, total sperm per ejaculate, percent prefreeze discards, percent postthaw sperm motility, and percent postthaw discards. Ejaculate volume, motility, total percent prefreeze discards, and percent postthaw discards were influenced negatively on the 6 very sick bulls. Percent postthaw discards were higher on all bulls treated with Dursban 44 for up to 6 mo post-treatment.

  12. 19 CFR 12.32 - Honeybees and honeybee semen.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Honeybees and honeybee semen. 12.32 Section 12.32 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY SPECIAL CLASSES OF MERCHANDISE Wild Animals, Birds, and Insects § 12.32 Honeybees and...

  13. 19 CFR 12.32 - Honeybees and honeybee semen.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 false Honeybees and honeybee semen. 12.32 Section 12.32 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY SPECIAL CLASSES OF MERCHANDISE Wild Animals, Birds, and Insects § 12.32 Honeybees and...

  14. TRIHALOMETHANE LEVELS IN HOME TAP WATER AND SEMEN QUALITY

    EPA Science Inventory

    Trihalomethane Levels in Home Tap Water and Semen Quality
    Laura Fenster, 1 Kirsten Waller, 2 Gayle Windham, 1 Tanya Henneman, 2 Meredith Anderson, 2 Pauline Mendola, 3 James W. Overstreet, 4 Shanna H. Swan5

    1California Department of Health Services, Division of Environm...

  15. 9 CFR 98.36 - Animal semen from Canada.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Animal semen from Canada. 98.36 Section 98.36 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EXPORTATION AND IMPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS IMPORTATION...

  16. 9 CFR 98.36 - Animal semen from Canada.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Animal semen from Canada. 98.36 Section 98.36 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EXPORTATION AND IMPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS IMPORTATION...

  17. 9 CFR 98.36 - Animal semen from Canada.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Animal semen from Canada. 98.36 Section 98.36 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EXPORTATION AND IMPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS IMPORTATION...

  18. 9 CFR 98.36 - Animal semen from Canada.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Animal semen from Canada. 98.36 Section 98.36 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EXPORTATION AND IMPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS IMPORTATION...

  19. 9 CFR 98.36 - Animal semen from Canada.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Animal semen from Canada. 98.36 Section 98.36 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EXPORTATION AND IMPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS IMPORTATION...

  20. 19 CFR 12.32 - Honeybees and honeybee semen.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 19 Customs Duties 1 2013-04-01 2013-04-01 false Honeybees and honeybee semen. 12.32 Section 12.32 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY SPECIAL CLASSES OF MERCHANDISE Wild Animals, Birds, and Insects § 12.32 Honeybees and...

  1. 19 CFR 12.32 - Honeybees and honeybee semen.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 false Honeybees and honeybee semen. 12.32 Section 12.32 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY SPECIAL CLASSES OF MERCHANDISE Wild Animals, Birds, and Insects § 12.32 Honeybees and...

  2. Semen quality in relation to biomarkers of pesticide exposure.

    PubMed Central

    Swan, Shanna H; Kruse, Robin L; Liu, Fan; Barr, Dana B; Drobnis, Erma Z; Redmon, J Bruce; Wang, Christina; Brazil, Charlene; Overstreet, James W

    2003-01-01

    We previously reported reduced sperm concentration and motility in fertile men in a U.S. agrarian area (Columbia, MO) relative to men from U.S. urban centers (Minneapolis, MN; Los Angeles, CA; New York, NY). In the present study we address the hypothesis that pesticides currently used in agriculture in the Midwest contributed to these differences in semen quality. We selected men in whom all semen parameters (concentration, percentage sperm with normal morphology, and percentage motile sperm) were low (cases) and men in whom all semen parameters were within normal limits (controls) within Missouri and Minnesota (sample sizes of 50 and 36, respectively) and measured metabolites of eight current-use pesticides in urine samples provided at the time of semen collection. All pesticide analyses were conducted blind with respect to center and case-control status. Pesticide metabolite levels were elevated in Missouri cases, compared with controls, for the herbicides alachlor and atrazine and for the insecticide diazinon [2-isopropoxy-4-methyl-pyrimidinol (IMPY)]; for Wilcoxon rank test, p = 0.0007, 0.012, and 0.0004 for alachlor, atrazine, and IMPY, respectively. Men from Missouri with high levels of alachlor or IMPY were significantly more likely to be cases than were men with low levels [odds ratios (ORs) = 30.0 and 16.7 for alachlor and IMPY, respectively], as were men with atrazine levels higher than the limit of detection (OR = 11.3). The herbicides 2,4-D (2,4-dichlorophenoxyacetic acid) and metolachlor were also associated with poor semen quality in some analyses, whereas acetochlor levels were lower in cases than in controls (p = 0.04). No significant associations were seen for any pesticides within Minnesota, where levels of agricultural pesticides were low, or for the insect repellent DEET (N,N-diethyl-m-toluamide) or the malathion metabolite malathion dicarboxylic acid. These associations between current-use pesticides and reduced semen quality suggest that

  3. Cryopreservation of Indian red jungle fowl (Gallus gallus murghi) semen.

    PubMed

    Rakha, B A; Ansari, M S; Akhter, S; Hussain, I; Blesbois, E

    2016-11-01

    The population of red jungle fowl is declining and needs special attention for its conservation with suitable approaches. For ex situ in vitro conservation of Indian red jungle fowl, establishment of semen cryobank is an appropriate option, for which an extender with adequate retrieval capacity for functional spermatozoa is required. Therefore, studies were designed to evaluate a wide range of extenders for cryopreservation of Indian red jungle fowl (Gallus gallus murghi) sperm to achieve maximal post-thawed semen quality and fertility. For this purpose, semen from eight mature cocks were collected, initially evaluated (percent sperm motility, volume and concentration), pooled, assessed for motility, plasma membrane integrity, viability and acrosome integrity, and divided into six aliquots for dilution (1:5; 37°C) in Beltsville poultry, red fowl extender, Lake, EK, Tselutin poultry and chicken semen extenders. Diluted semen was cooled from 37°C to 4°C @ -0.275°C/min. Glycerol (20%) was added to chilled semen, equilibrated for 10min, filled in 0.5mL French straws, kept over LN2 vapours for 10min and plunged into LN2 and stored at -196°C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were higher (P<0.05) in red fowl extender at 0, 2 and 4h of incubation post-thaw. After cryopreservation and post-thawing at 37°C the highest (P<0.05) recovery rates and absolute livability index was also recorded in red fowl extender that was thus used for further artificial insemination of cooled-diluted (Liquid) and cryopreserved sperm. The no. of fertilized eggs (Liquid, 20.6±0.4; Cryopreserved, 12.6±0.5), percent fertility (86.7±2.2; 57.2±3.9), no. of hatched chicks (18.2±0.8; 10.0±0.3), percent hatch (76.5±2.7; 45.3±2.2) and hatchability of fertilized eggs (88.3±3.4; 79.6±3.4) were higher with sperm respectively freshly cooled-diluted or cryopreserved in red fowl extender. However, the rates obtained with frozen-thawed sperm

  4. Ways to improve the biosecurity of bovine semen.

    PubMed

    de Ruigh, L; Bosch, J C; Brus, M C; Landman, B; Merton, J S

    2006-08-01

    Semen production and trade is a worldwide industry. A framework, based on international standards is awaiting international and national regulation. In the perspective of biosecurity of the final product, critical notes can be made according to the semen production regulation and product safety. Process description brings the obligatory health standards for the production bulls, collection and processing of semen, identification, registration, worldwide distribution and insemination into discussion. Test frequency, test quality and demands, way of sampling and test consistency can influence product safety. New scientific knowledge can influence the value of the regulation. Whether a country is free of notifiable disease should influence decisions regarding necessary tests for the production bulls. The biosecurity of the semen production process is influenced by several factors. The effectiveness of the antibiotics used is questionable. The extenders for cryopreservation added to the semen can affect product safety. The way materials and storage containers have to be disinfected must be clear. In modern industry, tracking and tracing is an important issue. Worldwide differences in ways of identification of straws do not benefit a proper identification and registration throughout the process. Regulation could help improve the transparency of production and trade. Before anything concerning biohazard is implemented in regulation, each rule should be thoroughly based on scientific research where possible and furthermore it must be possible to enforce the regulation. The effort it takes to enforce the regulation should be in balance with the benefit it provides. An approach to alter regulation quickly is advisable. To produce a safe product that is accepted for international trade is of vital interest for the survival of artificial insemination (AI) in cattle.

  5. Advanced fertility diagnosis in stallion semen using transmission electron microscopy.

    PubMed

    Pesch, Sandra; Bostedt, Hartwig; Failing, Klaus; Bergmann, Martin

    2006-02-01

    Routine semen analysis of stallions is based on light microscopy (LM). However, there are still a number of animals that are subfertile or even infertile not being identified with conventional semen analysis. The objective of this study was to investigate the suitability of transmission electron microscopy (TEM) for advanced fertility diagnosis in stallion. We examined ejaculates of 46 stallions with known fertility. Animals were divided into three different groups: group 1, fertile stallions (pregnant mares> or =70%, n=29); group 2, subfertile stallions (pregnant mares 10-69%, n=14); group 3, infertile stallions (pregnant mares<10%, n=3). Ejaculates were collected in spring 2002. Conventional semen analysis (volume, sperm concentration, motility, live:dead ratio and percentage of morphologically normal sperm) was immediately performed after semen collection. Ultrastructural analysis included the evaluation of 200 acrosomes, heads, midpieces and cross-sections of tails as well as 100 longitudinal sections of tails from every ejaculate. Using LM, we found a significant increase of morphological deviations from 24.5% (x ) in group 1 to 34.5% in group 2 and 73.5% in group 3. Using TEM, we found a significant increase of detached acrosomes from 6.1% in group 1 to 7.6% in group 2 and 21.4% in group 3. Deviations in tubule pattern were also increased (but not significant) from 2.7% in fertile and 2.8% in subfertile to 11.4% in infertile stallions as well as multiple tails from 1.9% in fertile to 2.0% in subfertile and 8.9% in infertile. Our data indicate that TEM is suitable for advanced fertility diagnostic in stallions, giving a connection between fertility and morphology. It suggests that the most likely reason for sub- and infertility in stallion in case of increased LM pathomorphology of semen are acrosomal alterations, especially detached acrosomes.

  6. Application and commercialization of flow cytometrically sex-sorted semen.

    PubMed

    Rath, D; Johnson, L A

    2008-07-01

    The current technology to sort X and Y chromosome bearing sperm population requires individual identification and selection of spermatozoa in a modified high-speed flow cytometer. For farm animal species, the technology is capable of producing sexed sperm at greater than 90% purity. However, only in the bovine, the technology has reached a developmental level that allows its commercial application. Meanwhile, the demand for female calves has grown rapidly, which encourages the demand for sex-sorted semen from high genetic value bulls. The success of the technology will depend mainly on the fertilizing capacity of the sorted spermatozoa, as this is the most affecting and economically relevant factor. To date, fertility is still variable and is quite dependent on post-sort processing. New processing techniques are under investigation and will likely be able to improve the fertility rates after AI with sex-sorted semen. It is of great importance to select the right bulls and to test the sorted samples on a routine basis. In addition to the demand for sex-sorted semen by the cattle industry, there is also a significant demand expressed by pig farmers. However, it is still unknown if the use of sex-sorted semen through commercial pig AI will be economically feasible. For the pig, the combination of in vitro fertilization with sexed semen and non-surgical embryo transfer is an alternative that merits further scientific attention. Recent developments in ovine AI and ET will make it very likely that commercial sheep industry will adopt the sexing technology in their breeding concepts.

  7. Long term storage of bee semen – A six month assessment of cryopreserved semen quality using motility as an index

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bee (Apis mellifera Linn.) spermatozoa is known for its ability to remain viable for at the least two years within the spermatheca as attested to by the queens’ continued laying of fertile eggs without remating over two to three (Woyke 1960; Winston 1987). However, the semen collected from the dron...

  8. Critical sources of bacterial contamination and adoption of standard sanitary protocol during semen collection and processing in Semen Station

    PubMed Central

    Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

    2015-01-01

    Aim: The present investigation was conducted to locate the critical sources of bacterial contamination and to evaluate the standard sanitation protocol so as to improve the hygienic conditions during collection, evaluation, and processing of bull semen in the Semen Station. Materials and Methods: The study compared two different hygienic procedures during the collection, evaluation and processing of semen in Central Semen Station, Anjora, Durg. Routinely used materials including artificial vagina (AV) inner liner, cone, semen collection tube, buffer, extender/diluter, straws; and the laboratory environment like processing lab, pass box and laminar air flow (LAF) cabinet of extender preparation lab, processing lab, sealing filling machine, and bacteriological lab were subjected to bacteriological examination in two phases of study using two different sanitary protocols. Bacterial load in above items/environment was measured using standard plate count method and expressed as colony forming unit (CFU). Results: Bacterial load in a laboratory environment and AV equipments during two different sanitary protocol in present investigation differed highly significantly (p<0.001). Potential sources of bacterial contamination during semen collection and processing included laboratory environment like processing lab, pass box, and LAF cabinets; AV equipments, including AV Liner and cone. Bacterial load was reduced highly significantly (p<0.001) in AV liner (from 2.33±0.67 to 0.50±0.52), cone (from 4.16±1.20 to 1.91±0.55), and extender (from 1.33±0.38 to 0) after application of improved practices of packaging, handling, and sterilization in Phase II of study. Glasswares, buffers, and straws showed nil bacterial contamination in both the phases of study. With slight modification in fumigation protocol (formalin @600 ml/1000 ft3), bacterial load was significantly decreased (p<0.001) up to 0-6 CFU in processing lab (from 6.43±1.34 to 2.86±0.59), pass box (from 12.13±2

  9. Effect of magnetized extender on sperm membrane integrity and development of oocytes in vitro fertilized with liquid storage boar semen.

    PubMed

    Lee, Sang-Hee; Park, Choon-Keun

    2015-03-01

    The objective of this study was to evaluate the effect of a magnetized extender on sperm membrane damage and development of oocytes in vitro fertilized with liquid storage boar semen. Before semen dilution, extender was flowed through a neodymium magnet (0, 2000, 4000 and 6000G) for 5min and collected semen was preserved for 168h at 18°C. In results, plasma membrane integrity with live sperm was significantly higher in semen treated with extenders magnetized at 4000G than sperm treated with extenders magnetized at 0G during semen preservation for 120-168h (p<0.05). In addition, acrosomal membrane damage was significantly lower in semen treated with extenders magnetized at 4000 and 6000G compared to 0 and 2000G during semen preservation for 168h (p<0.05). And mitochondrial membrane damage with all sperm was significantly lower in semen treated with extenders magnetized at 2000G than other groups during semen preservation for 168h. The ability of semen to achieve successful in vitro fertilization was also not significantly different among the groups during preservation. However, when the semen was preserved for 168h, the blastocyst formation rates were significantly higher at 6000G compared to 0 and 2000G (p<0.05). In conclusion, these results suggest that highly magnetized semen extender could protect the sperm membrane from damage, and improve the ability of rates of in vitro blastocyst development and magnetized semen diluter is beneficial for long liquid preservation of boar semen.

  10. Milk, caseinate and lactoferrin addition to equine semen cooling extenders.

    PubMed

    Martins, H S; Souza, M R; Penna, C F A M; da Silva, G C; Côrtes, S F; Stahlberg, R; Lagares, M A

    2016-11-01

    Cooled semen has been used routinely to prolong sperm viability until artificial insemination time. However, spermatozoa are subjected to oxidative stress. The aim of the present work was to investigate the protective and antioxidant effect of the milk proteins lactoferrin (Lf) and caseinate added to equine semen cooling extenders. Semen from six stallions was cooled at 5 °C after resuspension with C1) milk- and glucose-based, C2) 0.6% caseinate, C3) C2 + Lf 200 μg ml(-1) , C4) C2 + Lf 500 μg ml(-1) and C5) C2 + Lf 1000 μg ml(-1) extenders, and kept at 5 °C for 24 h. Sperm motility characteristics and intact membrane rates were not different among the treatments (P > 0.05). As a result of the cooling process, the nitrite concentration increased significantly in the cooled semen (69.6 ± 78.9 μm per ×10(6) spermatozoa) compared with the fresh semen (8.6 ± 1.9 μm per ×10(6) spermatozoa). In contrast, the H2 O2 concentrations were lower in the 0.6% caseinate extender (265.9 ± 221.3 μm per ×10(6) spermatozoa) than in the milk extender (430.9 ± 199.8 μm per ×10(6) spermatozoa, P < 0.05), showing an antioxidative effect of the caseinate compared with the milk. However, in all groups, hydrogen peroxide concentrations were similar to the undiluted fresh semen (332.8 ± 151.3 μm per ×10(6) spermatozoa). Caseinate showed to be as efficient as milk to protect equine-cooled spermatozoon.

  11. Studies on Freezing RAM Semen in Absence of Glycerol.

    NASA Astrophysics Data System (ADS)

    Abdelnaby, Abdelhady Abdelhakeam

    1988-12-01

    Glycerol is widely used as a major cryoprotective agent for freezing spermatozoa of almost all species. However, it reduces fertility of sheep inseminated cervically compared with intrauterine insemination. Studies were conducted to develop a method and procedure for freezing ram semen in the absence of glycerol. Post -thaw survival of ram spermatozoa frozen in the absence of glycerol was affected by time and temperature after collection and before dilution and time after dilution and before freezing. Increase in time at 5^ circC before or after dilution and before freezing increased both post-thaw motility and number of cells passing through Sephadex filter. A cold dilution method was developed. Slow cooling of fresh ram semen and diluting at 5^circ C 2-3 hr. after collection, then freezing 1 hr. after dilution improved both post-thaw motility and number of cells passing through Sephadex filter compared with immediate dilution at 30-37^circC after collection and freezing 3-4 hr. later (P < 0.05). An extender was developed to freeze ram semen in the absence of glycerol. An increase in post-thaw motility was obtained when semen was extended in TES titrated with Tris to pH 7.0 (TEST) and osmotic pressure of 375-400 mOsm/kg, containing 25-30% (v/v) egg yolk and 10% (v/v) maltose. A special device (boat) for freezing was constructed to insure the same height of the sample above LN _2 and thus the same freezing rate from freeze to freeze. Freezing of semen in 0.25cc straws at 5-10 cm above LN_2 (73.8 to 49.5 ^circC/min) yielded higher post-thaw motility than the rates resulted from freezing at 15 cm above LN_2 or 1 cm above LN _2. Faster Thawing in 37^ circC water for 30 sec. (7.8^ circC/sec.) increased post-thaw motility compared with slower thawing in 5 or 20^circ C water (P < 0.05). A lambing rate of 52.2% was obtained in one fertility trial conducted with ram semen frozen without glycerol and 17.1% in a second trial. One injection (IM) of 15 mg PGF_{2alpha}/ewe for

  12. Effect of antioxidant supplementation in semen extenders on semen quality and reactive oxygen species of chilled canine spermatozoa.

    PubMed

    Michael, A J; Alexopoulos, C; Pontiki, E A; Hadjipavlou-Litina, D J; Saratsis, P; Ververidis, H N; Boscos, C M

    2009-05-01

    The objective of this study was to evaluate quality of chilled dog semen processed with extenders containing various antioxidants. Single ejaculates from five dogs were always pooled and evaluated for concentration, sperm motility, progressive motility (RSF-movement), viability, acrosomal integrity and by the hypo-osmotic swelling (HOS)-test. Also, superoxide (O(2)(-)) production, hydroxyl radicals (OH) and total reactive oxygen species (tROS) were determined. Pooled semen was divided in seven aliquots (for control and test conditions), which were diluted to a final concentration of 67x10(6)spermatozoa/ml with TRIS-glucose-egg yolk extender with or without the following supplements: control (without antioxidants), vitamin C (0.5mM), N-acetyl-l-cysteine (NAC; 0.5mM), taurine (0.2mM), catalase (100u/ml), vitamin E (0.1mM) and 5-(4-dimethylamino-phenyl)-2-phenyl-penta-2,4-dienoic acid (B16; 0.1mM). The semen aliquots were chilled and preserved at 4 degrees C. Portions of chilled semen were removed at 24 and 72h, and semen quality was evaluated after rewarming. At 24h the mean (+/-S.E.M.) sperm motility was higher (p<0.001) when vitamin E, taurine and B16 were added in the extender, whereas more spermatozoa with RSF-movement were observed (p<0.001) in the vitamin E, catalase, B16 and taurine groups. Sperm viability was higher (p=0.040) in B16 and vitamin E groups and the percentage of swollen spermatozoa was higher (p=0.002) only in the B16 group. Acrosomal integrity and OH were not significantly influenced by any of the antioxidants tested. Superoxide production was significantly lower when vitamin C, B16 and vitamin E were added in semen extenders compared with the control (p=0.017). All antioxidant groups, except vitamin C and NAC, contained less tROS compared to the control group, but only the B16 group value differed significantly (p=0.05). At 72h sperm motility was higher (p<0.001) when vitamin E, catalase, B16, taurine and NAC were added in the extender. More

  13. Self-reported mobile phone use and semen parameters among men from a fertility clinic

    PubMed Central

    Lewis, Ryan C.; Mínguez-Alarcón, Lidia; Meeker, John D.; Williams, Paige L.; Mezei, Gabor; Ford, Jennifer B.; Hauser, Russ

    2017-01-01

    There is increasing concern that use of mobile phones, a source of low-level radio-frequency electromagnetic fields, may be associated with poor semen quality, but the epidemiologic evidence is limited and conflicting. The relationship between mobile phone use patterns and markers of semen quality was explored in a longitudinal cohort study of 153 men that attended an academic fertility clinic in Boston, Massachusetts. Information on mobile phone use duration, headset or earpiece use, and the body location in which the mobile phone was carried was ascertained via nurse-administered questionnaire. Semen samples (n=350) were collected and analyzed onsite. To account for multiple semen samples per man, linear mixed models with random intercepts were used to investigate the association between mobile phone use and semen parameters. Overall, there was no evidence for a relationship between mobile phone use and semen quality. PMID:27838386

  14. Self-reported mobile phone use and semen parameters among men from a fertility clinic.

    PubMed

    Lewis, Ryan C; Mínguez-Alarcón, Lidia; Meeker, John D; Williams, Paige L; Mezei, Gabor; Ford, Jennifer B; Hauser, Russ

    2017-01-01

    There is increasing concern that use of mobile phones, a source of low-level radio-frequency electromagnetic fields, may be associated with poor semen quality, but the epidemiologic evidence is limited and conflicting. The relationship between mobile phone use patterns and markers of semen quality was explored in a longitudinal cohort study of 153 men that attended an academic fertility clinic in Boston, Massachusetts. Information on mobile phone use duration, headset or earpiece use, and the body location in which the mobile phone was carried was ascertained via nurse-administered questionnaire. Semen samples (n=350) were collected and analyzed onsite. To account for multiple semen samples per man, linear mixed models with random intercepts were used to investigate the association between mobile phone use and semen parameters. Overall, there was no evidence for a relationship between mobile phone use and semen quality.

  15. A study on the effect of Pseudomonas aeruginosa in semen on bovine fertility.

    PubMed Central

    Eaglesome, M D; Garcia, M M; Bielanski, A B

    1995-01-01

    Two experiments were done to demonstrate whether the presence of Pseudomonas aeruginosa in bovine semen could affect fertilization and/or early embryonic development. In the first experiment, superovulated heifers were inseminated with semen naturally contaminated with P. aeruginosa (ADRI 102) or clean semen and seven day-old embryos were collected nonsurgically. The endometrium of treated heifers appeared more sensitive to the flush procedures. In experiment 2, heifers were inseminated at synchronized estrus with semen experimentally contaminated with P. aeruginosa (ADRI 102) and processed in the same way as commercial semen with antibiotics (gentamicin, lincomycin, spectinomycin and tylosin) or processed without antibiotics added. Embryos were recovered at slaughter seven days later. In general, there was no significant reduction in fertility or development of embryos in vitro as a result of relatively high numbers of P. aeruginosa in bovine semen. PMID:7704848

  16. Symmetry-breaking phase transitions in highly concentrated semen

    PubMed Central

    Creppy, Adama; Plouraboué, Franck; Praud, Olivier; Druart, Xavier; Cazin, Sébastien; Yu, Hui

    2016-01-01

    New experimental evidence of self-motion of a confined active suspension is presented. Depositing fresh semen sample in an annular shaped microfluidic chip leads to a spontaneous vortex state of the fluid at sufficiently large sperm concentration. The rotation occurs unpredictably clockwise or counterclockwise and is robust and stable. Furthermore, for highly active and concentrated semen, richer dynamics can occur such as self-sustained or damped rotation oscillations. Experimental results obtained with systematic dilution provide a clear evidence of a phase transition towards collective motion associated with local alignment of spermatozoa akin to the Vicsek model. A macroscopic theory based on previously derived self-organized hydrodynamics models is adapted to this context and provides predictions consistent with the observed stationary motion. PMID:27733694

  17. Symmetry-breaking phase-transitions in highly concentrated semen

    NASA Astrophysics Data System (ADS)

    Plouraboué, Franck; Creppy, Adama; Praud, Olivier; Druart, Xavier; Cazin, Sébastien; Yu, Hui; Degond, Pierre

    2015-11-01

    New experimental evidence of self-motion of a confined active suspension is presented. Depositing fresh semen sample in an annular shaped micro-fluidic chip leads to a spontaneous rotation motion of the fluid at sufficiently large sperm concentration. The rotation occurs unpredictably clockwise or counterclockwise and is robust and stable. Furthermore, for highly active and concentrated semen, richer dynamics can occur such as self-sustained or damped rotation oscillations. Experimental results obtained with systematic dilution provide a clear evidence of a phase transition toward collective motion associated with local alignment of spermatozoa akin to the Vicsek model. A macroscopic theory based on previously derived Self-Organized Hydrodynamics (SOH) models is adapted to this context and provides predictions consistent with the observed stationary motion.

  18. Immunological responses to semen in the female genital tract.

    PubMed

    Schuberth, H J; Taylor, U; Zerbe, H; Waberski, D; Hunter, R; Rath, D

    2008-11-01

    When spermatozoa, seminal plasma and semen extender reach the uterus and interact with local leukocytes and endometrial cells, several immune mechanisms are initiated which have immediate, mid-term and long-term effects on ovulation, sperm cell selection, fertilization and pregnancy success by assuring the acceptance of fetal tissues. This report gives an overview on relevant key immune mechanisms following roughly the time axis after insemination. Detailed knowledge regarding these mechanisms will aid maximizing reproductive efficiency in livestock production. In the future, the many species involved will require a more comparative approach, since evidence is growing that endometrial physiology and the response to varying amounts and compositions of seminal plasma, various semen extenders, and variable numbers of spermatozoa also provoke different immune responses.

  19. Bull breeding soundness, semen evaluation and cattle productivity.

    PubMed

    Chenoweth, P J; McPherson, F J

    2016-06-01

    The bull breeding soundness evaluation (BBSE) has evolved as a cost-effective veterinary procedure which provides benefits such as risk-reduction and improvements in strategic bull usage, herd fertility and economics. Semen evaluation is an important component of the BBSE when performed appropriately; a consideration that is increasingly addressed by third party andrology laboratories. The combination of competent physical/reproductive exams (including scrotal circumference measurements) and semen evaluations can contribute greatly to the fertility and economics of individual herds as well as adding to understanding of those factors which affect cattle fertility. Despite such advantages, there remain challenges in achieving full acceptance of BBSEs, particularly by the dairy industry and in developing countries.

  20. Preservation of mithun (Bos frontalis) semen at refrigeration temperature.

    PubMed

    Karunakaran, M; Dhali, A; Mech, A; Khate, K; Rajkhowa, C; Mishra, D P

    2007-10-01

    The objective of the present study was to investigate the possibility of preserving mithun (Bos frontalis) spermatozoa at refrigeration temperature using tris-egg yolk diluent. Semen samples were collected from four adult mithun bulls through rectal massage method. Good quality semen samples (n=30) were preserved at 4 degrees C using tris-egg yolk diluent for 72 h. Progressive motility, live spermatozoa count and morphological abnormalities were evaluated every 12 h until 72 h of preservation. The colour, consistency and mass activity of fresh semen samples were found to be creamy white, medium and 3+ to 4+ (5+ scale), respectively. The average (mean+/-S.E.) volume (ml), pH and spermatozoa concentration (10(6) ml(-1)) of fresh semen samples were found to be 0.6+/-0.01, 6.8+/-0.03 and 425+/-48, respectively. Progressive motility and live spermatozoa count were found to be less than 30% (P<0.01) after 48 h of storage. Head (P<0.05), midpiece (P<0.05), tail (P<0.01) and total (P<0.01) abnormalities were found to be increased significantly over the time of storage. It was observed that progressive motility and live spermatozoa count remained above 30% and 40%, respectively, until 36 h of storage. Simultaneously the percentage of morphologically abnormal spermatozoa was found to be significantly low until 36 h of storage. The results indicate that it is possible to preserve mithun spermatozoa at refrigeration temperature in tris-egg yolk diluent, which can be further used for artificial insemination within 36 h of storage.

  1. Semen parameters from 2002 to 2013 in Korea young population: A preliminary report

    PubMed Central

    Kim, Soon Ki; Kim, Yoo Seok; Cho, In-Chang

    2015-01-01

    Purpose To analyze the differences of semen parameters in Korean young population for three periods from 2002 to 2013. Materials and Methods A total of 516 semen samples were collected from Korean men presenting for infertility, varicoceles or other infectious problems for three periods from 2002 to 2012: January 2002-December 2003, January 2007-December 2008, and January 2012-December 2013. A standard World Health Organization procedure for semen analysis was performed for assessment of semen concentration, volume, motility, morphology, and pH. Results A total of 160, 162, 194 men constituted the study populations in 2002 to 2003, in 2007 to 2008, and in 2012 to 2013, respectively. The overall sperm parameter results suggested a statistically significant difference between 2002 to 2003 and 2012 to 2013 except pH. However, considering the data from 2007 to 2008, there were no trends in changes in overall semen parameters. Negative correlations were observed in all semen parameters with increasing age in all patients, except for pH. In addition, semen volume, motility, and morphology had higher negative correlation coefficients with age, from 2002 to 2013, serially. Conclusions There were no significant changes in the semen parameters of Korean men from 2002 to 2013. In addition, semen volume, motility, and morphology showed higher negative correlation coefficients with age from 2002 to 2013, serially. PMID:26682024

  2. Protective effects of l-carnitine on astheno- and normozoospermic human semen samples during cryopreservation.

    PubMed

    Zhang, Wei; Li, Feng; Cao, Haifeng; Li, Chuyan; Du, Congqi; Yao, Lingnv; Mao, Huan; Lin, Wenqin

    2016-04-01

    This study was conducted to determine the effects of l-carnitine (LC), as an antioxidant, in preventing spermatozoa damage during the freezing-thawing process in both astheno- and normozoospermic human semen samples. Seventy semen samples (37 asthenozoospermic and 33 normozoospermic) were involved in this study. Cryopreservation medium supplemented with 1.0 g/l LC was mixed with semen at a ratio of 1:1 (v/v). Controls were cryopreserved with freezing medium only. Assessment of motility, viability (VIA), mitochondrial membrane potential (MMP) and DNA fragmentation index (DFI) were performed on aliquots of fresh semen, frozen-thawed control and frozen-thawed LC treated samples. Supplementation of the cryopreservation medium with LC induced a significant improvement in post-thaw sperm parameters in both the asthenozoospermic and normozoospermic semen samples, compared with those of the control, regarding sperm fast forward motility, forward motility, total motility and VIA. LC showed better protective effects towards asthenozoospermia for DFI (F = 115.85, P < 0.01) and VIA (F = 67.14, P < 0.01) than did normozoospermic semen samples. We conclude that supplementation with LC prior to the cryopreservation process reduced spermatozoa cryodamage in both asthenozoospermic and normozoospermic semen samples. LC had better protective effects for asthenozoospermic human semen samples. Future research should focus on the molecular mechanism for and the different protective effects of LC between asthenozoospermic and normozoospermic semen samples during cryopreservation.

  3. "Tras de un Amoroso Lance" como Estructura Expresiva (The Poem, "Behind the Amorous Cast" as an Expressive Structure).

    ERIC Educational Resources Information Center

    Bratosevich, Nicolas

    1967-01-01

    An analysis of a poem by San Juan de la Cruz (St. John of the Cross), the sixteenth century Spanish mystic, identifies symbols and images, explains themes, and offers a synthesis of his structural patterns. The poem, "Tras de amoroso lance", deals with the theme of the search of the beloved (i.e., the soul) for the lover, and…

  4. Personality of semen donors and their social behaviour.

    PubMed

    Taus, L; Gerzová, J

    1991-01-01

    The authors examined, using three generally accepted methods, the personality structure of 80 semen donors (Cattell's 16-factor questionnaire, 16PF, Eysenck's personality questionnaire, EOD, and Leary's method of interpersonal diagnosis of personality). The donors were selected by means of the Questionnaire of semen donors. The group is subdivided into four sub-groups by the grade of education, i.e. university graduates, men with secondary and elementary education and university students. All are 20-40 years old. The authors describe the assembled results in different sub-groups and in the group as a whole and compare them mutually and with the standardized norm. With regard to the specificity of individual methods and their application the findings are summarized. The donors are balanced personalities, slightly extrovert, emotionally well developed with a realistic outlook. They have positive, sensitive relations with their environment an behaviour towards other people, they are considerate, careful and disciplined. They respect social norms as regards preservation of originality of personality. They have a slight tendency of sheltering behaviour, they wish to be somewhat more aggressive. No pathological phenomena were observed in the donors. Their intelligence is above average. They make a favourable impression with regard to the demand of mental health and transmission of genetic information. The authors evaluate favourably the Questionnaire for semen donors as the method for selection of donors.

  5. Evidence for decreasing quality of semen during past 50 years.

    PubMed Central

    Carlsen, E.; Giwercman, A.; Keiding, N.; Skakkebaek, N. E.

    1992-01-01

    OBJECTIVE--To investigate whether semen quality has changed during the past 50 years. DESIGN--Review of publications on semen quality in men without a history of infertility selected by means of Cumulated Index Medicus and Current List (1930-1965) and MEDLINE Silver Platter database (1966-August 1991). SUBJECTS--14,947 men included in a total of 61 papers published between 1938 and 1991. MAIN OUTCOME MEASURES--Mean sperm density and mean seminal volume. RESULTS--Linear regression of data weighted by number of men in each study showed a significant decrease in mean sperm count from 113 x 10(6)/ml in 1940 to 66 x 10(6)/ml in 1990 (p < 0.0001) and in seminal volume from 3.40 ml to 2.75 ml (p = 0.027), indicating an even more pronounced decrease in sperm production than expressed by the decline in sperm density. CONCLUSIONS--There has been a genuine decline in semen quality over the past 50 years. As male fertility is to some extent correlated with sperm count the results may reflect an overall reduction in male fertility. The biological significance of these changes is emphasised by a concomitant increase in the incidence of genitourinary abnormalities such as testicular cancer and possibly also cryptorchidism and hypospadias, suggesting a growing impact of factors with serious effects on male gonadal function. PMID:1393072

  6. Decreases in Human Semen Quality with Age Among Healthy Men

    SciTech Connect

    Eskenazi, B.; Wyrobek, A.J.; Kidd, S.A.; Moore, L.; Young, S.S.; Moore, D.

    2001-12-01

    The objective of this report is to characterize the associations between age and semen quality among healthy active men after controlling for identified covariates. Ninety-seven healthy, nonsmoking men between 22 and 80 years without known fertility problems who worked for or retired from a large research laboratory. There was a gradual decrease in all semen parameters from 22-80 years of age. After adjusting for covariates, volume decreased 0.03 ml per year (p = 0.001); sperm concentration decreased 2.5% per year (p = 0.005); total count decreased 3.6% per year of age (p < 0.001); motility decreased 0.7% per year (P < 0.001); progressive motility decreased 3.1% per year (p < 0.001); and total progressively motile sperm decreased 4.8% per year (p < 0.001). In a group of healthy active men, semen volume, sperm concentration, total sperm count, and sperm motility decrease continuously between 22-80 years of age, with no evidence of a threshold.

  7. Prospective surveillance of semen quality in the workplace

    SciTech Connect

    Schenker, M.B.; Samuels, S.J.; Perkins, C.; Lewis, E.L.; Katz, D.F.; Overstreet, J.W.

    1988-04-01

    We performed a prospective surveillance of semen quality among workers in the plant where 1,2-dibromo-3-chloropropane was first recognized as an occupational cause of impaired semen quality and of infertility. All male employees of the Agricultural Chemical Division were required to participate. Ninety-seven workers (92% participation) provided 258 semen samples over the 4 years of the program. Most samples were analyzed at the plant with a mini-laboratory designed for the study. Motility and shape measures were made objectively. Sixty-six subjects (68%) were non-azoospermic. Generalized multiple regression showed no significant predictors for any response, with the exception of the motility measures, which were reduced with longer times between ejaculation and assay. Between- and within-person standard deviations and correlations were calculated. Comparison of this population with fertile artificial insemination donors (16 men, 498 ejaculates) revealed generally higher ejaculate-to-ejaculate standard deviations in the worker samples. This is probably due to less well controlled conditions of sperm collection in the workplace setting. For cross-sectional studies, one ejaculate per worker is recommended as sufficient; for estimating an individual worker's mean, even three ejaculates may not provide enough precision.

  8. Fennel (Foeniculum vulgare) provides antioxidant protection for boar semen cryopreservation.

    PubMed

    Malo, C; Gil, L; Cano, R; González, N; Luño, V

    2012-05-01

    Boar semen is extremely vulnerable to cold shock and it is also sensitive to peroxidation due to the high content of unsaturated fatty acids in the plasma membrane. Antioxidants exert a protective effect on the plasma membrane of frozen boar sperm. Fennel has been shown to contain antioxidant substances. Therefore, this study was performed to evaluate the effect of different concentrations of fennel added to the freezing extender on boar semen quality and lipid peroxidation after thawing. Semen collected from four boars was cryopreserved in lactose-egg-yolk extender or in the same extender with varying concentration of fennel essences: low (LF); medium (MF); high (HF). Analysis of data clearly indicated that higher concentrations of fennel produced significant improvement in total motility. Moreover, when fennel was included in the extender, a dose-dependent tendency to increase sperm viability was observed. In contrast, the addition of fennel had no effect on acrosome integrity or hypoosmotic swelling test (HOST) compared with the control. Malondialdehyde (MDA) formation decreased significantly in fennel groups, yielding similar results for MF and HF. Fennel seems a new antioxidant for use in sperm cryopreservation, but its particular effects on sperm physiology must be further studied, especially the causes of motility stimulation and its effect on lipoxidation.

  9. Antioxidant supplements and semen parameters: An evidence based review

    PubMed Central

    Ahmadi, Sedigheh; Bashiri, Reihane; Ghadiri-Anari, Akram; Nadjarzadeh, Azadeh

    2016-01-01

    Many studies have focused on male infertility. There is limited evidence about the influence of nutrition on quality of semen. Approximately, 30-80% of infertility cases are caused by oxidative stress and decreased level of seminal total antioxidant capacity. This study was aimed to review the effects of oral antioxidant supplements on improving major semen parameters such as sperm concentration, motility, morphology, DNA damage, and fertility rate. Data were extracted from PubMed and Google scholar database by using the terms “antioxidant”, “multivitamin”, “carnitine”, “CoQ10”, “vitamin C”, “vitamin E”, “zinc”, “folic acid”, “N-acetyl cysteine” and “selenium” combined with “male infertility”, “semen”, and “sperm” to generate a set of relevant citations. Supplements such as CoQ10 and alpha-tocopherol significantly improve sperm count. Also, carnitine has positive effects on sperm motility and morphology. Simultaneous administration of vitamin E and vitamin C reduces the sperm DNA damage. However, in some studies, one or more factors have not changed substantially. In most of the studies, antioxidant supplementation improved the number, motility, morphology and sometimes DNA integrity of sperm. The present study showed that antioxidant supplements, especially a combination of antioxidants such as vitamin C, vitamin E, and CoQ10 intake can effectively improve semen parameters in infertile men. PMID:28066832

  10. Semen study of papaya workers exposed to ethylene dibromide

    SciTech Connect

    Ratcliffe, J.M.; Schrader, S.M.; Steenland, K.; Clapp, D.; Turner, T.

    1984-01-01

    A cross sectional semen and cytogenetic study was performed on male workers exposed to ethylene-dibromide (EDB) in the papaya fumigation industry in Hawaii. Semen analyses were conducted on 46 men in six fumigation facilities with an average length of employment of 5 years and airborne exposures to EDB ranging from 16 to 213 parts per billion. Statistically significant decreases in sperm count per ejaculate and the percentage of viable and motile sperm and increases in the proportion of specific morphological abnormalities were observed among exposed men when compared with controls. Semen volume and sperm concentration were also lower in the exposed group. No effect of exposure to EDB on sperm velocity, the overall proportion of sperm with normal morphology or YFF bodies was noted. The authors conclude that based on the decreases in sperm count, viability and motility and increases in certain types of morphological abnormalities among workers exposed to EDB, EDB may increase the risk of reproductive impairment in workers at exposure levels near the NIOSH recommended limit of 45 parts per billion and far below the current OSHA standard of 20 parts per million.

  11. Supplementing oregano essential oil to boar diet with strengthened fish oil: Effects on semen antioxidant status and semen quality parameters.

    PubMed

    Liu, Q; Duan, R J; Zhou, Y F; Wei, H K; Peng, J; Li, J L

    2017-02-22

    Previous research has shown benefits of dietary fish oil supplementation on semen quality of boars. However, little is known about how antioxidant protects lipid peroxidation on spermatozoa from n-3 polyunsaturated fatty acid (PUFA) addition. This study evaluated the effect of oregano essential oil (OEO) supplementation on semen antioxidant status and semen quality in boars fed a diet enriched with fish oil. Thirty-four mature boars of proven fertility, received daily 2.5 kg basal diet top-dressed with 45 g soybean oil and 15 g fish oil to meet the n-3 PUFA requirement of spermatozoa, randomly allocated to one of four groups supplemented with 100 mg α-tocopheryl acetate kg(-1) (control), or 250 or 500 or 750 mg OEO kg(-1) for 16 weeks. Semen was collected at weeks 0, 8, 12 and 16 for measurements of sperm production, motion characteristics, sperm α-tocopherol content, antioxidant enzyme activities, reactive oxygen species (ROS), DNA damage (8-hydroxydeoxyguanosine, 8-OHdG), lipoperoxidation (malondialdehyde, MDA) and seminal total antioxidant capacity (TAC). Sperm production and motion characteristics were similar (p > .05) among groups throughout the experimental week 16, but increased (p < .01) with experimental week. Although higher α-tocopherol content and superoxide dismutase (SOD) activities were in OEO group spermatozoa, feeding diet with 500 mg/kg OEO resulted in elevation in seminal TAC, decrease in sperm ROS, MDA and 8-OHdG than control group (p < .05). Overall, these results support the view that oregano essential oil has a positive effect on antioxidant capacity in boar when used fish oil.

  12. Semen analysis in chronic bacterial prostatitis: diagnostic and therapeutic implications

    PubMed Central

    Magri, Vittorio; Wagenlehner, Florian M E; Montanari, Emanuele; Marras, Emanuela; Orlandi, Viviana; Restelli, Antonella; Torresani, Erminio; Naber, Kurt G; Perletti, Gianpaolo

    2009-01-01

    The significance and diagnostic value of semen analysis in chronic bacterial prostatitis has been extensively debated and remains controversial. To investigate the diagnostic relevance of semen culture in the bacteriological workup of prostatitis patients, we retrospectively analyzed a clinical database of 696 symptomatic patients. All patients were routinely subjected to a four-glass test, followed by semen culture and analysis. This allowed to dissect from the database three different diagnostic scenarios, and to compare the 'two-glass' pre-/post- massage test and the standard 'four-glass' test with a 'five-glass' test (four-glass plus post-VB3 semen culture). The 'five-glass' test showed 3.6- or 6.5-fold increases in relative sensitivity and lesser reductions (−13.2% or −14.7%) in relative specificity for traditional uropathogens (TUs) compared with the four-glass or two-glass test, respectively. The area under the ROC curve and Jouden's index were increased, whereas positive and negative likelihood ratios were lower than comparators, indicating that the 'five-glass' assay may be superior in confirming the negative outcome of both standard tests. The five-, four-, and two-glass tests detected TUs (Enterobacteriaceae, Enterococci, etc.) in 120, 33, and 20 patients and unusual pathogens (Streptococci, other Gram-positive species, Mycoplasmata, and others) in 130, 56, and 45 patients, respectively. When patients were subjected to pharmacological treatment, including a combination of a fluoroquinolone and a macrolide, no differences in eradication rates were observed between groups diagnosed with different tests, irrespective of pathogen category. Eradication was associated with long-term sign/symptom remission; no significant intergroup differences in sign/symptom scores were observed throughout a 24-month off-therapy follow-up period. In conclusion, our data support the usefulness of semen analysis in the diagnostic workup of prostatitis patients when this

  13. Influence of chamber type integrated with computer-assisted semen analysis (CASA) system on the results of boar semen evaluation.

    PubMed

    Gączarzewicz, D

    2015-01-01

    The objective of the study was to evaluate the effect of different types of chambers used in computer-assisted semen analysis (CASA) on boar sperm concentration and motility parameters. CASA measurements were performed on 45 ejaculates by comparing three commonly used chambers: Leja chamber (LJ), Makler chamber (MK) and microscopic slide-coverslip (SL). Concentration results obtained with CASA were verified by manual counting on a Bürker hemocytometer (BH). No significant differences were found between the concentrations determined with BH vs. LJ and SL, whereas higher (p<0.01) values of this parameter were obtained with MK. Compared to MK and SL, significantly higher values were recorded in LJ for velocity (VCL and VAP) as well as amplitude of the lateral head displacement (ALH) and beat cross frequency (BCF), which was associated with significantly higher percentages of motile, progressively motile and rapidly progressive motile spermatozoa. Higher values for the linearity (LIN) and straightness (STR) of sperm movement were obtained for the analysis performed in MK and SL. In both these chambers, the results of all the linearity and kinetic parameters of sperm were similar (p>0.05). The results obtained show that CASA assessment of boar semen should account for the effect of counting chamber on the results of sperm motility and concentration, which confirms the need for further study on standardizing the automatic analysis of boar semen.

  14. Persistence of DNA from laundered semen stains: Implications for child sex trafficking cases.

    PubMed

    Brayley-Morris, Helen; Sorrell, Amber; Revoir, Andrew P; Meakin, Georgina E; Court, Denise Syndercombe; Morgan, Ruth M

    2015-11-01

    In sexual assault cases, particularly those involving internal child sex trafficking (ICST), victims often hide their semen-stained clothing. This can result in a lag time of several months before the items are laundered and subsequently seized during a criminal investigation. Although it has been demonstrated previously that DNA can be recovered from clothing washed immediately after semen deposition, laundered items of clothing are not routinely examined in ICST cases, due to the assumption that the time delay and washing would result in no detectable DNA. The aim of this study was to examine whether viable DNA profiles could be recovered from laundered semen stains where there has been a significant lag time between semen deposition from one or more individuals and one or more washes of the stained clothing. Items of UK school uniform (T-shirts, trousers, tights) were stained with fresh semen (either from a single donor or a 1:1 mixture from two donors) and stored in a wardrobe for eight months. Stained and unstained items (socks) were then washed at 30 °C or 60 °C and with non-biological or biological detergent. DNA samples extracted from the semen-stained sites and from the unstained socks were quantified and profiled. High quantities of DNA, (6-18 μg) matching the DNA profiles of the semen donors, were recovered from all semen-stained clothing that had been laundered once, irrespective of wash conditions. This quantity,and profile quality,did not decline significantly with multiple washes. The two donor semen samples yielded ∼ 10-fold more DNA from the T-shirts than from the trousers. This disparity resulted in the T-shirts yielding a ∼ 1:1 mixture of DNA from the two donors, whereas the trousers yielded a major DNA profile matching only that of the second donor. The quantities of DNA recovered from the unstained socks were an order of magnitude lower, with most of the DNA being attributable to the donor of the semen on the stained clothing within the

  15. Effect of species, breed, and age on bacterial load in bovine and bubaline semen

    PubMed Central

    Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

    2015-01-01

    Aim: The present study was conducted to investigate the effect of species, breed and age on bacterial load in fresh and frozen semen of Cattle and Buffalo bull. Materials and Methods: Present study covered 56 cow and 10 buffalo bulls stationed at Central Semen Station Anjora, Durg (Chhattisgarh). Impact of breeds on bacterial load in semen was assessed using six breeds of cattle viz. Sahiwal, Gir, Red Sindhi, Tharparkar, Jersey and Holstein Friesian (HF) cross. Cow bulls were categorized into four different groups based on their age (<4 years, 4-5 years, 5-6 years and > 6 years) to study variation among age groups. Bacterial load was measured in fresh and frozen semen samples from these bulls using the standard plate count (SPC) method and count was expressed as colony forming unit (CFU) per ml of semen. Results: Higher bacterial load was reported in fresh (2.36 × 104 ± 1943 CFU/ml) and frozen (1.00 × 10 ± 90 CFU/ml) semen of cow bulls as compared to buffalo bulls (1.95 × 104 ± 2882 and 7.75 × 102 ± 160 CFU/ml in fresh and frozen semen, respectively). Jersey bull showed significantly higher bacterial count (p < 0.05) both in fresh (4.07 × 104 ± 13927 CFU/ml) and frozen (1.92 × 103 ± 178 CFU/ml) semen followed by HF cross, Sahiwal, Gir, Red Sindhi and Tharparkar bull. Bulls aged < 4 years and more than 6 years yielded increased bacterial load in their semen. Although a minor variation was reported between species and among age groups, no significant differences were measured. Conclusion: Bacterial load in semen did not differ significantly between species and age groups; however significant variation was reported among different breeds. Bulls of Jersey breed showed significantly higher bacterial load in semen as compared to the crossbred and indigenous bull. PMID:27047115

  16. 9 CFR 98.38 - Restrictions on the importation of swine semen from the APHIS-defined European CSF region.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... swine semen from the APHIS-defined European CSF region. 98.38 Section 98.38 Animals and Animal Products... Certain Animal Semen § 98.38 Restrictions on the importation of swine semen from the APHIS-defined European CSF region. In addition to meeting all other applicable provisions of this part, swine...

  17. Detection of Ureaplasma spp. in semen samples from sheep in Brazil

    PubMed Central

    dos Santos, Sandra Batista; de Souza Neto, Orestes Luiz; de Albuquerque, Pedro Paulo Feitosa; da Rocha Mota, André; de Cássia Peixoto Kim, Pomy; de Moraes, Érica Paes Barreto Xavier; do Nascimento, Elmiro Rosendo; do Mota, Rinaldo Apareci

    2013-01-01

    A study was conducted to verify the presence of mycoplasmas and ureaplasmas DNA in sheep semen samples from the State of Pernambuco. The PCR assay was conducted of according with standard protocols with generic primers. Mollicutes DNA was detected in 26.0% and Ureaplasma spp. in 12.0% of semen samples. PMID:24516459

  18. A Review of the Phytochemistry and Pharmacological Activities of Raphani Semen

    PubMed Central

    Sham, Tung-Ting; Yuen, Ailsa Chui-Ying; Ng, Yam-Fung; Chan, Chi-On; Mok, Daniel Kam-Wah

    2013-01-01

    The dried ripe seed of Raphanus sativus L., commonly known as radish seed (or Raphani Semen), is used as traditional Chinese medicine (TCM) to treat constipation, chronic tracheitis, and hypertension. The major active compounds in Raphani Semen are alkaloids, glucosinolates, brassinosteroids, and flavonoids. Fatty acids are its main nutritional contents. Raphani Semen has been demonstrated to have beneficial effects on hypertension, obesity, diabetes mellitus, constipation, and cough. So far, there is no report about the adverse/toxic effects of this herb on humans. However, Raphani Semen processed by roasting was reported to exhibit some adverse effects on mice. Additionally, erucic acid, the main fatty acid in Raphani Semen, was shown to enhance the toxicity of doxorubicin. Thus, Raphani Semen has a potential risk of causing toxicity and drug interaction. In summary, Raphani Semen is a valuable TCM herb with multiple pharmacological effects. More studies on Raphani Semen could help better understand its pharmacological mechanisms so as to provide clear scientific evidence to explain its traditional uses, to identify its therapeutic potential on other diseases, and to understand its possible harmful effects. PMID:23935670

  19. [Evaluation of the quality of the human spermatozoon: comparison between spermatic DNA integrity and semen variables].

    PubMed

    Cruz, Ibis; Colmenares, Melisa; Berrueta-Carrillo, Leidith; Gomez-Perez, Roald; Montes, Henry; Berrueta, Lisbeth; Salmen, Siham; Osuna, Jesús Alfonso

    2010-03-01

    Semen analysis does not have an absolute predictive value on fertility, however it is a reflection of male fertility potential, which is related to its spermatozoa quality and other semen variables. Great variability in human semen parameters has been demonstrated within a single individual, an observation that could explain why a male with low semen quality can successfully fertilize an egg. Although conventional semen analysis, such as sperm concentration, motility and morphology, provide important information about the clinical status of male fertility, new procedures to predict the sperm functional capability have been developed in the last decade, such as analysis of nuclear DNA integrity, which have improved considerably the clinical diagnosis of male infertility, and increased the knowledge about spermatozoa function. DNA fragmentation consist in interruptions, both in single and double DNA strains, that frequently occur in sperm samples from infertile patients. We have conducted a clinical study in semen samples from patients who have attended the Andrology laboratory of the University of Los Andes, between March 2007 and March 2009. The aim of this study was to compare sperm DNA integrity, analyzed by flow cytometry, with traditional semen parameters. Our results show remarkable correlations between conventional human semen variables and sperm chromatin integrity, contributing to asses an integral evaluation of sperm quality allowing the analysis of its fertilizing potential in clinical studies.

  20. A PROTOCOL FOR THE SHORT-TERM STORAGE OF ATLANTIC STURGEON SEMEN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Short-term, liquid-phase storage trials were conducted in 2009 on Atlantic sturgeon semen obtained from captive males, held at the U.S. Fish and Wildlife Service, Northeast Fish Technology Center and wild males, collected ripe on the spawning grounds from the Hudson River. Semen samples collected, c...

  1. Ebola Virus RNA in Semen from an HIV-Positive Survivor of Ebola

    PubMed Central

    Rogers, Emerson; Baller, April; White, Stephen; Soka, Moses; Choi, Mary J.; Mahmoud, Nuha; Wasunna, Christine; Massaquoi, Moses; Kollie, Jomah; Dweh, Straker; Bemah, Philip; Ladele, Victor; Kpaka, Jonathan; Jawara, Mary; Mugisha, Margaret; Subah, Onyekachi; Faikai, Mylene; Bailey, Jeff A.; Rollin, Pierre; Marston, Barbara; Nyenswah, Tolbert; Gasasira, Alex; Knust, Barbara; Nichol, Stuart; Williams, Desmond

    2017-01-01

    Ebola virus is known to persist in semen of male survivors of Ebola virus disease (EVD). However, maximum duration of, or risk factors for, virus persistence are unknown. We report an EVD survivor with preexisting HIV infection, whose semen was positive for Ebola virus RNA 565 days after recovery from EVD. PMID:28287374

  2. Ebola Virus RNA in Semen from an HIV-Positive Survivor of Ebola.

    PubMed

    Purpura, Lawrence J; Rogers, Emerson; Baller, April; White, Stephen; Soka, Moses; Choi, Mary J; Mahmoud, Nuha; Wasunna, Christine; Massaquoi, Moses; Kollie, Jomah; Dweh, Straker; Bemah, Philip; Ladele, Victor; Kpaka, Jonathan; Jawara, Mary; Mugisha, Margaret; Subah, Onyekachi; Faikai, Mylene; Bailey, Jeff A; Rollin, Pierre; Marston, Barbara; Nyenswah, Tolbert; Gasasira, Alex; Knust, Barbara; Nichol, Stuart; Williams, Desmond

    2017-04-01

    Ebola virus is known to persist in semen of male survivors of Ebola virus disease (EVD). However, maximum duration of, or risk factors for, virus persistence are unknown. We report an EVD survivor with preexisting HIV infection, whose semen was positive for Ebola virus RNA 565 days after recovery from EVD.

  3. Sexed-semen usage for Holstein AI in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The dairy industry has used sexed-semen to reduce the birth of undesirable bull calves for over a decade. While the efficacy of sexed-semen has been determined experimentally, we sought to tabulate statistics on the generalized use of the technology in the US dairy herd and determine its effectivene...

  4. [Infrared spectrum analysis of admixture decoction of herba ephedrae with semen armeniacae amarum].

    PubMed

    Lin, Wen-Shuo; Chen, Rong; Guo, Shao-zhong; Lin, Ju-qiang; Feng, Shang-yuan; Li, Yong-zeng; Huang, Zu-fang; Cai, Yu-hui

    2008-12-01

    The infrared spectra of decoction of herba ephedra and semen armeniacae amarum and the mixed decoction of herba ephedra + semen armeniacae amarum were tested. The change in the the mixed decoction was discussed to study the relationship between herba ephedra and semen armeniacae amarum after decoction. The results showed that some absorption peaks of herba ephedra and semen armeniacae amarum were retained in the mixed decoction of herba ephedra + semen armeniacae amarum, such as 1402 and 1076 cm(-1), but some absorption peaks that never appear in the two ingredient spectra increased such as 1394 and 682 cm(-1). New absorption peaks were generated in the mixed decoction of herba ephedra + semen armeniacae amarum, such as 688 and 1187 cm(-1). It can be showed that there were differences in the chemistry environment of the various chemical groups in the three decoctions introduced above, with the variation in absorption peak position, and the biochemical structure of the material changed, possibly with some new chemical compositions created. Medical ingredients in the mixed decoction of herba ephedra + semen armeniacae amarum were not simply the addition of herba ephedra and semen armeniacae amarum based on studies of infrared spectrum of decoction, and the new notion of prescription spectroscopy was proposed.

  5. Evaluation of Lama glama semen viscosity with a cone-plate rotational viscometer.

    PubMed

    Casaretto, C; Martínez Sarrasague, M; Giuliano, S; Rubin de Celis, E; Gambarotta, M; Carretero, I; Miragaya, M

    2012-05-01

    Llama semen is highly viscous. This characteristic is usually evaluated subjectively by measuring the thread formed when carefully pippeting a sample of semen. The aims of this study were (i) to objectively determine and analyse llama semen viscosity, (ii) to compare semen viscosity between ejaculates of the same male as well as between different males, (iii) to study the correlation between viscosity and other semen characteristics and (iv) to evaluate the effect of collagenase on semen viscosity. Semen viscosity was evaluated using a cone-plate Brookfield rotational viscometer. A non Newtonian, pseudoplastic behaviour was observed in the 45 semen samples evaluated. Rheological parameters were determined obtaining the following results (mean ± SD): apparent viscosity at 11.5 s(-1): 46.71 ± 26.8 cpoise and at 115 s(-1): 12.61 ± 4.1 cpoise; structural viscosity (K) (dyne s cm(-2)): 2.18 ± 1.4 and coefficient of consistency (n): 0.45 ± 0.1. Statistical differences were found between different ejaculates of the same male for structural viscosity and apparent viscosity at 11.5 s(-1) (P < 0.01). Correlation was found only between coefficient of consistency (n) and sperm concentration (P < 0.01). Significant differences for coefficient of consistency (n) and viscosity at 115 s(-1) were found between samples incubated with and without collagenase (P < 0.05).

  6. Application of a quantitative 1H-NMR method for the determination of amygdalin in Persicae semen, Armeniacae semen, and Mume fructus.

    PubMed

    Tanaka, Rie; Nitta, Akane; Nagatsu, Akito

    2014-01-01

    A quantitative (1)H-NMR method (qHNMR) was used to measure the amygdalin content of Persicae semen, Armeniacae semen, and Mume fructus, in each of which amygdalin constitutes a major component. The purity of amygdalin was calculated from the ratio of the intensity of the amygdalin H-2 signal at δ 6.50 ppm in pyridine-d 5 to that of the hexamethyldisilane (HMD) signal at 0 ppm. The HMD concentration was corrected by the International System of Units (SI) traceability with certified reference material (CRM)-grade bisphenol A. qHNMR revealed the amygdalin contents to be 2.72 and 3.13% in 2 lots of Persicae semen, 3.62 and 5.19% in 2 lots of Armeniacae semen, and 0.23% in Mume fructus. Thus, we demonstrated the utility of this method for the quantitative analysis of crude drugs.

  7. Applications and cost benefits of sexed semen in pasture-based dairy production systems.

    PubMed

    Butler, S T; Hutchinson, I A; Cromie, A R; Shalloo, L

    2014-05-01

    Sexed semen technology is now commercially available in many countries around the world, and is primarily used in dairy cattle breeding. Sperm are sorted by flow cytometry on the basis of a 4% difference in DNA content between sperm containing X and Y chromosomes. Despite reliably producing a 90% gender bias, the fertility of the sexed semen product is compromised compared with conventional semen. The negative implications of the reduced fertility of sexed semen are amplified in seasonal systems of dairy production, as the importance of fertility is greater in these systems compared with year-round calving systems. A review of the literature indicates that conception rates (CR) to 1st service with frozen-thawed sexed semen are ~75% to 80% of those achieved with conventional frozen-thawed semen. Preliminary results from a large-scale field trial carried out in Ireland in 2013 suggest that significant improvements in the performance of sexed semen have been made, with CR of 87% of those achieved with conventional semen. The improved fertility of a sexed semen product that delivers a 90% gender bias has considerable implications for the future of breeding management in pasture-based dairy production systems. Sexed semen may facilitate faster, more profitable dairy herd expansion by increasing the number of dairy heifer replacements born. Biosecurity can be improved by maintaining a closed herd during the period of herd expansion. In a non-expansion scenario, sexed semen may be used to increase the value of beef output from the dairy herd. The replacement heifer requirements for a herd could be met by using sexed semen in the 1st 3 weeks of the breeding season, with the remaining animals bred to beef sires, increasing the sale value over that of a dairy bull calf. Alternatively, very short gestation sires could be used to shorten the calving interval. Market prices have a considerable effect on the economics of sexed semen use, and widespread use of sexed semen should

  8. Sequence-Specific Recognition and Cleavage of Telomeric Repeat (TTAGG)n by Endonuclease of Non-Long Terminal Repeat Retrotransposon TRAS1

    PubMed Central

    Anzai, Tomohiro; Takahashi, Hidekazu; Fujiwara, Haruhiko

    2001-01-01

    The telomere of the silkworm Bombyx mori consists of (TTAGG/CCTAA)n repeats and harbors a large number of telomeric repeat-specific non-long terminal repeat retrotransposons, such as TRAS1 and SART1. To understand how these retrotransposons recognize and integrate into the telomeric repeat in a sequence-specific manner, we expressed the apurinic-apryrimidinic endonuclease-like endonuclease domain of TRAS1 (TRAS1 EN), which is supposed to digest the target DNA, and characterized its enzymatic properties. Purified TRAS1 EN could generate specific nicks on both strands of the telomeric repeat sequence between T and A of the (TTAGG)n strand (bottom strand) and between C and T of the (CCTAA)n strand (top strand). These sites are consistent with insertion sites expected from the genomic structure of boundary regions of TRAS1. Time course studies of nicking activities on both strands revealed that the cleavages on the bottom strand preceded those on the top strand, supporting the target-primed reverse transcription model. TRAS1 EN could cleave the telomeric repeats specifically even if it was flanked by longer tracts of nontelomeric sequence, indicating that the target site specificity of the TRAS1 element was mainly determined by its EN domain. Based on mutation analyses, TRAS1 EN recognizes less than 10 bp around the initial cleavage site (upstream 7 bp and downstream 3 bp), and the GTTAG sequence especially is essential for the cleavage reaction on the bottom strand (5′. . . TTAGGTT ↓ AGG . . . 3′). TRAS1 EN, the first identified endonuclease digesting telomeric repeats, may be used as a genetic tool to shorten the telomere in insects and some other organisms. PMID:11113185

  9. Presence and Persistence of Zika Virus RNA in Semen, United Kingdom, 2016.

    PubMed

    Atkinson, Barry; Thorburn, Fiona; Petridou, Christina; Bailey, Daniel; Hewson, Roger; Simpson, Andrew J H; Brooks, Timothy J G; Aarons, Emma J

    2017-04-01

    Zika virus RNA has been detected in semen collected several months after onset of symptoms of infection. Given the potential for sexual transmission of Zika virus and for serious fetal abnormalities resulting from infection during pregnancy, information regarding the persistence of Zika virus in semen is critical for advancing our understanding of potential risks. We tested serial semen samples from symptomatic male patients in the United Kingdom who had a diagnosis of imported Zika virus infection. Among the initial semen samples from 23 patients, Zika virus RNA was detected at high levels in 13 (56.5%) and was not detected in 9 (39.1%); detection was indeterminate in 1 sample (4.4%). After symptomatic infection, a substantial proportion of men have detectable Zika virus RNA at high copy numbers in semen during early convalescence, suggesting high risk for sexual transmission. Viral RNA clearance times are not consistent and can be prolonged.

  10. Presence and Persistence of Zika Virus RNA in Semen, United Kingdom, 2016

    PubMed Central

    Atkinson, Barry; Thorburn, Fiona; Petridou, Christina; Bailey, Daniel; Hewson, Roger; Simpson, Andrew J.H.; Brooks, Timothy J.G.

    2017-01-01

    Zika virus RNA has been detected in semen collected several months after onset of symptoms of infection. Given the potential for sexual transmission of Zika virus and for serious fetal abnormalities resulting from infection during pregnancy, information regarding the persistence of Zika virus in semen is critical for advancing our understanding of potential risks. We tested serial semen samples from symptomatic male patients in the United Kingdom who had a diagnosis of imported Zika virus infection. Among the initial semen samples from 23 patients, Zika virus RNA was detected at high levels in 13 (56.5%) and was not detected in 9 (39.1%); detection was indeterminate in 1 sample (4.4%). After symptomatic infection, a substantial proportion of men have detectable Zika virus RNA at high copy numbers in semen during early convalescence, suggesting high risk for sexual transmission. Viral RNA clearance times are not consistent and can be prolonged. PMID:27997333

  11. Freezing African Elephant Semen as a New Population Management Tool

    PubMed Central

    Hermes, Robert; Saragusty, Joseph; Göritz, Frank; Bartels, Paul; Potier, Romain; Baker, Barbara; Streich, W. Jürgen; Hildebrandt, Thomas B.

    2013-01-01

    Background The captive elephant population is not self-sustaining and with a limited number of breeding bulls, its genetic diversity is in decline. One way to overcome this is to import young and healthy animals from the wild. We introduce here a more sustainable alternative method - importation of semen from wild bulls without removing them from their natural habitat. Due to the logistics involved, the only practical option would be to transport cryopreserved sperm. Despite some early reports on African elephant semen cryopreservation, the utility of this new population management tool has not been evaluated. Methodology/Principal Findings Semen was collected by electroejaculation from 14 wild African savanna elephant (Loxodonta africana) bulls and cryopreserved using the directional freezing technique. Sperm treatments evaluated included the need for centrifugation, the use of hen or quail yolk, the concentration of glycerol (3%, 5% or 7%) in the extender, and maintenance of motility over time after thawing. Our results suggest that dilution in an extender containing hen yolk and 7% glycerol after centrifugation best preserved post-thaw sperm motility when compared to all other treatments (P≤0.012 for all). Using this approach we were able to achieve after thawing (mean ± SD) 54.6±3.9% motility, 85.3±2.4% acrosome integrity, and 86.8±4.6% normal morphology with no decrease in motility over 1 h incubation at 37°C. Sperm cryopreserved during this study has already lead to a pregnancy of a captive female elephant following artificial insemination. Conclusions/Significance With working techniques for artificial insemination and sperm cryopreservation of both African and Asian elephants in hand, population managers can now enrich captive or isolated wild elephant populations without removing valuable individuals from their natural habitat. PMID:23483917

  12. Influence of boar breeds or hybrid genetic composition on semen quality and seminal plasma biochemical variables.

    PubMed

    Žaja, Ivona Žura; Samardžija, Marko; Vince, Silvijo; Majić-Balić, Ivanka; Vilić, Marinko; Đuričić, Dražen; Milinković-Tur, Suzana

    2016-01-01

    The enzyme concentrations of seminal plasma are important for spermatozoa metabolism and function in boars. The need has arisen for introducing a biochemical evaluation of semen, along with the usual standard semen analyses. There are no data on the influence of boar breeds on the seminal plasma biochemical variables investigated in this study. Therefore, the objective was to determine the influence of breed and hybrid genetic composition of boars on semen quality and seminal plasma biochemical variables. Semen samples of 27 boars (Swedish Landrace, German Landrace, Large White, Pietrain and Pig Improvement Company hybrid-PIC-hybrid), aged between 1.5 and 3 years, were collected. After evaluation of semen quality, the seminal plasma was separated from the spermatozoa by centrifugation of semen. The seminal plasma was subjected to spectrophotometric analysis to determine alkaline phosphatase (ALP), acid phosphatase (ACP), γ-glutamyltransferase (GGT), creatine kinase (CK) and lactate dehydrogenase (LDH) and to atomic absorption spectrophotometric analysis to measure the concentration of calcium and magnesium. Conventional semen quality variables differed depending on breed and PIC-hybrid genetic composition, though these differences were typically insignificant. In the seminal plasma, significant differences were determined in enzyme activity (ALP, GGT, CK and LDH) and in calcium concentration among boars of different breeds. There are, therefore, differences in semen quality and significant differences in the seminal plasma biochemical variables among boars of different breeds and PIC-hybrid genetic composition. The data and differences in semen variables detected in the present study provide knowledge for enhancing evaluation and monitoring of boar reproductive potential, semen quality and explain the potential causes of boar infertility.

  13. Shedding of Hepatitis C Virus in Semen of Human Immunodeficiency Virus-Infected Men

    PubMed Central

    Turner, Samuel S.; Gianella, Sara; Yip, Marcus J-S.; van Seggelen, Wouter O.; Gillies, Robert D.; Foster, Andrew L.; Barbati, Zachary R.; Smith, Davey M.; Fierer, Daniel S.

    2016-01-01

    Background. The epidemic of sexually transmitted hepatitis C virus (HCV) infection among human immunodeficiency virus (HIV)-infected men who have sex with men (MSM) has been documented for over a decade. Despite this, there is no consensus as to the risk factors for sexual acquisition of HCV in these men. Methods. We obtained paired semen and blood samples at 2-week intervals from HIV-infected MSM with recent and chronic HCV infection and quantified HCV in semen. Results. Hepatitis C virus was quantified in 59 semen specimens from 33 men. Hepatitis C virus was shed in 16 (27%) of semen specimens from 11 (33%) of the men. Median HCV viral load (VL) in semen was 1.49 log10 IU/mL. Hepatitis C virus VL in blood was significantly higher at the time of HCV shedding in semen than when HCV shedding in semen was not detected (P = .002). Furthermore, there was a significant correlation between the HCV VL in blood and semen overall (rs = 0.41; P = .001), and in the subgroup with recent HCV infection (rs = 0.37; P = .02), but not in the subgroup with chronic HCV infection (rs = 0.34; P = .1). Conclusions. One third of HIV-infected MSM coinfected with HCV shed HCV into their semen. Based on the HCV VL in semen in this study, an average ejaculate would deliver up to 6630 IU of virus into the rectum of the receptive partner. Therefore, our data strongly support that condoms should be used during anal intercourse among MSM to prevent transmission of HCV. PMID:27186582

  14. The in vitro effect of leptin on semen quality of water buffalo (Bubalus bubalis) bulls.

    PubMed

    Khaki, Amir; Batavani, Rooz Ali; Najafi, Gholamreza

    2013-01-01

    The purpose of this study was to evaluate the probable effects of leptin addition in different levels to the semen extender on sperm quality (motility and motility parameters, viability, sperm membrane integrity, and DNA damage). Semen specimens were evaluated immediately after leptin addition, equilibration time and after thawing the frozen semen. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 10, 20, 50, 100, and 200 ng mL(-1) leptin. The diluted semen was kept 4 hr in refrigerator to reach to the equilibration time and then packed in 0.5 mL French straws and frozen in liquid nitrogen. Our results showed that, in the fresh semen, no significant difference was observed in all sperm quality parameters evaluated among all of the examined leptin concentrations. Addition of 10 ng mL(-1) leptin into semen extender significantly preserved sperm motility, all of the motility parameters, and viability in equilibrated semen compared to that of control group. However, in vitro addition of 200 ng mL(-1) leptin, significantly decreased theses parameters. In the frozen thawed semen, all leptin concentrations decreased sperm motility and viability, but significant decrease was observed in concentrations of 100 and 200 ng mL(-1). Adding leptin to semen extender did not have any significant influence on sperm DNA damage and sperm membrane integrity in all examined groups. These findings suggest that in vitro addition of 10 ng mL(-1) leptin could preserve sperm motility and viability in cooled semen of buffaloes.

  15. Semen Bacterial Concentrations and HIV-1 RNA Shedding Among HIV-1–Seropositive Kenyan Men

    PubMed Central

    Srinivasan, Sujatha; Huang, Dandi; Ko, Daisy L.; Sanders, Eduard J.; Peshu, Norbert M.; Krieger, John N.; Muller, Charles H.; Coombs, Robert W.; Fredricks, David N.; Graham, Susan M.

    2017-01-01

    Introduction: HIV-1 is transmitted through semen from men to their sexual partners. Genital infections can increase HIV-1 RNA shedding in semen, but shedding also occurs in the absence of typical pathogens. We hypothesized that higher bacterial concentrations in semen would be associated with higher HIV-1 RNA levels. Methods: We analyzed semen samples from 42 HIV-1–seropositive Kenyan men using quantitative polymerase chain reaction (PCR) to assess bacterial concentrations and real-time PCR to measure HIV-1 RNA levels. Generalized estimation equations were used to evaluate associations between these 2 measures. Broad-range 16S rRNA gene PCR with pyrosequencing was performed on a subset of 13 samples to assess bacterial community composition. Results: Bacteria were detected in 96.6% of 88 samples by quantitative PCR. Semen bacterial concentration and HIV-1 RNA levels were correlated 0.30 (P = 0.01). The association between bacterial concentration and HIV-1 RNA detection was not significant after adjustment for antiretroviral therapy (ART) (adjusted odds ratio: 1.27, 95% CI: 0.84 to 1.91). Factors associated with semen bacterial concentration included insertive anal sex (adjusted beta 0.92, 95% CI: 0.12 to 1.73) and ART use (adjusted beta: −0.77, 95% CI: −1.50 to 0.04). Among 13 samples with pyrosequencing data, Corynebacterium spp., Staphylococcus spp., and Streptococcus spp. were most frequently detected. Conclusion: Most of these HIV-1–infected men had bacteria in their semen. ART use was associated with undetectable semen HIV-1 RNA and lower semen bacterial concentrations, whereas insertive anal sex was associated with higher bacterial concentrations. Additional studies evaluating the relationship between semen bacteria, inflammation, mucosal immunity, and HIV-1 shedding are needed to understand implications for HIV-1 transmission. PMID:27861240

  16. Leonardo da Vinci and the origin of semen

    PubMed Central

    Noble, Denis; DiFrancesco, Dario; Zancani, Diego

    2014-01-01

    It is well known that Leonardo da Vinci made several drawings of the human male anatomy. The early drawings (before 1500) were incorrect in identifying the origin of semen, where he followed accepted teaching of his time. It is widely thought that he did not correct this mistake, a view that is reflected in several biographies. In fact, he made a later drawing (after 1500) in which the description of the anatomy is remarkably accurate and must have been based on careful dissection. In addition to highlighting this fact, acknowledged previously in only one other source, this article reviews the background to Leonardo's knowledge of the relevant anatomy. PMID:27494016

  17. LEONARDO DA VINCI AND THE ORIGIN OF SEMEN.

    PubMed

    Noble, Denis; DiFrancesco, Dario; Zancani, Diego

    2014-12-20

    It is well known that Leonardo da Vinci made several drawings of the human male anatomy. The early drawings (before 1500) were incorrect in identifying the origin of semen, where he followed accepted teaching of his time. It is widely thought that he did not correct this mistake, a view that is reflected in several biographies. In fact, he made a later drawing (after 1500) in which the description of the anatomy is remarkably accurate and must have been based on careful dissection. In addition to highlighting this fact, acknowledged previously in only one other source, this article reviews the background to Leonardo's knowledge of the relevant anatomy.

  18. Dioxins in the semen of men with infertility.

    PubMed

    Galimova, E F; Amirova, Z K; Galimov, Sh N

    2015-10-01

    The purpose of the present study was to assess ejaculate contamination by polychlorinated dibenzo-p-dioxins/furans in male infertility. The database of 168 infertile and 49 fertile men was included in the study. Dioxin content was determined using gas chromatography/high-resolution mass spectrometry (GC/HRMS). In the ejaculate of infertile men, the content of dioxins and furans was 2.2-2.3 times higher than in fertile donors. The maximum level of the most toxic dioxin congener was detected in pathospermia. Contamination of semen of infertile men by polychlorinated dibenzo-p-dioxins/furans supports the hypothesis about the relationship between environmental factors and reproductive health.

  19. Laser researches on livestock semen and oocytes: A brief review

    PubMed Central

    Abdel-Salam, Z.; Harith, M.A.

    2014-01-01

    This article presents a brief review of the past and present literature pertinent to laser effects on sperm motility parameters, improvement of oocyte maturation and characterization of semen in livestock. The aim was, on one hand, to make the readers aware of such knowledge and on the other hand to trigger the interest of the animal reproduction scientific community in attempting some laser techniques that have not yet been fully exploited in the field of artificial insemination. With respect to the conventional methods, laser is a more sensitive and less costly technology that can be used for improving artificial insemination and embryo production system. Since 1980s, laser treatment came on the biological samples scene; its applications have continuously been developed thereafter. Exploitation of laser light by various researchers for improving the reproductive efficiency of sperm cells and the maturation rate in different livestock is demonstrated herein. Laser irradiation, in principal, can increase the production of adenosine triphosphate (ATP) and consequently increases the energy provided to the cell. Since sperm motility and oocyte maturation depend on the energy consumption, an increase in the energy supply to the cells will be of great importance. In addition, the authors also discuss the use of laser spectrochemical analytical techniques, such as laser induced breakdown spectroscopy (LIBS) and laser induced fluorescence (LIF), in characterization of semen samples. PMID:26257928

  20. Alkaline phosphatase in stallion semen: characterization and clinical applications.

    PubMed

    Turner, R M O; McDonnell, S M

    2003-06-01

    Significant amounts of alkaline phosphatase (AP) activity have been found in semen plasma from numerous species. In species in which the majority of semen plasma AP (SPAP) activity originates from the epididymis and testicle, SPAP activity can be used clinically as a marker to differentiate testicular origin azoospermia or oligospermia from ejaculatory failure. Information on SPAP activity in stallions to date has been limited. In this study, a standard clinical chemistry analyzer was used to determine AP activity in pre-ejaculatory fluid and ejaculates from groups of normal stallions. Additionally, accessory glands, epididymides, testicles and other components of the urogenital tract of normal stallions were assayed to determine which tissues contain SPAP activity. The results indicated that levels of AP activity are low in pre-ejaculatory fluid, but significantly higher in ejaculatory fluid from normal stallions. Spermatozoa were not a significant source of SPAP activity. High levels of SPAP activity were found in the testes and epididymides. These findings suggest that SPAP activity is a candidate for a sperm-independent marker for ejaculation in the stallion. Finally, AP activity was determined in ejaculatory fluid from a stallion with bilaterally blocked ampullae, both before and after relief of the blockage. While the blockage was present, AP activity in ejaculatory fluid was low. However, following relief of the blockage, AP activity in ejaculatory fluid rose dramatically, thus suggesting that AP activity will be useful as an inexpensive, simple clinical assay for differentiating ejaculatory failure or excurrent duct blockages from testicular origin azoospermia and oligospermia.

  1. Semen Characteristics of Purebred and Crossbred Male Rabbits

    PubMed Central

    El-Tarabany, Mahmoud Salah; El-Bayomi, Khairy; Abdelhamid, Tamer

    2015-01-01

    The aim of this investigation was to evaluate the semen quality traits of purebred male rabbits and their crosses under subtropical Egyptian conditions. A full 3 x 3 diallel crossing was performed for producing the first generation progeny of New Zealand White (N), Flander (F) and Rex (R) breeds. The highest ejaculate volume (p< 0.05) and percentage of live sperms (p<0.01) with the lowest percentage of sperm cell morphological abnormalities (p<0.05) had been recorded in the NF bucks. Moreover, they possessed positive estimates of direct heterosis for ejaculate volume, mass motility (Mm), individual motility (Im) and sperm cell concentration (SCC). On the contrary, pH had negative estimates of direct heterosis in all crosses and their reciprocal. Semen pH was negatively correlated with SCC (r = -0.18), Mm (r = -0.13) and Im (r = -0.23). In conclusion, the superiority of crossbreeding was particularly obvious in the New Zealand White x Flander males, which cumulated heterosis and favorable maternal effects of the Flander dams. PMID:26020961

  2. Impact of cell phone use on men's semen parameters.

    PubMed

    Gutschi, T; Mohamad Al-Ali, B; Shamloul, R; Pummer, K; Trummer, H

    2011-10-01

    The objective of the present retrospective study was to report our experience concerning the effects of cell phone usage on semen parameters. We examined 2110 men attending our infertility clinic from 1993 to October 2007. Semen analysis was performed in all patients. Serum free testosterone (T), follicle stimulating hormone (FSH), luteinising hormone (LH) and prolactin (PRL) were collected from all patients. The information on cell phone use of the patients was recorded and the subjects were divided into two groups according to their cell phone use: group A: cell phone use (n = 991); group B: no use (n = 1119). Significant difference was observed in sperm morphology between the two groups. In the patients of group A, 68.0% of the spermatozoa featured a pathological morphology compared to only 58.1% in the subjects of group B. Patients with cell phone usage showed significantly higher T and lower LH levels than those who did not use cell phone. No significant difference between the two groups was observed regarding FSH and PRL values. Our results showed that cell phone use negatively affects sperm quality in men. Further studies with a careful design are needed to determine the effect of cell phone use on male fertility.

  3. Effects of chronic boron exposure on semen profile.

    PubMed

    Korkmaz, Mehmet; Yenigün, Mehmet; Bakırdere, Sezgin; Ataman, Osman Yavuz; Keskin, Sıddık; Müezzinoğlu, Talha; Lekili, Murat

    2011-11-01

    The possible changes in semen quality were studied in men living in a boron mining area. The subjects in the boron group had exposure to boron at an average level of 6.5 mg/day, as determined by urinary analysis. The results obtained by the boron group were compared to those obtained for the control group whose subjects were living in the same geographical area but away from the boron region; average exposure level was 1.4 mg/day for this group. The semen samples were analyzed according to the recommendations of the World Health Organization. Boron levels were established in the water samples obtained from various locations in the study region. In the boron mining fields where the subjects in the boron group live, water samples contained boron in the range of 1.4-6.5 mg/L, while the values were <0.01 mg/L for the water samples obtained from the region where the subjects of the control group reside. No negative effects were found in the sperm samples obtained from the subjects of the boron group.

  4. Collection and preservation of pygmy hippopotamus (Choeropsis liberiensis) semen.

    PubMed

    Saragusty, J; Hildebrandt, T B; Bouts, T; Göritz, F; Hermes, R

    2010-09-01

    Knowledge about the reproduction of the endangered pygmy hippopotamus is almost non-existent. This study takes the first step toward changing this by devising a protocol for the collection, evaluation, and short-term preservation of semen of this endangered species. Semen was collected successfully from seven bulls by electroejaculation, using a specially designed rectal probe. Mean +/- SEM values of native sperm parameters from combined best fractions were: motility-80.0 +/- 4.1%, concentration-2421 +/- 1530 x 10(6) cells/mL, total collected cell number-759 +/- 261 x 10(6) cells, intact acrosome-87.8 +/- 1.2%, intact morphology-52.7 +/- 4.3%, and, for some, hypoosmotic swelling test-79.3 +/- 4.4% and seminal plasma osmolarity-297.5 +/- 3.3 mOsm. Seven different extenders were tested for sperm storage under chilling conditions: Berliner Cryomedium (BC), Biladyl, modification of Kenney modified Tyrode's medium (KMT), MES medium, Androhep((R)), boar M III() extender and Human Sperm Refrigeration Medium. While differences between males were apparent, the BC was consistently superior to all other extenders in sperm motility and facilitated storage for 7 d with up to 30% motility and some motility even after 3 weeks. With this knowledge in hand, the obvious two directions for future research are to conduct artificial insemination and to develop a technique for sperm cryopreservation.

  5. Effect of dialysis on quality characteristics of turkey semen during liquid storage.

    PubMed

    Iaffaldano, N; Meluzzi, A

    2003-08-01

    Low molecular weight substances such as zinc and peroxides are present in seminal plasma and are responsible for deleterious effects in stored semen. On the contrary, molecules larger than 50 kDa are beneficial to in-vitro storage of spermatozoa. Since the effects of different seminal plasma fractions in turkey semen are not completely known, the purpose of the study was to determine the effects of turkey semen dialysis with a 12-14 kDa cut-off on viability, hypo-osmotic membrane integrity, or sperm motility of turkey spermatozoa stored up to 48 h at 5 degrees C. Twelve pools of semen, each pool originating from four toms, were used. Each pool was divided into two aliquots, one of which was dialyzed while the other represented the control. Each semen aliquot was evaluated for sperm viability, membrane integrity and motility after 6, 24 and 48 h of in-vitro storage. Cold storage of turkey semen for 48 h significantly worsened (P<0.01) sperm viability, hypo-osmotic membrane integrity, and sperm motility index of both control and dialyzed samples. After 24 and 48 h sperm viability, membrane integrity and sperm motility index were better (P<0.01) in dialyzed semen compared to the control.

  6. Bacteriospermia and Its Impact on Basic Semen Parameters among Infertile Men

    PubMed Central

    Kandasamy, Balan; Jayachandran, Abiramy Lakshmy; Sathiyanarayanan, Sarasa; Tanjore Singaravelu, Vijayalakshmi; Krishnamurthy, Veeraraghavan; Elangovan, Vanithadevi

    2016-01-01

    Introduction. Semen analysis is considered as the surrogate marker for male fecundity while assessing infertile men. There are several reasons for altered semen quality and bacteriospermia could be one among them. Thereby the aim of our work is to study the semen culture and its impact on semen parameters among infertile men. Materials and Methods. Semen samples were collected from men attending infertility clinic. Semen parameters were analysed based on WHO guidelines. Also, samples were subjected to culture using standard bacteriological techniques. Results. A total of 85 samples were collected. A number of 47 (55.30%) had normal sperm count, 37 (43.50%) had oligozoospermia, and one (1.17%) had azoospermia. Teratozoospermia was the most common abnormality observed (81.17%) followed by asthenozoospermia (28.23%). The prevalence of bacteriospermia was 35.3%. Enterococcus faecalis (30%) was the most common organism isolated followed by Coagulase negative Staphylococcus (23.33%), Staphylococcus aureus (20%), and E. coli (10%). Other less frequently isolated organisms were Klebsiella pneumoniae (6.66%), Proteus sp. (6.66%), and Citrobacter sp. (3.33%). Conclusion. The presence of asymptomatic bacteriospermia did not correlate with abnormal semen parameters. PMID:26880908

  7. Dark-coloured semen in nonobstructive azoospermia: a report of four cases.

    PubMed

    Halder, A; Jain, M; Chaudhary, I; Kumar, G; Das, T; Gupta, Y K

    2014-04-01

    Dark-coloured semen is rarely observed in men with nonobstructive azoospermia in absence of genital tract bleeding. Dark-coloured semen is also observed rarely in men with spinal cord injury with or without genital tract bleeding. However, this condition has not been reported in the literature in absence of genital tract bleeding or spinal cord injury. Here, we report men with nonobstructive azoospermia with dark-coloured semen and without genital tract bleeding or spinal cord injury. The study included four subjects with dark-coloured semen. All the samples first investigated for the presence of red blood cells as well as haemoglobin. All the cases were examined in details with seminal, hormonal and genetic parameters. In addition, an elemental electron microscopy and inductively coupled plasma-atomic emission spectrometry evaluation for heavy metals was performed. The semen parameters showed normal volume, pH, leucocyte count and azoospermia or oligoazoospermia. Dark-coloured semen specimens contained neither red blood cells nor haeme pigment. Electron microscopy showed presence of platinum in all the cases, whereas inductively coupled plasma-atomic emission spectrometry revealed increased levels of lead, manganese and nickel in serum as compared to controls. The results indicate dark-coloured semen may be linked with heavy metals found in seminal cells.

  8. Determination of Sperm Sex Ratio in Bovine Semen Using Multiplex Real-time Polymerase Chain Reaction.

    PubMed

    Khamlor, Trisadee; Pongpiachan, Petai; Sangsritavong, Siwat; Chokesajjawatee, Nipa

    2014-10-01

    Gender selection is important in livestock industries; for example, female calves are required in the dairy industry. Sex-sorted semen is commonly used for the production of calves of the desired gender. However, assessment of the sex ratio of the sorted semen is tedious and expensive. In this study, a rapid, cost effective and reliable method for determining the sex ratio was developed using a multiplex real-time polymerase chain reaction (PCR) assay. In this assay, the X and Y chromosome-specific markers, i.e., bovine proteolipid protein (PLP) gene and sex-determining region Y (SRY) were simultaneously quantified in a single tube. The multiplex real-time PCR assay was shown to have high amplification efficiencies (97% to 99%) comparable to the separated-tube simplex real-time PCR assay. The results obtained from both assays were not significantly different (p>0.05). The multiplex assay was validated using reference DNA of known X ratio (10%, 50%, and 90%) as templates. The measured %X in semen samples were the same within 95% confidence intervals as the expected values, i.e., >90% in X-sorted semen, <10% in Y-sorted semen and close to 50% in the unsorted semen. The multiplex real-time PCR assay as shown in this study can thus be used to assess purity of sex-sorted semen.

  9. Season-induced variation in lipid composition is associated with semen quality in Holstein bulls.

    PubMed

    Argov-Argaman, N; Mahgrefthe, K; Zeron, Y; Roth, Z

    2013-05-01

    Season-induced variation in fatty acid and cholesterol composition in bovine semen has been associated with semen quality. Given the specific roles of the various semen compartments (seminal fluids, sperm head, and sperm tail) in fertilization, we hypothesized that environmental-stress-induced alterations in the lipid composition of a specific compartment might impair semen quality and sperm function. Semen samples were collected from five mature Holstein-Friesian bulls during the summer (August to September) and winter (December to January). Semen was evaluated by computerized sperm-quality analyzer, calibrated for bulls' semen, and centrifuged to separate the spermatozoa from the seminal fluids. The spermatozoal fraction was sonicated to separate the sperm head and tail compartments. Cold lipid extraction was performed with chloroform:methanol (2:1, vol/vol). Lipids were identified and quantified by gas chromatography. Seasonal variation was found in both physiological and structural parameters. The proportion of spermatozoa defined as morphologically normal was higher in the winter, with higher motility, progressive motility, and velocity relative to summer samples. Lipid composition within fractions varied between seasons with prominent impairment in the tail compartment, characterized by high saturated fatty acid, low polyunsaturated fatty acid, and low cholesterol concentrations during the summer. Given the association between alterations in lipid composition and reduced sperm motility and velocity during the summer, it is suggested that lipid composition might serve to predict sperm quality.

  10. Effect of contaminated preprocessed semen on fertilization rate and embryo quality in assisted reproductive techniques.

    PubMed

    Krissi, H; Orvieto, R; Ashkenazi, J; Gilboa, Y; Shalev, J; Moscovitch, I; Bar-Hava, I

    2004-02-01

    We aimed to identify the sources and prevalence of semen contamination from mastrubation and determine the effect of bacterospermia on fertilization rate and embryo quality in standard in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). This was a prospective controlled study, in an IVF unit of a university teaching hospital, of 93 consecutive couples undergoing IVF-embryo transfer cycles. We evaluated handwashing; semen collection and processing; and assisted reproductive technology using semen provided by masturbation. The main outcome measures were presence and type of micro-organisms in the semen samples and embryo culture medium; the effect of hand washing on rate of contamination; and the effect of semen contamination on fertilization rate and embryo quality. The first consecutive 52 men of the 93 couples were not instructed to wash their hands before masturbation, and the remainder were so instructed. Forty-nine semen cultures (94.2%) in the first group were contaminated compared to only 16 (39%) in the second (p < 0.016); 27 of the 65 positive cultures (41.5%) were contaminated by more than one organism. The most common contaminators were bacteria usually found on the skin. All but four embryo medium cultures were negative. There was no significant difference in fertilization rate and embryo quality by culture findings in either the IVF or the ICSI procedures. We found that a high percentage of manually obtained semen for standard IVF or ICSI procedures was contaminated, but this had no effect on fertilization rate and embryo quality.

  11. Semen inflammatory markers and Chlamydia trachomatis infection in male partners of infertile couples.

    PubMed

    Dehghan Marvast, L; Aflatoonian, A; Talebi, A R; Ghasemzadeh, J; Pacey, A A

    2016-09-01

    Previous studies have given conflicting results about the effect of generally infection and Chlamydia trachomatis on seminal ILs and semen parameters. The aim of this study was to investigate the relationship between semen quality and the level of seminal interleukins (ILs) in infertile couples with C. trachomatis. Blood, first void urine (FVU) and semen were obtained from 250 infertile men who had failed to conceive after 12 months of trying. Serological analysis for specific IgA, IgM and IgG antibodies to C. trachomatis in serum, the presence of C. trachomatis in FVU and semen sample and semen analysis were carried out. The main results are as follows: (i) elevated IL-6 and IL-8 are observed in C. trachomatis-positive men, but this is not significant and it varies by diagnostic method; and (ii) IL-6 and IL-8 levels were correlated with each other and the concentration of leucocytes, but IL-8 was correlated with semen volume and patient's age. This study showed that men with such an infection in FVU samples (PCR positive) had only lower semen volume compared with men without infection.

  12. Semen evaluation in four autochthonous wild raptor species using computer-aided sperm analyzer.

    PubMed

    Dogliero, Andrea; Rota, Ada; Lofiego, Renato; Mauthe von Degerfeld, Mitzy; Quaranta, Giuseppe

    2016-04-01

    At least 10 percent of the approximately 300 species of the order Falconiformes are listed as being globally threatened. The present work describes the seminal characteristics of three diurnal and one nocturnal raptor species. Semen was collected from clinically healthy Accipiter nisus (n = 1), Falco subbuteo (n = 6), and Falco tinnunculus (n = 5) adult males that were housed at the 'Centro Animali Non Convenzionali' of the Department of Veterinary Sciences of the University of Turin. The semen was collected after a period of recovery and before their release as well as from seven Bubo bubo males bred in captivity as part of a raptor conservation project. All the potential semen donors were trained in semen collection during the breeding season via a ritualized procedure. Ejaculation was achieved using a massaging technique. Each sample was evaluated for volume, degree of contamination, and spermatozoa concentration. The semen motility and kinetic parameters were assessed on diluted semen (modified tyroides albumin lactate pyruvate, pH 7.5, temperature 37.5 °C) using a computer-aided sperm analyzer. Semen collection was successful in all the diurnal species and in five B bubo individuals. The sperm motility and sperm kinetic parameters were very variable both among and within species. In contrast with previous studies that involved raptors bred in captivity and imprinted on humans, we worked with wild birds and attempted to overcome the problem of poor semen quality, which is strongly influenced by stress, by adopting a ritualized procedure that has never been reported for semen collection purposes.

  13. Effect of organic selenium on turkey semen quality during liquid storage.

    PubMed

    Dimitrov, S G; Atanasov, V K; Surai, P F; Denev, S A

    2007-08-01

    The aim of the present study was to evaluate the effect of dietary organic selenium on the turkey semen during storage. Twenty males (BUT, Big 6, 40 weeks of age) were divided into control (n=10) and experimental group (n=10). The turkeys in the both groups were fed with a commercial diet containing 0.1 ppm Se in the form of sodium selenite. The experimental birds were additionally supplied with 0.3 ppm organic Se in the form Sel-Plex (Alltech, Inc.). After 30 days of feeding, the semen samples were collected twice a week for the 3 weeks of the study and diluted 1+1(v/v) with TUR-2 diluent, and stored in a water bath (+10 to 15 degrees C) for 6 h. The percentage of motile spermatozoa, the sperm viability (live/dead spermatozoa), total lipids, phospholipids and total cholesterol were assessed in fresh and stored semen. The fertilizing ability of semen was assessed by artificial insemination of 30 hens per group with dose containing 200x10(6) spermatozoa weekly. After 6 h of semen storage, the motility of spermatozoa decreased significantly in the control group (by 8.7 relative percent, P<0.05) and only by four relative percent (P>0.05) in experimental group reflecting a protective effect of dietary Se supplementation. The proportion of live spermatozoa was higher in fresh semen and significantly lower in stored semen. The positive effect of Se supplementation was observed on the lipid composition of stored semen: the concentration of the total lipids and phospholipids in the seminal plasma from control group significantly increased, while in the experimental group remained constant. Better semen integrity in the experimental group was associated with an improved fertilizing ability of spermatozoa: the fertility rate of stored spermatozoa in the control group was 88%, while in the experimental group was 90.5%.

  14. Influence of Helicobacter pylori infection on levels of ghrelin and obestatin in human semen.

    PubMed

    Moretti, Elena; Collodel, Giulia; Campagna, Maria Stella; Franci, Maria Beatrice; Iacoponi, Francesca; Mazzi, Lucia; Figura, Natale

    2012-01-01

    Helicobacter pylori (HP) infection might have negative effects on the semen parameters of infertile men. We explored the possibility that this infection can influence systemic and seminal levels of ghrelin and obestatin, hormones mainly produced by the stomach. Ghrelin and obestatin exert many activities, including the regulation of reproductive biology, and are present in many organs and fluids, including human semen. In 78 men, we determined HP infection and cytotoxin-associated gene A protein (CagA) status by enzyme-linked immunosorbent assay and Western blotting, semen quality following World Health Organization guidelines, and ghrelin and obestatin levels in the blood stream (47 subjects) and semen by radioimmunoassay. Twenty-seven men (34.6%) were infected (HP+) and 11 out of 27 infected men (40.7%) were seropositive for CagA (CagA+). Sperm motility was significantly reduced in HP+/CagA+ men compared with HP+/CagA- men (P < .01). Ghrelin semen levels were decreased in HP+ men compared with uninfected individuals (P < .05), whereas they were increased in HP+/CagA+ men compared with HP+/CagA- subjects (P < .01). Ghrelin semen concentrations in HP+/CagA- men were lower than those measured in uninfected subjects (P < .001). Semen obestatin concentration was increased, in a nonsignificant manner, in HP+/CagA+ men. The obestatin levels were approximately 4 times higher than those of ghrelin in semen and approximately half the levels of ghrelin in serum specimens of all the analyzed groups. No significant differences were found in systemic levels of ghrelin and obestatin in HP+ to uninfected individuals. HP infection may influence the ghrelin seminal concentrations, probably as a response to a negative effect of infection on the semen quality.

  15. An Outbreak of Porcine Reproductive and Respiratory Syndrome Virus in Switzerland Following Import of Boar Semen.

    PubMed

    Nathues, C; Perler, L; Bruhn, S; Suter, D; Eichhorn, L; Hofmann, M; Nathues, H; Baechlein, C; Ritzmann, M; Palzer, A; Grossmann, K; Schüpbach-Regula, G; Thür, B

    2016-04-01

    An outbreak of porcine reproductive and respiratory syndrome virus (PRRSV) occurred in November 2012 in Switzerland (CH), traditionally PRRSV-free. It was detected after a German boar stud informed a semen importer about the detection of PRRSV during routine monitoring. Tracing of semen deliveries revealed 26 Swiss sow herds that had used semen from this stud after its last negative routine monitoring and 62 further contact herds. All herds were put under movement restrictions and examined serologically and virologically. As a first measure, 59 sows from five herds that had previously been inseminated with suspicious semen were slaughtered and tested immediately. Investigations in the stud resulted in 8 positive boars with recent semen deliveries to CH (Seven with antibodies and virus, one with antibodies only). In one boar out of six tested, virus was detected in semen. Of the 59 slaughtered sows, five from three herds were virus-positive. In one herd, the virus had spread, and all pigs were slaughtered or non-marketable animals euthanized. In the remaining herds, no further infections were detected. After confirmatory testings in all herds 3 weeks after the first examination gave negative results, restrictions were lifted in January 2013, and Switzerland regained its PRRSV-free status. The events demonstrate that import of semen from non-PRRS-free countries--even from negative studs--poses a risk, because monitoring protocols in boar studs are often insufficient to timely detect an infection, and infections of sows/herds occur even with low numbers of semen doses. The outbreak was eradicated successfully mainly due to the high disease awareness of the importer and because immediate actions were taken before clinical or laboratory diagnosis of a single case in the country was made. To minimize the risk of an introduction of PRRSV in the future, stricter import guidelines for boar semen have been implemented.

  16. Semen coagulum liquefaction, sperm activation and cryopreservation of capuchin monkey (Cebus apella) semen in coconut water solution (CWS) and TES-TRIS.

    PubMed

    Oliveira, Karol G; Miranda, Stefania A; Leão, Danuza L; Brito, Adriel B; Santos, Regiane R; Domingues, Sheyla F S

    2011-01-01

    The objectives of the present study were to test the effect of coconut water solution and TES-TRIS on the seminal coagulum liquefaction, sperm activation in fresh diluted semen, and on the cryopreservation of semen from capuchin monkeys (Cebus apella). Semen was collected from six males by electro-ejaculation, diluted in TES-TRIS or coconut water solution (CWS), and incubated at 35°C until the coagulated fraction of the semen was completely liquefied. In the experiment I, after liquefaction, samples were diluted in TES-TRIS or CWS, plus 6 and 10mM/mL of caffeine. Sperm motility and vigor were evaluated during 5h. For experiment II, after liquefaction, semen samples were extended in TES-TRIS (3.5% glycerol in the final solution) or CWS (2.5% glycerol in the final solution), cryopreserved and stored in liquid nitrogen for 1 week. The seminal coagulum was liquefied in (mean±SDM) 4.5±1.7 and 2.8±1.1h in TES-TRIS and CWS, respectively. Sperm were motile in TES-TRIS and CWS for 5.0±1.4 and 1.0±0.5h, respectively. The mean motility in this period was 38±22% (TES-TRIS) and 22.0±16.0 (CWS). Motility increased after caffeine addition only in samples diluted in CWS containing 6mM (22.5±16.0) or 10mM (28.0±19.0) caffeine. Post-thaw live sperm percentage was 26.2% in TES-TRIS and 13.2% in CWS. For cryopreservation of semen from C. apella TES-TRIS (3.5% glycerol) was more appropriate than CWS (2.5% glycerol). CWS+caffeine potentially increase sperm motility and may be useful in artificial insemination of fresh diluted semen.

  17. Cryopreserving turkey semen in straws and nitrogen vapour using DMSO or DMA: effects of cryoprotectant concentration, freezing rate and thawing rate on post-thaw semen quality.

    PubMed

    Iaffaldano, N; Di Iorio, M; Miranda, M; Zaniboni, L; Manchisi, A; Cerolini, S

    2016-04-01

    1. This study was designed to identify a suitable protocol for freezing turkey semen in straws exposed to nitrogen vapour by examining the effects of dimethylacetamide (DMA) or dimethylsulfoxide (DMSO) as cryoprotectant (CPA), CPA concentration, freezing rate and thawing rate on in vitro post-thaw semen quality. 2. Pooled semen samples were diluted 1:1 (v:v) with a freezing extender composed of Tselutin diluent containing DMA or DMSO to give final concentrations of 8% or 18% DMA and 4% or 10% DMSO. The semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen (LN2) surface (1, 5 and 10 cm) for 10 min. Semen samples were thawed at 4°C for 5 min or at 50°C for 10 s. After thawing, sperm motility, viability and osmotic tolerance were determined. 3. Cryosurvival of turkey sperm was affected by DMSO concentration. Freezing rate affected the motility of sperm cryopreserved using both CPAs, while thawing rates showed an effect on the motility of sperm cryopreserved using DMA and on the viability of sperm cryopreserved using DMSO. Significant interactions between freezing rate × thawing rate on sperm viability in the DMA protocol were found. 4. The most effective freezing protocol was the use of 18% DMA or 10% DMSO with freezing 10 cm above the LN2 surface and a thawing temperature of 50°C. An efficient protocol for turkey semen would improve prospects for sperm cryobanks and the commercial use of frozen turkey semen.

  18. Mechanisms of the harmful effects of bacterial semen infection on ejaculated human spermatozoa: potential inflammatory markers in semen.

    PubMed

    Fraczek, Monika; Kurpisz, Maciej

    2015-01-01

    The invasion of the male reproductive tract by microorganisms, and its subsequent consequences for sperm fertilizing potential, has been intensely discussed. The role of the bacteria that are responsible for the colonization and contamination of the male urogenital tract, rather than its infection, in diminished sperm parameters raises the most controversy. There are numerous premises suggesting that bacterial semen infection is associated with male infertility. However, the molecular mechanism by which the fertility is affected is complex and multifactorial, and still presents a puzzle. Some authors have suggested that direct interactions between bacteria and human spermatozoa facilitate sperm immobilization, affect sperm morphology, and thus weaken the ability of sperm to fertilize. On the other hand, the massive infiltration of activated leukocytes into the inflammatory site may be associated with impairment of sperm fertilizing potential, due to oxidative, apoptotic, and immune processes. This review presents current research trends and aims to summarize the present knowledge of semen inflammation and causative bacterial agents in the male urogenital tract, with its consequence on seminological parameters, and male fertility status.

  19. Infertility and venereal disease in cattle inseminated with semen containing bovine herpesvirus type 5.

    PubMed

    Kirkland, P D; Poynting, A J; Gu, X; Davis, R J

    2009-07-25

    A group of 20 cows and heifers experienced poor conception rates and probable ovarian dysfunction after being artificially inseminated. When first examined, some showed signs of vulvovaginitis, with pustular, ulcerative lesions consistent with a herpesvirus infection. They had had no contact with bulls during the current breeding season. Bovine herpesvirus type 5 (BHV-5) was isolated from samples of frozen semen from the batch that had been used for the artificial insemination programme. BHV-5 was also isolated from the semen of a second apparently healthy bull during routine screening of its semen.

  20. Determination of flavonoids in Semen Cuscutae by RP-HPLC.

    PubMed

    Ye, Min; Li, Yan; Yan, Yuning; Liu, Huwei; Ji, Xiuhong

    2002-05-15

    Flavonoids contents in 40 samples of Semen Cuscutae collected from areas all around China were investigated. Five principal flavonoids, quercetin 3-O-beta-D-galactoside-7-O-beta-D-glucoside, quercetin 3-O-beta-D-apiofuranosyl-(1-->2)-beta-D-galactoside, hyperoside, quercetin and kaempferol were analyzed simultaneously by using a reversed phase liquid chromatograph system with 0.025 M phosphoric acid-methanol as mobile phase. The recovery of the method was 97.0-102.9%, and all the flavonoids showed good linearity (r > or = 0.9990) in a relatively wide concentration range. The results indicated that contents of flavonoids in S. Cuscutae varied significantly from species to species, locality to locality, and parasiting host to host. Cuscuta australis contained a much higher content of kaempferol than C. chinensis, and few flavonoids were detected in C. japonica. The quality of S. Cuscutae can be evaluated according to the contents of flavonoids.

  1. Effect of selenium supplementation on semen characteristics of Brazil's ram.

    PubMed

    Piagentini, M; Silva, D C; Dell'Aqua, Cpf; Moya-Araujo, C F; Codognoto, V M; Ramos, A A; Oba, E

    2017-02-20

    The aim of this study was to investigate the effects of different concentrations of oral supplementation with selenium (Se) upon ram sperm parameters. Thirty rams managed in stall under intensive system were used and divided into five groups (six animals per group) as follows: control group (G1) mineral mixture supplementation without Se, group 2 (G2) mineral mixture supplemented with 5 mg/kg Se, group 3 (G3) supplemented with 10 mg/kg Se, group 4 (G4) supplemented with 15 mg/kg Se and group 5 (G5) supplemented with 20 mg/kg Se. For each group, there was an adjustment period of 14 days. The experimental period was 350 days. Every 56 days, the animals were weighed and semen samples were collected by electroejaculation. Semen analysis included volume, mass moviment, total motility, vigour, concentration and morphology. For plasmatic and acrosomal membrane integrity evaluation and mitochondrial membrane potential were used a combination of fluorescent probes. Differences between means values obtained by analysis of variance were verified by Tukey test with 5% probability. There was no statistical difference between treatment groups in relation to volume, mass moviment, total motility, vigour, concentration, plasma and acrosomal membrane integrity (p > .05). Sperm morphology was different between treatment groups, the G1 (0 mg of selenium) had the highest percentage of major defects (11.11 ± 1.11(a) ; p < .05). It was concluded that selenium decrease the percentage of sperm defects and did not directly influence on ram sperm volume, mass moviment, total motility, vigour, concentration and membrane integrity.

  2. Semen quality: variations among fathers and effects of moderate alcohol drinking.

    PubMed

    Cooper, Trevor G

    2015-01-01

    Semen analysis results from over 750 fathers in the USA demonstrated marked differences in the quality of semen from men at different locations and of different ethnic groups. Another paper failed to demonstrate any effects of moderate alcohol consumption during the week before provision of an ejaculate on semen quality and few on serum hormones, of over 8300 men in Europe and the USA. While these observations are interesting, the reasons for regional and ethnic differences in semen quality of fathers are unclear. Although, there was no attempt to confirm the participant-provided level of alcohol consumption, an increase in serum testosterone in the men at the higher end of alcohol intake is compatible with an alcohol effect on liver metabolism, although whether alcohol intake was the cause of higher testosterone, or men with higher androgen levels consume more alcohol, is not known.

  3. A novel method for semen collection and artificial insemination in large parrots (Psittaciformes).

    PubMed

    Lierz, Michael; Reinschmidt, Matthias; Müller, Heiner; Wink, Michael; Neumann, Daniel

    2013-01-01

    The paper described a novel technique for semen collection in large psittacines (patent pending), a procedure which was not routinely possible before. For the first time, a large set of semen samples is now available for analysis as well as for artificial insemination. Semen samples of more than 100 psittacine taxa were collected and analysed; data demonstrate large differences in the spermatological parameters between families, indicating an ecological relationship with breeding behaviour (polygamous versus monogamous birds). Using semen samples for artificial insemination resulted in the production of offspring in various families, such as Macaws and Cockatoos, for the first time ever. The present technique represents a breakthrough in species conservation programs and will enable future research into the ecology and environmental factors influencing endangered species.

  4. Critical appraisal of conventional semen analysis in the context of varicocele.

    PubMed

    Kruger, Thinus

    2016-01-01

    Varicocele is present in approximately 15% of men, and, although it is the most commonly diagnosed cause of male infertility, nearly two-thirds of men with varicoceles remain fertile. It was decided to make use of the current evidence obtained from the previous meta-analyses between 2004 and 2015 as well as available articles covering this field, preferably randomized controlled articles dealing with the topic of semen analysis before and after repair. Two important meta-analyses were discussed as well as other articles dealing with the topic of semen analysis before and after varicocelectomy. The evidence suggests that all semen parameters improve after varicocele repair. Based on the available evidence, it is clear that there is a benefit in treating men with a palpable varicocele. One can expect that all semen parameters will improve within 3 months after repair.

  5. Semen characteristics and diabetes mellitus: significance of insulin in male infertility.

    PubMed

    García-Díez, L C; Corrales Hernandez, J J; Hernandez-Diaz, J; Pedraz, M J; Miralles, J M

    1991-01-01

    A study was made of semen quality and serum hormonal profiles (FSH, LH, prolactin, testosterone) of patients with type I diabetes mellitus. Semen parameters and levels of prolactin and testosterone were significantly altered in the diabetic state. The concentration of insulin in serum and seminal plasma and the serum levels of FSH, LH, and testosterone were measured in 80 men classified in the following groups: fertile subjects, infertile normoglycemic subjects, subjects with carbohydrate intolerance, and excretory and secretory azoospermic subjects. In all groups, seminal insulin concentrations were higher than those obtained in serum. The hormone appears to freely cross the blood-testis barrier, there to be concentrated in the semen. The levels of insulin in serum and seminal plasma did not correlate with semen parameters and are not suitable markers of seminal quality. For unknown reasons, the concentrations of insulin in seminal plasma were lower in the subjects suffering from carbohydrate intolerance.

  6. A novel method for semen collection and artificial insemination in large parrots (Psittaciformes)

    PubMed Central

    Lierz, Michael; Reinschmidt, Matthias; Müller, Heiner; Wink, Michael; Neumann, Daniel

    2013-01-01

    The paper described a novel technique for semen collection in large psittacines (patent pending), a procedure which was not routinely possible before. For the first time, a large set of semen samples is now available for analysis as well as for artificial insemination. Semen samples of more than 100 psittacine taxa were collected and analysed; data demonstrate large differences in the spermatological parameters between families, indicating an ecological relationship with breeding behaviour (polygamous versus monogamous birds). Using semen samples for artificial insemination resulted in the production of offspring in various families, such as Macaws and Cockatoos, for the first time ever. The present technique represents a breakthrough in species conservation programs and will enable future research into the ecology and environmental factors influencing endangered species. PMID:23797622

  7. Effect of organic and inorganic forms of selenium in diets on turkey semen quality.

    PubMed

    Slowińska, M; Jankowski, J; Dietrich, G J; Karol, H; Liszewska, E; Glogowski, J; Kozłowski, K; Sartowska, K; Ciereszko, A

    2011-01-01

    The effects of Se supplementation and its organic or inorganic form on semen quantitative parameters (ejaculate volume, sperm concentration, and total number of sperm) and biochemical parameters of seminal plasma (protein concentration, acid phosphatase activity, superoxide dismutase activity, and total antioxidant capacity) were investigated over a 25-wk reproductive season. Additionally, DNA fragmentation and motility characteristics of turkey spermatozoa were measured. The parameters of turkey semen in relation to yellow semen syndrome were also determined. Twenty-four males (Big 6) were divided into 3 experimental groups differing in form of Se supplementation (no Se supplementation, 0.3 mg/kg of inorganic Se from sodium selenite and 0.3 mg/kg of organic Se from Sel-Plex, Alltech Inc., Nicholasville, KY). Dietary Se supplementation enhanced the sperm concentration and total number of sperm and did not influence the antioxidative properties of turkey seminal plasma and most biochemical parameters. Only seminal plasma acid phosphatase activity was increased in turkeys fed inorganic Se. The main sperm DNA fragmentation parameters were not affected by dietary Se. The highest percentage of motile spermatozoa (85%) was recorded for the semen of turkeys fed organic Se. Values of the biochemical parameters (acid phosphatase, superoxide dismutase, total antioxidant capacity) of seminal plasma increased during the reproductive season. Yellow semen was characterized by increased biochemical parameters and decreased spermatozoa motility characteristics. However, the percentage of motile spermatozoa did not differ between white and yellow semen. Organic Se seemed to be the preferred form of diet supplementation in comparison with inorganic Se. Biochemical parameters of semen and spermatozoa motility parameters appear to be useful for evaluating the effect of age on semen quality. Monitoring the DNA fragmentation of spermatozoa at the end of the reproductive season could be

  8. The Association Between Testis Cancer and Semen Abnormalities Before Orchiectomy: A Systematic Review

    PubMed Central

    Burner, Elizabeth; Parikh, Pooja M.; Beroukhim Kay, Dorsa; Hays, Krystal

    2014-01-01

    Purpose: Testicular germ cell tumors (TGCT) are the most common solid organ malignancy in young men. It is a largely curable disease, so the extent to which it affects quality of life—including male fertility—is important. Abnormal semen analysis is highly predictive of male infertility. We conducted a systematic review of published studies that reported pre-orchiectomy semen parameters (as a surrogate for fertility) in TGCT patients to evaluate the association between TGCT and semen abnormalities before orchiectomy. Methods: We conducted a systematic review of peer-reviewed publications reporting semen parameters before orchiectomy in adult patients diagnosed with TGCT. Further, we assessed the association between TGCT and semen abnormalities that may lead to infertility. Results: We applied MeSH search terms to four online databases (PubMed, Cochrane Reviews, Web of Science, and Ovid), resulting in 701 potentially relevant citations. After conducting a three-stage screening process, six articles were included in the systematic review. For each study, the participants' data and the study's quality and risk of bias were assessed and described. All studies showed semen abnormalities—including count, motility, and morphology—in men with TGCT prior to orchiectomy. Conclusions: TGCT is associated with semen abnormalities before orchiectomy. This review shows an increase in abnormal semen parameters among men with TGCT even outside the treatment effects of orchiectomy, radiation, or chemotherapy. To improve long-term quality of life, these findings should be considered when counseling patients on future fertility and sperm banking during discussions about treatment and prognosis for TGCT. PMID:25538860

  9. The Association Between Testis Cancer and Semen Abnormalities Before Orchiectomy: A Systematic Review.

    PubMed

    Djaladat, Hooman; Burner, Elizabeth; Parikh, Pooja M; Beroukhim Kay, Dorsa; Hays, Krystal

    2014-12-01

    Purpose: Testicular germ cell tumors (TGCT) are the most common solid organ malignancy in young men. It is a largely curable disease, so the extent to which it affects quality of life-including male fertility-is important. Abnormal semen analysis is highly predictive of male infertility. We conducted a systematic review of published studies that reported pre-orchiectomy semen parameters (as a surrogate for fertility) in TGCT patients to evaluate the association between TGCT and semen abnormalities before orchiectomy. Methods: We conducted a systematic review of peer-reviewed publications reporting semen parameters before orchiectomy in adult patients diagnosed with TGCT. Further, we assessed the association between TGCT and semen abnormalities that may lead to infertility. Results: We applied MeSH search terms to four online databases (PubMed, Cochrane Reviews, Web of Science, and Ovid), resulting in 701 potentially relevant citations. After conducting a three-stage screening process, six articles were included in the systematic review. For each study, the participants' data and the study's quality and risk of bias were assessed and described. All studies showed semen abnormalities-including count, motility, and morphology-in men with TGCT prior to orchiectomy. Conclusions: TGCT is associated with semen abnormalities before orchiectomy. This review shows an increase in abnormal semen parameters among men with TGCT even outside the treatment effects of orchiectomy, radiation, or chemotherapy. To improve long-term quality of life, these findings should be considered when counseling patients on future fertility and sperm banking during discussions about treatment and prognosis for TGCT.

  10. Comparison of two dilution rates on canine semen quality after cryopreservation in a coconut water extender.

    PubMed

    de Cássia Soares Cardoso, Rita; Silva, Alexandre Rodrigues; da Silva, Lúcia Daniel Machado

    2006-05-01

    The objective of the present study was to evaluate the effect of sperm dilution (one part semen:one part extender or at 200 x 10(6) spermatozoa/mL) using a coconut water extender on the post-thaw sperm quality. Twelve ejaculates were collected from six dogs. Semen was divided into two aliquots, one for dilution one part semen:one part extender (group 1) and another for a concentration of 200 x 10(6) spermatozoa/mL (group 2). Semen was initially extended at 37 degrees C at a proportion of one part semen:half part extender (1:1/2) for group 1 (A-fraction). For group 2, the volume for a concentration of 200 x 10(6) spermatozoa/mL was calculated and a half of this volume was used for the initial dilution (A-fraction, 37 degrees C). Coconut water extender containing 20% egg yolk was used for this initial dilution in both groups. After dilution, the semen was cooled for 40 min in a thermal box (15 degrees C) and for 30 min in a refrigerator. The other half of the extender (B-fraction) containing egg yolk and glycerol (12%) was added to semen in both groups. Subsequently, the final concentration of glycerol in the extender was 6%. Ejaculates were frozen in 0.25 mL straws 5 cm above the surface of liquid nitrogen and stored at -196 degrees C. After 1 week, straws were thawed at 37 degrees C for 1 min and the microscopic criteria were evaluated. The dilution method had no influence on sperm motility, vigor and normal spermatozoa (71.4 compared with 67.7%). There was no effect of dog, ejaculate within male on post-thaw semen quality. Moreover, there was not a male x treatment interaction. Both treatments were efficient in preserving sperm quality.

  11. Germination and colonization success of Gonyostomum semen (Raphidophyceae) cysts after dispersal to new habitats

    PubMed Central

    Sassenhagen, Ingrid; Sefbom, Josefin; Godhe, Anna; Rengefors, Karin

    2015-01-01

    Colonization of new habitats through dispersal of phytoplankton cysts might be limited, if resident populations outcompete invaders during germination. We reciprocally transferred Gonyostomum semen (Raphidophyceae) cysts from three lakes into native and foreign waters originating from the respective habitats. Germination rate and germling growth were impacted by water origin, but there was no preference for native water. Gonyostomum semen's ability to germinate in different conditions might explain its expansion in northern Europe. PMID:26412910

  12. Analysis of breed effects on semen traits in light horse, warmblood, and draught horse breeds.

    PubMed

    Gottschalk, Maren; Sieme, Harald; Martinsson, Gunilla; Distl, Ottmar

    2016-05-01

    In the present study, systematic effects on semen quality traits were investigated in 381 stallions representing 22 breeds. All stallions were used for AI either at the Lower Saxon National Stud Celle or the North Rhine-Westphalian National Stud Warendorf. A total of 71,078 fresh semen reports of the years 2001 to 2014 were edited for analysis of gel-free volume, sperm concentration, total number of sperm, progressive motility, and total number of progressively motile sperm. Breed differences were studied for warmblood and light horse breeds of both national studs (model I) and for warmblood breeds and the draught horse breed Rhenish German Coldblood from the North Rhine-Westphalian National stud (model II) using mixed model procedures. The fixed effects of age class, year, and month of semen collection had significant influences on all semen traits in both analyses. A significant influence of the horse breed was found for all semen traits but gel-free volume in both statistical models. Comparing warmblood and light horse stallions of both national studs, we observed highest sperm concentrations, total numbers of sperm, and total numbers of progressively motile sperm in Anglo-Arabian stallions. The draught horse breed Rhenish German Coldblood had the highest least squares means for gel-free volume, whereas all other investigated semen traits were significantly lower in this breed compared to the warmblood stallions under study. The variance components among stallions within breeds were significant for all semen traits and accounted for 40% to 59% of the total variance. The between-breed-variance among stallions was not significant underlining the similar size of the random stallion effect in each of the horse breeds analyzed here. In conclusion, breed and stallion are accounting for a significant proportion of the variation in semen quality.

  13. Semen quality in adult male survivors 5 years after the 2008 Wenchuan earthquake.

    PubMed

    Chen, X M; Chen, S M; Yue, H X; Lin, L; Wu, Y B; Liu, B; Jiang, M; Ma, Y X

    2016-12-01

    The influence of the Wenchuan earthquake on semen quality of adult male survivors is unclear. We investigated the semen quality included 673 male survivors from the worse-affected counties in the earthquake between Aug 2008 and July 2013. Semen parameters including pH, volume, concentration, motility and morphology were measured according to the World Health Organization (WHO) criteria. Kruskal-Wallis analysis of variance was used to examine the statistical differences between years, and a logistic regression was used to analyse the impacts caused by earthquake on the changes of semen quality. We found the medians (5th and 95th) were 2.5 ml (0.6-5.5) for semen volume, 59.0 × 10(6)  ml(-1) [(13.0-133.0)] × 10(6)  ml(-1) for semen concentration, 46% (13-64%) for sperm progressive motility and 3.0% (0-17.5%) for normal morphology for adult male survivors. Semen concentration, the percentage of sperm progressive motility, total motility and sperm normal morphology were all decreased in the first 3 years, and the differences among years 1, 2 and 3 were significant except the percentage of sperm progressive motility (P < 0.05). The casualties and heavy housing damage caused by earthquake had a negative effect on semen quality. The main findings will provide further diagnosis and therapy basis of male fertility by data, for affected populations in the earthquake.

  14. In vivo adverse effects of alpha-tocopherol on the semen quality of male bucks.

    PubMed

    Majid, A; Qureshi, M S; Khan, R U

    2015-10-01

    Oxidative stress has detrimental effects on semen quality during spermatogenesis and semen processing for artificial insemination. This work was conducted to study the effect of different levels of vitamin E on the semen traits, oxidative status and trace minerals in Beetal bucks. Thirty-six bucks of similar body weight and age (1 year) were randomly divided into four groups. One group was kept as control with no supplementation (group 1), and the others were supplemented with 200 (group 2), 400 (group 3) and 800 IU (group 4) vitamin E/animal/day for 2 months. At the end of the experiment, semen samples were collected and evaluated. Seminal plasma was separated to study the concentration of superoxide dismutase (SOD), glutathione peroxidase (GPx), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and trace minerals (Zn, Cu, Mn and Fe). Group 3 showed significantly higher (p < 0.05) semen volume and per cent motility and lower dead sperm percentage compared to control group. Superoxide dismutase, GPx, Zn, Cu and Mn were higher in the same group. The level of AST decreased in group 3 without any change on the concentration of ALT. It is suggested that vitamin E at the rate of 400 IU/buck/day supported higher semen volume, per cent motility, per cent live spermatozoa, antioxidants (SOD, GPx) and trace mineral levels (Zn, Cu, Mn) in the seminal plasma. The increased supplementation from 0 to 400 showed a general increasing trend in improving semen quality. However, the dose of 800 IU/kg had no useful effect in further improving the semen quality.

  15. Cotinine concentrations in semen, urine, and blood of smokers and nonsmokers.

    PubMed Central

    Vine, M F; Hulka, B S; Margolin, B H; Truong, Y K; Hu, P C; Schramm, M M; Griffith, J D; McCann, M; Everson, R B

    1993-01-01

    Cotinine levels in the semen, urine, and blood of 88 male smokers and nonsmokers, aged 18 to 35, were analyzed via radioimmunoassay. Detectable cotinine levels were found in all three body fluids, and cotinine levels in all three fluids were highly correlated. Cotinine levels in semen and blood were of similar magnitude; cotinine levels in urine were an order of magnitude or more higher. In all three fluids, cotinine levels increased with an increase in cigarette smoke exposure. PMID:8363014

  16. Human papillomavirus proteins are found in peripheral blood and semen Cd20+ and Cd56+ cells during Hpv-16 semen infection

    PubMed Central

    2013-01-01

    Background Human papillomavirus (HPV) currently represents an important risk factor for cancer development and infertility in humans. Whilst binding of HPV to spermatozoa has been associated with male infertility, an investigation about the presence of HPV-DNA in non-spermatozoal semen cells is lacking. Previous findings documented the presence of HPV in peripheral blood leukocytes. The aim of this study was to investigate the expression of HPV markers in semen and blood leukocytes during HPV-16 infection. Methods A total of 32 subjects, 16 patients affected by HPV-16 semen infection and 16 controls, were evaluated in our andrological centre and enrolled in the study. Semen non-spermatozoal cells from all subjects were isolated and evaluated for the expression of HPV-16 markers (DNA and L1, E6 proteins) and further characterized for their molecular phenotype. Analogue determination was performed on peripheral blood mononuclear cells. Results The presence of HPV-DNA by FISH analysis in a round cell population from semen, confirmed to be CD45+ leukocytes, was observed. These HPV-DNA containing-cells also displayed HPV-16-E6 and HPV-16-L1 viral proteins and, upon further investigation, were found to be CD20+ and CD56+, likely phenotypes of B cells and natural killer cells (NK) respectively. In 25% of the patient group, a very small population of peripheral blood mononuclear cells was found to be positive for HPV-DNA via FISH. These cells displayed the CD20+ and CD56+ phenotype alike. None of the control subjects displayed HPV-DNA in either semen or peripheral blood. Conclusion Considering the role of CD20+ and CD56+ cell populations in the antiviral immune response, the detection of HPV markers on leukocytes may reflect the presence of virus particles within the endosomal compartment. However, the presence of HPV markers in circulating mononuclear cells raise concerns about the risk of developing cancers to distal organs. PMID:24341689

  17. Effect of prolonged freezing of semen on exosome recovery and biologic activity.

    PubMed

    Welch, Jennifer L; Madison, Marisa N; Margolick, Joseph B; Galvin, Shannon; Gupta, Phalguni; Martínez-Maza, Otoniel; Dash, Chandravanu; Okeoma, Chioma M

    2017-03-24

    Exosomes are important vehicles of intercellular communication that shape host responses to physiologic, tumorigenic, and pathogenic conditions. The composition and function of exosomes are dynamic and depends on the state and condition of the cellular source. In prior work, we found that semen exosomes (SE) from healthy donors who do not use illicit drugs potently inhibit HIV-1. Following semen donation, specimens are either used immediately or frozen for use at a later time. It has been shown that short-term freezing of semen has no effect on SE-mediated HIV-1 inhibition. However, the effect of illicit drugs and prolonged freezing on SE bioactivity is unknown. Here, we show preservation of SE physical properties, (morphology, concentration, intensity/size) irrespective of illicit drug use or duration of semen freezing. Interestingly, illicit drugs and prolonged freezing decreased the levels of SE-bound CD63/CD9 and acetylcholinesterase activity respectively. Furthermore, we show differential effects of illicit drug use and prolonged freezing on SE-mediated HIV-1 inhibition. Our results highlight the importance of the source of SE and condition of semen storage on SE content and function. In-depth evaluation of donor drug-use and duration of semen storage on SE cargo and bioactivity will advance our understanding of SE composition and function.

  18. Effect of organic and inorganic selenium supplementation on semen quality and blood enzymes in buffalo bulls.

    PubMed

    El-Sharawy, Mohamed; Eid, Entsar; Darwish, Samy; Abdel-Razek, Ibrahim; Islam, Md Rashedul; Kubota, Kaiyu; Yamauchi, Nobuhiko; El-Shamaa, Ibrahim

    2016-11-21

    The present study aimed to evaluate the effect of organic and inorganic selenium (Se) supplementation on semen quality and blood serum profiles of buffalo bulls. Nine mature buffalo bulls were divided into three groups: control (non-supplemented); organic Se (10 mg Sel-Plex®/head twice weekly) and inorganic Se (10 mg sodium selenite/head twice weekly). Semen was collected twice a week for 3 months during Se supplementation. Semen properties were evaluated from fresh ejaculate. Moreover, fructose concentration, aspartate and alanine transaminase (AST and ALT) activities, total protein and total cholesterol were assayed in seminal plasma. Additionally AST, ALT, testosterone and Se levels were determined in the blood serum. Results showed that Se supplementation significantly (P < 0.05) influences the semen parameters during 3 months of treatment. Organic Se significantly (P < 0.05) increased the percentage of viable sperms compared to inorganic Se and the control group. Fructose concentration was significantly higher (P < 0.05) in the seminal plasma of organic Se-treated bulls. Serum testosterone and Se concentrations were significantly (P < 0.05) increased in the Se supplemented groups than the control group. In conclusion, Se supplementation improved the parameters of buffalo bull semen and more precisely, organic Se was more effective for the improvement of semen quality and some blood components than inorganic Se.

  19. Toxoplasma gondii transmission by artificial insemination in sheep with experimentally contaminated frozen semen.

    PubMed

    Consalter, Angélica; Silva, Andressa F; Frazão-Teixeira, Edwards; Matos, Luis F; de Oliveira, Francisco C R; Leite, Juliana S; Silva, Franciele B F; Ferreira, Ana M R

    2017-03-01

    Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 10(7) tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep.

  20. What affects fertility of sexed bull semen more, low sperm dosage or the sorting process?

    PubMed

    Frijters, A C J; Mullaart, E; Roelofs, R M G; van Hoorne, R P; Moreno, J F; Moreno, O; Merton, J S

    2009-01-01

    Until now it has been unclear to what extent the reduced fertility with sexed semen in the dairy industry is caused by too few sperm per AI dose, or by the effect of flow cytometric sorting, which is the established procedure for sexing semen. Therefore, we evaluated the effects of low sperm numbers per dose with and without sorting on non-return rates after 56 days (NRR 56); in addition, we evaluated the effects of bulls, in order to further optimize use of sexed semen. Based on results of using sexed semen from seven Holstein bulls, an overall numerical decline of 13.6% in NRR 56 was observed (P<0.05). About two-thirds of this decline (8.6%) was due to the low dose (P<0.05), and a third (5.0%) due to the process of sorting (P<0.05). The effect of low dosage and sorting differed among bulls. We observed a sex ratio of 91.6% females for sexed semen from the first 131 calves born. Currently the best way to increase fertility of sexed semen is by closely monitoring fertility so that the highest fertility bulls are used, and by improving farm animal management. However, to make substantial progress, more in depth studies are needed on the sexing technology, especially on aspects such as sorting procedures and sperm dosage.

  1. Reducing Inter-Laboratory Differences between Semen Analyses Using Z Score and Regression Transformations

    PubMed Central

    Leushuis, Esther; Wetzels, Alex; van der Steeg, Jan Willem; Steures, Pieternel; Bossuyt, Patrick M.M.; van Trooyen, Netty; Repping, Sjoerd; van der Horst, Frans A.L.; Hompes, Peter G.A. Hompes; Mol, Ben Willem J.; van der Veen, Fulco

    2016-01-01

    Background Standardization of the semen analysis may improve reproducibility. We assessed variability between laboratories in semen analyses and evaluated whether a transformation using Z scores and regression statistics was able to reduce this variability. Materials and Methods We performed a retrospective cohort study. We calculated between-laboratory coefficients of variation (CVB) for sperm concentration and for morphology. Subsequently, we standardized the semen analysis results by calculating laboratory specific Z scores, and by using regression. We used analysis of variance for four semen parameters to assess systematic differences between laboratories before and after the transformations, both in the circulation samples and in the samples obtained in the prospective cohort study in the Netherlands between January 2002 and February 2004. Results The mean CVBwas 7% for sperm concentration (range 3 to 13%) and 32% for sperm morphology (range 18 to 51%). The differences between the laboratories were statistically significant for all semen parameters (all P<0.001). Standardization using Z scores did not reduce the differences in semen analysis results between the laboratories (all P<0.001). Conclusion There exists large between-laboratory variability for sperm morphology and small, but statistically significant, between-laboratory variation for sperm concentration. Standardization using Z scores does not eliminate between-laboratory variability. PMID:26985342

  2. Endocrinological, biophysical, and biochemical parameters of semen collected via masturbation versus sexual intercourse.

    PubMed

    Sofikitis, N V; Miyagawa, I

    1993-01-01

    In clinical programs of assisted reproduction involving infertile males, it is essential to obtain semen of maximum quality. To evaluate ways of achieving this objective, and to assess the fertilizing capacity of the sperm, six semen samples were collected from each of 38 infertile men via masturbation. Six more samples were then collected from each man at sexual intercourse using a semen collection device (SCD). We confirmed that the volume of seminal plasma, total sperm count, sperm motility, and percentage of morphologically normal spermatozoa were significantly higher in samples collected at intercourse than masturbation, as reported previously. In addition, the markers of the secretory function of the prostate and the outcome of sperm function tests (hypoosmotic swelling test, acrosin assay, and sperm penetration assay) were significantly higher for the samples collected at intercourse. There were no significant differences in markers of the secretory function of the seminal vesicles and epididymis between the samples. The improved spermatozoal parameters in the samples collected at intercourse may reflect a higher prostatic secretory function at that time. There were no significant differences in the serum concentrations of gonadotropins, or in the serum or seminal plasma concentrations of testosterone, before or after masturbation or sexual intercourse. Therefore, the differences in prostatic secretory function and semen parameters may not be attributed to differences in hormonal levels. Semen collection during intercourse using an SCD appears to be the method of choice for selecting semen samples for artificial insemination.

  3. Semen parameters can be predicted from environmental factors and lifestyle using artificial intelligence methods.

    PubMed

    Girela, Jose L; Gil, David; Johnsson, Magnus; Gomez-Torres, María José; De Juan, Joaquín

    2013-04-01

    Fertility rates have dramatically decreased in the last two decades, especially in men. It has been described that environmental factors as well as life habits may affect semen quality. In this paper we use artificial intelligence techniques in order to predict semen characteristics resulting from environmental factors, life habits, and health status, with these techniques constituting a possible decision support system that can help in the study of male fertility potential. A total of 123 young, healthy volunteers provided a semen sample that was analyzed according to the World Health Organization 2010 criteria. They also were asked to complete a validated questionnaire about life habits and health status. Sperm concentration and percentage of motile sperm were related to sociodemographic data, environmental factors, health status, and life habits in order to determine the predictive accuracy of a multilayer perceptron network, a type of artificial neural network. In conclusion, we have developed an artificial neural network that can predict the results of the semen analysis based on the data collected by the questionnaire. The semen parameter that is best predicted using this methodology is the sperm concentration. Although the accuracy for motility is slightly lower than that for concentration, it is possible to predict it with a significant degree of accuracy. This methodology can be a useful tool in early diagnosis of patients with seminal disorders or in the selection of candidates to become semen donors.

  4. Fertility and uterine hemodynamic in cows after artificial insemination with semen assessed by fluorescent probes.

    PubMed

    Oliveira, Bruna Marcele; Arruda, Rubens Paes; Thomé, Helder Esteves; Maturana Filho, Milton; Oliveira, Guilherme; Guimarães, Carina; Nichi, Marcílio; Silva, Luciano Andrade; Celeghini, Eneiva Carla Carvalho

    2014-09-15

    Fluorescent probes (propidium iodide, Hoechst 33342, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin, and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide) were used to simultaneously evaluate the integrity of plasma and acrosomal membranes as well as mitochondrial membrane function in cryopreserved bovine semen and to verify its influence on fertility and postinsemination uterine vascularization. One hundred eighty-two Nellore cows were distributed for artificial insemination (AI) using semen batches separated according to the cell percentage presenting intact plasma membrane, intact acrosome, and high mitochondrial function (IPIAH): group G (44.5% IPIAH, n = 68), group M (23.0% IPIAH, n = 56), and group R (8.5% IPIAH, n = 58). The uterine hemodynamic was evaluated by Doppler sonogram in three periods: 30 hours before AI, 4 and 24 hours after AI were considered the resistance index and the uterine vascularization score. The pregnancy rate of group G (64.7%) was greater (P > 0.05) compared with group R (36.2%), but both did not differ from group M (50.0%). There was no effect (P > 0.05) of semen quality on uterine vascularization. Greater vascularization was noticed 4 hours after AI than 30 hours before and 24 hours after AI. Semen evaluation using fluorescent probes contributes to predicting fertilizing potential of semen. The use of semen with less percentage of IPIAH sperm does not alter uterine hemodynamic in cows.

  5. Collection, analysis and cryopreservation of semen from Malayan gaur (Bos gaurus hubbacki): A preliminary study

    PubMed Central

    Iswadi, M.I.; Ann, Z.F.; Hafiz, M.M.; Hafiz, M.D.; Fahrul, F.J.; Hajarian, H.; Wahid, H.; Zawawi, I.; Khairiah, M.S.; Mazni, O.A.

    2012-01-01

    The Malayan gaur (Bos gaurus hubbacki) or Seladang is classified as vulnerable by the International Union for Conservation of Nature and Natural Resources (IUCN). The Malayan gaur is mainly distributed in the tropical woodlands of Peninsular Malaysia and Southern Thailand. The aim of this study was to collect, analyze and cryopreserve the semen of wild Malayan gaur. Transrectal massage (TM) and electroejaculation (EEJ) technique was applied in semen collection of the Malayan gaur. The semen was then cryopreserved in liquid nitrogen using slow freezing technique. Makler counting chamber was used to evaluate sperm concentration and motility, while the sperm viability and morphology of fresh and post-thaw sperm was determined using eosin-nigrosin staining protocol. As a result, we have successfully collected the Malayan gaur semen using EEJ technique. Sperm motility, viability and morphological changes of the post-thaw semen of Malayan gaur were found undesirable due to the complication of the cryopreservation process. On the basis of current study it can be concluded that Malayan gaur bulls semen can be obtain by EEJ with no evidence of rectal trauma. Optimization of the process of cryopreservation for Malayan gaur sperm is needed to maintain the cryoviability of the good sperm quality. The data generated in this study would be useful in conservation of genetic diversity program for Malayan gaur. PMID:26623302

  6. Effect of prolonged freezing of semen on exosome recovery and biologic activity

    PubMed Central

    Welch, Jennifer L.; Madison, Marisa N.; Margolick, Joseph B.; Galvin, Shannon; Gupta, Phalguni; Martínez-Maza, Otoniel; Dash, Chandravanu; Okeoma, Chioma M.

    2017-01-01

    Exosomes are important vehicles of intercellular communication that shape host responses to physiologic, tumorigenic, and pathogenic conditions. The composition and function of exosomes are dynamic and depends on the state and condition of the cellular source. In prior work, we found that semen exosomes (SE) from healthy donors who do not use illicit drugs potently inhibit HIV-1. Following semen donation, specimens are either used immediately or frozen for use at a later time. It has been shown that short-term freezing of semen has no effect on SE-mediated HIV-1 inhibition. However, the effect of illicit drugs and prolonged freezing on SE bioactivity is unknown. Here, we show preservation of SE physical properties, (morphology, concentration, intensity/size) irrespective of illicit drug use or duration of semen freezing. Interestingly, illicit drugs and prolonged freezing decreased the levels of SE-bound CD63/CD9 and acetylcholinesterase activity respectively. Furthermore, we show differential effects of illicit drug use and prolonged freezing on SE-mediated HIV-1 inhibition. Our results highlight the importance of the source of SE and condition of semen storage on SE content and function. In-depth evaluation of donor drug-use and duration of semen storage on SE cargo and bioactivity will advance our understanding of SE composition and function. PMID:28338013

  7. The impact of body mass index on semen parameters in infertile men.

    PubMed

    Alshahrani, S; Ahmed, A-F; Gabr, A H; Abalhassan, M; Ahmad, G

    2016-12-01

    This hospital-based, prospective study was conducted to evaluate the relationship between body mass index (BMI) and various semen parameters in infertile men. A total of 439 men presented for infertility evaluation were assessed by basic infertility evaluation measures including semen analysis and BMI calculation. The main outcome measure was the relationship between BMI groups [BMI: 18.5-24.9 kg/m(2) (normal weight), 25-29.9 kg/m(2) (overweight) and ≥30 kg/m(2) (obese)] and different semen parameters [volume, concentration, motility and morphology]. The mean BMI was 29.67 ± 5.89. Most of patients (82.91%) were overweight or obese. The 3 BMI groups were comparable in semen parameters (P > 0.05). BMI had a negative correlation with various semen parameters. However, this correlation was significant only with sperm concentration (P = 0.035). We concluded that sperm concentration was the only semen parameter which showed significant reduction with higher BMI in infertile men.

  8. Presence of aerobic micro-organisms and their influence on basic semen parameters in infertile men.

    PubMed

    Filipiak, E; Marchlewska, K; Oszukowska, E; Walczak-Jedrzejowska, R; Swierczynska-Cieplucha, A; Kula, K; Slowikowska-Hilczer, J

    2015-09-01

    Urogenital tract infections in males are one of the significant etiological factors in infertility. In this prospective study, 72 patients with abnormal semen parameters or any other symptoms of urogenital tract infection were examined. Semen analysis according to the WHO 2010 manual was performed together with microbial assessment: aerobic bacteria culture, Chlamydia antigen test, Candida culture, Ureaplasma and Mycoplasma-specific culture. In total, 69.4% of semen samples were positive for at least one micro-organism. Ureaplasma sp. was the most common micro-organism found in 33% of semen samples of infertile patients with suspected male genital tract infection. The 2nd most common micro-organisms were Enterococcus faecalis (12.5%) and Escherichia coli (12.5%), followed by Staphylococcus aureus (7%), Chlamydia trachomatis (7%) and Candida sp. (5.6%). Generally, bacteria were sensitive to at least one of the antibiotics tested. No statistically significant relationship was observed between the presence of aerobic micro-organisms in semen and basic semen parameters: volume, pH, concentration, total count, motility, vitality and morphology.

  9. Seasonal effects of semen collection and artificial insemination on dairy cow conception.

    PubMed

    Haugan, T; Reksen, O; Gröhn, Y T; Kommisrud, E; Ropstad, E; Sehested, E

    2005-11-01

    The effects of four seasons of semen collection and of artificial insemination on conception in dairy cows were studied. The solstices and equinoxes (December, March, June and September) defined the beginning and/or end of each season. Semen was collected from 973 progeny-test bulls over 8 years at the two Norwegian AI stations at 60.8 degrees N and 63.4 degrees N where artificial light was used to provide a minimum photoperiod of 10 h/day. The effect of using semen of elite bulls during progeny testing and after selection as elite sires also was investigated. Norwegian Red (NRF) cows were inseminated over a 7-year period using progeny test semen and over the last 4 years of the same period using the semen of the elite sires. The probability of conception to only first inseminations for cows up to, and including, the fifth lactation was assessed by 56-day non-return rate (56d NRR) and calving rate. Two data sets were analysed which excluded cows culled within 270 days of AI or included such cows as non-calving. The reasons for culling were categorised as those for fertility problems or all other reasons. Semen was used for AI irrespective of the season in which it had been collected. Season of semen collection did not affect 56d NRR but calving rate was significantly higher (by 0.5-0.8%, approximately; P < 0.01) for semen collected in the December-March period, when photoperiod was increasing, than at other times of the year. The season in which AI was performed showed a peak of 56d NRR in spring for heifers (P < 0.01) and in summer for parous animals (P < 0.01). For calving rate, however, no seasonal peak was found in heifers, whereas pluriparous cows had much higher calving rates in summer and autumn/early winter than late winter and spring (P < 0.01). Semen of elite sires resulted in higher calving rates by 0.5 (NS) to 1.9% (P < 0.01) when used after selection than when used during progeny testing. The difference between the calving rate achieved when the semen

  10. Semen parameters in fertile US men: the Study for Future Families.

    PubMed

    Redmon, J B; Thomas, W; Ma, W; Drobnis, E Z; Sparks, A; Wang, C; Brazil, C; Overstreet, J W; Liu, F; Swan, S H

    2013-11-01

    Establishing reference norms for semen parameters in fertile men is important for accurate assessment, counselling and treatment of men with male factor infertility. Identifying temporal or geographic variability in semen quality also requires accurate measurement of semen parameters in well-characterized, defined populations of men. The Study for Future Families (SFF) recruited men who were partners of pregnant women attending prenatal clinics in Los Angeles CA, Minneapolis MN, Columbia MO, New York City NY and Iowa City IA. Semen samples were collected on site from 763 men (73% White, 15% Hispanic/Latino, 7% Black and 5% Asian or other ethnic group) using strict quality control and well-defined protocols. Semen volume (by weight), sperm concentration (hemacytometer) and sperm motility were measured at each centre. Sperm morphology (both WHO, 1999 strict and WHO, 1987) was determined at a central laboratory. Mean abstinence was 3.2 days. Mean (median; 5th-95th percentile) values were: semen volume, 3.9 (3.7; 1.5-6.8) mL; sperm concentration, 60 (67; 12-192) × 10(6) /mL; total sperm count 209 (240; 32-763) × 10(6) ; % motile, 51 (52; 28-67) %; and total motile sperm count, 104 (128; 14-395) × 10(6) respectively. Values for sperm morphology were 11 (10; 3-20) % and 57 (59; 38-72) % normal forms for WHO (1999) (strict) and WHO (1987) criteria respectively. Black men had significantly lower semen volume, sperm concentration and total motile sperm counts than White and Hispanic/Latino men. Semen parameters were marginally higher in men who achieved pregnancy more quickly but differences were small and not statistically significant. The SFF provides robust estimates of semen parameters in fertile men living in five different geographic locations in the US. Fertile men display wide variation in all of the semen parameters traditionally used to assess fertility potential.

  11. Diagnosis of Partial Retrograde Ejaculation in Non-Azoospermic Infertile Men with Low Semen Volume

    PubMed Central

    Mieusset, Roger; Walschaerts, Marie; Isus, François; Almont, Thierry; Daudin, Myriam

    2017-01-01

    In non-azoospermic patients with low semen volume (LSV), looking for partial retrograde ejaculation (PRE) by searching sperm in the postejaculatory urine (PEU) is required. The use of a retro-ejaculatory index (R-ratio) was suggested to define PRE, but none of the studies indicated a specific threshold above which PRE must be considered. Our objective was to propose a threshold value for the R-ratio as indicative of PRE in patients with LSV selected to be devoid of any known causes or risk factors for retrograde ejaculation or LSV. Among our data base (2000–2009) including 632 patients with PEU, 245 male patients from infertile couples who had had a first semen analysis with LSV (< 2mL) and a second semen analysis associated with PEU, were selected on the previous criteria. A prospective control group was randomly constituted (2007–2008) of 162 first consulting male patients from infertile couples, with a normal semen volume (≥ 2mL) on a first semen analysis and who accepted to collect PEU with their usual second semen analysis, selected on the previous criteria. To define an R-ratio threshold indicative of PRE, we used a ROC curve analysis and a regression tree based on a classification and regression tree (CART) algorithm. Of the 245 LSV patients, 146 still presented low semen volume (< 2 mL) on the second semen analysis. From the use of the CART algorithm, two low (1.5% and 2.8%) and two high R-values (7.1% and 8.3%) were defined, according to the lower reference limit for semen volume of 2.0 mL (WHO 1999) or 1.5 mL (WHO 2010) respectively. As only one or no patient with normal semen volume was observed above the two high R-values, we suggest an R-value higher than the range of [7.1–8.3]% as indicative of PRE until confirmation by a prospective multicenter study. PMID:28060836

  12. Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope.

    PubMed

    Nel-Themaat, L; Gómez, M C; Damiani, P; Wirtu, G; Dresser, B L; Bondioli, K R; Lyons, L A; Pope, C E; Godke, R A

    2007-01-01

    Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 degrees C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10 degrees C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross-bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that eland cells were from the semen donor. Eland epithelial cells were transferred into eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.

  13. The effect of mark enhancement techniques on the subsequent detection of semen/spermatozoa.

    PubMed

    Simmons, Rory; Deacon, Paul; Phillips, Darren J; Farrugia, Kevin

    2014-11-01

    Fingermarks, footwear marks, blood and semen are amongst the most commonly encountered types of evidence at crime scenes. Previous work has extensively investigated fingermark and blood enhancement techniques and a sequence developed to maximise evidence recovery; however, there is limited research as to the effect of these techniques on the subsequent detection of body fluids such as semen. In this study, seven fingermark and blood enhancement techniques (e.g. powder suspension, cyanoacrylate fuming and acid violet 17) were employed followed by the subsequent detection of semen/spermatozoa. Other variables included in the study were the use of two substrates (white ceramic tiles and grey laminate flooring), a depletion series and ageing periods of 1, 7, 14 and 28 days. The effect these techniques had on the subsequent detection of semen was assessed by visual and fluorescence examination followed by presumptive and confirmatory testing for semen and spermatozoa. The results found that protein stains (acid violet 17 and acid yellow 7) caused a loss in presumptive test reactivity; however, sperm heads were still observed using microscopic examination after extraction and staining. The use of black magnetic powder, Bluestar(®) Forensic Magnum luminol, Lumicyano™ 4% and cyanoacrylate fuming followed by basic yellow 40 staining did not hinder subsequent presumptive and confirmatory tests for semen and sperm heads. Powder suspension caused a loss in both presumptive test reactivity and sperm heads from the substrate. In general, the enhancement techniques resulted in the improved visualisation of the semen stains under white and violet/blue light. The results from this study aim to provide a strategy to maximise evidence recovery and improve efficiency in an integrated forensic approach.

  14. Encapsulation of sex sorted boar semen: sperm membrane status and oocyte penetration parameters.

    PubMed

    Spinaci, Marcella; Chlapanidas, Theodora; Bucci, Diego; Vallorani, Claudia; Perteghella, Sara; Lucconi, Giulia; Communod, Ricardo; Vigo, Daniele; Galeati, Giovanna; Faustini, Massimo; Torre, Maria Luisa

    2013-03-01

    Although sorted semen is experimentally used for artificial, intrauterine, and intratubal insemination and in vitro fertilization, its commercial application in swine species is still far from a reality. This is because of the low sort rate and the large number of sperm required for routine artificial insemination in the pig, compared with other production animals, and the greater susceptibility of porcine spermatozoa to stress induced by the different sex sorting steps and the postsorting handling protocols. The encapsulation technology could overcome this limitation in vivo, protecting and allowing the slow release of low-dose sorted semen. The aim of this work was to evaluate the impact of the encapsulation process on viability, acrosome integrity, and on the in vitro fertilizing potential of sorted boar semen. Our results indicate that the encapsulation technique does not damage boar sorted semen; in fact, during a 72-hour storage, no differences were observed between liquid-stored sorted semen and encapsulated sorted semen in terms of plasma membrane (39.98 ± 14.38% vs. 44.32 ± 11.72%, respectively) and acrosome integrity (74.32 ± 12.17% vs. 66.07 ± 10.83%, respectively). Encapsulated sorted spermatozoa presented a lower penetration potential than nonencapsulated ones (47.02% vs. 24.57%, respectively, P < 0.0001), and a significant reduction of polyspermic fertilization (60.76% vs. 36.43%, respectively, polyspermic ova/total ova; P < 0.0001). However, no difference (P > 0.05) was observed in terms of total efficiency of fertilization expressed as normospermic oocytes/total oocytes (18.45% vs. 15.43% for sorted diluted and sorted encapsulated semen, respectively). The encapsulation could be an alternative method of storing of pig sex sorted spermatozoa and is potentially a promising technique in order to optimize the use of low dose of sexed spermatozoa in vivo.

  15. Liquid semen storage in elephants (Elephas maximus and Loxodonta africana): species differences and storage optimization.

    PubMed

    Kiso, Wendy K; Brown, Janine L; Siewerdt, Frank; Schmitt, Dennis L; Olson, Deborah; Crichton, Elizabeth G; Pukazhenthi, Budhan S

    2011-01-01

    Artificial insemination plays a key role in the genetic management of elephants in zoos. Because freshly extended semen is typically used for artificial insemination in elephants, it has become imperative to optimize conditions for liquid storage and semen transport. The objectives of this study were to examine the interactions between different extenders and storage temperatures on sperm total motility, progressive motility, and acrosomal integrity in Asian (Elephas maximus) and African (Loxodonta africana) elephants. Ejaculates were collected by rectal massage, diluted using a split-sample technique in 5 semen extenders: TL-Hepes (HEP), Modena (MOD), Biladyl (BIL), TEST refrigeration medium (TES), and INRA96 (INR), maintained at 35°C, 22°C, or 4°C. At 0, 4, 6, 12, and 24 hours, aliquots were removed and assessed for sperm total motility, progressive motility, and acrosomal integrity. After 24 hours of storage, African elephant spermatozoa exhibited greater longevity and higher values in sperm quality parameters compared with those of Asian elephants. In both species, semen storage at 35°C resulted in a sharp decline in all sperm quality parameters after 4 hours of storage, whereas storage at 22°C and 4°C facilitated sperm survival. In Asian elephants, MOD and HEP were most detrimental, whereas BIL, TES, and INR maintained motility up to 12 hours when spermatozoa were cooled to 22°Cor4°C. In African elephants, there were no differences among extenders. All media maintained good sperm quality parameters at 22°C or 4°C. However, although MOD, BIL, and INR were most effective at lower temperatures, HEP and TES maintained sperm motility at all storage temperatures. This study demonstrated sperm sensitivity to components of various semen extenders and storage temperatures and offers recommendations for semen extender choices for liquid semen storage for both Asian and African elephants.

  16. Total Antioxidant Capacity and Lipid Peroxidation in Semen of Patient with Hyperviscosity

    PubMed Central

    Layali, Issa; Tahmasbpour, Eisa; Joulaei, Manijeh; Jorsaraei, Seyed Gholam Ali; Farzanegi, Parvin

    2015-01-01

    Semen hyperviscosity (SHV) is one of the factors involved in deficiency in sperm function. This research aimed to evaluate seminal plasma total antioxidant capacity (TAC) and malondialdehyde (MDA) levels in infertile patients with hyperviscous and non-hyperviscous semen samples to understand whether hyperviscous semen is associated with oxidative damage in infertile subjects. In this cross sectional study, 59 semen samples were provided by fertile (n=12) individuals as control, infertile patients with normal viscosity (n=25) and infertile patients with hyperviscosity (n=22). After semen parameters examination, semen viscosity was studied by glass pipettes. Seminal plasma TAC and MDA levels were measured by ferric reducing of antioxidant power (FRAP) and thiobarbituric acid reaction (TBAR) methods, respectively. A probability less than 0.05 was considered statistically significant throughout the article. The mean of sperm parameters including: counts, motility and normal morphology in patients with hyperviscosity were significantly lower than those in non-hyperviscosity patients (p<0.05, p<0.01 and p<0.001, respectively). The mean of seminal plasma TAC value in seminal plasma of non-hyperviscosity patients (1710.31 ± 458.67 µmol/l) was significantly (p<0.01) higher than that of hyperviscosity group (1230.25 ± 352 µmol/l). A trend toward a higher mean of seminal plasma MDA value was estimated for hyperviscous group compared with non-hyperviscous (1.01 ± 0.41 nmol/ml vs. 0.94 ± 0.28 nmol/l); however, it was nonsignificant. Hyperviscous semen impairs seminal plasma TAC which is eventually associated with sperm membrane lipid peroxidation. PMID:25685746

  17. Naturally and stimulated levels of reactive oxygen species in cooled stallion semen destined for artificial insemination.

    PubMed

    Johannisson, A; Lundgren, A; Humblot, P; Morrell, J M

    2014-10-01

    The decrease in foaling rates after artificial insemination with cooled semen warrants the search for new predictors of fertility. The objectives were to investigate levels of naturally occurring reactive oxygen species (ROS) in cooled, stored stallion semen doses for artificial insemination (AI), and their relationship with parameters of semen quality and with pregnancy rate. Semen was collected from warmblood stallions (n=15) and used to prepare commercial semen doses for AI. Sperm quality was evaluated after cooled transport to the laboratory overnight. The results were correlated with observed foaling and pregnancy rates. Hydroethidine and dichlorodihydrofluorescein diacetate were used as indicators for the ROS superoxide and hydrogen peroxide, respectively. Sperm morphology, motility, plasma membrane integrity and chromatin integrity were also evaluated. These variables were correlated with each other and with pregnancy rates. We found a high inter-individual variation in the ROS levels between stallions. The proportion of live, hydrogen peroxide-negative spermatozoa was correlated with progressive motility, whereas live hydrogen peroxide-negative spermatozoa and chromatin damage were negatively correlated, indicating that low levels of hydrogen peroxide were correlated with good chromatin integrity. The percentage of dead hydrogen peroxide-positive sperm was negatively related to the foaling rate. The negative relationships were stronger when combining results from both assays for ROS. These results for stored semen samples indicate that high individual variation exists for superoxide and hydrogen peroxide measurements, and that ROS status can influence sperm quality. Thus, ROS may be some of the factors influencing fertility. Moreover, combinations of ROS variables improved the correlation with fertility, indicating the usefulness of including these variables in a future model for prediction of the fertility of a semen sample.

  18. Evaluation of sperm DNA damage in bulls by TUNEL assay as a parameter of semen quality.

    PubMed

    Takeda, Kumiko; Uchiyama, Kyoko; Kinukawa, Masashi; Tagami, Takahiro; Kaneda, Masahiro; Watanabe, Shinya

    2015-01-01

    Sperm DNA damage affects the conception rate resulting from human assisted reproduction technology. The objective of this study was to adapt the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to provide a quality parameter for bull semen based on the detection of sperm DNA damage. Fresh semen was collected from two Japanese Black bulls (A, B) several times over the course of a year, and the percentage of TUNEL-positive spermatozoa (sperm TUNEL index) was determined. Individual differences in semen were detected using the sperm TUNEL index in these bulls (P < 0.01). The sperm TUNEL index of cryopreserved semen obtained from test-mated Japanese Black (n = 30, including two bulls with a conception rate lower than 10%) and Holstein (n = 34) bulls were analyzed. The average sperm TUNEL index and conception rate resulting from artificial insemination (AI) were 4.7% and 55.7% for Japanese Black, and 4.9% and 39.5% for Holstein, respectively. A weak negative correlation between sperm TUNEL index and conception rate was observed in Holstein bulls (P < 0.05). Semen samples from six bulls with more than 10% sperm TUNEL index were studied, and these samples showed low sperm viability. However, semen resulting in a very low conception rate did not have a high sperm TUNEL index. Although it would be difficult to predict a low conception rate resulting from AI using the sperm TUNEL index alone, the index can be used as an additional parameter to provide a more comprehensive description of semen quality.

  19. Temperature management during semen processing: Impact on boar sperm quality under laboratory and field conditions.

    PubMed

    Schulze, M; Henning, H; Rüdiger, K; Wallner, U; Waberski, D

    2013-12-01

    Freshly collected boar spermatozoa are sensitive to a fast reduction in temperature because of lipid phase transition and phase separation processes. Temperature management during semen processing may determine the quality of stored samples. The aim of this study was to evaluate the influence of isothermic and hypothermic semen processing protocols on boar sperm quality under laboratory and field conditions. In the laboratory study, ejaculates (n = 12) were first diluted (1:1) with Beltsville Thawing Solution (BTS) at 32 °C, then processed either with isothermic (32 °C) or hypothermic (21 °C) BTS, stored at 17 °C, and assessed on days 1, 3, and 6. Temperature curves showed that 150 minutes after the first dilution, semen doses of both groups reached the same temperature. Two-step hypothermic processing resulted in lower sperm motility on days 1 and 6 (P < 0.05). Concomitantly, hypothermally processed samples contained less membrane intact sperm on days 3 and 6 (P < 0.05). Using AndroStar Plus extender instead of BTS reduced the negative effect of hypothermic processing. In the field study, 15 semen samples from each of 23 European artificial insemination studs were evaluated as part of an external quality control program. Semen quality based on motility, membrane integrity, mitochondrial activity, and a thermoresistance test was higher for stations using one-step isothermic dilutions (n = 7) compared with artificial insemination centers using two-step hypothermic protocols (n = 16). Both studies show that chilling injury associated with hypothermic dilution results in lower quality of stored boar semen compared with isothermic dilution and that the type of semen extender affects the outcomes.

  20. High resolution melt curve analysis based on methylation status for human semen identification.

    PubMed

    Fachet, Caitlyn; Quarino, Lawrence; Karnas, K Joy

    2017-03-01

    A high resolution melt curve assay to differentiate semen from blood, saliva, urine, and vaginal fluid based on methylation status at the Dapper Isoform 1 (DACT1) gene was developed. Stains made from blood, saliva, urine, semen, and vaginal fluid were obtained from volunteers and DNA was isolated using either organic extraction (saliva, urine, and vaginal fluid) or Chelex(®) 100 extraction (blood and semen). Extracts were then subjected to bisulfite modification in order to convert unmethylated cytosines to uracil, consequently creating sequences whose amplicons have melt curves that vary depending on their initial methylation status. When primers designed to amplify the promoter region of the DACT1 gene were used, DNA from semen samples was distinguishable from other fluids by a having a statistically significant lower melting temperature. The assay was found to be sperm-significant since semen from a vasectomized man produced a melting temperature similar to the non-semen body fluids. Blood and semen stains stored up to 5 months and tested at various intervals showed little variation in melt temperature indicating the methylation status was stable during the course of the study. The assay is a more viable method for forensic science practice than most molecular-based methods for body fluid stain identification since it is time efficient and utilizes instrumentation common to forensic biology laboratories. In addition, the assay is advantageous over traditional presumptive chemical methods for body fluid identification since results are confirmatory and the assay offers the possibility of multiplexing which may test for multiple body fluids simultaneously.

  1. Biochemical and physiological characteristics of semen of sex-reversed female rainbow trout (Oncorhynchus mykiss, Walbaum).

    PubMed

    Nynca, Joanna; Kuźmiński, Henryk; Dietrich, Grzegorz J; Hliwa, Piotr; Dobosz, Stefan; Liszewska, Ewa; Karol, Halina; Ciereszko, Andrzej

    2012-01-01

    This works studies the biochemical (protein concentration, osmolality, antitrypsin activity, lactate dehydrogenase activity) and physiological characteristics (sperm motility characteristics) of semen of sex-reversed female rainbow trout (n=42) obtained with the application of 11β-hydroksyandrostendione for sex reversal. All data were arbitrarily divided into three classes depending on the percentage of sperm motility: I XX<25%; II XX 25-50% and III XX>50%. The average percentage of sperm motility was 18±7% n=12 (group I XX); 42±6% n=15 (group II XX) and 65±12% n=15 for group III XX, respectively) to link the values of semen parameters to the maturation stage of semen. Semen from 12 normal males of the same age was used as a reference group. Sperm concentration as well as protein concentration, osmolality, antitrypsin activity, and lactate dehydrogenase activity in seminal plasma of sex-reversed females were higher compared with the values obtained for normal male rainbow trout. The values of these parameters declined with the increasing percentage of sperm motility toward values established for normal males. The fertilization success of semen (3×10(6) spermatozoa/egg) of sex-reversed females was very high (above 90%) for both the percentage of eyed embryos and hatched larvae and was related to sperm motility classes. Correlations between the quality parameters of sex-reversed females semen corresponded to those established previously for the semen of normal male rainbow trout. Antitrypsin activity, lactate dehydrogenase, protein concentration, and osmolality were found to be characteristic of seminal plasma of sex-reversed females. The maturity of sex-reversed female spermatozoa seems to be associated with the decline in the values of those parameters toward the values characteristic for seminal plasma of normal males.

  2. An Overview of a 2004 Ron Brown Cruise to Study Saharan Dust: The NCAS Science Plan for CTAPS: Caracterizacion Tras-Atlantico del Polvo del Sahara

    NASA Astrophysics Data System (ADS)

    Morris, V.

    2003-12-01

    Global transport of dust has been acknowledged as a significant factor in atmospheric radiative balance, atmospheric oxidizing capacity, the deposition of limiting nutrients into the upper ocean, transport of fungi and microorganisms, and in the indirect aerosol effect via cloud modification. In early spring 2004, a group of scientists from NOAA, Howard University, the University of Puerto Rico - Mayagez, and several international partners will take part in a research cruise aboard the Ron Brown to study the impacts and microphysical evolution of Saharan dust during its trans-Atlantic transport on the local atmosphere and marine boundary layer. The fundamental purpose of this research cruise is to provide a set of critical measurements to characterize the impacts of Saharan dust aerosol transport across the Atlantic Ocean. The preliminary science plan and goals of the cruise will be presented and discussed as well as some background measurements taken over the past two years in the Caribbean and Canary Islands.

  3. Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility.

    PubMed

    Campanholi, Suzane Peres; Monteiro, Fabio Morato; Ribeiro Dias, Erika Aline; Mercadante, Maria Eugênia Zerlotti; de Paz, Claudia Cristina Paro; Dell'Aqua Junior, José Antonio; Papa, Frederico Ozanam; Dell'Aqua, Camila de Paula Freitas; Vantini, Roberta; Garcia, Joaquim Mansano

    2017-02-01

    Cryopreservation of bull semen is a common biotechnology procedure in cattle breeding. However, when the ejaculate is obtained by electroejaculation, wide variation is observed in the sperm/seminal plasma (SP) ratio that can affect the freezability of semen in this species. The removal of SP may improve the quality of frozen bull semen. The objective of this study was to evaluate the effect of SP removal from the ejaculate on the cryopreservation of semen from 38 Nellore bulls collected by electroejaculation. After collection, the ejaculate was divided into three aliquots: (1) control (N) diluted to a concentration of 60 × 10(6) spermatozoa/mL and frozen with SP; (2) centrifugation (C) at ×600g for 10 minutes and the pellet resuspended and frozen at the same concentration as N; and (3) filtration (F) through SpermFilter and sperm recovered and frozen at the same concentration as N. After thawing, sperm kinetics, plasma and acrosome membrane integrity, mitochondrial membrane potential, oxidative stress, and in vitro fertility were evaluated. Statistical analysis was performed using the SAS 9.2 package, and differences were considered significant when P < 0.05. Higher average path velocity and straight-line velocity were observed in the groups submitted to SP removal compared to the control group (P < 0.01). In contrast, filtered samples exhibited higher beat cross frequency, straightness, and linearity compared to the other groups. Plasma membrane integrity was reduced when SP was removed, but lower oxidative stress was observed in groups C and F (34.91 ± 2.95% and 31.63 ± 2.95%, respectively) compared to group N (57.39 ± 2.95%). However, the percentage of hatched blastocysts was similar in the N and F groups (21.22 ± 1.05% and 24.00 ± 1.05%, respectively) and higher compared to group C (18.83 ± 1.05%). In conclusion, removal of SP by centrifugation for bull semen freezing reduced the rate of in vitro-produced embryos, whereas filtration of

  4. Effect of widespread and limited use of sexed semen on genetic progress and reproductive performance of dairy cows.

    PubMed

    Khalajzadeh, S; Nejati-Javaremi, A; Mehrbani Yeganeh, H

    2012-09-01

    Stochastic simulation was used for studying the impacts of sexed semen on genetic progress and reproductive performance of dairy cows. Three strategies were compared: WSS (use unsexed semen in cows and heifers), SSH (use sexed semen in heifers and unsexed semen in cows) and SSCH (use sexed semen in both cows and heifers). Conception rate (CR) of unsexed semen was considered to be 35% and 65% in cows and heifers, respectively. CR of sexed semen was considered to be 15 (20% in cows and 50% in heifers), 10, 5 and 0 percentage points lower than unsexed semen. Thus, four subschemes were compared under SSCH (SSCH15, SSCH10, SSCH5, SSCH0) and SSH (SSH15, SSH10, SSH5, SSH0). Moreover, the effect was studied in four distinct paths of selection: active sires (AS), young bulls (YB), bull dams (BD) and milking cows (CW). The average genetic superiority of CW was 12% and 9.5% in SSCH15 and SSH15 strategies relative to a base scheme, respectively. The average genetic superiority of CW was 19% and 10.5% in SSCH0 and SSH0, respectively. Regression analysis showed that genetic superiority of CW increased significantly, that is, 0.5% and 0.1% per every 1% increase in CR in SSCH and SSH, respectively. The result showed that there is a significant difference between genetic superiority of cows in SSCH and SSH schemes. Widespread and limited use of sexed semen in commercial dairy herds resulted in a large genetic advantage in CW. The genetic advantage of gender control was minimal in the selection paths of AS, YB and BD. Open days and services per conception reached to 153 v. 125 days and 5 v. 2.86 under SSCH15 compared with WSS. The age at first calving increased from 774 to 790 days in SSH15 and SSCH15 strategies. Mean of parities decreased to 2.26 v. 2.42 by using sexed semen. The widespread use of sexed semen increased the age average of cows in all parities. Sexed semen increased selection intensity in the CW path, and this contributed to the genetic merit of future cows. On the

  5. Effective freezing rate for semen cryopreservation in endangered Mediterranean brown trout (Salmo trutta macrostigma) inhabiting the Biferno river (South Italy).

    PubMed

    Iaffaldano, Nicolaia; Di Iorio, Michele; Manchisi, Angelo; Esposito, Stefano; Gibertoni, Pier Paolo

    2016-10-01

    This study was designed to determine: (i) the in vitro effects of different freezing rates on post-thaw semen quality of Mediterranean brown trout (Salmo trutta macrostigma) from the Biferno river; and (ii) the in vivo fertilization and hatching percentage of freezing rate giving rise to the best post-thaw semen quality. Pooled semen samples were diluted 1:3 (v:v) in a freezing extender composed of 300 mM glucose, 10% egg yolk and 10% dimethyl sulfoxide (DMSO). The extended semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen surface (1, 5 or 10 cm) for 10 min to give three different freezing rates. Semen samples were thawed at 30°C for 10 s. The variables assessed after thawing were sperm motility, duration of motility and viability. Our results clearly indicate a significant effect of freezing rate on post-thaw semen quality. Semen frozen 5 cm above the liquid nitrogen surface showed the best quality after freezing/thawing. Based on these in vitro data, 2 groups of 200 eggs were fertilized with fresh semen or semen frozen 5 cm above the liquid nitrogen surface. Fertilization and hatching rates recorded for eggs fertilized with frozen semen were significantly lower (25.4% and 22.5%, respectively) than the ones obtained using fresh semen (87.8% and 75.5%, respectively). An effective freezing protocol will allow for the creation of a sperm cryobank to recover the original population of Mediterranean brown trout in the Biferno river.

  6. Altered expression of KLC3 may affect semen parameters

    PubMed Central

    Kargar- Dastjerdy, Pegah; Tavalaee, Marziyeh; Salehi, Mansoor; Falahati, Mojtaba; Izadi, Tayebeh; Nasr Esfahani, Mohammad Hossein

    2016-01-01

    Background: KLC3 protein as a member of the kinesin light-chain protein family plays an important role in spermatogenesis, during formation of mitochondrial sheath in the mid piece of the sperm tail. Objective: This study for the first time aims to compare the expression of the KLC3 gene between fertile and infertile individuals. Materials and Methods: Semen samples were collected from 19 fertile individuals who were selected from embryo-donor volunteers and 57 infertile individuals who had abnormal sperm parameters according to world health organization criteria. Sperm parameters using computer assisted sperm analysis and the quantitative KLC3-gene expression using the real-time PCR method were measured. Results: Our results revealed a significant correlations between sperm concentration with relative expression of KLC3 only in infertile groups (r=0.45, p=0.00). A significant correlation was not found between KLC3 expression and sperm motility; however, the relative expression of KLC3 was significantly higher in asthenozoospermic compared to non-asthenozoospermic individuals. Conclusion: Low expression of KLC3 may result in improper function of midpiece, which has important function in sperm motility. The results of this study show that aberrant expression of KLC3 might be associated with phenomena like oligozoospermia and asthenozoospermia. This article is extracted from student’s thesis. PMID:27141544

  7. Electroejaculation and semen buffer evaluation in the microbat Carollia perspicillata.

    PubMed

    Fasel, Nicolas Jean; Helfenstein, Fabrice; Buff, Samuel; Richner, Heinz

    2015-03-15

    Scientific interests and conservation needs currently stress the necessity to better understanding bat reproductive biology. In this study, we present the first, safe, inexpensive, and reliable method to obtain sperm from a microbat species (Carollia perspicillata) by electroejaculation. This method revealed to be highly efficient (100% success rate). We obtained ejaculates composed of two characteristically different fractions. We compared three buffers and recommend using an Earle's balanced salt solution as a semen extender. Earle's balanced salt solution provided significant repeatable measure of swimming ability (intraclass correlation coefficient: 0.74, P < 0.01) and proportion of motile sperms (intraclass correlation coefficient: 0.08, P = 0.01) and allowed sperm to maintain optimal swimming capacity over time. None of the buffers could dissolve all the coagulated sperm. Although the trypsin buffer freed a larger fraction of spermatozoa in the ejaculate, it impaired swimming ability without improving motility, viability, and stamina. We thus argue that the sperm population analyzed with Earle's balanced salt solution is a representative of the ejaculate. Finally, we found that the mean sperm velocity of C perspicillata (78.8 μm/s) is lower than that predicted by regressing sperm velocity on relative testes mass, a proxy of sperm competition. The question as to whether C perspicillata is an outsider for sperm velocity, or whether bats evolved yet another unique mechanism to cope with sperm competition deserves more investigations.

  8. Chronic epididymitis: impact on semen parameters and therapeutic options.

    PubMed

    Haidl, G; Allam, J P; Schuppe, H-C

    2008-04-01

    Chronic inflammatory conditions of the genital tract are frequently encountered in male fertility problems. The diagnosis, however, is hampered by a mostly asymptomatic course of the disease as well as inappropriate definitions and unspecific diagnostic criteria. With regard to their impact on male reproductive function, epididymitis seems to be more relevant than inflammation/infection of the prostate and/or seminal vesicles. Chronic epididymitis may result in reduced sperm count and motility. Impaired sperm motility because of epididymal dysfunction is frequently associated with an atypical staining behaviour of sperm tails. In many cases of chronic epididymitis, the number of leukocytes in the ejaculate is below the threshold of 10(6) per ml; therefore, consideration of additional markers of inflammation such as granulocyte elastase, pro-inflammatory cytokines (e.g. interleukin-6 or 8) or reactive oxygen species is helpful for establishing the diagnosis. Besides changes in the conventional sperm parameters, alterations in DNA integrity have been observed. Positive effects of antiphlogistic/antibiotic treatment on semen quality have been reported; however, controlled prospective studies are still lacking.

  9. Free and conjugated estrogens and androgens in stallion semen.

    PubMed

    Lemazurier, Emmanuel; Moslemi, Safa; Sourdaine, Pascal; Desjardins, Isabelle; Plainfosse, Bruno; Seralini, Gilles-Eric

    2002-02-01

    The steroid content of semen from a total of 11 mature fertile stallions was studied during two breeding seasons and one winter. The levels of free and conjugated substrates (testosterone and androstenedione), and products (estradiol and estrone), of aromatase were measured by radioimmunoassay with a validated method. The results were seasonally and monthly highly variable with characteristic peaks. The concentrations of free and conjugated estrogens were always higher in the gel-free ejaculate than in the gel except in one subfertile stallion used as comparison. Furthermore, the steroid production and the maximal resulting aromatase activity, estimated by the estrogens/androgens ratio, peaked in April-May and June. The breeding season (spring and summer) presents a clear estrogenic profile with estrogens/androgens ratios higher in contrast to the nonbreeding period (autumn and winter). The involvement of estrogens in the regulation of reproduction and equine spermatogenesis is discussed, and estrogens production and thus equine aromatase is proposed as a strong marker of testicular endocrine function.

  10. Sperm head phenotype and male fertility in ram semen.

    PubMed

    Maroto-Morales, A; Ramón, M; García-Álvarez, O; Montoro, V; Soler, A J; Fernández-Santos, M R; Roldan, E R S; Pérez-Guzmán, M D; Garde, J J

    2015-12-01

    Although there is ample evidence for the effects of sperm head shape on sperm function, its impact on fertility has not been explored in detail at the intraspecific level in mammals. Here, we assess the relationship between sperm head shape and male fertility in a large-scale study in Manchega sheep (Ovis aries), which have not undergone any selection for fertility. Semen was collected from 83 mature rams, and before insemination, head shapes were measured for five parameters: area, perimeter, length, width, and p2a (perimeter(2)/2×π×area) using a computer-assisted sperm morphometric analysis. In addition, a cluster analysis using sperm head length and p2a factor was performed to determine sperm subpopulations (SPs) structure. Our results show the existence of four sperm SPs, which present different sperm head phenotype: SP1 (large and round), SP2 (short and elongated), SP3 (shortest and round), and SP4 (large and the most elongated). No relationships were found between males' fertility rates and average values of sperm head dimensions. However, differences in fertility rates between rams were strongly associated to the proportion of spermatozoa in an ejaculate SP with short and elongated heads (P < 0.001). These findings show how the heterogeneity in sperm head shape of the ejaculate has an effect on reproductive success, and highlight the important role of modulation of the ejaculate at the intraspecific level.

  11. Semen quality in captive Houbara bustard, Chlamydotis undulata undulata.

    PubMed

    Wishart, G J; Lindsay, C; Staines, H J; McCormick, P

    2002-01-01

    Semen quality in captive-bred Houbara bustards, Chlamydotis undulata undulata, was assessed during three consecutive breeding seasons. In any one season, sperm quality, in terms of the proportion of eosin-permeable spermatozoa and of spermatozoa with abnormally large nuclei, varied among individual males, but not among their ejaculates. Neither the proportion of spermatozoa with large nuclei, nor those permeable to eosin were related to the total sperm output of males. The fertilizing ability of males was related to their mean seasonal proportion of eosin-permeable spermatozoa, but not the proportion of spermatozoa with large nuclei. The ranking of males on the basis of the proportion of spermatozoa with large nuclei in their ejaculates was significantly positively correlated between seasons, although ranking on the basis of sperm eosin-permeability was not. The cause or consequence of producing spermatozoa with large nuclei (and excess DNA) remains to be elucidated, but appears to be a trait that is characteristic of houbara bustard males that is maintained between breeding seasons.

  12. Sperm motility patterns in Andalusian donkey (Equus asinus) semen: effects of body weight, age, and semen quality.

    PubMed

    Dorado, J; Acha, D; Gálvez, M J; Ortiz, I; Carrasco, J J; Díaz, B; Gómez-Arrones, V; Calero-Carretero, R; Hidalgo, M

    2013-04-15

    The aims of this study were to (1) identify sperm subpopulations with specific motion characteristics in fresh Andalusian donkey ejaculates; (2) evaluate the effects of individual donkey and ejaculates within the same donkey on the distribution of the subpopulations found; and (3) explore the relationship between the age and the body weight of donkey donors, the sperm quality parameters, and the sperm subpopulations structure. Sixty ejaculates from 12 Andalusian donkeys (five ejaculates per donkey), ranging in age from 4 to 15 years, were collected. Immediately after collection, sperm characteristics (volume, sperm concentration, objective sperm motility, and sperm morphology) were assessed. Donkeys were evaluated for body weight. Significant (P < 0.05) correlations were established between the body weight of the donkeys and the pH (r = -0.52), sperm motility (percentage of motile spermatozoa: r = -0.31; percentage of progressive motile spermatozoa: r = -0.34), and total sperm abnormalities (r = 0.38). The correlations of the age with the measures of semen quality were low and not significant (P > 0.05). A multivariate clustering procedure separated 65,342 motile spermatozoa into four subpopulations: subpopulation 1, consisting of slow and nonprogressive spermatozoa (15.4%), subpopulation 2, consisting of moderately slow but progressive spermatozoa (35.9%), subpopulation 3, consisting of highly active but nonprogressive spermatozoa (18.5%), and subpopulation 4, consisting of highly active and progressive spermatozoa (30.2%). The distribution of these subpopulations varied significantly (P < 0.05) according to several parameters such as the individual donkey, the ejaculate of the same donkey, the total motility, and the overall sperm concentration. Our results show the existence of four well-defined motile sperm subpopulations in Andalusian donkey ejaculates, and suggest a high heterogeneity in the ejaculate structure in donkey. The relationship between the

  13. Effect of dietary selenium and vitamin E on ganders' response to semen collection and ejaculate characteristics.

    PubMed

    Jerysz, Anna; Lukaszewicz, Ewa

    2013-06-01

    Compared to other domestic bird species, geese exhibit the lowest reproductive efficiency (poor semen quality, low egg production, and poor fertility and hatchability rates). From an economic perspective, it is a necessity of improve these reproductive traits. Studies have demonstrated that the essential trace element-selenium-plays key roles in testicular development and the maintenance of spermatogenesis. The aim of the present study was to determine the effect of feed supplementation with organic selenium and vitamin E on ganders' response to manual semen collection and semen quality. Sixteen 3-year-old White Koluda ganders were randomly divided into two groups. The control group was provided commercial feed while the experimental group was provided with the same commercial feed supplemented with selenium (0.3 mg/kg) and vitamin E (100 mg/kg). The response of individual ganders from both groups to manual semen collection and the quality of the semen collected were evaluated. The supplements increased (P ≤ 0.05) the frequency and decreased the time interval of a complete ejaculatory response of the ganders to manual semen collections (82.7 % supplement vs. 73.5 % control). Males from the supplemented group had significantly higher (P ≤ 0.01; P ≤ 0.05) ejaculate volumes, sperm concentrations, and percentages of viable sperm and lower percentages of immature sperm (spermatids). Lipids peroxidation, expressed in terms of the malondialdehyde concentration, was lower (P ≤ 0.01) in semen of the supplemented group (0.172 nmol/50 × 10(6)) as compared to the controls (0.320 nmol/50 × 10(6)). Moreover, the duration of the reproductive period of the ganders in the experimental group was 1 week longer. The results show that supplemental dietary selenium and vitamin E improved both the ganders' response to manual semen collection and semen quality. We conclude that such feed supplementation could lead to greater economic benefits

  14. Successful preservation of capercaillie (Tetrao urogallus L.) semen in liquid and frozen states.

    PubMed

    Kowalczyk, Artur; Łukaszewicz, Ewa; Rzońca, Zenon

    2012-03-15

    Experiments on semen collection and preservation were undertaken by Wrocław University of Environmental and Life Sciences and Forestry Wisła, Poland to assist in the protection of the capercaillie (Tetrao urogallus L.) and to create an ex situ in vitro cryobank. Semen was collected from 11 captive-bred males, using dorsoabdominal massage. Ejaculates once obtained were diluted 3-fold at room temperature with EK diluent and then a number of them were stored at 4 °C for 18, 24, and 48 hours, while the remaining ejaculates were equilibrated with 6% dimethylacetamide and frozen by pipetting, drop-by-drop directly onto a liquid nitrogen surface. Frozen pellets were thawed at 60 °C in a water bath after 4 to 28 mo of storage. In total, 103 individually collected ejaculates (54 stored as liquid and 49 frozen in liquid nitrogen) were of appropriate value for further processing. The volume of ejaculates varied from 30 to 240 μL; spermatozoa concentration from 70 × 10(6) mL(-1) to 1950 × 10(6) mL(-1). The total amount of live spermatozoa in the fresh semen varied from 85.3% to 99.0%, of which from 41.1% to 85.3% were morphologically normal. Among morphologically abnormal forms, bulb-head (5.6% to 36.0%) and midpiece deformations (1.3% to 16.6%) were the most frequent. Dilution and semen storage up to 24 h at 4 °C did not affect the semen quality, as far as motility and sperm morphology are concerned. A significant (P < 0.05) decrease in total live (94.9 vs. 91.7%) and live normal cells (66.4 vs. 56.7%) was observed after 48 h. About 30% to 40% of spermatozoa remained motile. Cryopreservation significantly decreased (P < 0.05) the total number of live and live normal spermatozoa however, in relation to the fresh semen, their average content was 44.1% and 37.4%, respectively. Significant (P < 0.05) individual differences were observed in the quality of the fresh, liquid stored and the frozen-thawed semen assessed in terms of spermatozoa motility and morphology. After a

  15. Artificial insemination field data on the use of sexed and conventional semen in nulliparous Holstein heifers.

    PubMed

    Healy, A A; House, J K; Thomson, P C

    2013-03-01

    This study investigated conception rates and other reproductive outcomes achieved with artificial insemination (AI) of nulliparous Holstein heifers using sexed and conventional semen in a commercial Australian dairy herd in central western New South Wales from January 2004 to April 2009. Retrospective data from on-farm records of 9,870 inseminations of 4,456 heifers were analyzed using several mixed models to assess the effect of temperature and humidity surrounding breeding, insemination sire, artificial insemination technician, service number, and heifer weight and age at breeding on reproductive traits (conception rates, sex ratios, gestation length, and abortion and stillbirth rates). Semen was used from 15 sexed sires and 41 unsexed sires. Sexed semen was primarily used at first and second service. Empirical conception rates of 31.6 and 39.6% were achieved for sexed and unsexed semen respectively, whereas model-based predictions were lower, at 21.3 and 32.1%. Conception rates were significantly affected by insemination sire, sex-sorting, heifer age at breeding, temperature and humidity surrounding breeding, service number, and AI technician. Sexed semen yielded 86% heifers, compared with 48% for conventional semen. Significant predictors of calf sex included semen sexing, gestation length, and insemination sire. Twinning rate was high, at 3.6% for both semen types, and gestation length and heifer weight at breeding were significant predictors of twinning. Abortion rates for sexed and unsexed conceptions were similar at 6.1 and 6.5%, respectively, and were affected by heifer age at breeding. Stillbirth rate was affected by calf sex, twinning, gestation length, and AI technician; semen sorting, age at breeding, and temperature and humidity were marginally significant predictors. No abnormalities were observed in the development of offspring, except for a marginally higher stillbirth rate for sexed calves, a finding that needs further investigation. Many

  16. Effect of seminal plasma vesicular structures in canine frozen-thawed semen.

    PubMed

    Goericke-Pesch, S; Hauck, S; Failing, K; Wehrend, A

    2015-12-01

    Membrane vesicles (MVs) in the ejaculate have been identified in various species and are considered to affect membrane fluidity due to their characteristic molecular composition. Addition of MV to human frozen semen has been shown to improve post-thaw motility. Similarly, a beneficial effect has been suggested for frozen equine semen. As post-thaw canine semen quality varies widely between dogs, the aim of our study was to test for the effect of addition of canine MV on post-thaw semen quality in dogs. Semen samples from 10 male dogs were purified from MV and prepared for freezing. In experiment 1, three groups were compared: sperm frozen (1) with MV (S1); (2) without MV, but MV added immediately after thawing (S2); and (3) without MV (C). Semen analysis included computer-assisted sperm analysis of motility parameters immediately after thawing (t0), after 10 (t10) and 30 minutes (t30), % living sperm, % membrane intact, % morphologically normal sperm (all t0 and t30). Computer-assisted sperm analysis motility distance and velocity parameters (all P < 0.05) and % living sperm (P < 0.001) were significantly affected by treatment with a temporary increase of distance and velocity parameters at t0 to t10, but a significant decrease of the aforementioned parameters at t30 in samples with MV. In experiment 2, different MV protein concentrations added after thawing were compared: 0.05 mg, 0.1 mg, and 0.2 mg/mL. Computer-assisted sperm motility analysis was performed at t0, t10, and t30. No differences between MV concentrations were identified, only a significant interaction between effect of treatment and time for progressive motility (P < 0.01). Our study identified a short-term beneficial effect of canine MV on post-thaw distance and velocity parameters, whereas at t30 progressive motility, motility parameters and % living sperm were reduced in samples with MV compared to C. The results point to species-specific differences regarding the MV effect on frozen

  17. Simple and Effective Methods of Freezing Capercaillie (Tetrao urogallus L.) Semen

    PubMed Central

    Kowalczyk, Artur; Łukaszewicz, Ewa

    2015-01-01

    A continuous decline in the number and range of capercaillie (Tetrao urogallus L.) in many European countries can be observed, mostly due to habitat destruction by human activity, unecological forestry management, and increased density of natural predators. Ex situ in vitro gene banks provide a unique opportunity to preserve the genetic material for future generations. Simple and effective cryopreservation methods for capercaillie semen are discussed. Semen was collected from seven males kept in the Capercaillie Breeding Centre at Forestry Wisła in Poland. Within five minutes after collection, ejaculates were diluted with EK diluent, then divided into two parts, and subjected to two freezing procedures: in pellets and in straws. In fresh semen, ejaculate clearness, viscosity, color and volume, as well as sperm concentration, motility and morphology, were evaluated, while in frozen-thawed semen only motility and morphology of sperm were determined. Fertilizing ability of thawed semen was examined for samples frozen in straws. Significant (P<0.05) differences between individual males were found in relation to the majority of fresh semen traits: ejaculate volume averaged 102.1 µL (varying from 49.0 to 205.0); average sperm concentration was 632.5 x106 mL-1 (178.8–1257.1); percentage of live normal cells varied from 39.2 to 70.3% (58.7% on an average); percentage of motile cells ranged from 76.0 to 85.7%) and motility parameters were male dependent, as well. Both cryopreservation methods had a negative effect on morphology and motility of frozen-thawed semen; however, the straw method yielded 60.7% and the pellet method 42.5% of live cells in total in thawed semen (P<0.05), while the number of live normal (intact) cells was similar (22.4 and 22.2%, respectively). Egg fertility varied between 77.8 and 91.7% (average 84.4%). Both freezing procedures seem to be effective in obtaining acceptable viability and high fertilizing potency of thawed sperm and can be used to

  18. Cryopreservation of bull semen shipped overnight and its effect on post-thaw sperm motility, plasma membrane integrity, mitochondrial membrane potential and normal acrosomes.

    PubMed

    Anzar, M; Kroetsch, T; Boswall, L

    2011-06-01

    In the Canadian Animal Genetic Resource Program, bull semen is donated in frozen or fresh (diluted) states. This study was designed to assess the cryopreservation of diluted bull semen shipped at 4°C overnight, and to determine the post-thaw quality of shipped semen using different straw volumes and freezing rates. Semen was collected from four breeding bulls (three ejaculates per bull). Semen was diluted in Tris-citric acid-egg yolk-glycerol (TEYG) extender, cooled to 4°C and frozen as per routine (control semen). After cooling to 4°C, a part of semen was removed and shipped overnight to the research laboratory via express courier (shipped semen). Semen was packaged in 0.25 or 0.5 ml straws and frozen in a programmable freezer using three freezing rates, i.e., -10, -25 or -40°C/min. Control semen was also shipped to the research laboratory. Post-thaw sperm motility characteristics were assessed using CASA, and post-thaw sperm plasma membrane, mitochondrial membrane potential and normal acrosomes were assessed using flow cytometry. Post-thaw sperm quality was greater in shipped semen as compared to control (P<0.001). The shipped semen packaged in 0.25 ml straws had better post-thaw sperm quality than in 0.5 ml straws (P<0.001). Freezing rate had no effect on post-thaw sperm quality. In conclusion, bull semen can be shipped overnight for subsequent cryopreservation and gene banking. Overnight shipping of semen was found advantageous for bull semen cryopreservation. Semen packaging in 0.25 ml straws yielded better post-thaw quality than 0.5 ml straws.

  19. Successful ram semen cryopreservation with lyophilized egg yolk-based extender.

    PubMed

    Alcay, Selim; Berk Toker, M; Gokce, Elif; Ustuner, Burcu; Tekin Onder, N; Sagirkaya, Hakan; Nur, Zekariya; Kemal Soylu, M

    2015-10-01

    The aim of this study was to evaluate the effects of lyophilized egg yolk extender on ram semen cryopreservation. Ejaculates with a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. Sperm were diluted to final concentration of 1/5 (semen/extender) in lyophilized egg yolk or fresh egg yolk extenders using two-step dilution method. The equilibrated semen was frozen in 0.25 mL straws. Semen samples were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) at three time points: after dilution with extender A, equilibration and post-thaw. The results showed that freezing and thawing procedures (dilution, equilibration and thawing) had negative effects on motility (P<0.001), plasma membrane integrity (P<0.001), acrosome integrity (P<0.001) and DNA integrity (P<0.001). In the study, there were no significant differences between lyophilized and fresh egg yolk extenders when comparing motility, plasma membrane integrity, acrosome integrity and DNA integrity between groups. In conclusion, lyophilized egg yolk extender provided similar cryoprotective effects with fresh egg yolk extender to cryopreserve ram semen.

  20. Zinc and iron concentration and SOD activity in human semen and seminal plasma.

    PubMed

    Marzec-Wróblewska, Urszula; Kamiński, Piotr; Lakota, Paweł; Szymański, Marek; Wasilow, Karolina; Ludwikowski, Grzegorz; Kuligowska-Prusińska, Magdalena; Odrowąż-Sypniewska, Grażyna; Stuczyński, Tomasz; Michałkiewicz, Jacek

    2011-10-01

    The aim of the present study was to measure zinc (Zn) and iron (Fe) concentration in human semen and superoxide dismutase (SOD) activity in seminal plasma and correlate the results with sperm quality. Semen samples were obtained from men (N = 168) undergoing routine infertility evaluation. The study design included two groups based on the ejaculate parameters. Group I (n = 39) consisted of males with normal ejaculate (normozoospermia), and group II (n = 129) consisted of males with pathological spermiogram. Seminal Zn and Fe were measured in 162 samples (group I, n = 38; group II, n = 124) and SOD activity in 149 samples (group I, n = 37; group II, n = 112). Correlations were found between SOD activity and Fe and Zn concentration, and between Fe and Zn concentration. SOD activity was negatively associated with volume of semen and positively associated with rapid progressive motility, nonprogressive motility, and concentration. Negative correlation was stated between Fe concentration and normal morphology. Mean SOD activity in seminal plasma of semen from men of group I was higher than in seminal plasma of semen from men of group II. Fe concentration was higher in teratozoospermic males than in males with normal morphology of spermatozoa in group II. Our results suggest that Fe may influence spermatozoa morphology.

  1. The use of maca (Lepidium meyenii) to improve semen quality: A systematic review.

    PubMed

    Lee, Myeong Soo; Lee, Hye Won; You, Sooseong; Ha, Ki-Tae

    2016-10-01

    The aim of this review was to assess the evidence for the effectiveness of maca (Lepidium meyenii) in improving semen quality. We searched 11 databases from their inception to March 2016 and included all clinical trials on the improvement of semen quality parameters in infertile and healthy men, regardless of the study design or the type of maca. The risk of bias for each study was assessed using the Cochrane criteria. The selection of studies, data extraction, and validation were performed independently by the first two authors. Discrepancies were resolved through discussion by the same two authors. Five studies - 3 randomized clinical trials (RCTs) and 2 uncontrolled observational studies (UOSs) - met all of the inclusion criteria. One RCT found favorable effects of maca on sperm mobility in infertile men. The two other RCTs showed positive effects of maca on several semen quality parameters in healthy men. The two UOSs also suggested favorable effects of maca on semen quality. The results of our systematic review provide suggestive evidence for the effectiveness of maca in improving semen quality. However, the total number of trials, the total sample size, and the risk of bias of the included studies prevent the drawing firm conclusions. More rigorous studies are warranted.

  2. Effect of uremia on semen quality and reproductive function in humans.

    PubMed

    Xu, Longgen; Xu, Huiming; Zhu, Xiaofeng; Zhang, Junrong; Ma, Meili; Shi, Xiaomin

    2012-01-01

    In this study, we sought to evaluate the effect of uremia on semen quality and reproductive function in humans. For this purpose, 53 end-stage uremic patients were randomly selected. The semen samples were produced by masturbation. Fertility index (FI) was calculated according to the following formula: sperm density (×10(6)/ml) × sperm motility (%) × normal sperm morphology rate (% per 10,000). The semen samples of uremic patients were compared with those of fertile and infertile males. The results show that three patients failed to produce semen. There were no sperm found in four semen samples. The sperm motility, survival rate, sperm density, and normal sperm morphology rate of the remaining 46 patients were found to be significantly lower than those of controls. The uremic patients had the FI of 0.68(2.08) which was obviously lower than that of fertile 7.7(13.51) and infertile 4.13(5.77) males. It was, therefore, concluded that uremia caused a significant decline in sperm quality and reproductive function which resulted in consequential infertility in humans.

  3. Embryo production and possible species preservation by nuclear transfer of somatic cells isolated from bovine semen.

    PubMed

    Liu, Jie; Westhusin, Mark; Long, Charles; Johnson, Gregory; Burghardt, Robert; Kraemer, Duane

    2010-12-01

    Somatic cells in semen are a potential source of nuclei for nuclear transfer to produce genetically identical animals; this is especially important when an animal has died and the only viable genetic material available is frozen semen. Usefulness of somatic cells obtained from fresh (cultured) and frozen (isolated, not cultured) bovine semen for nuclear transfer was evaluated. Twelve ejaculates were collected from nine bulls representing three breeds: Charolais, Brahman, and crossbred Rodeo bull. All samples were processed immediately and cell growth was obtained from seven of the twelve ejaculates (58.3%). Cells from three bulls (with the best growth rates) were evaluated by optical microscopy and used in cloning experiments. In culture, these cells exhibited classic epithelial morphology and expressed cytokeratin and vimentin, indicating they were of epithelial origin. When cells from the three bulls were used as donor cells, 15.9% (18/113), 34.5% (29/84), and 14.4% (13/90) of the fused embryos developed into blastocysts, respectively. Of the blastocyst stage embryos, 38.9% (7/18), 72.4% (21/29), and 61.5% (8/13) hatched, respectively. Somatic cells isolated (not cultured) from frozen bovine semen were also used in the cloning experiments. Although cleavage occurred, no compact morulae or blastocysts were obtained. In conclusion, epithelial cell growth was obtained from fresh bovine ejaculates with relatively high efficiency. Somatic cells from semen can be used as nucleus donors to produce cloned blastocyst-stage embryos.

  4. Low lead environmental exposure alters semen quality and sperm chromatin condensation in northern Mexico.

    PubMed

    Hernández-Ochoa, Isabel; García-Vargas, Gonzalo; López-Carrillo, Lizbeth; Rubio-Andrade, Marisela; Morán-Martínez, Javier; Cebrián, Mariano E; Quintanilla-Vega, Betzabet

    2005-01-01

    We evaluated environmental-lead (Pb) effects on semen quality and sperm chromatin, considering Pb in seminal fluid (PbSF), spermatozoa (PbSpz), and blood (PbB) as exposure biomarkers in urban men (9.3 microg/dL PbB). Several individuals (44%) showed decreases in sperm quality; sperm concentration, motility, morphology and viability associated negatively with PbSpz, whereas semen volume associated negatively with PbSF. Multiple linear regression estimated PbSF and PbSpz thresholds for alterations in semen quality. Forty-eight percent of samples showed high values of nuclear chromatin condensation (NCD) positively associated with PbSF and zinc in spermatozoa (ZnSpz). ZnSpz values were higher than in fertile men. These results suggest that Pb may affect sperm chromatin by altering sperm Zn availability. PbB was not associated with semen quality or NCD, suggesting that Pb in semen compartments assesses better the amount of Pb in the reproductive tract; therefore, these are better biomarkers to evaluate toxicity at low Pb-exposure levels.

  5. Genetic parameters and breeding values for semen characteristics in Hanoverian stallions.

    PubMed

    Labitzke, D; Sieme, H; Martinsson, G; Distl, O

    2014-08-01

    The objectives of this study were to show whether semen traits of 30 Hanoverian stallions regularly used in AI may be useful for breeding purposes. Semen characteristics were studied using 15 149 ejaculates from 30 Hanoverian stallions of the State Stud Celle of Lower Saxony. Semen samples were collected between 2005 and 2009. Traits analysed were gel-free volume, sperm concentration, total and motile sperm number and progressive motility. A linear multivariate animal model was employed to estimate heritabilities and permanent environmental variances for stallions. The same model was used to predict breeding values for all traits simultaneously. Heritabilities were high for gel-free volume (h(2) = 0.43) and moderate for total number of sperm (h(2) = 0.29) and progressive motility (h(2) = 0.20). Gel-free volume, sperm concentration and total number of sperm were genetically negatively correlated with progressive motility. The effect of the permanent environment for stallions accounted for 9-55% of the trait variance. The total variance among stallions explained 37-69% of the trait variance. The average reliabilities of the breeding values were 0.43-0.76 for the 30 Hanoverian stallions. In conclusion, the study could demonstrate large effects of stallions, routinely employed in a breeding programme, on semen characteristics analysed here. We could demonstrate that estimated breeding values (EBV) with sufficient high reliabilities can be predicted using data from these stallions and these EBV are useful in horse breeding programmes to achieve genetic improvement in semen quality.

  6. 9 CFR 85.10 - Interstate movement of swine semen and swine embryos for insemination of or implantation into swine.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... swine embryos for insemination of or implantation into swine. 85.10 Section 85.10 Animals and Animal... and swine embryos for insemination of or implantation into swine. Swine semen and swine embryos moved... collection of the semen or embryos or were members of a qualified pseudorabies negative herd, and had...

  7. 9 CFR 85.10 - Interstate movement of swine semen and swine embryos for insemination of or implantation into swine.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... swine embryos for insemination of or implantation into swine. 85.10 Section 85.10 Animals and Animal... and swine embryos for insemination of or implantation into swine. Swine semen and swine embryos moved... collection of the semen or embryos or were members of a qualified pseudorabies negative herd, and had...

  8. 9 CFR 85.10 - Interstate movement of swine semen and swine embryos for insemination of or implantation into swine.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... swine embryos for insemination of or implantation into swine. 85.10 Section 85.10 Animals and Animal... and swine embryos for insemination of or implantation into swine. Swine semen and swine embryos moved... collection of the semen or embryos or were members of a qualified pseudorabies negative herd, and had...

  9. 9 CFR 85.10 - Interstate movement of swine semen and swine embryos for insemination of or implantation into swine.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... swine embryos for insemination of or implantation into swine. 85.10 Section 85.10 Animals and Animal... and swine embryos for insemination of or implantation into swine. Swine semen and swine embryos moved... collection of the semen or embryos or were members of a qualified pseudorabies negative herd, and had...

  10. 9 CFR 85.10 - Interstate movement of swine semen and swine embryos for insemination of or implantation into swine.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... swine embryos for insemination of or implantation into swine. 85.10 Section 85.10 Animals and Animal... and swine embryos for insemination of or implantation into swine. Swine semen and swine embryos moved... collection of the semen or embryos or were members of a qualified pseudorabies negative herd, and had...

  11. Assessment of extent of apoptosis and DNA defragmentation in chilled semen of stallions during the breeding season.

    PubMed

    Krakowski, L; Obara, J; Wąchocka, A; Piech, T; Bartoszek, P; Kostro, K; Tatara, M R

    2013-10-01

    The objective of the study was to assess apoptosis and DNA defragmentation in equine semen diluted and chilled to +4°C. Semen was collected from nine fertile stallions, including four Arabian thoroughbreds and five coldbloods. Examinations were carried out immediately after semen collection (0) and at five storage times (24, 48, 72, 96 and 120 h). The basic semen evaluation was performed in terms of volume, sperm concentration, viable sperm percentage, progressive motility and morphology. Using flow cytometry, DNA defragmentation and cell membrane integrity of spermatozoa were determined. The results of basic tests did not demonstrate significant differences amongst stallions, except for progressive sperm motility, which was significantly higher (p < 0.05) in the semen of Arabian stallions. In the semen of the same stallions, a significant decrease in the percentage of alive spermatozoa was observed at 72, 96 and 120 h of storage, whereas a significant increase in the number of spermatozoa with DNA defragmentation was found after 24 h storage. In the semen of coldblood stallions, significantly reduced live spermatozoa percentage was observed at 96 and 120 h, while increased DNA defragmentation was observed at 48 h. These findings demonstrated that the semen of Arabian stallions chilled to +4°C retained original characteristics until 24 h of storage, whereas in coldbloods, these were preserved up to 48 h of storage.

  12. Cryopreservation of canine semen after cold storage in a Neopor box: effect of extender, centrifugation and storage time.

    PubMed

    Hidalgo, M; Portero, J M; Demyda-Peyrás, S; Ortiz, I; Dorado, J

    2014-07-05

    The aim of this work was to assess the combined effect of sperm centrifugation, semen extender and storage time before freezing on post-thaw sperm quality and freezability on chilled stored canine semen in a Neopor box. Sperm parameters evaluated were total and progressive sperm motility by Computer-Assisted Sperm Analysis (CASA) and sperm viability and acrosome integrity using a triple fluorescent stain. Sperm quality and freezability indexes were also studied. First, the effect of centrifugation and two commercial extenders from Minitübe (Biladyl A and CaniPRO Freeze A) was evaluated in chilled semen after 24 and 45 hours of cold storage. No significant differences were observed between treatments in almost all the sperm parameters assessed. Secondly, chilled semen was frozen after 24 and 45 hours of cold storage in a Neopor box. The best results were obtained when semen was centrifuged, chilled with CaniPRO Freeze A and then frozen after 24 hours of cold storage, showing no differences in both post-thaw sperm quality and freezability in comparison with semen immediately frozen after collection. In conclusion, dog semen centrifuged after collection and extended with CaniPRO Freeze can be frozen after 24 hours of cold storage in a Neopor box, obtaining similar results to semen immediately frozen after collection.

  13. Selenium status and GSH-Px activity in semen and blood of boars at different ages used for artificial insemination.

    PubMed

    Lasota, B; Błaszczyk, B; Seremak, B; Udała, J

    2004-10-01

    This study was performed to determine the relationship between selenium (Se) content and Se-dependent glutathione peroxidase (GSH-Px) activity in blood and semen, and semen quality of boars at different age used in an artificial insemination (AI) station. Routine macroscopic and microscopic analyses of semen quality were accompanied by measurement of Se content and GSH-Px activity in blood and semen. The Se concentration in blood plasma, seminal fluid and spermatozoa was measured by fluorometric method, the GSH-Px activity by a method based on NADPH-coupled reaction. A total of 155 ejaculates and 58 blood samples were investigated. The results of this study showed that there was no direct relationship between the Se content and GSH-Px activity in blood plasma and semen, and semen quality of sexual matured boars. The mechanisms controlling Se content and GSH-Px activity in blood and semen seem to be independent. The age of boars as a differentiating factor for Se content and GSH-Px activity in blood and semen is possible. It is concluded that a determination of Se status and/or GSH-Px activity in organism before Se supplementation is indicated.

  14. Intravaginal artificial insemination in bitches using frozen/thawed semen after dilution in powdered coconut water (ACP-106c).

    PubMed

    Uchoa, D C; Silva, T F P; Mota Filho, A C; Silva, L D M

    2012-12-01

    The aim of this study was to evaluate powdered coconut water extender (ACP-106c; ACP Serviços Tecnológicos Ltda, ACP Biotecnologia, Fortaleza, Ceará, Brazil) as a diluent for freezing dog semen and the fertility after vaginal insemination of semen frozen therein. Ten ejaculates were collected from five dogs, evaluated fresh, diluted in ACP-106c, 10% egg yolk and 6% glycerol, cooled and frozen. In the first phase of the study, straws with frozen semen were thawed and immediately subjected to the same analysis as the fresh semen and, in addition, to Computer-Assisted Semen Analysis (CASA). In phase 2, 10 bitches that had been subjected to natural breeding during a preceding oestrous cycle were vaginally inseminated with thawed semen that had been re-diluted in ACP-106c. After thawing, a mean of 77% sperm motility was obtained through subjective analysis and 77.3% through CASA. Following artificial insemination, a 60% pregnancy rate was observed, resulting in a 50% parturition rate and a mean litter size of 3.4 (SEM 0.6), with 47.1% males and 52.9% females. ACP-106c can be successfully used for freezing canine semen, and vaginal deposition of such semen yields similar pregnancy rates to those reported in other studies.

  15. Quality assessment of wild Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) semen under conditions of short-term storage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Short-term storage trials were conducted with Atlantic sturgeon semen collected from a total of nine wild males during the 2008 and 2009 spawning seasons on the Hudson River. Semen samples were kept refrigerated (4 plus or minus 1 degree C) and stored in different gaseous atmospheres and storage ext...

  16. Cryopreservation of turkey semen: effect of breeding line and freezing method on post-thaw sperm quality, fertilization, and hatching

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cryopreservation methods for poultry semen are not reliable for germplasm preservation, especially for turkeys, where fertility rates from frozen/thawed semen are particularly low. The objective was to evaluate cryopreservation methods for effectiveness in promoting cryosurvival and post-thaw funct...

  17. Visual Detection of Brucella spp. in Spiked Bovine Semen Using Loop-Mediated Isothermal Amplification (LAMP) Assay.

    PubMed

    Prusty, Bikash R; Chaudhuri, Pallab; Chaturvedi, V K; Saini, Mohini; Mishra, B P; Gupta, Praveen K

    2016-06-01

    Several pathogens including Brucella spp. are shed in semen of infected bulls and can be transmitted to cows through contaminated semen during artificial insemination. The present study reports omp2a and bcsp31 gene based loop-mediated isothermal amplification (LAMP) assays for detection of Brucella genomic DNA in semen from infected bulls. The positive results could be interpreted visually by change in colour of reaction mixture containing hydroxyl naphthol blue (HNB) dye from violet to sky blue. LAMP assays based on omp2a and bcsp31 could detect as little as 10 and 100 fg of B. abortus S19 genomic DNA, respectively. Sensitivity of omp2a and bcsp31 LAMP assays for direct detection of organisms in bovine semen was 2.28 × 10(1) CFU and 2.28 × 10(2) CFU of B. abortus S19 in spiked bovine semen, respectively. The omp2a LAMP assay was found equally sensitive to TaqMan probe based real-time PCR and 100 times more sensitive than conventional PCR in identifying Brucella in spiked semen. The diagnostic applicability of the omp2a LAMP assay was evaluated with seventy-nine bovine semen samples and results were re-evaluated through TaqMan probe based real-time PCR and conventional PCR. Taken together, the omp2a LAMP assay is easy to perform, rapid and sensitive in diagnosis of Brucella spp. in bovine semen.

  18. Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR.

    PubMed

    Gimenes, Fabrícia; Medina, Fabiana Soares; Abreu, André Luelsdorf Pimenta de; Irie, Mary Mayumi Taguti; Esquiçati, Isis Baroni; Malagutti, Natália; Vasconcellos, Vinícius Rodrigo Bulla; Discacciati, Michele Garcia; Bonini, Marcelo Gialluisi; Maria-Engler, Silvya Stuchi; Consolaro, Marcia Edilaine Lopes

    2014-01-01

    Sexually transmitted diseases (STDs) may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR) assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV) -1 and -2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV) and genotypes by single PCR (sPCR) in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%), sensitivity (100.00%), specificity (99.70%), positive (96.40%) and negative predictive values (100.00%) and accuracy (99.80%). The prevalence of STDs was very high (55.3%). Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks.

  19. Effect of semen collection method on sperm motility of gray wolves (Canis lupus) and domestic dogs (C. l. familiaris).

    PubMed

    Christensen, Bruce W; Asa, Cheryl S; Wang, Chong; Vansandt, Lindsey; Bauman, Karen; Callahan, Margaret; Jens, Jackie K; Ellinwood, N Matthew

    2011-09-15

    Genetic management of Mexican gray wolves includes semen banking, but due to the small number of animals in the population and handling restrictions, improvements in semen collection and cryopreservation rely on results from studies of domestic dogs. Semen collection from wolves requires anesthesia and electroejaculation, which introduce potentially important variables into species comparisons, as dog semen is typically collected manually from conscious animals. To investigate possible effects of collection method on semen quality, we compared semen collection by the traditional manual method and by electroejaculation (EE) in a group of dogs (n = 5) to collection by EE only in wolves (n = 7). Samples were divided into two aliquots: neat or diluted in Tris/egg yolk extender, with motility evaluated at intervals up to 24 h. There were no differences (P > 0.10) in sperm motility in either neat or extended samples at 24 h from EE dogs and wolves, although motility of the wolf neat samples declined more rapidly (P < 0.05). However, there were differences (P < 0.01) between EE and manually collected dog semen in motility at 24 h, in both the neat and extended samples. Therefore, general motility patterns of dog and wolf semen collected by EE were similar, especially when diluted with a Tris/egg yolk extender, but sperm collected from dogs by EE did not maintain motility as long as manually collected samples, perhaps related to the longer exposure of EE samples to more prostate fluid.

  20. Sensitive Simultaneous Detection of Seven Sexually Transmitted Agents in Semen by Multiplex-PCR and of HPV by Single PCR

    PubMed Central

    de Abreu, André Luelsdorf Pimenta; Irie, Mary Mayumi Taguti; Esquiçati, Isis Baroni; Malagutti, Natália; Vasconcellos, Vinícius Rodrigo Bulla; Discacciati, Michele Garcia; Bonini, Marcelo Gialluisi; Maria-Engler, Silvya Stuchi; Consolaro, Marcia Edilaine Lopes

    2014-01-01

    Sexually transmitted diseases (STDs) may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR) assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV) −1 and −2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV) and genotypes by single PCR (sPCR) in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%), sensitivity (100.00%), specificity (99.70%), positive (96.40%) and negative predictive values (100.00%) and accuracy (99.80%). The prevalence of STDs was very high (55.3%). Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks. PMID:24921247

  1. Differences in ability of jennies and mares to conceive with cooled and frozen semen containing glycerol or not.

    PubMed

    Vidament, Marianne; Vincent, Pierrick; Martin, François-Xavier; Magistrini, Michele; Blesbois, Elisabeth

    2009-05-01

    A suitable method for the cryopreservation of donkey semen would be very valuable for the ex situ management of genetic diversity in this species. This report uses a variety of observation and trials to evaluate the effect of cryoprotectants in per-cycle pregnancy rates (PC) in equids females (jennies (donkey) and mares (horse)). This was explored by (1) comparing the results of insemination of jennies and mares with cooled or frozen donkey semen, (2) examining the possible toxic effect of the cryoprotectant (CPA) glycerol in these two species and (3) studying alternative solutions. Donkey and horse semen was either used immediately, or cooled according to some steps of the pre-freezing procedure or frozen and thawed. The pre-freezing procedure included semen dilution, centrifugation, resuspension in milk or in INRA82+2% egg yolk+various % CPA (expressed as final concentrations in extended semen (v/v)) and then cooling to 4 degrees C. PC was similar in mares and jennies inseminated with donkey semen cooled to 4 degrees C in milk. However, the PC was significantly higher in mares than in jennies when donkey semen was frozen with 2.2% glycerol (36%, n=50 cycles vs. 11%, n=38 cycles; P<0.01). Increasing the concentrations of glycerol (0, 2.2, 3.5, 4.8%) before cooling stallion semen resulted in a progressive decrease in mare PC (87, 53, 53, 13% (n=15 cycles for each concentration); P<0.0001). The addition of 2.2% glycerol before cooling donkey semen decreased the PC measured in jennies to 0. The replacement of glycerol by 2% dimethylformamide increased the fertility obtained in jennies with cooled donkey semen (PC: 67%, n=12 cycles) but did not increase the fertility obtained with frozen-thawed donkey semen (PC: 11%, n=28 cycles with dimethylformamide vs. 0%, n=16 cycles with glycerol). In conclusion, this study clearly shows that the ability of jennies to conceive after AI with donkey frozen semen is lower than that of mares. Glycerol affects the fertility of donkey

  2. Artificial insemination of red deer (Cervus elaphus) with frozen-thawed wapiti semen.

    PubMed

    Haigh, J C; Bowen, G

    1991-09-01

    Semen collected from wapiti (Cervus elaphus) in Canada in 1983 was frozen in two extenders. In 1988, the semen was used to inseminate 200 red deer hinds on 2 farms in New Zealand. Oestrus was synchronized in the hinds with progesterone-impregnated intravaginal devices (CIDR); 200 iu pregnant mares' serum gonadotrophin was given to each hind on Day 11. The CIDRs were removed on Day 12 at 20/h, as the numbers of the hinds were recorded. On Day 14, 54-56 h after CIDR removal, the hinds were brought into the yards in the same batches and laparoscopically inseminated. Semen from three sires was used. The overall conception rate was 51%. Gestation length ranged from 239 to 247 days. One hind was lost at calving, 3 calves had to be hand raised and there were 2 neonatal calf deaths.

  3. Oxidative and glycolytic metabolism of semen components by washed guinea pig spermatozoa.

    PubMed

    Frenkel, G; Peterson, R N; Freund, M

    1975-02-01

    The concentration of several potentially metabolizable substances in guinea pig semen and the ability of these substances to support ATP synthesis and the motility of guinea pig sperm have been determined. Both glucose and fructose were present in high concentration in semen and were equipotent at the concentration tested in maintaining high levels of ATP and a high rate of motility. Lactic and pyruvic acids also supported a high rate of sperm motility but maintained lower levels of ATP. These constituents of guinea pig semen, as well as the metabolites alpha-glycerophosphate, succinic acid, and beta-hydroxybutyric acid, are oxidized at unusually high rates. The active oxidative metabolism of guinea pig sperm is compared with that of human sperm which is primarily glycolytic.

  4. Race Differentiation Based on Raman Spectroscopy of Semen Traces for Forensic Purposes.

    PubMed

    Muro, Claire K; Lednev, Igor K

    2017-04-03

    Several novel methods to determine externally visible characteristics of body fluid donors have been developed in recent years. These tests can help forensic investigators make predictions about the appearance of a suspect or victim, such as their sex, race, hair color, or age. While their potential benefit is undeniable, these methods destroy the physical evidence in the process. Raman spectroscopy has recently been used as a nondestructive technique to test for many of these characteristics. Here, we present the results from a study to determine the race of semen donors. Using Raman spectroscopy and multivariate data analysis, we were able to build a statistical model that accurately identified the race of all 18 semen donors in the calibration data set, as well as seven additional external validation donors. These results demonstrate Raman spectroscopy's potential to differentiate Caucasian and Black semen donors using chemometrics.

  5. Lead exposure and semen quality among traffic police in Arequipa, Peru.

    PubMed

    Eibensteiner, Lynn; Del Carpio Sanz, Ada; Frumkin, Howard; Gonzales, Carla; Gonzales, Gustavo F

    2005-01-01

    This study examined lead exposure (n = 43) and semen quality (n = 18) among traffic police officers in Arequipa, Peru, where leaded gasoline is used. Blood lead (PbB) was measured using atomic absorption spectrophotometry, and semen was analyzed following World Health Organization (WHO) protocol. Mean PbB was 48.5 microg/dL. Although current PbB was associated with declines in several semen parameters (sperm morphology, concentration and total number of sperm), only sperm motility and viability differed significantly between the < or = 40 microg/dL and > 40 microg/dL categories, and decreased with increasing PbB in simple linear regression. Traffic police are an indicator group for excessive ambient lead exposure, and these results support earlier findings on the male reproductive toxicity of lead. The results should be interpreted cautiously since the numbers were small and the analysis was unable to control for all potential confounders due to incomplete data.

  6. Genome-wide association study for semen quality traits in German Warmblood stallions.

    PubMed

    Gottschalk, Maren; Metzger, Julia; Martinsson, Gunilla; Sieme, Harald; Distl, Ottmar

    2016-08-01

    We performed a genome-wide association study for semen quality traits in 139 German Warmblood stallions. Stallions were genotyped using the Illumina equine SNP50 Beadchip. Traits analysed were de-regressed estimated breeding values (EBVs) for gel-free volume, sperm concentration, total number of sperm, progressive motility and the total number of progressively motile sperm. The GWAS revealed 29 SNPs on 12 different chromosomes as genome-wide significantly associated with semen quality traits. For ten genomic regions we could retrieve candidate genes influencing stallion fertility. Among the candidate genes, we could find the genes encoding cysteine-rich secretory proteins (CRISP1, CRISP2 and CRISP3). This was the first GWAS in horses performed for semen quality traits.

  7. Single Layer Centrifugation Can Be Scaled-Up Further to Process up to 150 mL Semen

    PubMed Central

    Morrell, J. M.; van Wienen, M.; Wallgren, M.

    2011-01-01

    Single-Layer centrifugation has been used to improve the quality of sperm samples in several species. However, where stallion or boar semen is to be used for AI, larger volumes of semen have to be processed than for other species, thus limiting the effectiveness of the original technique. The objective of the present study was to scale up the SLC method for both stallion and boar semen. Stallion semen could be processed in 100 mL glass tubes without a loss of sperm quality, and similarly, boar semen could be processed in 200 mL and 500 mL tubes without losing sperm quality. The results of these preliminary studies are encouraging, and larger trials are underway to evaluate using these methods in the field. PMID:23738111

  8. A commercial box for dog semen transport: What happens inside when the environmental temperature is increasing?

    PubMed

    Cunha, I C N; Henning, H; Urhausen, C; Beyerbach, M; Günzel-Apel, A R

    2014-06-10

    Environmental temperatures may influence the temperature inside commercial transport boxes during semen shipment and thereby storage conditions of diluted dog semen. To evaluate the temperature changes inside boxes and their influence on sperm quality, split semen samples (n=8) were placed in Neopor boxes(®) exposed for 48h to room temperature (RT) (Box 1), 40°C for 6h and then kept at RT (Box 2) or 40°C (Box 3). A fourth subsample was kept at 4-5°C in a refrigerator (control). Inside Box 1 temperature initially decreased to <3°C before it stabilized at 7-8°C, while in Box 2 no decrease occurred and temperature was at 7-8°C for 48 h. Temperature inside Box 3 was at 14-15°C for 24h and, thereafter, increased to 36.1°C. Analysis of sperm motility (CASA) and viability (PI and FITC-PNA) after 24 and 48 h revealed marked sensitivity of dog spermatozoa to temperature fluctuations (Box 1). A constant storage temperature of 7-8°C (Box 2) provided the most desirable semen quality in terms of motility, viability, as well as osmotic resistance when samples were stored for 48 h. Furthermore, results indicate that during 24h preservation a storage temperature of 14-16°C may provide optimum conditions for maintenance of sperm viability and function. An increase of the inside temperature to >30°C (Box 3) resulted in an almost complete loss in sperm integrity. In conclusion, results suggest a revision of current recommendations for storage temperature of diluted dog semen. Boxes for semen transport should be prepared depending on the expected environmental temperatures.

  9. Computer aided boar semen motility analysis for cereulide detection in different food matrices.

    PubMed

    Rajkovic, Andreja; Uyttendaele, Mieke; Debevere, Johan

    2007-02-28

    Computer Aided Semen Analysis (CASA) study of the boar semen motility has been demonstrated to be an appropriate assay for detection of cereulide (Bacillus cereus emetic toxin). Application of the boar semen bio-assay to detect cereulide directly in foods requires investigation of potential interference of food components, preservatives and other microbial and chemical food contaminants with the bio-assay. Current study provides evidence that none of included Staphylococcus aureus enterotoxins A, B, C and D nor B. cereus Hemolysin BL (HBL) and non-hemolytic enterotoxin (NHE) and three mycotoxins (Sterigmatocystin, Fumonisin B1 and Patulin) exhibited a toxic impact on semen progressive motility. Aflatoxin M1, M3 and zearalenone impaired semen motility only at concentrations (0.004 mg ml(-1), 0.1 mg ml(-1) and 10 mg ml(-1), respectively) much higher than those found in foods and those permitted by legislation, in comparison to cereulide which induces motility cease at concentrations lower than 20 ng ml(-1). Ten commonly used preservatives, namely potassium sorbate, sodium benzoate, (DL) malic acid, citric acid, (L+) tartaric acid, acetic acid, (DL) lactic acid, (L+) ascorbic acid, sodium chloride and sucrose induced no cease in spermatozoa motility even at preservative concentrations higher than permitted by legislation. Dioxins, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and acrylamide had no acute effect on spermatozoa motility at concentrations of 500 and 10,000 mg ml(-1), respectively. Robustness of computer aided boar semen motility analysis, tested with 14 different foods inoculated with cereulide producing B. cereus, showed distinct cereulide production in seven samples (although B. cereus growth to counts higher than 8 log CFU g(-1) was noted in 11 samples), in amounts close to those reported in foodborne outbreaks. Test evaluation in 33 samples suspected to hold cereulide showed actual cereulide presence in ten samples and no interference of food matrix

  10. A volatile inhibitor immobilizes sea urchin sperm in semen by depressing the intracellular pH

    SciTech Connect

    Johnson, C.H.; Clapper, D.L.; Winkler, M.M.; Lee, H.C.; Epel, D.

    1983-08-01

    Sea urchin spermatozoa are normally immotile in semen, but motility can be initiated by increasing gas flow over the semen--for example, by blowing N2 gas over a thin layer of semen. This result indicates that sperm motility is not O2 limited and suggests that seminal fluid contains a volatile inhibitor of motility which is responsible for the paralysis of sperm in semen. This inhibitor might be carbon dioxide, which reversibly immobilizes sperm. /sup 31/P-NMR measurements of pH show that the sperm intracellular pH (pHi) increases by 0.36 pH unit upon dilution of semen into seawater. Since previous studies have shown that this magnitude of pH increase is sufficient to trigger sperm motility, we suggest that the volatile inhibitor is inhibiting sperm motility in semen by depressing the pHi. A simple hypothesis that explains these observations is that the volatile motility inhibitor is CO/sub 2/, which could acidify pHi as a diffusable weak acid. In this regard, sperm diluted into seawater release acid, and this acid release is related to the pHi increase and motility initiation. In fact, nearly half of the acid released by sperm upon dilution is volatile and may therefore be due to CO/sub 2/ efflux. Most of the acid, however, cannot be attributed to CO/sub 2/ release because it is not volatile. Thus, when sperm are diluted into seawater, they raise their pHi by releasing CO/sub 2/ and protons from the cytoplasm into the surrounding seawater.

  11. Influence of semen collection method on sperm cryoresistance in small ruminants.

    PubMed

    Jiménez-Rabadán, P; Soler, A J; Ramón, M; García-Álvarez, O; Maroto-Morales, A; Iniesta-Cuerda, M; Fernández-Santos, M R; Montoro, V; Pérez-Guzmán, M D; Garde, J J

    2016-04-01

    Semen collection for cryopreservation is a key step for small ruminant conservation programs. While in these species semen is mainly collected via artificial vagina (AV), electroejaculation (EE) provides a viable alternative for untrained males. Herein we investigated the effect of semen collection method on post-thaw sperm quality by comparing two small ruminant species, sheep and goats. Semen from Blanca-Celtibérica bucks and Manchega rams was collected by AV and EE on the same day and cryopreserved using a standard protocol. At thawing, sperm motion parameters were evaluated by CASA, whereas membrane stability (YO-PRO-1), sperm viability (propidium iodide, PI) and mitochondrial activity (Mitotracker Deep Red) were analyzed using flow cytometry. The semen collection method negatively affected post-thaw sperm quality in bucks but not in rams. Thus, in bucks, post-thaw sperm motility was higher for samples collected by AV as compared to those obtained via EE. Similarly, post-thaw sperm parameters evaluated by flow cytometry were worse for buck samples collected by EE than those collected by AV in the same species, or than ram samples regardless of collection method. These results suggest that ovine and caprine spermatozoa have a different response to the cryopreservation process depending upon the semen collection method used. We hypothesize that the EE procedure may lead to changes in the composition of the ejaculate in bucks that would make spermatozoa more susceptible to the cryopreservation process, whereas this procedure would have had no effect on ram spermatozoa. This assumption requires further investigation.

  12. [Effects of captopril on the male reproductive organs and various semen parameters of rabbits].

    PubMed

    Köhler-Samouilidis, G; Schmidt-Adamopoulou, B; Samouilidis, S; Papaioannou, N; Kotsaki-Kovatsi, V P

    1997-06-01

    The effect of the administration of captopril on the concentrations of Zn, Cu, Mg and Ca into different organs, on their histological structure and several semen parameters of male rabbits was studied. For 9 weeks 6.5 mgs captopril/kg b.w. were administered daily to 7 months old White New Zealand rabbits p.o. Semen samples were collected at the beginning of the experiment and after 4 and 9 weeks. The animals were sacrificed 9 weeks after the beginning of the experiment and organ samples were collected for histological examination and for the determination of the Zn, Cu, Mg and Ca concentrations in several tissues and the semen samples. The absolute and relative weight of the right and left testes of the test animals revealed a tendency for increase. Absolute and relative weight of the right epididymis and the relative weight of the left epididymis were significantly increased. The concentration of Zn in the blood, of Cu and Ca in the epididymis and of Mg in the testes of the test animals were significantly decreased. A significant increase was observed of the Cu and Mg concentrations in the adrenals. In the semen Cu concentration was significantly increased 9 weeks after the beginning of the experiment. Mg concentration was significantly decreased 9 weeks as compared with 4 weeks after the beginning of the experiment. Histological examination of tissue specimens of brain, liver, kidney, adrenal glands, testes, epididymis, ductus deferens and seminal vesicles from all experimental animals didn't reveal any remarkable lesion under the light microscope. The other semen parameters like volume, motility, sperm number and morphology had not changed. As the values of alcalic and acid phosphatase and ASAT in the semen samples showed many variations, statistical analysis could not be performed.

  13. The Relationship between the Amount of Saturated Fat Intake and Semen Quality in Men

    PubMed Central

    Dadkhah, Hajar; Kazemi, Ashraf; Nasr-Isfahani, Mohammad-Hossien; Ehsanpour, Soheila

    2017-01-01

    Introduction: Infertility in men is one of the current problems of human society. Some studies suggest that the metabolic status of a man, including cholesterol intake, closely correlates with sperm function. The aim of this study was to evaluate the correlation between saturated fat intake and semen quality among men referring to the Isfahan Fertility and Infertility Center, Iran. Materials and Methods: This cross-sectional study was conducted on all men who referred to the Isfahan Fertility and Infertility Center. The study population consisted of 120 men selected through simple random sampling. The data collection tool consisted of a two-part questionnaire including a demographic and anthropometric characteristics form and the Food Frequency Questionnaire (FFQ). The FFQ consists of 168 items that assess food and nutrient intake for 3 months. The reliability and validity of the instruments were confirmed in previous studies. Semen analysis was performed using computer-aided semen analysis (CASA) method. Descriptive statistics and logistic regression test were used to analyze the data. Results: Results showed that the chances of having semen volume of higher than 1.5 ml (normal volume) increased by 27.5% for every 1 g increase in total fat (CI: 1.11–1.46) (P = 0.001) and reduced 38% for every 1 g increase in saturated fat (CI: 0.42–0.90, P = 0.010). No association was found between semen quality and intake of dietary fats. Conclusions: Based on the findings of this study and the prevalence of infertility in recent decades, changes in diet and saturated fatty acids intake may improve semen quality.

  14. Cytokines in the blood and semen of infertile patients

    PubMed Central

    Havrylyuk, Anna; Chopyak, Valentyna; Boyko, Yaryna; Kril, Iryna

    2015-01-01

    Cytokines have been important mediators of the immunity and can be involved in numerous processes in the male genital tract including acting as immunomodulatory elements within the male gonad. The aims of this study were: 1) to detect pro- and anti-inflammatory cytokine levels in the control group and subgroups of infertile men; and 2) to set up the practical recommendations concerning determination of cytokine levels for the male infertility diagnosis. Observations were performed in a group of 82 men: healthy controls (n = 27) and infertile patients (n = 55). The male infertility group was further subdivided into patients with: varicocele (n = 22), idiopathic infertility (n = 13) and partners of couples with recurrent spontaneous abortion (RSA; n = 20). Semen analysis was determined following WHO criteria. The cytokine interleukin 1β (IL-1β), IL-6, IL-10, IL-18; tumor necrosis factor α (TNF-α), interferon g (IFN-g) and transforming growth factor β1 (TGF-β1) contents in serum and seminal plasma were determined by quantitative ELISA. An interesting marker of male infertility appears to be TGF-β1 (blood) significantly elevated in idiopathically infertile males and in the RSA group. Besides elevated TGF-β1 in a group of idiopathic infertility significantly elevated IL-10, IL-18, IFN-g (blood) and statistically decreased IL-1β while increased IFN-g were revealed in seminal plasma compared to healthy controls. We may postulate novel cytokine micropatterns for patients with different background of infertility. Therefore, circulating cytokines: IL-1β, IL-10, IL-18, TGF-β1, IFN-g and IL-1β, IFN-g and TGF-β1 in seminal plasma should be extended in evaluation of specific types of male infertility. PMID:26648778

  15. Fatty acids profiling reveals potential candidate markers of semen quality.

    PubMed

    Zerbinati, C; Caponecchia, L; Rago, R; Leoncini, E; Bottaccioli, A G; Ciacciarelli, M; Pacelli, A; Salacone, P; Sebastianelli, A; Pastore, A; Palleschi, G; Boccia, S; Carbone, A; Iuliano, L

    2016-11-01

    Previous reports showed altered fatty acid content in subjects with altered sperm parameters compared to normozoospermic individuals. However, these studies focused on a limited number of fatty acids, included a short number of subjects and results varied widely. We conducted a case-control study involving 155 patients allocated into four groups, including normozoospermia (n = 33), oligoasthenoteratozoospermia (n = 32), asthenozoospermia (n = 25), and varicocoele (n = 44). Fatty acid profiling, including 30 species, was analyzed by a validated gas chromatography (GC) method on the whole seminal fluid sample. Multinomial logistic regression modeling was used to identify the associations between fatty acids and the four groups. Specimens from 15 normozoospermic subjects were also analyzed for fatty acids content in the seminal plasma and spermatozoa to study the distribution in the two compartments. Fatty acids lipidome varied markedly between the four groups. Multinomial logistic regression modeling revealed that high levels of palmitic acid, behenic acid, oleic acid, and docosahexaenoic acid (DHA) confer a low risk to stay out of the normozoospermic group. In the whole population, seminal fluid stearic acid was negatively correlated (r = -0.53), and DHA was positively correlated (r = 0.65) with sperm motility. Some fatty acids were preferentially accumulated in spermatozoa and the highest difference was observed for DHA, which was 6.2 times higher in spermatozoa than in seminal plasma. The results of this study highlight complete fatty acids profile in patients with different semen parameters. Given the easy-to-follow and rapid method of analysis, fatty acid profiling by GC method can be used for therapeutic purposes and to measure compliance in infertility trials using fatty acids supplements.

  16. Semen analysis of military personnel associated with military duty assignments.

    PubMed

    Weyandt, T B; Schrader, S M; Turner, T W; Simon, S D

    1996-01-01

    A collaborative study between the U.S. Army Biomedical Research and Development Laboratory (USABRDL) and the National Institute for Occupational Safety and Health (NIOSH) was designed to assess fecundity of male artillery soldiers with potential exposures to airborne lead aerosols. Potential exposure assessment was based upon information provided in an interactive questionnaire. It became apparent from extensive questionnaire data that many soldiers in the initial control population had potentially experienced microwave exposure as radar equipment operators. As a result, a third group of soldiers without potential for lead or microwave exposures, but with similar environmental conditions, was selected as a comparison population. Blood hormone levels and semen analyses were conducted on artillerymen (n = 30), radar equipment operators (n = 20), and the comparison group (n = 31). Analysis of the questionnaire information revealed that concern about fertility problems motivated participation of some soldiers with potential artillery or microwave exposures. Although small study population size and the confounding variable of perceived infertility limit the reliability of the study, several statistically significant findings were identified. Artillerymen who perceived a possible fertility concern demonstrated lower sperm counts/ejaculate (P = 0.067) and lower sperm/mL (P = 0.014) than the comparison group. The group of men with potential microwave exposures demonstrated lower sperm counts/mL (P = 0.009) and sperm/ejaculate (P = 0.027) than the comparison group. Variables used to assess endocrine, accessory sex gland, and sperm cell function were not different than the comparison group. Additional studies, incorporating larger numbers of individuals, should be performed in order to more optimally characterize potential lead and microwave exposure effects on male fecundity.

  17. The Semen pH Affects Sperm Motility and Capacitation.

    PubMed

    Zhou, Ji; Chen, Li; Li, Jie; Li, Hongjun; Hong, Zhiwei; Xie, Min; Chen, Shengrong; Yao, Bing

    2015-01-01

    As the chemical environment of semen can have a profound effect on sperm quality, we examined the effect of pH on the motility, viability and capacitation of human sperm. The sperm in this study was collected from healthy males to avoid interference from other factors. The spermatozoa cultured in sperm nutrition solution at pH 5.2, 6.2, 7.2 and 8.2 were analyzed for sperm total motility, progressive motility (PR), hypo-osmotic swelling (HOS) rate, and sperm penetration. Our results showed that these parameters were similar in pH 7.2 and 8.2 sperm nutrition solutions, but decreased in pH 5.2 and 6.2 solutions. The HOS rate exhibited positive correlation with the sperm total motility and PR. In addition, the sperm Na(+)/K(+)-ATPase activity at different pHs was measured, and the enzyme activity was significantly lower in pH 5.2 and 6.2 media, comparing with that in pH 8.2 and pH 7.2 solutions. Using flow cytometry (FCM) and laser confocal scanning microscopy (LCSM) analysis, the intracellular Ca2(+ )concentrations of sperm cultured in sperm capacitation solution at pH 5.2, 6.2, 7.2 and 8.2 were determined. Compared with that at pH 7.2, the mean fluorescence intensity of sperm in pH 5.2 and 6.2 media decreased significantly, while that of pH 8.2 group showed no difference. Our results suggested that the declined Na(+)/K(+)-ATPase activity at acidic pHs result in decreased sperm movement and capacitation, which could be one of the mechanisms of male infertility.

  18. The relationship between sperm quality in cool-shipped semen and embryo recovery rate in horses.

    PubMed

    Love, C C; Noble, J K; Standridge, S A; Bearden, C T; Blanchard, T L; Varner, D D; Cavinder, C A

    2015-12-01

    The relationship between the quality of cool-shipped stallion semen and fertility has not been adequately described. This study evaluated sperm quality of cool-shipped semen from 459 ejaculates (N = 130 stallions) that were used for insemination of 196 embryo donor mares (n = 496 estrous cycles). Embryo recovery rate (ERR; %) increased, as all sperm measures (e.g., motility, viability, DNA quality, morphology, concentration, and total number) increased. Threshold values are reported for each sperm quality measure (e.g., total sperm motility ≥ 65%) that separate two ERR groups (e.g., average: ∼50% ERR; high: ∼65% ERR).

  19. Studies on liquefaction and storage of ejaculated dromedary camel (Camelus dromedarius) semen.

    PubMed

    Wani, N A; Billah, M; Skidmore, J A

    2008-12-01

    The purpose of this study was to evaluate seminal liquefaction and quality of ejaculated camel semen during storage in different extenders at room (23 degrees C) and refrigeration (4 degrees C) temperature. Semen was collected using an artificial vagina and diluted immediately (1:1), using a split-sample technique, in five extenders [(1) Tris-tes egg yolk, (2) Tris-lactose egg yolk, (3) citrate egg yolk, (4) sucrose egg yolk and (5) Tris-fructose egg yolk], while one fraction was kept without an extender to act as control. The semen was transported to the lab at 37 degrees C, in a portable incubator within half an hour, and thereafter liquefaction of semen was monitored every 15 min. After complete liquefaction of the semen it was evaluated for sperm concentration and morphology and then was extended to a final ratio of 1:3. Aliquots of each semen sample were then stored at refrigeration and room temperature. The average volume of an ejaculate was 4.3+/-0.4 mL and it had a very viscous consistency. The average concentration of spermatozoa was 230.4+/-10.7 x 10(6)mL(-1) and the proportion of spermatozoa with protoplasmic droplets averaged 1.02+/-0.2, while 2.7+/-0.6 and 9.7+/-2.9% had mid-piece and tail abnormalities, respectively. All extended semen samples liquefied within 1.5h at 37 degrees C, however, there was slow liquefaction in the sample without an added extender (control). Best liquefaction was observed in Tris-lactose extender followed by Tris-fructose and citrate egg yolk diluents whereas in the other two extenders there was head-to-head agglutination of the spermatozoa. There was no difference in the initial motility of the spermatozoa in extenders 1-5 after its liquefaction, however, after 24 and 48 h of storage a higher proportion of spermatozoa were motile in extenders 1, 2 and 4 (P<0.05) at both the temperatures. There was a gradual decline in viability of the spermatozoa in all extenders at both the temperatures, although, a high portion of the

  20. Trials of the detection of semen and vaginal fluid RNA using the genome profiling method.

    PubMed

    Takasaka, Tomokazu; Sakurada, Koichi; Akutsu, Tomoko; Nishigaki, Koichi; Ikegaya, Hiroshi

    2011-09-01

    The identification of sperm at the scene of a sexual crime is important evidence that can be used to prove that a crime took place. We used the new genome profiling (GP) method in this study to identify sperm and vaginal fluid from RNA extracted from bodily fluids. We randomly amplified genes via a PCR approach from these semen and vaginal fluid samples and performed temperature gradient gel electrophoresis between 15-65°C. We identified specific species identification dots (spiddos) for semen and vaginal fluid. The results showed that the GP method is effective for the identification of bodily fluids at the scene of a sex crime.

  1. Strategies to improve the fertility of fresh and frozen donkey semen.

    PubMed

    de Oliveira, José Victor; Oliveira, Pedro Victor de Luna Freire; Melo e Oña, Cely Marini; Guasti, Priscilla Nascimento; Monteiro, Gabriel Augusto; Sancler da Silva, Yamê Fabres Robaina; Papa, Patrícia de Mello; Alvarenga, Marco Antônio; Dell'Aqua Junior, Jose Antonio; Papa, Frederico Ozanam

    2016-04-15

    Fertility rates of donkey semen in jennies are lower compared to mares. The aims of this study were to evaluate different sperm cryopreservation methods and insemination strategies to improve the fertility of donkey semen in jennies. Three experiments were performed: (1) the comparison of two freezing methods of donkey semen (conventional method and automated method); (2) the determination of a suitable insemination dose of fresh donkey semen for jennies and mares; and (3) the influence of the semen deposition site on fertility of jennies inseminated with frozen donkey semen. For experiment 1, no differences were observed in total motility, angular velocity, curvilinear velocity, straight-line velocity, and plasma membrane integrity between samples frozen with the conventional (Styrofoam box) and the automated method (TK 4000C). However, the automated method provided higher values of progressive motility and rapid cells in frozen-thawed samples in comparison with the conventional method (P < 0.05). For experiment 2, mares were bred using 500 × 10(6) fresh sperm (M); and jennies using 1 × 10(9) (J1) or 500 × 10(6) fresh sperm (J5). Pregnancy rates in M, J1, and J5 were 93% (14/15), 73% (11/15), and 40% (6/15), respectively. When using different insemination doses, 500 × 10(6) or 1 × 10(9) sperm, no significant difference was observed in pregnancy rates of mares (M, 14/15) and jennies (J1, 11/15). Furthermore, there was no significant difference between the two insemination doses in jennies. However, with an insemination dose of 500 × 10(6) fresh sperm, the pregnancy rates were significantly higher in mares (M, 14/15) than in jennies (J5, 6/15; P < 0.05). For experiment 3, the inseminations were carried out in the uterine body (UB) or in the uterine horn of jennies with frozen-thawed donkey semen. No pregnancies were achieved with inseminations performed in the UB (0/12). The pregnancy rate for uterine horn group was 28.26% (13/46) and thus

  2. The preservability of turkey semen quality during liquid storage in relation to strain and age of males.

    PubMed

    Iaffaldano, N; Manchisi, A; Rosato, M P

    2008-12-01

    It is difficult to maintain turkey semen quality after in vitro liquid storage and the problem is worsened by animal aging. Little is currently known about the effects of both reproductive period and strain on the preservability of qualitative characteristics of turkey semen during liquid storage. The purpose of this study was to evaluate the effect of the reproductive period of two commercial turkey strains on semen quality changes during in vitro storage for upto 48 h at 5 degrees C. Two different periods were considered: first period from 32 to 40 weeks of age and the second one from 44 to 52 weeks. Turkey males from either British United Turkeys (BUT) Big-6 line and Hybrid Large White line (Hybrid) were used. Semen pools of each tom strain were diluted with Beltsville Poultry Semen Extender (BPSE) and the motility, viability and membrane integrity of sperm were evaluated at 3, 24 and 48 h of liquid storage at 5 degrees C. The sperm concentration was significantly affected by period (P<0.01) and strain (P<0.05), with best values in first period and in the Hybrid semen. Besides also the motility, viability and membrane integrity during 48 h of storage were better (P<0.05) in the first period compared to the second one for both strains, particularly in Hybrid semen. During storage it was clearly shown in the first period that Hybrid sperm worsened more than the BUT one: in spite of the motility and viability values were at first (3h) higher (P<0.05) in Hybrid semen, after 48 h of storage the motility did not show any significant difference between strains while the viability resulted even better (P<0.05) in BUT semen. In the second period, although the semen quality decreased during the storage with a similar trend for both strains, better (P<0.05) values were found in BUT semen. Our results indicated that the reproductive period affected the quality of turkey semen in a different manner according to the strain. Moreover BUT semen showed a better in vitro storage

  3. Desmosterol, the main sterol in rabbit semen: distribution among semen subfractions and its role in the in vitro spermatozoa acrosome reaction and motility

    PubMed Central

    Mourvaki, Evangelia; Cardinali, Raffaella; Roberti, Rita; Dal Bosco, Alessandro; Castellini, Cesare

    2010-01-01

    Sterols are essential components of the cell membrane lipid bilayer that include molecules such as cholesterol and desmosterol, which are significantly found in the spermatozoa of several animal species. However, the presence of desmosterol in rabbit semen has never been investigated. The aims of this study were to characterize the sterol composition of subfractions of ejaculated rabbit semen and evaluate the in vitro effects of sterol on the spermatozoa acrosome reaction and motility. Two sterols, occurring prevalently in the free form (94.3%), were identified in whole semen collected from 10 fertile New Zealand White rabbits, specifically desmosterol (58.5% of total sterols) and cholesterol (35.9% of total sterols). Desmosterol was the predominant sterol found in all subfractions of rabbit semen, varying from 56.7% (in the prostatic secretory granules, PSGs) to 63.8% (in the seminal plasma). Spermatozoa contained an intermediate proportion of desmosterol (59.8%), which was asymmetrically distributed between the heads (52.0% of the total content of sterols) and the tails (81.8%). Results showed that both desmosterol and cholesterol can be transferred from the PSGs to the spermatozoa and are equally effective in inhibiting in vitro spermatozoa capacitation at a concentration higher than 1 mg L−1. In contrast, neither desmosterol nor cholesterol had a significant effect on spermatozoa motility. Thus, it was concluded that, the various fractions of rabbit seminal fluid differ from each other in sterol composition and quantity, probably due to their different functional properties, and these fractions may undergo significant sterol changes depending on the stage of spermatozoa capacitation. PMID:20729867

  4. Semen alterations in porcine rubulavirus-infected boars are related to viral excretion and have implications for artificial insemination.

    PubMed

    Solís, M; Ramírez-Mendoza, H; Mercado, C; Espinosa, S; Vallejo, V; Reyes-Leyva, J; Hernández, J

    2007-12-01

    Porcine rubulavirus (PoRV), also known as blue eye disease (BED) of swine, causes respiratory and reproductive problems in pigs at several developmental stages. To study the effect of PoRV infection on semen production, five boars were infected with 1 x 10(6) TCID(50)/ml of PoRV strain PAC-3 and evaluated for 59 days post inoculation (DPI). Infected boars developed reproductive tract pathology that included swelling of the testes and epididymides. Analysis of the semen showed that the infection had little effect on semen production in four animals, but semen from one boar showed severe alterations in sperm concentration, motility, and morphology. When motility was analyzed in BTS-diluted semen after 24, 48, or 72 h, alterations were detected in all boars. Furthermore, viral antigen was detected in semen, the seminal plasma fraction, or sperm fraction from all boars. These results showed that PoRV is excreted via semen and, therefore, artificial insemination is a potential route of dissemination.

  5. Effect of oral administration of Tribulus terrestris extract on semen quality and body fat index of infertile men.

    PubMed

    Salgado, R M; Marques-Silva, M H; Gonçalves, E; Mathias, A C; Aguiar, J G; Wolff, P

    2016-07-12

    Male fertility can be evaluated through complete semen analysis. Plants belonging to the Tribulus genus are known for their role in enhancing sex hormone levels and semen quality. The aim of this study was to evaluate the effects of T. terrestris on semen quality and physiological parameters. Sixty-five men with abnormal semen evaluation were included in this study, in which they were prescribed with oral administration of Androsten(®) (250 mg of Tribulus terrestris dried extract per capsule). Body fat percentage, lean muscle mass gain, fluctuation in steroid hormone levels and all semen parameters were analysed during the period of treatment. The results demonstrated that decrease in the percentage of body fat and increase in lean mass were significant, as well as increase in dihydrotestosterone levels. Complete semen analysis evaluated at the end of treatment showed significant enhancement in sperm concentration, motility and liquefaction time. Protodioscin, the main phytochemical agent of the Tribulus genus, acts on sertoli cells, germ cell proliferation and growth of seminiferous tubules. This component is known to convert testosterone into dihydrotestosterone, which plays important roles in male attributes. Our results indicate the therapeutic use of Tribulus terrestris by men presenting altered semen parameters, and/or undergoing infertility treatment.

  6. The use of an alternative light source to detect semen in clinical forensic medical practice.

    PubMed

    Lincoln, C A; McBride, P M; Turbett, G R; Garbin, C D; MacDonald, E J

    2006-05-01

    One of the primary aims of forensic examination in sexual offences is to detect and recover biological material that will link the offender with the complainant. One potentially valuable method by which trace biological evidence may be identified in other forensic settings is via the use of an Alternate Light Source (ALS). The aim of this study was to determine whether or not there was any potential benefit in using an ALS as an adjunct in sexual assault examinations to aid the detection of forensically relevant areas on the body which are not identifiable on visual inspection for sampling. We present two case reports, which illustrate the potential value of using an ALS in clinical forensic medical practice as an adjunct in sexual assault examinations to detect potentially forensically useful areas of skin to sample for semen. Prior to introducing the ALS into our clinical forensic medical practice, we undertook a number of simple laboratory studies to determine a protocol for its use. Semen is known to fluoresce using an ALS at a wavelength of 450 nm. Although we did not conduct a rigorous scientific evaluation of the technique, we evaluated the use of an ALS to detect semen on a range of inanimate surfaces as well as human skin. On all surfaces, visibility of fluorescence was increased by reduced distance of light source from the surface and increased concentration of semen on the surface, but was not noticeably affected by the angle at which the light source was held in relation to the surface.

  7. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... compulsorily notifiable disease; and (ii) An effective surveillance and monitoring system for scrapie is in... Database as part of the Scrapie Program in the United States. Imported semen may be further distributed to..., import permit, declaration, and any other accompanying documents. (Approved by the Office of...

  8. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... compulsorily notifiable disease; and (ii) An effective surveillance and monitoring system for scrapie is in... Database as part of the Scrapie Program in the United States. Imported semen may be further distributed to..., import permit, declaration, and any other accompanying documents. (Approved by the Office of...

  9. The use of forensic tests to distinguish blowfly artifacts from human blood, semen, and saliva.

    PubMed

    Durdle, Annalisa; Mitchell, R John; van Oorschot, Roland A H

    2015-03-01

    This study investigated whether routinely used forensic tests can distinguish 3-day-old or 2-week-old fly artifacts, produced after feeding on human blood, semen, or saliva, from the biological fluid. Hemastix(®) , Hemident(™) , and Hemascein(™) were unable to distinguish blood from artifacts. Hemastix(®) returned false positives from negative controls. ABAcard(®) Hematrace(®) and Hexagon OBTI could distinguish blood from 3-day-old artifacts, but not 2-week-old artifacts. Phadebas(®) and SALIgAE(®) were unable to distinguish saliva from artifacts. RSID(™) -Saliva was able to distinguish saliva from 3-day-old artifacts, but not 2-week-old artifacts. Semen tests Seminal Acid Phosphatase, RSID(™) -Semen, and ABAcard(®) p30 were all able to distinguish semen from 3-day-old artifacts, but not 2-week-old artifacts. The tests investigated cannot be relied upon to distinguish artifacts from biological fluids. However, if an artifact is identified by its morphology, a positive result may indicate which biological fluid the fly consumed, and this knowledge may prove useful for investigators searching for DNA.

  10. A comparative evaluation of semen parameters in pre- and post-Hurricane Katrina human population.

    PubMed

    Baran, Caner; Hellstrom, Wayne J; Sikka, Suresh C

    2015-01-01

    A natural disaster leading to accumulation of environmental contaminants may have substantial effects on the male reproductive system. Our aim was to compare and assess semen parameters in a normospermic population residing in the Southern Louisiana, USA area pre- and post-Hurricane Katrina. We retrospectively evaluated semen analyses data (n = 3452) of 1855 patients who attended the Tulane University Andrology/Fertility Clinic between 1999 and 2013. The study inclusion criteria were men whose semen analyses showed ≥ 1.5 ml volume; ≥15 million ml -1 sperm concentration; ≥39 million total sperm count; ≥40% motility; >30% morphology, with an abstinence interval of 2-7 days. After the inclusion criteria applied to the population, 367 normospermic patients were included in the study. Descriptive statistics and group-based analyses were performed to interpret the differences between the pre-Katrina (Group 1, 1999-2005) and the post-Katrina (Group 2, 2006-2013) populations. There were significant differences in motility, morphology, number of white blood cell, immature germ cell count, pH and presence of sperm agglutination, but surprisingly there were no significant differences in sperm count between the two populations. This long-term comparative analysis further documents that a major natural disaster with its accompanied environmental issues can influence certain semen parameters (e.g., motility and morphology) and, by extension, fertility potential of the population of such areas.

  11. Efficacious long-term cooling and freezing of Sapajus apella semen in ACP-118(®).

    PubMed

    Leão, D L; Miranda, S A; Brito, A B; Lima, J S; Santos, R R; Domingues, S F S

    2015-08-01

    The objectives of the present study were to test the effect of coconut water solution (CWS), TES-TRIS and ACP-118(®) on the seminal cooling and cryopreservation of semen from capuchin monkeys (Sapajus apella). Semen was collected from six males by electro-ejaculation, diluted in TES-TRIS, CWS or ACP-118(®), and maintained at 4°C for 28h. Semen was subsequently evaluated (Experiment I) or cryopreserved in the presence of different glycerol concentrations (3%, 5% or 7%) (Experiment II). ACP-118(®) was the preferred extender to preserve sperm motility and viability after 28h incubation at 4°C. Cooled sperm were successfully frozen-thawed in a medium containing 3% glycerol. After thawing, sperm retained the capacity to fertilize oocytes and zygotes were obtained. In conclusion, ACP-118(®) can be effectively and efficiently used as extender for the cooling of S. apella semen. Furthermore, cryopreservation using ACP-118(®) by adding 3% glycerol is suitable to maintain sperm morphology and the capacity of these cells to fertilize in vitro.

  12. Inbreeding depression on semen quality in Austrian dual-purpose simmental bulls.

    PubMed

    Maximini, L; Fuerst-Waltl, B; Gredler, B; Baumung, R

    2011-02-01

    Using pedigree data, the inbreeding coefficients of 715 Austrian dual-purpose Simmental (Fleckvieh) bulls stationed in two artificial insemination (AI) centres in Upper and Lower Austria were calculated and incorporated in statistical models for the analysis of semen quality. Five semen quality parameters (volume, concentration, motility, number of spermatozoa per ejaculate and percentage of viable spermatozoa) of approximately 30,000 ejaculates, used in two separate data sets, were investigated. The mixed model included the fixed effects age class of the bull, bull handler, semen collector, month and year of collection, number of collection per bull and day, time interval since last collection, the linear continuous effect of the inbreeding coefficient of the bull, interactions between age class and month, and age class and interval since last collection, respectively, as well as the random effect of the bull and the random residual effect. Non-linear effects of inbreeding were significant for motility only. Despite the quite low inbreeding coefficients (mean 1.3%), all semen quality traits showed inbreeding depression, in four of the five traits significantly in at least one of the data sets. The magnitude of inbreeding depression was small, which might partly be caused by the low inbreeding levels and a potential pre-selection of the bulls in the AI centres. However, monitoring of inbreeding depression on fertility traits is recommended to avoid unrecognized deterioration of such traits.

  13. LONGITUDINAL STUDY OF SEMEN QUALITY AFTER INTERMITTENT EXPOSURE TO AIR POLLUTION

    EPA Science Inventory

    LONGITUDINAL STUDY OF SEMEN QUALITY AFTER INTERMITTENT EXPOSURE TO AIR POLLUTION. J. Rubes*, D. Zudova*, Veterinary Research Institute, Brno, CR, S.G. Selevan*, US EPA/ORD/NCEA, Washington, DC, D.P. Evenson, South Dakota State University, Brookings, SD, and S.D. Perreault, US ...

  14. Attempted semen collection using the massage technique in blue-fronted Amazon parrots (Amazona aestiva aestiva).

    PubMed

    Della Volpe, Angelique; Volker, Schmidt; Krautwald-Junghanns, Maria-Elisabeth

    2011-03-01

    The purpose of this study was to establish a technique for collecting semen from blue-fronted Amazon parrots (Amazona aestiva aestiva) and to evaluate the samples that were collected. The massage method is the most common technique used to collect semen in birds and has been proven successful in several psittacine species; however, collection attempts in larger parrots have been unsatisfactory. Six blue-fronted Amazon parrot males, 3 paired with hens and 3 unpaired, were used in this study. The semen collection technique was revised to allow collection from individual birds by a single person. Semen collection was attempted from the 6 parrots on 52-56 occasions, which totaled 330 single attempts. Nineteen ejaculates were collected, and each bird produced at least 1 ejaculate that contained spermatozoa. Large ranges of sample volume (1-15.4 microL), sperm quality (motility = 2%-60%; live:dead ratio = 2:198 to 185:15), sperm concentration (0.79-3.3 x 10(6) sperm/mL), and contamination rate (0%-100%) were observed. Measured parameters did not appear to be significantly impacted by birds being paired or kept singly. Because of the relatively short acclimation period, the birds appeared to be sexually inactive for the majority of the study. Further research using sexually active birds will be necessary to determine standard spermatological parameters and verify the success of the methodology used here.

  15. Evaluation of semen quality in patients with malignancies referred for sperm banking before cancer treatment.

    PubMed

    Amirjannati, N; Sadeghi, M; Hosseini Jadda, S H; Ranjbar, F; Kamali, K; Akhondi, M A

    2011-10-01

    Different cancer treatments such as chemotherapy and radiotherapy can lead to azoospermia and even sterility for an unknown period. Whether the type of cancer could affect semen quality or not is under debate. In this study, we have reviewed semen parameters of men with cancer who deposited their sperm samples at the Avicenna Research Institute tissue bank before undergoing cytotoxic treatment. This descriptive retrospective study examined 73 cases referred to sperm bank, because of malignancy, prior to initiation of cancer treatments including chemotherapy and radiotherapy. The data recorded were age, marital status, reproductive history, semen analysis reports and cancer history of the patients. Semen samples were analysed according to recommendations of the World Health Organization (1999) before freezing. Results of the analysis showed that 71.2% (52) of patients had oligozoospermia, 93.2% (68) teratozoospermia and 86.3% (63) asthenozoospermia. Different groups of cancer patients did not show any differences in oligozoospermia, teratozoospermia and asthenozoospermia. Impaired spermatogenesis even prior to cancer treatment indicates the importance of fertility preservation. As the majority of patients had suitable specimens for freezing and assisted reproduction, sperm banking is recommended to be performed promptly and before any treatment, especially surgery.

  16. Methylation patterns in fetal tissues generated from gilts inseminated with fresh or cryopreserved semen

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Environmental influences, such as pollutants, climate, or diet, can alter the epigenetic configuration of gametes. The objective of our study was to evaluate differences in methylation activity of fetal placenta and liver from porcine pregnancies derived from fresh or frozen/thawed semen. Thirty cyc...

  17. Hepatoprotective effects of hoveniae semen cum fructus extracts in ethanol intoxicated mice

    PubMed Central

    Cho, Ilje; Kim, Joowan; Jung, Jaijun; Sung, Soohyun; Kim, Jongkyu; Lee, Namju; Ku, Saekwang

    2016-01-01

    [Purpose] The objective of this study was to evaluate the hepatoprotective effects of Hoveniae Semen Cum Fructus extract in ethanol induced hepatic damages. [Methods] Hepatic damages were induced by oral administration of ethanol and then Hoveniae Semen Cum Fructus extract was administered. [Results] Following Hoveniae Semen Cum Fructus extract administration, body and liver weights were increased, while aspartate aminotransferase, alanine aminotransferase, albumin, γ-glutamyl transferase, and triglyceride levels in the serum, triglyceride contents, tumor necrosis factor -α level, cytochrome (CY) P450 2E1 activity in the liver and mRNA expression of hepatic lipogenic genes, and Nitrotyrosine and 4-HNE-immunolabelled hepatocytes were decreased. However, mRNA expression of genes involved in fatty acid oxidation was increased. Also, as a protective mechanism for hepatic antioxidant defense systems, decreased liver MDA contents, increased glutathione contents, increased dismutase and catalase activities were observed when compared to the ethanol control. [Conclusion] Hoveniae Semen Cum Fructus extract favorably protected against liver damages, mediated by its potent anti-inflammatory and anti-steatosis properties through the augmentation of the hepatic antioxidant defense system by NF-E2-related factor-2 activation, and down-regulation of the mRNA expression of hepatic lipogenic genes or up-regulation of the mRNA expression of genes involved in fatty acid oxidation. PMID:27298813

  18. ANEUPLOIDY AND CHROMOSOME BREAKAGE IN SWIM-UP VERSUS UNPROCESSED SEMEN FROM TWENTY HEALTHY MEN

    EPA Science Inventory

    Toxicologic and epidemiologic studies have investigated a number of factors believed to induce cytogenetic damage in human sperm cells in order to estimate heritable risk to future generations. Most of these studies, however, have not enriched research semen specimens for fertil...

  19. Effect of cholesterol-loaded-cyclodextrin on sperm viability and acrosome reaction in boar semen cryopreservation.

    PubMed

    Lee, Yong-Seung; Lee, Seunghyung; Lee, Sang-Hee; Yang, Boo-Keun; Park, Choon-Keun

    2015-08-01

    This study was undertaken to examine the effect of cholesterol-loaded-cyclodextrin (CLC) on boar sperm viability and spermatozoa cryosurvival during boar semen cryopreservation, and methyl-β-cyclodextrin (MBCD) was treated for comparing with CLC. Boar semen treated with CLC and MBCD before freezing process to monitor the effect on survival and capacitation status by flow cytometry with appropriate fluorescent probes. Sperm viability was higher in 1.5mg CLC-treated sperm (76.9±1.01%, P<0.05) than un-treated and MBCD-treated sperm before cryopreservation (58.7±1.31% and 60.3±0.31%, respectively). For CTC patterns, F-pattern was higher in CLC treated sperm than MBCD-treated sperm, for B-pattern was higher in CLC-treated sperm than fresh sperm (P<0.05). For AR pattern (an acrosome-reacted sperm) was lower in CLC-treated sperm than MBCD-treated sperm (P<0.05). Moreover, we examined in vitro development of porcine oocytes after in vitro fertilization using CLC-treated frozen-thawed semen, in which CLC treatment prior to freezing and thawing increased the development of oocytes to blastocyst stage in vitro. In conclusion, CLC could protect the viability of spermatozoa from cryodamage prior to cryopreservation in boar semen.

  20. FREQUENCY OF ANEUPLOID SPERMATOZOA STUDIED BY MULTICOLOR FISH IN SERIAL SEMEN SAMPLES

    EPA Science Inventory

    Frequency of aneuploid spermatozoa studied by multicolor FISH in serial semen samples

    M. Vozdova1, S. D. Perreault2, O. Rezacova1, D. Zudova1 , Z. Zudova3, S. G. Selevan4, J. Rubes1,5
    1Veterinary Research Institute, Brno, Czech Republic; 2U.S. Environmental Protection A...

  1. ANALYSIS OR THE POTENTIAL SPERM BIOMARKER, SP22, IN HUMAN SEMEN

    EPA Science Inventory

    ANALYSIS OF THE POTENTIAL SPERM BIOMARKER SP22 IN HUMAN SEMEN
    Rebecca A. Morris Ph.D.1, Gary R. Klinefelter Ph.D.1, Naomi L. Roberts 1, Juan D. Suarez 1,
    Lillian F. Strader 1, Susan C. Jeffay 1 and Sally D. Perreault Ph.D.1

    1 U.S. EPA / ORD / National Health a...

  2. Use of Aloe vera-based extender for chilling and freezing collared peccary (Pecari tajacu) semen.

    PubMed

    Souza, A L P; Lima, G L; Peixoto, G C X; Silva, A M; Oliveira, M F; Silva, A R

    2016-05-01

    As an alternative for the conservation of collared peccary semen, this research aims at evaluating the use of Aloe vera (AV) extract as a cryoprotectant for semen chilling and freezing. Five ejaculates were divided in two aliquots that were diluted in Tris plus egg yolk (EY; 20%) or AV extract (20%) and chilled at 5 °C. In both treatments, an adequate semen conservation was achieved and values closer to 40% motile sperm with viability and osmotic response ranging from 20% to 40%, and normal morphology of 80% were found after 36 hours of storage. Moreover, 12 other ejaculates were diluted in Tris plus EY (20%) or AV extract (5, 10, or 20%) and glycerol (3%). Samples were frozen in liquid nitrogen and thawed after 1 week. After thawing, all the treatments containing EY or AV provided similar values for sperm morphology, viability, osmotic response, membrane integrity, sperm motility, amplitude of lateral head, beat cross frequency, and rapid, low, and static subpopulations, but the highest values for straightness and the lowest values for curvilinear velocity were found using 20% AV (P < 0.05). In conclusion, we found that AV extract at a 20% concentration could be used as an alternative substitute to EY in the formulation of Tris extenders for collared peccaries' semen chilling or freezing.

  3. SEMEN QUALITY AND REPRODUCTIVE HEALTH OF YOUNG CZECH MEN EXPOSED TO SEASONAL AIR POLLUTION

    EPA Science Inventory

    Semen quality and reproductive health of young Czech men exposed to seasonal air pollution.

    Selevan SG, Borkovec L, Slott VL, Zudova Z, Rubes J, Evenson DP, Perreault SD.

    U.S. Environmental Protection Agency, Washington, DC 20460, USA.

    This study of male repr...

  4. A comparative evaluation of semen parameters in pre- and post-Hurricane Katrina human population

    PubMed Central

    Baran, Caner; Hellstrom, Wayne J; Sikka, Suresh C

    2015-01-01

    A natural disaster leading to accumulation of environmental contaminants may have substantial effects on the male reproductive system. Our aim was to compare and assess semen parameters in a normospermic population residing in the Southern Louisiana, USA area pre- and post-Hurricane Katrina. We retrospectively evaluated semen analyses data (n = 3452) of 1855 patients who attended the Tulane University Andrology/Fertility Clinic between 1999 and 2013. The study inclusion criteria were men whose semen analyses showed ≥ 1.5 ml volume; ≥15 million ml-1 sperm concentration; ≥39 million total sperm count; ≥40% motility; >30% morphology, with an abstinence interval of 2–7 days. After the inclusion criteria applied to the population, 367 normospermic patients were included in the study. Descriptive statistics and group-based analyses were performed to interpret the differences between the pre-Katrina (Group 1, 1999–2005) and the post-Katrina (Group 2, 2006–2013) populations. There were significant differences in motility, morphology, number of white blood cell, immature germ cell count, pH and presence of sperm agglutination, but surprisingly there were no significant differences in sperm count between the two populations. This long-term comparative analysis further documents that a major natural disaster with its accompanied environmental issues can influence certain semen parameters (e.g., motility and morphology) and, by extension, fertility potential of the population of such areas. PMID:25677132

  5. Analysis of semen parameters during 2 weeks of daily ejaculation: a first in humans study

    PubMed Central

    Benson, Aaron D.; Frederick, Luke; Leader, Benjamin; Tirado, Edna; Feustel, Paul; Kontio, James; McAsey, Mary; Köhler, Tobias S.

    2016-01-01

    Background Timed and frequent intercourse around the time of female ovulation is recommended to improve conception. Although a significant number of articles have examined how the length of abstinence affects these semen analysis, the effects of frequent (daily) ejaculation has not been rigorously studied. Methods Twenty normal men were recruited for daily ejaculation over 14 consecutive days, after a 3–5 days abstinence period. Semen samples were collected at the beginning of the study (day 1) and then on days 3, 7 and 14. In addition to the standard semen analysis, markers of sperm DNA quality were assessed. Results The mean age of men completing the study was 25 years (range, 23–33 years). Significant decreases were observed in mean semen volume, total motile count (TMC) and sperm concentration during the study period without significant changes in motility or morphology. A large initial change in ejaculate volume, TMC and sperm concentration provided the primary difference in these values over the study period, with a plateau in values after this initial decrease (after study day 3). Metrics of DNA integrity did not change in a statistically or clinically meaningful way during the study period. Conclusions While a small study, this represents the most extensive examination of sperm quality with daily ejaculation. These findings generally support an approach of a short period of abstinence followed by daily copulation around ovulation to maximize the number of sperm available and optimize conception. PMID:27785432

  6. Cervicovaginal fluid and semen block the microbicidal activity of hydrogen peroxide produced by vaginal lactobacilli

    PubMed Central

    2010-01-01

    Background H2O2 produced by vaginal lactobacilli is believed to protect against infection, and H2O2-producing lactobacilli inactivate pathogens in vitro in protein-free salt solution. However, cervicovaginal fluid (CVF) and semen have significant H2O2-blocking activity. Methods We measured the H2O2 concentration of CVF and the H2O2-blocking activity of CVF and semen using fluorescence and in vitro bacterial-exposure experiments. Results The mean H2O2 measured in fully aerobic CVF was 23 ± 5 μM; however, 50 μM H2O2 in salt solution showed no in vitro inactivation of HSV-2, Neisseria gonorrhoeae, Hemophilus ducreyii, or any of six BV-associated bacteria. CVF reduced 1 mM added H2O2 to an undetectable level, while semen reduced 10 mM added H2O2 to undetectable. Moreover, the addition of just 1% CVF supernatant abolished in vitro pathogen-inactivation by H2O2-producing lactobacilli. Conclusions Given the H2O2-blocking activity of CVF and semen, it is implausible that H2O2-production by vaginal lactobacilli is a significant mechanism of protection in vivo. PMID:20482854

  7. Relationship between self-reported residential indoor remodeling and semen quality: a case-control study.

    PubMed

    Miao, Mao-Hua; Li, Zheng; Li, De-Kun; Yan, Bei; Liang, Hong; Zhi, Er-Lei; Du, Hong-Wei; Yuan, Wei

    2015-01-01

    The present study examined the association between residential indoor remodeling and poor semen quality. Sperm donors aged 18-45 years old were recruited in Shanghai, China. Semen specimens were collected and analyzed. An in-person interview was conducted to obtain information on the history of indoor remodeling and potential confounders. A total of 70 participants with abnormal semen quality (case group) and 68 controls were examined. A total of 20 subjects reported indoor remodeling in the recent 24 months, and among them 17 subjects reported indoor remodeling in the recent 12 months. Compared with participants with no history of indoor remodeling, participants with a history of indoor remodeling in the recent 24 months were more than three times as likely to have poor sperm quality (adjusted odds ratio = 3.8, 95% confidence interval: 1.3-12.0) after controlling for potential confounders. The association was strengthened when the analysis was restricted to those who had indoor remodeling in the recent 12 months. Our findings provide preliminary evidence that indoor remodeling has an adverse effect on semen quality.

  8. Occurrence of "Haemophilus somnus" in bovine semen and in the prepuce of bulls and steers.

    PubMed Central

    Humphrey, J D; Little, P B; Barnum, D A; Doig, P A; Stephens, L R; Thorsen, J

    1982-01-01

    Haemophilus somnus was isolated from 40 of 79 unprocessed bovine semen samples, 14 of 23 preputial washings of bulls and three of eight preputial washings of steers. The results indicate nonvenereal colonization of the male urogenital tract. It is suggested that dissemination of H. somnus from the urogenital tract may be of significance in the epizootiology of H. somnus associated diseases. PMID:7093816

  9. Revisiting the assessment of semen viscosity and its relationship to leucocytospermia.

    PubMed

    Flint, M; du Plessis, S S; Menkveld, R

    2014-10-01

    With infertility challenges posing an obstacle to many couples, the extension of variables to assess male fertility is an important line of research. At the Reproductive Biology Unit where the study was undertaken, a considerable proportion of male patient's seeking fertility assessment presented with hyperviscous semen samples and elevated concentrations of leucocytes. Despite viscosity being included as part of a routine spermiogram, it raises a considerable amount of concern as it is assessed semiquantitatively. The study was undertaken to evaluate the quantification of semen viscosity in centipoise (cP) and to investigate whether a correlation exists between hyperviscosity and leucocytospermia. A total of 200 semen samples were assessed from a sample cohort of two population groups: 162 male patients undergoing fertility assessment and 38 volunteer donors. Semen viscosity was determined by measuring the filling time of a capillary-loaded Leja chamber and quantifying the viscosity in cP. Leucocytes were identified histochemically with a leucocyte peroxidase test. The viscosity when quantified in cP was significantly higher in the peroxidase positive sample group (9.01 ± 0.49 vs. 7.39 ± 0.23 cP; P < 0.005). The introduction of a more accurate method of quantifying viscosity may possibly help to identify, diagnose and treat patients suffering from leucocytospermia to ultimately enhance their fertility potential.

  10. Cadmium, Chromium, and Copper Concentration plus Semen-Quality in Environmental Pollution Site, China

    PubMed Central

    LI, Yan; GAO, Qiaoyan; LI, Mingcai; LI, Mengyang; GAO, Xueming

    2014-01-01

    Abstract Background The environmental pollution is one of the factors contributing to the decrease of sperm quality for human beings. The aim of this study was to assess cadmium (Cd), chromium (Cr), and copper (Cu) concentration of man in environmental pollution site, and explore relationships between men exposure to Cd, Cr, and Cu and semen-quality parameters in environmental pollution site. Methods Ninety five men were recruited through pollution area and controls in 2011. We measured semen quality using Computer-aided Semen Quality Analysis, and Cd, Cr, and Cu levels in seminal plasma using Graphite Gurnace Atomic Absorption Spectroscopy. Spearman rank correlation analysis was used to evaluate the correlation between Cd, Cr and Cu concentration in seminal plasma and semen quality. Results The mean of seminal plasma Cd, Cr, and Cu values in pollution area was higher than the controls. Seminal plasma Cr values displayed a significant negative correlation with total motility and normomorph sperm rate. Seminal plasma Cu values also displayed a negative correlation with normomorph sperm rate. Conclusions Male reproductive health may be threatened by environmental pollution, and it may be influence local population diathesis. PMID:26060677

  11. Dynamics of Ebola RNA persistence in semen: report from the Postebogui cohort in Guinea.

    PubMed

    Subtil, Fabien; Delaunay, Charlotte; Keita, Alpha Kabinet; Sow, Mamadou; Touré, Abdoulaye Ii; Leroy, Sandrine; Msellati, Philippe; Magassouba, N'Fally; Baize, Sylvain; Raoul, Hervé; Ecochard, René; Barry, Moumié; Delaporte, Eric; Etard, Jean-François

    2017-03-14

    This study modeled the presence of EBOV-RNA in the semen of male Ebola survivors participating in the Postebogui study in Guinea. The median time of RT-PCR negativity was 46.4 days after symptom onset (95% CI: [11; 82.6]). The results emphasize the importance of the WHO recommendations for survivors' management.

  12. 9 CFR 98.33 - Ports designated for the importation of certain animal semen.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Ports designated for the importation of certain animal semen. 98.33 Section 98.33 Animals and Animal Products ANIMAL AND PLANT HEALTH..., Washington. (c) Mexican border ports. The following land border ports are designated as having...

  13. 9 CFR 98.35 - Declaration, health certificate, and other documents for animal semen.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... broker, the origin of the animal semen, the number, breed, species, and purpose of the importation, the... national government of the region of origin, or issued by a veterinarian designated or accredited by the national government of the region of origin and endorsed by a full-time salaried veterinary officer of...

  14. Effects of cell phone use on semen parameters: Results from the MARHCS cohort study in Chongqing, China.

    PubMed

    Zhang, Guowei; Yan, Huan; Chen, Qing; Liu, Kaijun; Ling, Xi; Sun, Lei; Zhou, Niya; Wang, Zhi; Zou, Peng; Wang, Xiaogang; Tan, Lu; Cui, Zhihong; Zhou, Ziyuan; Liu, Jinyi; Ao, Lin; Cao, Jia

    2016-05-01

    Epidemiological and experimental evidence for detrimental effects of cell phone use on semen quality is still equivocal. And that recruiting participants from infertility clinic not from general population may raise the possibility of a selection bias. To investigate effects of cell phone use on semen parameters in a general population,We screened and documented the cell phone use information of 794 young men from the Male Reproductive Health in Chongqing College students (MARHCS) cohort study in 2013, followed by 666 and 568 in 2014 and 2015, respectively. In the univariate regression analyses, we found that the daily duration of talking on the cell phone was significantly associated with decreased semen parameters, including sperm concentration [β coefficient=-6.32% per unit daily duration of talking on the cell phone (h); 95% confidence interval (CI), -11.94, -0.34] and total sperm count (-8.23; 95% CI, -14.38, -1.63) in 2013; semen volume (-8.37; 95% CI, -15.93, -0.13) and total sperm count (-16.59; 95% CI, -29.91, -0.73) in 2015]. Internet use via cellular networks was also associated with decreased sperm concentration and total sperm counts in 2013 and decreased semen volume in 2015. Multivariate analyses were used to adjust for the effects of potential confounders, and significant negative associations between internet use and semen parameters remained. Consistent but nonsignificant negative associations between talking on the cell phone and semen parameters persisted throughout the three study years, and the negative association was statistically significant in a mixed model that considered all three years of data on talking on the cell phone and semen quality. Our results showed that certain aspects of cell phone use may negatively affect sperm quality in men by decreasing the semen volume, sperm concentration, or sperm count, thus impairing male fertility.

  15. Effects of pH during liquid storage of goat semen on sperm viability and fertilizing potential.

    PubMed

    Liu, Chang-He; Dong, Hai-Bo; Ma, Dong-Li; Li, You-Wei; Han, Dong; Luo, Ming-Jiu; Chang, Zhong-Le; Tan, Jing-He

    2016-01-01

    A specific problem in goat semen preservation is the detrimental effect of seminal plasma on sperm viability in extenders containing yolk or milk. Thus, the use of chemically defined extenders will have obvious advantages. Although previous studies indicate that the initial pH of an extender is crucial to sustain high sperm motility, changes in extender pH during long-term semen storage have not been observed. Monitoring extender pH at different times of semen storage and modeling its variation according to nonlinear models is thus important for protocol optimization for long-term liquid semen preservation. The present results showed that during long-term liquid storage of goat semen, both sperm motility and semen pH decreased gradually, and a strong correlation was observed between the two. Whereas increasing the initial extender pH from 6.04 to 6.25 or storage with stabilized pH improved, storage with artificially lowered pH impaired sperm motility. Extender renewal improved sperm motility by maintaining a stable pH. Sperm coating with chicken (Gallus gallus) egg yolk improved motility by increasing tolerance to pH decline. A new extender (n-mZAP) with a higher buffering capacity was formulated, and n-mZAP maintained higher sperm motility, membrane integrity and acrosome intactness than the currently used mZAP extender did. Goat semen liquid-stored for 12 d in n-mZAP produced pregnancy and kidding rates similar to those obtained with freshly collected semen following artificial insemination. In conclusion, maintenance of a stable pH during liquid semen storage dramatically improved sperm viability and fertilizing potential.

  16. A data-driven search for semen-related phenotypes in conception delay

    PubMed Central

    Patel, C. J.; Sundaram, R.; Buck Louis, G. M.

    2016-01-01

    SUMMARY Sperm count, morphology, and motility have been reported to be predictive of pregnancy, although with equivocal basis prompting some authors to question the prognostic value of semen analysis. To assess the utility of including semen quality data in predicting conception delay or requiring >6 cycles to become pregnant (referred to as conception delay), we utilized novel data-driven analytic techniques in a pre-conception cohort of couples prospectively followed up for time-to-pregnancy. The study cohort comprised 402 (80%) male partners who provided semen samples and had time-to-pregnancy information. Female partners used home pregnancy tests and recorded results in daily journals. Odds ratios (OR), false discovery rates, and 95% confidence intervals (CIs) for conception delay (time-to-pregnancy > 6 cycles) were estimated for 40 semen quality phenotypes comprising 35 semen quality endpoints and 5 closely related fecundity determinants (body mass index, time of contraception, lipids, cotinine and seminal white blood cells). Both traditional and strict sperm phenotype measures were associated with lower odds of conception delay. Specifically, for an increase in percent morphologically normal spermatozoa using traditional methods, we observed a 40% decrease in conception delay (OR = 0.6, 95% CI = 0.50, 0.81; p = 0.0003). Similarly, for an increase in strict criteria, we observed a 30% decrease in odds for conception delay (OR = 0.7, 95% CI = 0.52, 0.83; p = 0.001). On the other hand, an increase in percent coiled tail spermatozoa was associated with a 40% increase in the odds for conception delay (OR = 1.4, 95% CI = 1.12, 1.75; p = 0.003). However, our findings suggest that semen phenotypes have little predictive value of conception delay (area under the curve of 73%). In a multivariate model containing significant semen factors and traditional risk factors (i.e. age, body mass index, cotinine and ever having fathered a pregnancy), there was a modest

  17. Diagnosis and effects of urine contamination in cooled-extended stallion semen.

    PubMed

    Ellerbrock, R; Canisso, I; Feijo, L; Lima, F; Shipley, C; Kline, K

    2016-04-15

    Urospermia is known to affect semen quality in many mammals, including stallions. Determinations of semen pH and creatinine and urea concentrations have been used to diagnose urine contamination in raw stallion semen. Unfortunately, practitioners suspecting urine contamination in cooled-shipped samples have no proven means to confirm the presence of urine. Therefore, the objectives of this study were (1) to assess the effects of urine contamination on sperm motility of extended fresh and cooled-stored stallion semen, (2) to evaluate the usefulness of semen color, odor, pH, and creatinine and urea concentrations for urospermia diagnosis, and (3) to evaluate the accuracy of a commercial blood urea nitrogen test strip in diagnosing urine contamination in extended-cooled stallion semen. Thirty-seven ejaculates were obtained from 11 stallions with no history of urospermia before division into 5 mL aliquots, and contamination with stallion urine. Each resulting sample was assessed for sperm motility, color, odor, pH, creatinine, and urea nitrogen concentration using both a semiquantitative test strip (Azostix), and a quantitative automated analyzer before and after cooling for 24 hour. Sperm motility parameters, pH, and creatinine and urea concentrations were analyzed using mixed models. Urine contamination decreased total and progressive motility in all samples before and after cooling (P < 0.05). Mean control total motility was 80% at 0 hour and 67% at 24 hours, whereas urine-contaminated samples ranged from 30% to 71% at 0 hour and 27% to 61% at 24 hours. Control mean urea (29 mg/dL) and creatinine (0.6 mg/dL) concentrations were significantly different (P < 0.05) from all urine-contaminated samples (158 mg/dL and 11.6 mg/dL, respectively) at 0 hour. Similarly, control mean urea (8 mg/dL) and creatinine (0.9 mg/dL) concentrations were significantly different than all urine-contaminated samples at 24 hours. Odor assessment presented moderate sensitivity (65

  18. The Usefulness of Selected Physicochemical Indices, Cell Membrane Integrity and Sperm Chromatin Structure in Assessments of Boar Semen Sensitivity

    PubMed Central

    Wysokińska, A.; Kondracki, S.; Iwanina, M.

    2015-01-01

    The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II (p≤0.05). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower (p≤0.05), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors

  19. Influence of cool storage before freezing on the quality of frozen-thawed semen samples in dogs.

    PubMed

    Santana, M; Batista, M; Alamo, D; González, F; Niño, T; Cabrera, F; Gracia, A

    2013-02-01

    The aim of this study was to determinate the semen quality of frozen-thawed samples that were chilled for up to 2 days before freezing. The ejaculates (n = 18) from six dogs were collected, pooled and divided into six aliquots. The first aliquot (C, control) was frozen in liquid nitrogen using a conventional protocol to reach a final concentration of 100 × 10(6) spermatozoa/ml, 20% egg yolk and 5% glycerol. The remaining five aliquots were diluted with a chilled extender (Tris-glucose and 20% egg yolk) and cooled at 4 °C as follows: R1, the semen was cooled for 1 h; R6, the semen was cooled for 6 h; R12, the semen was cooled for 12 h; R24, the semen was cooled for 24 h and R48, the semen was cooled for 48 h. After the chilling period, a second extender was added (Tris-glucose, 20% egg yolk, 10% glycerol and Equex at 1%) to reach a final composition similar to aliquot C, and then, the semen samples (R1, R6, R12, R24 and R48) were frozen in liquid nitrogen. The post-thaw sperm quality was assessed in 30 straws from each experimental group. After freezing-thawing, the total sperm motility (approximately 60-70%) in the semen chilled for up to 48 h did not show any differences from the samples frozen by the conventional cryopreservation method (63.2%). No significant differences were detected in the percentages of abnormal sperm cells among the fresh semen, the control group and the frozen samples after the different cooling times. Finally, the post-thaw percentages of damaged acrosomes showed a very uniform distribution, with mean values ranging between 7% and 10.5%. The results clearly demonstrated that cooling the semen up to 48 h before freezing did not produce a decrease in the semen quality when was compared with semen frozen by a traditional procedure.

  20. Contribution of semen trait selection, artificial insemination technique, and semen dose to the profitability of pig production systems: A simulation study.

    PubMed

    Gonzalez-Pena, Dianelys; Knox, Robert V; Rodriguez-Zas, Sandra L

    2016-01-15

    The economic impact of selection for semen traits on pig production systems and potential interaction with artificial insemination (AI) technique and semen dose remains partially understood. The objectives of this study were to compare the financial indicators (gross return, net profit, cost) in a three-tier pig production system under one of two selection strategies: a traditional strategy including nine paternal and maternal traits (S9) and an advanced strategy that adds four semen traits (S13). Maternal traits included the number of pigs born alive, litter birth weight, adjusted 21-day litter weight, and the number of pigs at 21 days, and paternal traits included days to 113.5 kg, back fat, average daily gain, feed efficiency, and carcass lean percentage. The four semen traits included volume, concentration, progressive motility of spermatozoa, and abnormal spermatozoa. Simultaneously, the impact of two AI techniques and a range of fresh refrigerated semen doses including cervical AI with 3 × 10(9) (CAI3) and 2 × 10(9) (CAI2) sperm cells/dose, and intrauterine AI with 1.5 × 10(9) (IUI1.5), 0.75 × 10(9) (IUI0.75), and 0.5 × 10(9) (IUI0.5) sperm cells/dose were evaluated. These factors were also evaluated using a range of farrowing rates (60%-90%), litter sizes (8-14 live-born pigs), and a selected semen collection frequency. The financial impact of the factors was assessed through simulation of a three-way crossbreeding system (maternal nucleus lines A and B and paternal nucleus line C) using ZPLAN. The highest return on investment (profit/cost) of boars was observed at 2.33 collections/wk (three periods of 24 hours between collections). Under this schedule, a significant (P < 0.0001) interaction between the selection strategy and the AI technique-dose combination was identified for the gross return; meanwhile, significant (P < 0.0001) additive effects of the selection strategy and AI technique-dose combination were observed for the net

  1. Rhodiola sacra aqueous extract (RSAE) improves biochemical and sperm characteristics in cryopreserved boar semen.

    PubMed

    Zhao, H-W; Li, Q-W; Ning, G-Z; Han, Z-S; Jiang, Z-L; Duan, Y-F

    2009-03-15

    Although Rhodiola sacra aqueous extract (RSAE) has been used in many studies as an antioxidant, its effects on semen characteristics and its antioxidant properties during cryopreservation of boar sperm have never been evaluated. Semen was collected from five Duroc boars (2-4-year-old) twice weekly and frozen-thawed in extender with RSEA. Motion characteristics were assessed with a computer-aided semen analysis (CASA) system, whereas other sperm quality end points were assessed by routine methods. The effective concentration of RSEA in extender ranged from 4 to 8mg/L and the effect of RSEA on sperm quality was better in glycerol-free extender than extender containing glycerol (P<0.05). In frozen-thawed boar semen, there was a direct correlation (P<0.05) between RSEA concentration and glutathione (GSH) concentrations, mitochondrial activity, and hypoosmotic swelling test (HOST), and an inverse correlation (r=-0.982, P<0.05) between RSEA concentration and malondialdehyde (all end points were significantly higher at 6mg/L than in the control group). In summary: (i) the effective concentration of RSEA in extender ranged from 4 to 8mg/L; (ii) the effect of RSEA on sperm quality was better in extender without glycerol; and (iii) there was a significant correlation between RSEA concentrations and concentrations of GSH and MAD in frozen-thawed boar semen (antioxidant effects of RSEA were concentration-dependent). Further studies are needed to define the active ingredient in RSEA that protects boar sperm against ROS.

  2. Assessment of the vaginal residence time of biomarkers of semen exposure☆,☆☆,★

    PubMed Central

    Thurman, Andrea; Jacot, Terry; Melendez, Johan; Kimble, Thomas; Snead, Margaret; Jamshidi, Roxanne; Wheeless, Angie; Archer, David F.; Doncel, Gustavo F.; Mauck, Christine

    2016-01-01

    Objective The primary objective of this pilot study is to determine and compare the residence time in the vagina of biomarkers of semen exposure for up to 15 days post exposure. The biomarkers are prostate-specific antigen (PSA), Y chromosome DNA, the sex determining region of the Y chromosome (SRY) and testis-specific protein Y-encoded 4 (TSPY4). The secondary objectives are to determine if biomarker concentrations differed between intercourse and inoculation groups, to establish whether the sampling frequency post exposure affected biomarker concentrations and decay profile and to determine if biomarker concentrations in vaginal swabs obtained by the participant at home were similar to swabs obtained by the nurse in the clinic. Study design We randomized healthy women to unprotected intercourse (n=17) versus vaginal inoculation with the male partner’s semen in the clinic (n=16). Women were then further randomized to have vaginal swabs obtained at either 7 or 4 time points after semen exposure, up to 15 days post exposure, either obtained at home by the participant or in the clinic by the research nurse. Results PSA and SRY were markers of recent semen exposure. TSPY4 was detectable in approximately 50% of participants at 15 days post exposure. Unprotected intercourse resulted in significantly higher concentrations of select biomarkers. Sampling frequency and home versus clinic sampling had no significant effect on biomarker concentrations. Conclusions Objective biomarkers of recent or distant semen exposure may have great utility for verifying protocol compliance in a variety of clinical trials. PMID:27259675

  3. TRIS-lecithin extender supplemented with antioxidant catalase for chilling of canine semen.

    PubMed

    Beccaglia, M; Anastasi, P; Chigioni, S; Luvoni, G C

    2009-07-01

    The aims were to evaluate the suitability of a non-commercial TRIS-lecithin (LC) extender and the effect of different concentrations of catalase (CAT) on motility, capacitation status (Chlortetracycline-assay) and zona pellucida (ZP) binding capacity of canine spermatozoa stored at +5 degrees C for 4 days. The sperm-rich fractions of the ejaculates of four stud dogs were divided into four aliquots. After centrifugation, sperm pellets were diluted (200 x 10(6) sperm/ml) in TRIS buffer, citric acid, fructose, antibiotics, supplemented with 20% egg yolk (TRIS-EY) or 0.04% soybean lecithin (TRIS-LC) with CAT (150 or 450 UI/ml) or without CAT, and then preserved at 5 degrees C for 4 days. The results showed that LC is a valid alternative to EY for chilling canine semen, as similar rates of motility, number of uncapacitated spermatozoa and of spermatozoa binding the oocyte ZP were obtained in semen chilled in TRIS-LC or TRIS-EY. Different concentrations of CAT in a TRIS-LC based extender did not improve the quality of semen after chilling. However, a concentration of 150 UI/ml CAT resulted in an increased number of spermatozoa bound to the oocyte ZP, after 4 days of chilling when compared to semen chilled with TRIS-EY and TRIS-LC. In conclusion, an animal protein-free extender with soybean LC, as a replacement of EY, is suitable for 4 days chilling of canine spermatozoa, but the addition of CAT does not improve general semen quality except for a slight effect on sperm-ZP binding.

  4. Improvement of stored turkey semen quality as a result of He-Ne laser irradiation.

    PubMed

    Iaffaldano, N; Meluzzi, A; Manchisi, A; Passarella, S

    2005-02-01

    Two experiments were carried out to evaluate the effects of He-Ne laser irradiation at various energy doses on the quality of stored turkey semen. Four semen pools were used in Experiment 1. Each pool was divided into 10 aliquots, nine of which were irradiated with energy doses ranging from 0.144 to 10.8 J/cm2 while the tenth one was not irradiated (control). Each sample was evaluated for motility immediately after irradiation, 24 and 48 h later. Energy doses ranging from 3.24 to 5.4 J/cm2 had higher (P <0.01) sperm motility index (SMI) value compared to the control and samples irradiated with lower and higher laser doses. The energy dose of 3.96 J/cm2 was selected for Experiment 2 to obtain further insight on its effects on turkey sperm preservation for up to 60 h. Each pool of four semen was divided into two aliquots: one represented the control and the other one was irradiated with He-Ne laser at an energy dose of 3.96 J/cm2. Each sample was evaluated for motility and viability immediately after irradiation and then at 12 h intervals up to 60 h. The cell energy charge was also measured by HPLC. Exposure to 3.96 J/cm2 increased the SMI and viability of turkey semen stored for 60 h compared to the control (P <0.05). The cell energy charge of irradiated samples was 200% higher than in the control. Laser irradiation increased the longevity of stored turkey spermatozoa, and might be a useful technique to enhance semen quality in long-term storage.

  5. The use of semen evaluation and assisted reproduction in Spix's macaws in terms of species conservation.

    PubMed

    Fischer, Dominik; Neumann, Daniel; Purchase, Cromwell; Bouts, Tim; Meinecke-Tillmann, Sabine; Wehrend, Axel; Lierz, Michael

    2014-01-01

    The Spix's macaw (Cyanopsitta spixii) is the rarest parrot on earth. The remaining captive population consists of 79 individuals. Captive propagation is ongoing to increase the number of individuals for future reintroduction back into the wild. Unfortunately, from 2004 to 2012, only 33 chicks hatched from 331 eggs. Semen evaluation and assisted reproduction might help to overcome this problem. Therefore, a recently developed electro-stimulated semen collection technique was used in Spix's macaws. Semen collection was successful in 39 of 78 attempts in 10 out of 17 males. Examination of the semen included evaluation of volume, color, consistency, contaminations and pH, as well as determination of motility, viability, morphology, concentration, and total count of spermatozoa. The median volume of semen samples was 5.6 µl. On average, 34.7 ± 21.9% (median 30%) of the sperm were motile and 23.1 ± 22.1% (median 16.5%) were progressively motile. In addition to spermatozoa, round cells were detected in the samples. Median sperm concentration was 15,500/µl (range 500-97,500/µl) and median viability was 50% (range 5-87%). Morphological examination revealed in 26.5% normal spermatozoa, high numbers of malformations of the head (50.2%) and tail region (20.5%), with 29% of all sperm showing multiple abnormalities. Artificial insemination was performed in three females; two eggs laid after artificial insemination had spermatozoa present on the perivitelline layer, suggesting the possible success of the insemination technique. Although no fertilization could be demonstrated, these preliminary results are promising, as they indicate that assisted reproduction might be a tool for species conservation in the Spix's macaw.

  6. Oxidation-reduction potential of semen: what is its role in the treatment of male infertility?

    PubMed Central

    Agarwal, Ashok; Roychoudhury, Shubhadeep; Bjugstad, Kimberly B.; Cho, Chak-Lam

    2016-01-01

    The diagnosis of male infertility relies largely on conventional semen analysis, and its interpretation has a profound influence on subsequent management of patients. Despite poor correlation between conventional semen parameters and male fertility potential, inclusion of advanced semen quality tests to routine male infertility workup algorithms has not been widely accepted. Oxidative stress is one of the major mediators in various etiologies of male infertility; it has deleterious effects on spermatozoa, including DNA damage. Alleviation of oxidative stress constitutes a potential treatment strategy for male infertility. Measurement of seminal oxidative stress is of crucial role in the identification and monitoring of patients who may benefit from treatments. Various tests including reactive oxygen species (ROS) assay, total antioxidant capacity (TAC) assay or malondialdehyde (MDA) assay used by different laboratories have their own drawbacks. Oxidation-reduction potential (ORP) is a measure of overall balance between oxidants and antioxidants, providing a comprehensive measure of oxidative stress. The MiOXSYS™ System is a novel technology based on a galvanostatic measure of electrons; it presents static ORP (sORP) measures with static referring to the passive or current state of activity between oxidants and antioxidants. Preliminary studies have correlated sORP to poor semen qualities. It is potentially useful in prognostication of assisted reproductive techniques outcomes, screening of antioxidants either in vivo or during IVF cycles, identification of infertile men who may benefit from treatment of oxidative stress, and monitoring of treatment success. The simplified laboratory test requiring a small amount of semen would facilitate clinical application and research in the field. In this paper, we discuss the measurement of ORP by the MiOXSYS System as a real-time assessment of seminal oxidative stress, and argue that it is a potential valuable clinical test

  7. Broiler breeder semen quality as affected by trace minerals in vitro.

    PubMed

    Barber, S J; Parker, H M; McDaniel, C D

    2005-01-01

    Research has shown that trace elements, such as Se, Mn, and Zn, can alter reproductive functions. The aim of the current study was to evaluate the sperm quality index (SQI) and sperm viability as affected by various levels and sources of Se, Mn, and Zn when added in vitro to broiler breeder semen. In vitro treatments consisted of the following sources and levels of minerals: Control, no minerals added to sperm; seleno L-methionine, 4 levels ranging from 8.78 to 7,896 microg/L; sodium selenite, 4 levels ranging from 8.78 to 7,896 microg/L; MnSO4, 8 levels ranging from 6,500 to 65,000 mg/L; Zn 180 (Zinpro Corporation), 4 levels ranging from 0.65 to 650 mg/L; and ZnSO4, 4 levels ranging from 0.65 to 650 mg/L. The addition of 7,896 microg of sodium selenite/L to semen was detrimental to sperm motility. Also, MnSO4 adversely affected SQI and sperm viability at concentrations of 6,500 mg/L and greater. Sperm viability was decreased when 650 mg/L of Zn 180 was added to semen. Sperm motility was depressed by exposure to Zn 180 at 650 mg/L and ZnSO4 at 65 and 650 mg/L. Our results suggest that these trace minerals must act at the reproductive tissue level during spermatogenesis to improve semen quality. Direct in vitro application of these elements to semen appears to be detrimental to spermatozoa.

  8. Sex ratio determination in bovine semen: a new approach by quantitative real time PCR.

    PubMed

    Parati, K; Bongioni, G; Aleandri, R; Galli, A

    2006-12-01

    Sex preselection of livestock offspring in cattle represents, nowadays, a big potential for genetic improvement and market demand satisfaction. Sperm sorting by flow cytometer provides a powerful tool for artificial insemination and production of predefined sexed embryos but, an accurate verification of the yield of sperm separation remains essential for a field application of this technique or for improvement and validation of other related semen sexing technologies. In this work a new method for the determination of the proportion of X- and Y-bearing spermatozoa in bovine semen sample was developed by real time PCR. Two sets of primers and internal TaqMan probes were designed on specific X- and Y-chromosome genes. To allow a direct quantification, a standard reference was established using two plasmid cDNA clones (ratio 1:1) for the specific gene targets. The method was validated by a series of accuracy, repeatability and reproducibility assays and by testing two sets of sorted and unsorted semen samples. A high degree of accuracy (98.9%), repeatability (CV=2.58%) and reproducibility (CV=2.57%) was shown. The results of X- and Y-sorted semen samples analysed by real time PCR and by flow cytometric reanalysis showed no significant difference (P>0.05). The evaluation of X-chromosome bearing sperms content in unsorted samples showed an average of 51.11+/-0.56% for ejaculates and 50.17+/-0.58% for the commercial semen. This new method for quantification of the sexual chromosome content in spermatozoa demonstrated to be rapid and reliable, providing a valid support to the sperm sexing technologies.

  9. Collection and evaluation of semen from the three-toed sloth (Bradypus tridactylus).

    PubMed

    Peres, M A; Benetti, E J; Milazzotto, M P; Visintin, J A; Miglino, M A; Assumpção, M E O A

    2008-10-01

    Sloths (Bradypus sp.) are extremely sensitive animals that suffer with the destruction and fragmentation of forests. They present a low population growth rate and need to be further studied for the preservation of the specie. Thus, the aim of this study was to establish an efficient semen collection protocol as well as characterize sperm concentration, motility and morphology in order to contribute with information about the reproductive traits of this specie, which has never been described in the literature before. For that, nine Bradypus tridactylus males were captured during the wet season and six during the dry season, in Manaus (AM), located in the north region of Brazil, semen was collected by electroejaculation with shocks given in sequences of progressive intensities (minimum 20mA and maximum 60mA). All animals ejaculated small volumes of semen and in some of them, the volume ejaculated was not enough for a complete spermiogram. Physical characteristics observed on the collections of the wet season were different from those seen in the specimen collected in the dry season. Motility an vigor was very low and did not show forward progression, only oscillatory movement. After Spermac stain, spermatozoa presented a wide variety of defects; however, the differences in morphology were not significant between seasons. The morphology assessed by scanning electron microscopy shows that the head in both groups could be elongated, short or could have a base narrower than the apex and the midpiece narrowed abruptly, forming a nip in its transition to the tail. Although further studies are necessary to verify our preliminary findings concerning seasonal variation in sperm quality, these results demonstrate that semen can be safely collected from sloths by electroejaculation and provide the first reports of semen characteristics in this species.

  10. Urinary Concentrations of Benzophenone-Type Ultra Violet Light Filters and Semen Quality

    PubMed Central

    Buck Louis, Germaine M.; Chen, Zhen; Kim, Sungduk; Sapra, Katherine J.; Bae, Jisuk; Kannan, Kurunthachalam

    2015-01-01

    Objective To assess benzophenone-type ultra violet (UV) filter concentrations, chemicals used in sunscreen and personal care products, and semen endpoints. Design Cohort. Setting 16 counties in Michigan and Texas Participants 413 men provided semen and urine samples, 2005–2009. Five UV filters were quantified (ng/mL) in urine using liquid chromatography-triple-quadrupole mass spectrometry: BP-1 (2,4-dihydroxybenzophenone), BP-2 (2,2′,4,4′-tetrahydroxybenzophenone), BP-3 (2-hydroxy-4-methoxybenzophenone), BP-8 (2,2′-dihydroxy-4-methoxybenzophenone), and 4-OH-BP (4-hydroxybenzophenone). Using linear regression, beta coefficients (β) and 95% confidence intervals (CIs) for each chemical dichotomized at the 75th percentile and Box-Cox transformed semen endpoint were estimated, after adjusting for age, BMI, cotinine, season, and site. Interventions None. Main Outcome Measures 35 semen endpoints. Results BP-2 was associated with diminished sperm concentration (β=−0.74; 95% CI −1.41, −0.08), straight (β=−4.57; 95% CI −8.95, −0.18) and linear movement (β=−3.15; 95% CI −6.01, −0.30), more immature (β=0.38; 95% CI 0.15, 0.62) sperm, and a decreased percentage of other tail abnormalities (β=−0.16; 95% CI −0.31, −0.01). BP-8 was associated with decreased hypo-osmotic swelling (β=−2.57; 95% CI −4.86, −0.29) and higher acrosome area (β=1.14; 95% CI 0.01, 2.26). No associations were observed for BP-1, BP-3 or 4OH-BP. Conclusion The findings suggest that specific UV filters may be associated with some aspects of semen endpoints, but await future corroboration. PMID:26253817

  11. Is photometry an accurate and reliable method to assess boar semen concentration?

    PubMed

    Camus, A; Camugli, S; Lévêque, C; Schmitt, E; Staub, C

    2011-02-01

    Sperm concentration assessment is a key point to insure appropriate sperm number per dose in species subjected to artificial insemination (AI). The aim of the present study was to evaluate the accuracy and reliability of two commercially available photometers, AccuCell™ and AccuRead™ pre-calibrated for boar semen in comparison to UltiMate™ boar version 12.3D, NucleoCounter SP100 and Thoma hemacytometer. For each type of instrument, concentration was measured on 34 boar semen samples in quadruplicate and agreement between measurements and instruments were evaluated. Accuracy for both photometers was illustrated by mean of percentage differences to the general mean. It was -0.6% and 0.5% for Accucell™ and Accuread™ respectively, no significant differences were found between instrument and mean of measurement among all equipment. Repeatability for both photometers was 1.8% and 3.2% for AccuCell™ and AccuRead™ respectively. Low differences were observed between instruments (confidence interval 3%) except when hemacytometer was used as a reference. Even though hemacytometer is considered worldwide as the gold standard, it is the more variable instrument (confidence interval 7.1%). The conclusion is that routine photometry measures of raw semen concentration are reliable, accurate and precise using AccuRead™ or AccuCell™. There are multiple steps in semen processing that can induce sperm loss and therefore increase differences between theoretical and real sperm numbers in doses. Potential biases that depend on the workflow but not on the initial photometric measure of semen concentration are discussed.

  12. PrP(Sc) detection and infectivity in semen from scrapie-infected sheep.

    PubMed

    Rubenstein, Richard; Bulgin, Marie S; Chang, Binggong; Sorensen-Melson, Sharon; Petersen, Robert B; LaFauci, Giuseppe

    2012-06-01

    A scrapie-positive ewe was found in a flock that had been scrapie-free for 13 years, but housed adjacent to scrapie-positive animals, separated by a wire fence. Live animal testing of the entire flock of 24 animals revealed seven more subclinical scrapie-positive ewes. We hypothesized that they may have contracted the disease from scrapie-positive rams used for breeding 4 months prior, possibly through the semen. The genotypes of the ewe flock were highly scrapie-susceptible and the rams were infected with the 'Caine' scrapie strain having a short incubation time of 4.3-14.6 months in sheep with 136/171 VQ/VQ and AQ/VQ genotypes. PrP(Sc) accumulates in a variety of tissues in addition to the central nervous system. Although transmission of prion diseases, or transmissible spongiform encephalopathies, has been achieved via peripheral organ or tissue homogenates as well as by blood transfusion, neither infectivity nor PrP(Sc) have been found in semen from scrapie-infected animals. Using serial protein misfolding cyclic amplification followed by a surround optical fibre immunoassay, we demonstrate that semen from rams infected with a short-incubation-time scrapie strain contains prion disease-associated-seeding activity that generated PrP(Sc) in sPMCA (serial protein misfolding cyclic amplification). Injection of the ovinized transgenic mouse line TgSShpPrP with semen from scrapie-infected sheep resulted in PrP(Sc)-seeding activity in clinical and, probably as a result of the low titre, non-clinical mouse brain. These results suggest that the transmissible agent, or at least the seeding activity, for sheep scrapie is present in semen. This may be a strain-specific phenomenon.

  13. Increased conception rates in beef cattle inseminated with nanopurified bull semen.

    PubMed

    Odhiambo, John F; DeJarnette, J M; Geary, Thomas W; Kennedy, Chelsey E; Suarez, Susan S; Sutovsky, Miriam; Sutovsky, Peter

    2014-10-01

    Aberrant sperm phenotypes coincide with the expression of unique sperm surface determinants that can be probed by objective, biomarker-based semen analysis and targeted as ligands for semen purification. This study evaluated a nanoparticle-based magnetic purification method that removes defective spermatozoa (∼30% of sample) from bull semen and improves sperm sample viability and fertilizing ability in vitro and in vivo. Two types of nanoparticles were developed: a particle coated with antibody against ubiquitin, which is present on the surface of defective spermatozoa, and a particle coated with the lectin peanut agglutinin, which binds to glycans exposed by acrosomal damage. In a 2 yr artificial insemination field trial with 798 cows, a conception rate of 64.5% ± 3.7% was achieved with a 10 × 10(6) sperm dose of peanut agglutinin-nanopurified spermatozoa, comparable to a control nonpurified full dose of 20 × 10(6) spermatozoa per dose (63.3% ± 3.2%) and significantly higher than a 10 × 10(6) sperm dose of nonpurified control semen (53.7% ± 3.2%; P < 0.05). A total of 466 healthy calves were delivered, and no negative side effects were observed in the inseminated animals or offspring. Because the method is inexpensive and can be fully integrated in current protocols for semen cryopreservation, it is feasible for use in the artificial insemination industry to improve fertility with reduced sperm dosage inseminations. Spermatology will benefit from nanopurification methodology by gaining new tools for the identification of candidate biomarkers of sperm quality such as binder of sperm protein 5 (BSP5), described in the present study.

  14. Heavy Metal Level in Human Semen with Different Fertility: a Meta-Analysis.

    PubMed

    Sun, Jiantao; Yu, Guangxia; Zhang, Yucheng; Liu, Xi; Du, Chuang; Wang, Lu; Li, Zhen; Wang, Chunhong

    2017-03-01

    There are conflicting reports on the heavy metal levels in human semen with different fertilities. The purpose of this analysis is to merge and analyze the differences of heavy metal lead (Pb), cadmium (Cd), zinc (Zn), and copper (Cu) levels in male semen with normal and low fertilities. All documents in both Chinese and English were collected from the PubMed, Web of Science, and Chinese National Knowledge Infrastructure (CNKI) database from inception date to February 19, 2016. We have used RevMan software (version 5.2) for the meta-analysis and Stata software (version 12.0) for the meta-regression and sensitivity analyses. A total of 20 literatures were included in the study. The results of the meta-analysis indicate a significant difference between fertility with three metal ions (Pb, Cd, Zn) while no significant difference with copper, detailed as follows: (i) 10 studies on the lead concentrations with a standardized mean difference (SMD) = 2.07, 95 %CI (0.97, 3.17), P < 0.01; (ii) 13 studies on the cadmium concentrations with an SMD = 0.75, 95 %CI (0.44, 1.07), P < 0.01; (iii) 8 studies on the concentrations of zinc with an SMD = -0.61, 95 %CI (-1.08, -0.14), P < 0.01; and (iv) 9 studies on the copper concentrations with an SMD = 0.42, 95 %CI (-0.29, 1.13), P = 0.247. The results indicate that the men with low fertility have higher semen Pb and Cd levels and lower semen Zn levels; more studies are needed to indicate the association of the semen copper level with fertility.

  15. Development of soya milk extender for semen cryopreservation of Karan Fries (crossbreed cattle).

    PubMed

    Singh, V K; Singh, A K; Kumar, R; Atreja, S K

    2013-01-01

    Egg yolk based semen extenders are used widely, with the potential risk of xenobiotic contamination. This study was designed to develop a soya milk based extender to substitute egg yolk based extender for bovine semen cryopreservation. In the first experiment soya milk was prepared from fresh soya bean (Glycine max). Concentration of soya milk in tris based extender was standardized based on quality parameters of spermatozoa during liquid preservation at 5°C up to 72 h and compared with egg yolk tris (EYT) extender. Sperm in soya milk tris (SMT) extender with 25 percent soya milk showed no significant (P > 0.05) differences in all the quality parameters like motility, viability, membrane integrity and acrosome integrity, as compared to sperm in EYT extender up to 72h in liquid dilution. In the second experiment the Karan Fries semen was cryopreserved in SMT extender with 25 percent soya milk (selected from the first experiment) using different concentration of glycerol, as cryoprotectant, ranging from 6-7 percent with a difference of 0.2 percent to standardize optimum concentration based on post thaw motility of spermatozoa. Glycerol at a final concentration of 6.4 percent was found to be the best among all. Further, semen samples were split and cryopreserved in newly developed SMT extender containing 6.4 percent glycerol and compared with conventional EYT extender for post thaw sperm quality parameters and degree of cryocapacitation. There were no significant (P > 0.05) differences between sperm in EYT extender and SMT extender for post thaw motility, viability, membrane integrity, acrosome integrity and cryocapacitation. In conclusion, the newly developed SMT extender maintained comparable semen quality as compared to EYT extender hence it can.

  16. Satellite test of the isotropy of the one-way spe ed of light using ExTRAS

    NASA Technical Reports Server (NTRS)

    Wolf, Peter

    1995-01-01

    A test of the second postulate of special relativity, the universality of the speed of light, using the ExTRAS (Experiment on Timing Ranging and Atmospheric Sounding) payload to be flown on board a Russian Meteor-3M satellite (launch date January 1997) is proposed. The propagation time of a light signal transmitted from one point to another without reflection would be measured directly by comparing the phases of two hydrogen maser clocks, one on board and one on the ground, using laser or microwave time transfer systems. An estimated uncertainty budget of the proposed measurements is given, resulting in an expected sensitivity of the experiment of delta c/c is less than 8xl0(exp -10) which would be an improvement by a factor of approximately 430 over previous direct measurements and by a factor of approximately 4 over the best indirect measurement. The proposed test would require no equipment additional to what is already planned and so is of inherently low-cost. It could be carried out by anyone having access to a laser or microwave ground station and a hydrogen maser.

  17. Successful artificial insemination in the koala (Phascolarctos cinereus) using extended and extended-chilled semen collected by electroejaculation.

    PubMed

    Allen, Camryn D; Burridge, Michelle; Mulhall, Sarah; Chafer, Mandy L; Nicolson, Vere N; Pyne, Michael; Zee, Yeng Peng; Jago, Sophia C; Lundie-Jenkins, Geoff; Holt, William V; Carrick, Frank N; Curlewis, Jonathan D; Lisle, Allan T; Johnston, Stephen D

    2008-04-01

    Artificial insemination in the koala using chilled, electroejaculated semen provides for a marked improvement in the reproductive and genetic management of captive koala colonies in Australia and internationally, and makes available the option of using semen collected from wild populations to expand restricted gene pools. Dilution of koala semen for artificial insemination is complicated because koalas are induced ovulators, and it is thought that ovulating factors are present in the semen, so that semen extension for preservation purposes might be anticipated to result in a failure to induce ovulation. The first two experiments of this study were designed to determine whether artificial insemination using undiluted, extended, and extended-chilled semen collected by electroejaculation was capable of inducing a luteal phase and/or the production of pouch young. In Experiment 1, 1 ml undiluted electroejaculated semen, 2 ml diluted (1:1) semen, and 1 ml diluted (1:1) semen resulted in seven of nine, six of nine, and six of nine koalas showing a luteal phase, respectively; four pouch young were produced in each treatment. A second artificial insemination experiment was conducted in which 2 ml diluted (1:1) semen was administered in three groups of nine koalas. The first group received semen that had been collected and diluted immediately without chilling, the second group was deposited with semen stored chilled for 24 h, and the final group received semen that had been chilled for 72 h. In the first group, five females had a luteal phase, but none became pregnant. In group 2, two of the five females that had a luteal phase gave birth, whereas in group 3, four of the six females that had a luteal phase produced pouch young. In addition, experiment 3 was conducted to determine whether it was possible to produce pouch young by naturally mating koalas that were in the latter stages of their behavioral estrus; this information is important to the logistics of transporting

  18. Molecular Detection of Chlamydia trachomatis and Other Sexually Transmitted Bacteria in Semen of Male Partners of Infertile Couples in Tunisia: The Effect on Semen Parameters and Spermatozoa Apoptosis Markers

    PubMed Central

    Sellami, Hanen; Znazen, Abir; Sellami, Afifa; Mnif, Hela; Louati, Nour; Zarrouk, Soumaya Ben; Keskes, Leila; Rebai, Tarek; Gdoura, Radhouane; Hammami, Adnene

    2014-01-01

    This study was undertaken to determine the prevalence of Chlamydia trachomatis, Mycoplasmas, and Ureaplasmas in semen samples of the male partners of infertile couples and to investigate whether Chlamydia trachomatis could initiate apoptosis in human spermatozoa. A total of 85 males partners of infertile couples undergoing routine semen analysis according to World Health Organization guidelines were included. Specimens were examined for the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum and Ureaplasma parvum by Real time PCR (qPCR). Semen specimens were analysed for the appearance of apoptotic markers (sperm DNA fragmentation, activated caspase 3 levels, mitochondrial membrane potential (ΔΨm)) using flow cytometry. C. trachomatis, N. gonorrhoeae, U. urealyticum, M genitalium were detected in semen samples of 13 (15.2%), 5 (5.8%), 5 (5.8%) and 3 (3.5%) male partners of infertile couples, respectively. M. hominis and U. parvum were detected in semen sample of only one patient (1.1%). The semen of infertile men positive for C. trachomatis showed lower mean of semen count and lower rapid progressive motility (category [a]) of spermatozoa compared to uninfected men with statistically significances (p = 0.02 and p = 0.04, respectively). Flow cytometry analyses demonstrated a significant increase of the mean rate of semen with low ΔΨm and caspase 3 activation of infertile men positive for C. trachomatis compared to uninfected men (p = 0.006 and p = 0.001, respectively). DNA fragmentation was also increased in sperm of infertile men positive for C. trachomatis compared to uninfected men but without statistical significances (p = 0.62). Chlamydial infection was associated to loss of ΔΨm and caspase 3activation. Thus, C. trachomatis infection could be incriminated in apoptosis induction of spermatozoa. These effects may explain the negative direct impact of C. trachomatis

  19. The relationship between male BMI and waist circumference on semen quality: data from the LIFE study

    PubMed Central

    Eisenberg, Michael L.; Kim, Sungduk; Chen, Zhen; Sundaram, Rajeshwari; Schisterman, Enrique F.; Buck Louis, Germaine M.

    2014-01-01

    STUDY QUESTION What is the relationship between body size, physical activity and semen parameters among male partners of couples attempting to become pregnant? SUMMARY ANSWER Overweight and obesity are associated with a higher prevalence of low ejaculate volume, sperm concentration and total sperm count. WHAT IS KNOWN ALREADY Higher BMI is associated with impaired semen parameters, while increasing waist circumference (WC) is also associated with impaired semen parameters in infertile men. STUDY DESIGN, SIZE, DURATION Data from the Longitudinal Investigation of Fertility and the Environment (LIFE) Study were utilized. The LIFE study is a population-based prospective cohort of 501 couples attempting to conceive in two geographic areas (Texas and Michigan, USA) recruited in 2005–2009. Couples were recruited from four counties in Michigan and 12 counties in Texas to ensure a range of environmental exposures and lifestyle characteristics. In person interviews were conducted to ascertain demographic, health and reproductive histories followed by anthropometric assessment. PARTICIPANTS/MATERIALS, SETTING, METHODS We categorized BMI (kg/m2) as <25.0 (underweight and normal), 25.0–29.9 (overweight) 30.0–34.9 (obese, class I) and ≥35 (obese, class II) for analysis. Data were available for analysis in 468 men (93% participation), with a mean ± SD age of 31.8 ± 4.8 years, BMI of 29.8 ± 5.6 kg/m2 and WC of 100.8 ± 14.2 cm. The majority of the cohort (82%) was overweight or obese with 58% reporting physical activity <1 time/week. The median sperm concentration for the men in the cohort was 60.2 M/ml with 8.6% having oligospermia (<15 M/ml). MAIN RESULTS AND THE ROLE OF CHANCE When examining semen parameters, ejaculate volume showed a linear decline with increasing BMI and WC (P < 0.01). Similarly, the total sperm count showed a negative linear association with WC (P < 0.01). No significant relationship was seen between body size (i.e. BMI or WC) and semen

  20. Preliminary examination of polymorphisms of GSTM1, GSTT1, and GSTZ1 in relation to semen quality

    EPA Science Inventory

    Background: Environmental, lifestyle, and occupational exposures on semen quality have been investigated in epidemiological studies with inconsistent results. Genetic factors involved in toxicant activation and detoxification have been examined in relation to the risk of outcomes...

  1. Evaluation of bison (Bison bison) semen from Yellowstone National Park, Montana, USA, bulls for Brucella abortus shedding.

    PubMed

    Frey, Rebecca K; Clarke, P Ryan; McCollum, Matt P; Nol, Pauline; Johnson, Kammy R; Thompson, Brent D; Ramsey, Jennifer M; Anderson, Neil J; Rhyan, Jack C

    2013-07-01

    To determine if bison (Bison bison) bulls from Yellowstone National Park (YNP), Montana, USA, shed an infective dose of Brucella abortus in semen, 50 YNP bulls were captured on public lands in Montana during the winter and early spring (April-May) of 2010 and 2011. The bulls were immobilized, and blood and semen samples were collected for serology and Brucella culture. Thirty-five bulls (70%) were antibody-positive, and B. abortus was cultured from semen in three (9%) of the 35 antibody-positive or suspect bulls, though not at concentrations considered an infective dose. Eight bulls (six antibody-positive, two negative) had palpable lesions of the testes, epididymides, or seminal vesicles consistent with B. abortus infection. Breeding soundness exams and semen analysis suggested that antibody-positive bulls were more likely to have nonviable ejaculate (8/35; 23%) than bulls without detectable antibody (2/15; 13%).

  2. Rapid detection of bovine herpesvirus 1 in the semen of infected bulls by a nested polymerase chain reaction assay.

    PubMed Central

    Masri, S A; Olson, W; Nguyen, P T; Prins, S; Deregt, D

    1996-01-01

    A nested polymerase chain reaction (PCR) assay was developed for the detection of bovine herpesvirus 1 (BHV-1) in bovine semen and compared with the virus isolation method. When extended semen, commonly used in the bovine artificial insemination industry, was inoculated with BHV-1, the PCR assay detected BHV-1 DNA in semen inoculated at 0.25-2.5 TCID50 per 0.5 mL. In contrast, the lower limit of detection for virus isolation was 250 TCID50 of BHV-1 inoculated in 0.5 mL of extended semen. These methods were also used to detect BHV-1 in the semen of four bulls which were experimentally infected with BHV-1. All infected bulls demonstrated balanitis at 3 d post-inoculation (DPI) and severe balanoposthitis at 4 DPI. BHV-1 was detected in raw semen by virus isolation and PCR at 2 DPI, before balanitis was evident. For virus isolation, the last day that BHV-1 was detected during primary infection was 7 DPI for two bulls and 9 and 11 DPI for the other two bulls. In contrast, PCR detected BHV-1 in the bulls' semen until 14 or 18 DPI. For individual animals, PCR detected BHV-1 during primary infection for at least 1-10 d longer than virus isolation. Reactivation of BHV-1 from latency without the presence of visible lesions was promoted twice by two series of 5 d dexamethasone injections. For the first series of dexamethasone treatments, a positive virus isolation result was obtained on the 5th d of treatment for only one bull. In contrast, two bulls demonstrated evidence of viral reactivation on this day by PCR. All bulls shed BHV-1 in semen on d 4 after dexamethasone treatment, as evidenced by positive virus isolation and PCR results. One bull was still PCR positive 13 d later. For the second series of dexamethasone treatments, a small amount of virus was isolated from semen collected on d 3 or 4 after treatment for two bulls but not from the other two bulls. In contrast, semen samples from all bulls were PCR positive for either or both of these 2 d. In total, from 80 semen

  3. Efficiency of different extenders on cooled semen collected during long and short day length seasons in Martina Franca donkey.

    PubMed

    Contri, Alberto; De Amicis, Ippolito; Veronesi, Maria Cristina; Faustini, Massimo; Robbe, Domenico; Carluccio, Augusto

    2010-07-01

    Artificial insemination with cooled semen is routine in equids because of its good fertility rates and relatively low costs. In several donkey breeds, especially in restricted populations, the use of cooled semen could be seen as the best way of improving reproductive performance and avoiding excessive inbreeding. Furthermore, most jennies have ovulatory estrous throughout the year, and thus, cooled semen could also be used during short day length season. The aims of this study were to evaluate the effects of different extenders on sperm quality during cooling in the Martina Franca breed, and to verify the preservation of cooled semen collected during long day length (May-June) and short day length (November-December) seasons. Three ejaculates were collected at 10-day intervals from each of six jackasses during both May-June and again in November-December time periods. Each ejaculate was cooled in INRA96 or E-Z Mixin at a low cooling rate and evaluated daily over a 120-h preservation time. The results showed a significant extender influence on preservation time in both periods. Semen diluted with INRA96 maintained a progressive motility of 36% and a straightness of 89% at 120h, whereas semen extended with E-Z Mixin had a mean progressive motility of 32% and a straightness of 81% at 48h during the May-June period. Despite having the same initial characteristics, semen collected during the short day length season had a higher rate of decline in semen quality during storage at 5 degrees C with E-Z Mixin.

  4. Effect of hypoxia by intermittent altitude exposure on semen characteristics and testicular morphology of male rhesus monkeys

    NASA Astrophysics Data System (ADS)

    Saxena, D. K.

    1995-09-01

    Semen characteristics and testicular morphology of rhesus monkeys were studied on exposure to a simulated high altitude of 4411 m for 21 days. There was a partially reversible decrease in the semen volume, sperm count and sperm motility, as well as an elevation of pH and fructose concentration. These changes were associated with degeneration of the germinal epithelium and spermatogenic arrest at the end of third week of exposure which had not recovered even 3 weeks after the exposure.

  5. Ejaculate collection efficiency and post-thaw semen quality in wild-caught Griffon vultures from the Sardinian population.

    PubMed

    Madeddu, Manuela; Berlinguer, Fiammetta; Ledda, Massimo; Leoni, Giovanni G; Satta, Valentina; Succu, Sara; Rotta, Andrea; Pasciu, Valeria; Zinellu, Angelo; Muzzeddu, Marco; Carru, Ciriaco; Naitana, Salvatore

    2009-02-19

    This study aimed to test the feasibility of a programme of semen collection and cryopreservation in Griffon vultures. Four wild-caught individuals kept in captivity because of unrecoverable traumas were used. Semen collection attempts were made twice a week during three consecutive reproductive seasons (December - March) using the abdominal massage method. Ejaculation was successfully induced between late January and late February. Semen collection efficiency was rather low (27.9%) and it did not vary among individuals (p > 0.05). No differences were found in ejaculate volumes (12.5 +/- 9.1 microl), spermatozoa concentration (28.4 +/- 30.9 million cells/ml) and viability (61.3 +/- 13.9%) among the 4 vultures. ATP values differed among the four vultures (p < 0.001); B showed higher nucleotide concentration than both C and D, while it did not differ form A, whose values were higher compared with D. After freezing and thawing, semen in vitro viability, DNA integrity and ATP intracellular concentration were determined. Spermatozoa viability after thawing did not differ among the four individuals (52.6 +/- 5.8 in A, 53.4 +/- 4.6 in B, 50.4 +/- 3.2 in C, 42.5 +/- 2.7 in D), but it decreased significantly compared to fresh semen (p < 0.05). During 4 hrs in vitro culture, spermatozoa collected from B maintained over time a higher viability in vitro when compared to A, C and D. As evaluated by the comet assay method, DNA fragmentation after freezing and thawing did not differ in the 4 vultures. ATP concentration in frozen/thawed semen was significantly lower than in fresh semen (p < 0.0001). This study indicates that semen cryopreservation can be considered as a useful tool in the conservation of Griffon vulture genetic resources, but further studies are needed to optimize this technique.

  6. Introducing and validation of SYBR Green Real-Time PCR method to determinate sex ratio in bovine semen.

    PubMed

    Maleki, Adham Fani; Moussavi, AliReza Heravi; Nassiri, Mohammad Reza; Tahmoorespur, Mojtaba; Vakili, Seyed Alireza

    2013-07-01

    Flow cytometry is a widely used application for validating the accuracy of sperm sexing. However, this method is relatively expensive and requires considerable technical support. An alternative method employing simpler technology at low cost could be suitable for the evaluation of bovine semen in laboratories with low budgets. We used a SYBR Green Real-Time PCR assay to determinate sex ratio in bovine semen. The PLP and SRY genes were amplified to isolate the specific fragments of X- and Y-chromosome sequences, respectively. Two certified standard curves were obtained using two plasmids containing PLP and SRY amplicons. Our results show no significant difference in semen sex ratio in unsorted semen (54.7±0.52% X and 47.6±0.60% Y). However, significant difference was observed in X/Y-sorted semen (93.3±0.08% X and 91.4±0.06% Y-sperm), as compared to the expected ratio in unsorted semen or the post-sorting reanalysis data. The evolution of X-chromosome bearing sperm content in unsorted samples showed an average of 52.6 for ejaculates and 51.8 for the commercial semen. In order to confirm our results, the accuracy, repeatability and reproducibility of the method were tested resulting in 98.2% accuracy, repeatability of CV=5.59% and reproducibility of CV=5.40%. Thus, this method is demonstrated to be a reliable and inexpensive way to test sexual chromosome content in semen samples.

  7. Semen molecular and cellular features: these parameters can reliably predict subsequent ART outcome in a goat model

    PubMed Central

    Berlinguer, Fiammetta; Madeddu, Manuela; Pasciu, Valeria; Succu, Sara; Spezzigu, Antonio; Satta, Valentina; Mereu, Paolo; Leoni, Giovanni G; Naitana, Salvatore

    2009-01-01

    Currently, the assessment of sperm function in a raw or processed semen sample is not able to reliably predict sperm ability to withstand freezing and thawing procedures and in vivo fertility and/or assisted reproductive biotechnologies (ART) outcome. The aim of the present study was to investigate which parameters among a battery of analyses could predict subsequent spermatozoa in vitro fertilization ability and hence blastocyst output in a goat model. Ejaculates were obtained by artificial vagina from 3 adult goats (Capra hircus) aged 2 years (A, B and C). In order to assess the predictive value of viability, computer assisted sperm analyzer (CASA) motility parameters and ATP intracellular concentration before and after thawing and of DNA integrity after thawing on subsequent embryo output after an in vitro fertility test, a logistic regression analysis was used. Individual differences in semen parameters were evident for semen viability after thawing and DNA integrity. Results of IVF test showed that spermatozoa collected from A and B lead to higher cleavage rates (0 < 0.01) and blastocysts output (p < 0.05) compared with C. Logistic regression analysis model explained a deviance of 72% (p < 0.0001), directly related with the mean percentage of rapid spermatozoa in fresh semen (p < 0.01), semen viability after thawing (p < 0.01), and with two of the three comet parameters considered, i.e tail DNA percentage and comet length (p < 0.0001). DNA integrity alone had a high predictive value on IVF outcome with frozen/thawed semen (deviance explained: 57%). The model proposed here represents one of the many possible ways to explain differences found in embryo output following IVF with different semen donors and may represent a useful tool to select the most suitable donors for semen cryopreservation. PMID:19900288

  8. Ejaculate collection efficiency and post-thaw semen quality in wild-caught Griffon vultures from the Sardinian population

    PubMed Central

    Madeddu, Manuela; Berlinguer, Fiammetta; Ledda, Massimo; Leoni, Giovanni G; Satta, Valentina; Succu, Sara; Rotta, Andrea; Pasciu, Valeria; Zinellu, Angelo; Muzzeddu, Marco; Carru, Ciriaco; Naitana, Salvatore

    2009-01-01

    This study aimed to test the feasibility of a programme of semen collection and cryopreservation in Griffon vultures. Four wild-caught individuals kept in captivity because of unrecoverable traumas were used. Semen collection attempts were made twice a week during three consecutive reproductive seasons (December – March) using the abdominal massage method. Ejaculation was successfully induced between late January and late February. Semen collection efficiency was rather low (27.9%) and it did not vary among individuals (p > 0.05). No differences were found in ejaculate volumes (12.5 +/- 9.1 μl), spermatozoa concentration (28.4 +/- 30.9 million cells/ml) and viability (61.3 +/- 13.9%) among the 4 vultures. ATP values differed among the four vultures (p < 0.001); B showed higher nucleotide concentration than both C and D, while it did not differ form A, whose values were higher compared with D. After freezing and thawing, semen in vitro viability, DNA integrity and ATP intracellular concentration were determined. Spermatozoa viability after thawing did not differ among the four individuals (52.6 +/- 5.8 in A, 53.4 +/- 4.6 in B, 50.4 +/- 3.2 in C, 42.5 +/- 2.7 in D), but it decreased significantly compared to fresh semen (p < 0.05). During 4 hrs in vitro culture, spermatozoa collected from B maintained over time a higher viability in vitro when compared to A, C and D. As evaluated by the comet assay method, DNA fragmentation after freezing and thawing did not differ in the 4 vultures. ATP concentration in frozen/thawed semen was significantly lower than in fresh semen (p < 0.0001). This study indicates that semen cryopreservation can be considered as a useful tool in the conservation of Griffon vulture genetic resources, but further studies are needed to optimize this technique. PMID:19228408

  9. Deleterious effects of cypermethrin on semen characteristics and testes of dwarf goats (Capra hircus).

    PubMed

    Ahmad, Maqbool; Hussain, Ijaz; Khan, Ahrar; Najib-ur-Rehman

    2009-07-01

    This study was carried out on 30 male dwarf goats to determine the effects of cypermethrin (CY) on semen characteristics and at tissue level in testes. Animals were divided randomly into five equal groups and each group was dipped with 0%, 0.1%, 0.4%, 0.8% or 1.6% CY, on days 0 and 15. The semen was collected at day 0, then fortnightly till 75 days and evaluated for physical characteristics, sperm morphology and concentration. None of the parameters studied showed any statistical difference on days 0 and 15 of sample collection. From day 30, effect of CY on semen characteristics started to appear and was more pronounced from days 45 to 75. Significantly (P<0.01) decreased ejaculatory volume, motility percentage, mass activity, and concentration of spermatozoa were observed in treated groups as compared to control. With the treatment of CY, pH of semen became more alkaline. Semen color changed from creamy (control) to milky white to straw color in treated bucks. Percentage of abnormal spermatozoa (tailless, bent tails, coiled tailed) was much higher in treated animals than in control. Dead spermatozoa increased significantly (P<0.01) in the treatment groups from days 30 to 75 as compared to the control group. All these changes were dose dependent, being less with low CY dose and more even extensive with high CY dose. Slight to moderate improvement in the above parameters was observed at day 75 nearly in all treated groups. Grossly decreased weight of testis and cyanotic epididymides were observed in bucks treated with 1.6% CY. Histopathologically, degenerative changes and loss of spermatogonia, spermatocyte, Sertoli cells, spermatids, and spermatozoa in seminiferous tubules were also dose dependent. It was concluded that CY caused dose-dependent effects on all parameters studied. High doses of CY (0.8% and 1.6% solution) affected the parameters on semen characteristics, though this effect seems to be transient as improvement in these parameters was observed at day

  10. Mitochondrial Biomarkers Reflect Semen Quality: Results from the MARCHS Study in Chongqing, China

    PubMed Central

    Zhang, Guowei; Wang, Zhi; Ling, Xi; Zou, Peng; Yang, Huan; Chen, Qing; Zhou, Niya; Sun, Lei; Gao, Jianfang; Zhou, Ziyuan; Cao, Jia; Ao, Lin

    2016-01-01

    Unexplained infertility requires that more sensitive and mechanism-based biomarkers should be developed and used independently of or in addition to conventional semen parameters for an infertility diagnosis. In the present study, semen samples were collected from young men participating in the Male Reproductive Health in Chongqing College students (MARCHS) cohort study in the follow-up stage in 2014. Conventional semen parameters were measured in all 656 participants, whereas sperm mitochondrial membrane potential (MMP), mitochondrial DNA copy number (mtDNAcn), mtDNA integrity and apoptotic parameters were measured among 627, 386, 362, and 628 participants, respectively. We found that sperm MMP was significantly positively correlated with all of conventional semen parameters including semen volume (r = 0.090, p = 0.025), sperm concentration (r = 0.301, p<0.01), total sperm count (r = 0.324, p<0.01), and progressive motility (r = 0.399, p<0.01); sperm MMP was also negatively correlated with Annexin V+ sperm (r = -0.553, p<0.01); mtDNAcn was significantly negatively correlated with sperm concentration (r = -0.214, p<0.01), total sperm count (r = -0.232, p<0.01), and progressive motility (r = -0.164, p = 0.01); mtDNA integrity was also significantly positively correlated with sperm concentration (r = 0.195, p<0.01), total sperm count (r = 0.185, p<0.01), and progressive motility (r = 0.106, p = 0.043). After adjusting for potential confounders, these relationships remained significant. Furthermore, we explored the potential effects of lifestyles on such mitochondrial biomarkers and found that the current drinkers displayed a higher level of sperm MMP; additionally, mt DNAcn was increased with age. The results indicated that certain mitochondrial biomarkers could serve as predictors of semen quality in a general population, and the study provides a baseline for the effects of population characteristics and lifestyles on such mitochondrial markers. PMID:28006017

  11. Cloned embryos from semen. Part 2: intergeneric nuclear transfer of semen-derived eland (Taurotragus oryx) epithelial cells into bovine oocytes.

    PubMed

    Nel-Themaat, Liesl; Gómez, Martha C; Pope, C Earle; Lopez, Monica; Wirtu, Gemechu; Jenkins, Jill A; Cole, Alex; Dresser, Betsy L; Bondioli, Kenneth R; Godke, Robert A

    2008-03-01

    The production of cloned offspring by nuclear transfer (NT) of semen-derived somatic cells holds considerable potential for the incorporation of novel genes into endangered species populations. Because oocytes from endangered species are scarce, domestic species oocytes are often used as cytoplasts for interspecies NT. In the present study, epithelial cells isolated from eland semen were used for intergeneric transfer (IgNT) into enucleated bovine oocytes and compared with bovine NT embryos. Cleavage rates of bovine NT and eland IgNT embryos were similar (80 vs. 83%, respectively; p > 0.05); however, development to the morula and blastocyst stage was higher for bovine NT embryos (38 and 21%, respectively; p < 0.0001), than for eland IgNT embryos (0.5 and 0%, respectively). DNA synthesis was not observed in either bovine NT or eland IgNT cybrids before activation, but in 75 and 70% of bovine NT and eland igNT embryos, respectively, cell-cycle resumption was observed at 16 h postactivation (hpa). For eland IgNT embryos, 13% had > or = 8 cells at 84 hpa, while 32% of the bovine NT embryos had > or = 8 cells at the same interval. However, 100 and 66% of bovine NT and eland IgNT embryos, respectively, that had > or = 8 cells synthesized DNA. From these results we concluded that (1) semen-derived epithelial cell nuclei can interact and be transcriptionally controlled by bovine cytoplast, (2) the first cell-cycle occurred in IgNT embryos, (3) a high frequency of developmental arrest occurs before the eight-cell stage in IgNT embryos, and (4) IgNT embryos that progress through the early cleavage stage arrest can (a) synthesize DNA, (b) progress through subsequent cell cycles, and (c) may have the potential to develop further.

  12. Cloned embryos from semen. Part 2: Intergeneric nuclear transfer of semen-derived eland (Taurotragus oryx) epithelial cells into bovine oocytes

    USGS Publications Warehouse

    Nel-Themaat, L.; Gomez, M.C.; Pope, C.E.; Lopez, M.; Wirtu, G.; Jenkins, J.A.; Cole, A.; Dresser, B.L.; Bondioli, K.R.; Godke, R.A.

    2008-01-01

    The production of cloned offspring by nuclear transfer (NT) of semen-derived somatic cells holds considerable potential for the incorporation of novel genes into endangered species populations. Because oocytes from endangered species are scarce, domestic species oocytes are often used as cytoplasts for interspecies NT. In the present study, epithelial cells isolated from eland semen were used for intergeneric transfer (IgNT) into enucleated bovine oocytes and compared with bovine NT embryos. Cleavage rates of bovine NT and eland IgNT embryos were similar (80 vs. 83%, respectively; p > 0.05); however, development to the morula and blastocyst stage was higher for bovine NT embryos (38 and 21%, respectively; p < 0.0001), than for eland IgNT embryos (0.5 and 0%, respectively). DNA synthesis was not observed in either bovine NT or eland IgNT cybrids before activation, but in 75 and 70% of bovine NT and eland igNT embryos, respectively, cell-cycle resumption was observed at 16 h postactivation (hpa). For eland IgNT embryos, 13% had ???8 cells at 84 hpa, while 32% of the bovine NT embryos had ???8 cells at the same interval. However, 100 and 66% of bovine NT and eland IgNT embryos, respectively, that had ???8 cells synthesized DNA. From these results we concluded that (1) semen-derived epithelial cell nuclei can interact and be transcriptionally controlled by bovine cytoplast, (2) the first cell-cycle occurred in IgNT embryos, (3) a high frequency of developmental arrest occurs before the eight-cell stage in IgNT embryos, and (4) IgNT embryos that progress through the early cleavage stage arrest can (a) synthesize DNA, (b) progress through subsequent cell cycles, and (c) may have the potential to develop further. ?? 2008 Mary Ann Liebert, Inc.

  13. Bioactivity-Guided Fractionation Identifies Amygdalin as a Potent Neurotrophic Agent from Herbal Medicine Semen Persicae Extract

    PubMed Central

    Yang, Chuanbin; Zhao, Jia; Cheng, Yuanyuan; Li, Xuechen; Rong, Jianhui

    2014-01-01

    Herbal medicine Semen Persicae is widely used to treat blood stasis in Chinese medicine and other oriental folk medicines. Although little is known about the effects of Semen Persicae and its active compounds on neuron differentiation, our pilot study showed that Semen Persicae extract promoted neurite outgrowth in rat dopaminergic PC12 cells. In the present study, we developed a bioactivity-guided fractionation procedure for the characterization of the neurotrophic activity of Semen Persicae extract. The resultant fractions were assayed for neurite outgrowth in PC12 cells based on microscopic assessment. Through liquid-liquid extraction and reverse phase HPLC separation, a botanical glycoside amygdalin was isolated as the active compound responsible for the neurotrophic activity of Semen Persicae extract. Moreover, we found that amygdalin rapidly induced the activation of extracellular-signal-regulated kinase 1/2 (ERK1/2). A specific ERK1/2 inhibitor PD98059 attenuated the stimulatory effect of amygdalin on neurite outgrowth. Taken together, amygdalin was identified as a potent neurotrophic agent from Semen Persicae extract through a bioactivity-guided fractional procedure. The neurotrophic activity of amygdalin may be mediated by the activation of ERK1/2 pathway. PMID:25050339

  14. Bioactivity-guided fractionation identifies amygdalin as a potent neurotrophic agent from herbal medicine Semen Persicae extract.

    PubMed

    Yang, Chuanbin; Zhao, Jia; Cheng, Yuanyuan; Li, Xuechen; Rong, Jianhui

    2014-01-01

    Herbal medicine Semen Persicae is widely used to treat blood stasis in Chinese medicine and other oriental folk medicines. Although little is known about the effects of Semen Persicae and its active compounds on neuron differentiation, our pilot study showed that Semen Persicae extract promoted neurite outgrowth in rat dopaminergic PC12 cells. In the present study, we developed a bioactivity-guided fractionation procedure for the characterization of the neurotrophic activity of Semen Persicae extract. The resultant fractions were assayed for neurite outgrowth in PC12 cells based on microscopic assessment. Through liquid-liquid extraction and reverse phase HPLC separation, a botanical glycoside amygdalin was isolated as the active compound responsible for the neurotrophic activity of Semen Persicae extract. Moreover, we found that amygdalin rapidly induced the activation of extracellular-signal-regulated kinase 1/2 (ERK1/2). A specific ERK1/2 inhibitor PD98059 attenuated the stimulatory effect of amygdalin on neurite outgrowth. Taken together, amygdalin was identified as a potent neurotrophic agent from Semen Persicae extract through a bioactivity-guided fractional procedure. The neurotrophic activity of amygdalin may be mediated by the activation of ERK1/2 pathway.

  15. Peptidase activities in the semen from the ductus deferens and uterus of the neotropical rattlesnake Crotalus durissus terrificus.

    PubMed

    Marinho, Camila Eduardo; Almeida Santos, Selma Maria; Yamasaki, Simone Cristina; Silveira, Paulo Flavio

    2009-07-01

    To understand the role of peptidases in seminal physiology of Crotalus durissus terrificus, intra- and inter-seasonal activity levels of acid (APA), basic (APB), puromycin-sensitive (APN-PS) and puromycin-insensitive neutral (APN-PI), cystyl (CAP), dipeptidyl-IV (DPPIV), type-1 pyroglutamyl (PAP-I) and prolyl-imino (PIP) aminopeptidases as well as prolyl endopeptidase (POP) were evaluated in soluble (SF) and/or membrane-bound (MF) fractions of semen collected from the ductus deferens of the male reproductive tract and from the posterior portion of the uterus. Seminal APB, PIP and POP were detected in SF, while other peptidases were detected in SF and MF. Only the convoluted posterior uterus in winter and autumn had semen. Relative to other examined peptidases, in general, APN-PI, APN-PS and APB activities were predominant in the semen from the uterus and throughout the year in the semen from the ductus deferens, suggesting their great relevance in the seminal physiology of C. d. terrificus. The levels of peptidase activities in the ductus deferens semen varied seasonally and were different from those of semen in the uterus, suggesting that their modulatory actions on susceptible peptides are integrated to the male reproductive cycle events and spermatozoa viability of this snake.

  16. A comparative study on the cryogenic preservation of semen from the sandhill crane and the domestic fowl

    USGS Publications Warehouse

    Sexton, T.J.; Gee, G.F.; Watson, P.F.

    1978-01-01

    SYNOPSIS: Recent findings on the cryogenic preservation of semen from the crane, Grus canadensis pratensis and the domestic fowl, Gallus domesticus, are compared. Highest levels of post-thaw motility for crane semen (55%) were obtained when semen was diluted 1:1 with the Beltsville Poultry Semen Extender (BPSE) and held for 30 min at 5 C before it was equilibrated with 4% dimethyl sulfoxide (DMSO) for 15 min. In contrast, post-thaw motility for fowl spermatozoa was highest (80%) when semen was diluted 1:3 with BPSE and held for 60 min at 5 C before it was equilibrated with 4% DMSO for 60 min. Post-thaw motility of spermatozoa of both species was highest when the following freezing rates were used: l C per min from +5 to -20 C, 50 C per min from -20 to -80 C, then plunging into liquid nitrogen which resulted in a rate of 160 C per min from -80 to -196 C. One of four crane eggs resulting from insemination with frozen-thawed semen was fertile, whereas 27 of 55 fowl eggs were fertile, but this difference may have been due largely to fewer spermatozoa being inseminated into the female crane than into the fowl.

  17. Association of Smoking With Semen Quality and µ-Calpain Level in Normospermia: A Case-Control Study

    PubMed Central

    Ashtary-Larky, Damoon; Ghaffari, Mohammad Ali; Noorbehbahani, Mozhgan; Alipour, Meysam

    2016-01-01

    Objective: Calpains are a family of Ca2+ dependent proteases. There is some evidence that calpains involved in fusion process that occurs between spermatozoa and the oocyte. The current study aimed to investigate the association of smoking with semen quality and µ-calpain level. Materials and methods: This case-control study was conducted on 117 normospermia males between June 2013 and march 2014 in Jahad Laboratory in ahvaz, Iran. The semen samples were collected from male smokers (n = 50) and non-smokers (n = 67). We divided these participants as light, moderate, or heavy smokers based on their cigarettes per day (CPD). ELISA assays were used to measure µ-calpain concentration. All semen samples were analyzed according to World Health Organization guidelines. Results: The analysis of semen showed the volume, concentration, motility and morphology of semen were significantly lower among the smoker men than the non-smoker men. Also this significant difference was observed based on the number (light, moderate and heavy smokers) and duration (short term and long term smoker) of smoking. Although, showed no significant difference between µ-calpain of smoker men and non-smoker men. CPD showed negatively correlation with semen volume, concentration, motility and morphology of sperm. Conclusion: Sperm quality was negatively correlated with CPD and duration of smoking. However, there is no significant correlation between smoking and µ-calpain concentration. PMID:27385969

  18. Improved fertility in gilts and sows after artificial insemination of frozen-thawed boar semen by supplementation of semen extender with caffeine and CaCl2.

    PubMed

    Yamaguchi, Shoichiro; Funahashi, Hiroaki; Murakami, Tetsuya

    2009-12-01

    Supplementation of semen extender with caffeine and CaCl(2) for artificial insemination (AI) of fresh spermatozoa has been demonstrated to reduce recruitment of uterine polymorphonuclear leukocytes (PMNs) and the activity of phagocytosis. Here, we determined if addition of caffeine and CaCl(2) to semen extender improves the fertility of frozen-thawed boar semen. In experiment 1, gilts were cervically inseminated twice with frozen-thawed boar spermatozoa (25 x 10(8) cells per dose) suspended in Modena solution (n=7) or modified Beltsville Thawing Solution supplemented with caffeine and CaCl(2) (BCC, n=7). The gilts were slaughtered 4 h later, and their oviducts and uterine horns plus the body of the uterus were flushed to recover PMNs and non-phagocytosed spermatozoa. There was no difference in the total number of uterine PMNs between gilts inseminated with Modena solution and those inseminated with BCC (3.8 x 10(8) vs. 1.5 x 10(8) cells, respectively); however, the total number of uterine spermatozoa was higher when gilts were inseminated with BCC (40.6 x 10(6) cells) compared with those inseminated with Modena solution (1.4 x 10(6) cells, P<0.05). In experiment 2, gilts and sows were subjected to intrauterine insemination twice with frozen-thawed spermatozoa suspended (25 x 10(8) sperm per dose) in Modena (n=21) or BCC (n=21). The overall pregnancy and farrowing rates were higher in females inseminated with BCC (71.4 and 61.9%, respectively) compared with those inseminated with Modena solution (38.1 and 28.6%, respectively, P<0.05). However, no significant difference in litter size of piglets was observed between treatments (7.2 +/- 1.6 piglets for Modena solution vs. 8.2 +/- 0.9 piglets for BCC solution). In conclusion, we demonstrated that use of BCC solution for frozen-thawed boar semen produced better pregnancy and farrowing rates following AI than Modena solution, probably by reducing the phagocytosis of spermatozoa.

  19. Genetic characterization of HIV-1 from semen and blood from clade C-infected subjects from India and effect of therapy in these body compartments

    PubMed Central

    Shen, Chengli; Ding, Ming; Craigo, Jodi K.; Tarwater, Patrick; Chatterjee, Ramdas; Roy, Pratima; Guha, Subhasish K; Saha, Bibhuti; Modak, Dolonchapa; Neogi, Dhruba; Chen, Yue; Gupta, Phalguni

    2010-01-01

    Biologic and genetic differences between HIV-1 clade C in India and clade B in US suggest that the effect of anti-viral therapy in various body compartments may differ between these two clades. We examined the effect of therapy viral loads in semen and blood of HIV-1-clade C infected subjects from India and evaluated whether HIV-1 in the semen is different from that in blood in these subjects. HIV-1 RNA was detected in semen and blood at all stages of the disease. Viral loads in semen and blood were strongly correlated with each other, but not with the CD4+ T cell count. Antiviral treatment reduced viral load drastically in blood and semen within one month of post therapy. Genetic characterization of HIV-1 in the semen and blood demonstrated that they were highly compartmentalized. These data have important implications of sexual transmission of HIV-1 in clade C HIV-1 infected subjects. PMID:20231027

  20. Sperm Morphological Features Associated with Chronic Chagas Disease in the Semen of Experimentally Infected Dogs

    PubMed Central

    Rodríguez-Morales, Olivia; Pedro-Martínez, Elvia; Hernández-Pichardo, José Ernesto; Alejandre-Aguilar, Ricardo; Aranda-Fraustro, Alberto; Graullera-Rivera, Verónica; Arce-Fonseca, Minerva

    2014-01-01

    The presence of trypanosomatids in the reproductive systems of different mammals (causing genital lesions in the acute stage of the disease) may predispose the animals to low semen quality. However, there are no studies examining the alterations in the sperm morphological features in the chronic stage of Trypanosoma cruzi infection. Knowledge of these aspects is important to understand the other ways of transmission of the Chagas disease. Progressive motility, mass motility, concentration, and sperm morphology of 84 ejaculates of dogs that were chronically infected with T. cruzi were evaluated. Most of the findings were consistent with the reference values and with those obtained from healthy control dogs. The scrotal circumference was not correlated with spermatozoa concentration in the infected animals. In conclusion, the T. cruzi Ninoa (MHOM/MX/1994/Ninoa) strain does not cause significant alterations in the semen quality of dogs experiencing chronic Chagas disease (at concentrations of 5 × 104 to 1 × 106 parasites per animal). PMID:25114010

  1. Effects of whole-body, ionizing radiation on the semen in beagle dogs

    SciTech Connect

    Day, P.A.

    1988-01-01

    Six beagle dogs were exposed to a total dose of 183 R of gamma radiation at a dose rate of 1 R/day, while three other dogs were exposed to a single dose of 100 R. Weekly semen analysis was performed on all irradiated dogs plus four nonirradiated dogs. Semen volume, sperm concentration, total sperm count, sperm motility and sperm head morphometry were examined. Dogs exposed to chronic radiation showed a severe decline in sperm numbers, detected after seven weeks of exposure. Sperm concentration and total sperm count were the first parameters affected and were the only parameters consistently affected. The dogs exposed to 100 R as a single dose, did not show a significant decline in sperm numbers. During a 36 week recovery period, the chronically irradiated dogs did show a slight increase in sperm numbers, but they never approached pre-exposure levels.

  2. Concentrations of morphologically normal, motile spermatozoa: Mg, Ca and Zn in the semen of infertile men.

    PubMed

    Pandy, V K; Parmeshwaran, M; Soman, S D; Dacosta, J C

    1983-03-01

    Semen from infertile men (n = 23) has been compared with that of control subjects (n = 25). Whereas the concentrations of morphologically normal, motile sperms, Mg, Ca and Zn fell within the acceptable limits for all the control subjects, only two infertile men qualified by all five parameters. Of the patient group, seven were abnormal on all counts; sperm motility, Mg and Zn were low in 16, Ca in 19 and abnormal morphology was encountered in 12. Since there was no linear correlation between any two parameters, it is possible that each factor may singly or jointly influence the physiological integrity of the spermatozoa. The results are discussed from a consideration of pathological manifestations known to occur in deficiency of these trace elements à propos their role in determining the fertility index of the semen.

  3. A new method for evaluating stallion sperm viability and mitochondrial membrane potential in fixed semen samples.

    PubMed

    Peña, F J; Ball, B A; Squires, E L

    2016-12-29

    Multiparametric assessment of stallion sperm quality using flow cytometry can be a useful adjunct in semen evaluation; however, the availability of flow cytometers in veterinary practice is limited. The ability to preserve and transport sperm samples for later flow cytometric analysis using fixable probes would potentially facilitate this process. In the current study, we validated the combination of live/dead Zombie Green(®) (a fixable dye used to assess live and dead sperm) and MitoTracker Deep Red(®) (used to assess mitochondrial membrane potential). The assay was validated against classic, non-fixable, membrane assays (SYBR-14/PI). Our results demonstrated the feasibility of the assay. In conclusion, stained and fixed semen samples stored for 72 h obtained equivalent results to the exam on the same day; this new protocol shall facilitate the wider use of flow cytometry in stallion andrology in the future. © 2016 International Clinical Cytometry Society.

  4. Is there a relationship between cell phone use and semen quality?

    PubMed

    Fejes, I; Závaczki, Z; Szöllosi, J; Koloszár, S; Daru, J; Kovács, L; Pál, A

    2005-01-01

    This study was conducted to determine a possible relationship between regular cell phone use and different human semen attributes. The history-taking of men in our university clinic was supplemented with questions concerning cell phone use habits, including possession, daily standby position and daily transmission times. Semen analyses were performed by conventional methods. Statistics were calculated with SPSS statistical software. A total of 371 were included in the study. The duration of possession and the daily transmission time correlated negatively with the proportion of rapid progressive motile sperm (r = -0.12 and r = -0.19, respectively), and positively with the proportion of slow progressive motile sperm (r = 0.12 and r = 0.28, respectively). The low and high transmitter groups also differed in the proportion of rapid progressive motile sperm (48.7% vs. 40.6%). The prolonged use of cell phones may have negative effects on the sperm motility characteristics.

  5. Longitudinal follow-up of Zika virus RNA in semen of a traveller returning from Barbados to the Netherlands with Zika virus disease, March 2016.

    PubMed

    Reusken, Chantal; Pas, Suzan; GeurtsvanKessel, Corine; Mögling, Ramona; van Kampen, Jeroen; Langerak, Thomas; Koopmans, Marion; van der Eijk, Annemiek; van Gorp, Eric

    2016-06-09

    We report the longitudinal follow-up of Zika virus (ZIKV) RNA in semen of a traveller who developed ZIKV disease after return to the Netherlands from Barbados, March 2016. Persistence of ZIKV RNA in blood, urine, saliva and semen was followed until the loads reached undetectable levels. RNA levels were higher in semen than in other sample types and declined to undetectable level at day 62 post onset of symptoms.

  6. Concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation to the occurrence of pathological spermatozoa.

    PubMed

    Massányi, P; Trandzik, J; Nad, P; Koreneková, B; Skalická, M; Toman, R; Lukac, N; Halo, M; Strapak, P

    2004-01-01

    In this study the concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation of these metals to spermatozoa morphology was investigated. Analysis by atomic absorption spectrophotometry showed that copper concentration was significantly higher (p<0.0001) in ram semen in comparison with bull semen. The zinc concentration was higher in bull semen in comparison with ram semen. The iron and cadmium concentrations in the semen were similar. Higher concentration of lead was found in ram semen. Higher levels of nickel were found in ram semen in comparison with bulls. In bull semen 11.79+/-4.88% of pathological spermatozoa was found. Higher occurrence of pathological spermatozoa was in ram semen (17.17+/-3.76) in comparison with the semen of bulls. Separated tail, tail torso, and knob twisted tail were the most frequent forms of pathological spermatozoa in both species. Correlation analysis in bulls showed high positive relation between iron and zinc (r = 0.72), nickel and separated tail (r = 0.76), separated tail and tail torso (r = 0.71), tail torso and total number of pathological spermatozoa (r=0.72), and between tail ball and total number of pathological spermatozoa (r = 0.78). In rams high positive correlation between cadmium and lead (r=0.98), nickel and separated tail (r=0.77), separated tail and total number of pathological spermatozoa (r=0.69), knob twisted tail and retention of cytoplasmic drop (r=0.78), and between knob twisted tail and other pathological spermatozoa (r = 0.71) was found. High negative correlation in ram semen was observed between copper and nickel (r=0.71), copper and separated tail (r=0.70), and between iron and tail torso (r=0.67). The results suggest that the studied metals have a direct effect on spermatozoa quality.

  7. Metabolic changes in droplet vitrified semen of wild endangered Persian sturgeon Acipenser persicus (Borodin, 1997).

    PubMed

    Abed-Elmdoust, Amirreza; Farahmand, Hamid; Mojazi-Amiri, Bagher; Rafiee, Gholamreza; Rahimi, Ruhollah

    2017-03-21

    Comparative quantitative metabolite profiling can be used for better understanding of cell functions and dysfunctions in particular circumstances such as sperm banking which is an important approach for cryopreservation of endangered species. Cryopreservation techniques have some deleterious effects on spermatozoa which put the obtained results in controversy. Therefore, in the present study, quantitative (1)H NMR (Nuclear Magnetic Resonance) based metabolite profiling was conducted to evaluate metabolite changes related to energetics and some other detected metabolites in vitrified semen of critically endangered wild Acipenser persicus. The semen was diluted with extenders containing 0, 5, 10, and 15 μM of fish antifreeze protein (AFP) type III as a cryoprotectant. Semen-extenders were vitrified and stored for two days. Based on post-thaw motility duration and motility percentage assessments, two treatments with 10 μM and 0 μM of AFP had the highest and the lowest motility percentages respectively and they were objected to (1)H NMR spectroscopy investigations in order to reveal the extremes of the metabolites dynamic range. Univariate (ANOVA) and multivariate (PCA) analysis of the resulting metabolic profiles indicated significant changes (P > 0.05) in metabolites. The level of some metabolites including acetate, adenine, creatine, creatine phosphate, lactate, betaine, sarcosine, β-alanine and trimethylamine N-oxide significantly decreased in vitrified semen while some others such as creatinine, guanidinoacetate, N, N-dimethylglycine, and glycine significantly increased. There were also significant differences between vitrified treatments in levels of creatine, creatine phosphate, creatinine, glucose, guanidinoacetate, lactate, N, N-dimethylglycine, and glycine, suggesting how fish AFP type III can be effective as a cryoprotectant.

  8. Should We Perform Semen Analysis, DNA Fragmentation, and Hypo-osmotic Swelling Tests together?

    PubMed Central

    Hasanzadeh Keshteli, Samaneh; Farsi, Mir Mehrdad; Khafri, Soraya

    2016-01-01

    Semen analysis, sperm DNA fragmentation (SDF) and hypo-osmotic swelling test (HOST) are usually performed for the evaluation of sperm fertilizing ability. There are some debates over the necessity of SDF and HOST incorporation in male infertility work-up.Semen of 77 men was evaluated by SDF and HOST through three semen analyses. Sperm parameters were arranged into different categories: <5%, 5-15%, >15% for normal morphology; <50%, 50-70%, >70 % for motility; and <10, 10-20, 21-34, 35-50, >50 million/ml for concentration. SDF analysis was performed and values under 30% were assumed to be normal. Normal range of HOST was considered to be >60%.Only normal sperm morphology had significant relationship with DF rate (P<0.001). Normal morphology, motility, and concentration of sperms had significant relationship with HOST (P<0.001, 0.05, and <0.003,respectively). There was a significant negative correlation between sperm morphology and DF rate. The correlations between sperm parameters and percentage of HOST were significantly positive (r: 0.44, 0.19, and 0.32 for morphology, motility, and concentration, respectively). Receiver operating characteristic curve (ROC) showed that sperm morphology is a strong predictor of the rate of DF and HOST (accuracy: 0.74‚ and 0.81, respectively). The best sperm morphology cut off point for DF and HOST rate prediction was 4.5% and 5.5%, respectively.Sperm morphology had significant correlation with DF rate and HOST and is supposed to be a predictor for these tests. Performing these three tests collectively for evaluation of semen samples would not be necessarily required in all cases.

  9. Development of extender and techniques for frozen turkey semen. 1. Development.

    PubMed

    Graham, E F; Nelson, D S; Schmehl, M K

    1982-03-01

    An extender for turkey semen with a frozen-thawed recovery of greater than or equal to 50% motile spermatozoa and a vigorous swirl was developed. The effect of glucose, a comparison of 10 different sugars in the extender, the use of sodium acetate and potassium acetate, the ratio of dimethylsulfoxide to ethylene glycol, the percent total cryoprotectant, equilibration time, and osmotic pressure were all tested. Replacement of approximately half of the extender with an iso-osmolar glucose solution yielded higher percentages of motile spermatozoa with both fresh and frozen-thawed semen samples. Replacement of glucose with other carbohydrates did not enhance recovery of frozen-thawed semen. The proportion of sodium and potassium acetates to TESNaK2 in the extender had no effect on motility. Approximately equal proportions of ethylene glycol to dimethylsulfoxide, with a final concentration of 11.2% after dilution, produced at or near optimal recovery of spermatozoal motility after freezing. Twenty to 90 min equilibration times before freezing yielded significantly higher recovery of motile sperm postthaw than shorter periods. A wide range of extender osmotic pressures were compatible with recovery of spermatozoal motility postthaw. The final extender consisted of 4.70 g sodium acetate, 3.39 g potassium acetate 9.23 g TES, .353 g sodium hydroxide, .492 g potassium hydroxide, 32.00 g glucose, H2O q.s. 1000 ml, 370 mOsm/kg, pH 7.2. Ethylene glycol and dimethylsulfoxide were added (1:1 ratio) for a final concentration of 11.2% cryoprotectant after dilution. Semen was held undiluted for 6 min, extended 1:4 at 22 C, and equilibrated for 30 min at 0 C before pellet freezing.

  10. Conventional and fluorescent based semen quality assessment in Karan Fries bulls

    PubMed Central

    Panmei, A.; Gupta, A. K.; Shivahre, P. R.; Bhakat, M.; Upadhyay, A.

    2015-01-01

    Aim: The present study was carried out on semen ejaculates of 15 Karan Fries (KF) bulls maintained at Artificial Breeding Research Centre, National Dairy Research Institute, Karnal, India with an objective to evaluate the relationship between the conventional and fluorescent based semen quality analysis of the bulls. Materials and Methods: A total of 96 ejaculates were collected from 15 KF (Holstein Friesian [HF] crossbred) bulls. Semen were evaluated for color, volume, mass activity (MA) and percentage of individual motility (IM), sperm concentration, percent live spermatozoa, hypo-osmotic swelling test and acrosome integrity, chromatin integrity, sperm viability, and membrane integrity. Data were analyzed using SPSS software package for descriptive analysis. The correlation between rankings of sires based on conventional and fluorescent semen parameters were calculated by Spearman’s rank correlation coefficient. Results: The average ejaculates volume (ml), sperm concentration (106/ml), MA, IM (%), live (%), morphological abnormalities (%), host (%), acrosome integrity (%), chromomycin A3 (CMA3) (%), SYBR-PI (%), and fluorescent isothiocyanate-peanut agglutinin (FITC-PNA) (%) were 4.57±0.36, 1162.98±97.93, 2.95±0.09, 60.8±1.22, 71.41±2.10, 9.31±1.15, 65.5±1.81, 86.6±1.59, 3.53±0.43, 65.39±2.23 and 74.47±2.53, respectively. Rank correlations were found to be significant for SYBR-PI and FITC-PNA with most of the parameters evaluated by conventional methods. Overall, among conventional criteria, IM revealed ranking of bulls almost similar to that of fluorescent criteria. Conclusion: Overview of our results indicated that, among conventional criteria, MA and IM revealed ranking of bulls almost similar to that of fluorescent criteria. PMID:27047025

  11. Semen quality detection using time of flight and acoustic wave sensors

    SciTech Connect

    Newton, M. I.; Evans, C. R.; Simons, J. J.; Hughes, D. C.

    2007-04-09

    The authors report a real-time technique for assessing the number of motile sperm in a semen sample. The time of flight technique uses a flow channel with detection at the end of the channel using quartz crystal microbalances. Data presented suggest that a simple rigid mass model may be used in interpreting the change in resonant frequency using an effective mass for the sperm.

  12. Semen characteristics and selected biochemical markers of canine seminal plasma in various seasons of the year.

    PubMed

    Strzeżek, R; Szemplińska, K; Filipowicz, K; Kordan, W

    2015-01-01

    The aim of this study was to evaluate the influence of season on selected qualitative semen characteristics and biochemical markers of canine seminal plasma. Whole ejaculates were collected from 5 crossbred dogs aged 2-8 years. The study covered a period of one year divided into four seasons: spring (March, April, May), summer (June, July, August), autumn (September, October, November) and winter (December, January, February). Semen samples were subjected to macroscopic and microscopic analyses to determine semen volume, total sperm counts and sperm morphology parameters. The study also involved the determination of sperm motility parameters (CASA system), sperm plasma membrane integrity (SPMI, fluorescent staining SYBR-14/PI), sperm mitochondrial membrane potential (MMP, fluorescent staining JC-1/PI) and the ATP content of sperm cells. Total protein content (TPC) and the activity of alkaline phosphatase (AP) and acid phosphatase (AcP) were determined in biochemical analyses of seminal plasma. No significant differences in ejaculate volume, SMPI or ATP content of sperm cells were observed between seasons. The highest total sperm counts were reported in ejaculates acquired in summer and autumn. The lowest MMP values were determined in summer ejaculates. No significant differences in sperm motility (MOT) were observed throughout the experiment, but ejaculates collected in autumn and winter were characterized by the highest progressive motility (PMOT). AP activity and TPC were not significantly affected by season. However, AcP activity levels were significantly lower in autumn than in the remaining seasons. Seasonal variations in the analyzed macroscopic and microscopic parameters of ejaculates and biochemical markers of seminal plasma did not exert a clear negative effect on the quality of canine semen.

  13. Selenium supplementation does not affect testicular selenium status or semen quality in North American men.

    PubMed

    Hawkes, Wayne Chris; Alkan, Zeynep; Wong, Kenneth

    2009-01-01

    Selenium (Se) is essential for sperm function and male fertility, but high Se intake has been associated with impaired semen quality. We reported previously a decrease in sperm motility in men fed high-Se foods, but we could not rule out the influence of other environmental and dietary factors. We now report on a randomized, controlled study on the potential adverse effects of Se supplementation on semen quality in 42 free-living men administered Se (300 microg/d) as high-Se yeast for 48 weeks. Semen analysis was performed 4 times before treatment began, then twice each week during treatment at 6, 12, 24, 36, and 48 weeks, and then after treatment at 72 and 96 weeks. Blood samples were collected 3 times before treatment and at each subsequent visit. Se concentration increased 61% in blood plasma and 49% in seminal plasma. However, Se supplementation had no effect on sperm Se, serum androgen concentrations, or sperm count, motility, progressive velocity, or morphology. We observed progressive decreases in serum luteinizing hormone, semen volume, and sperm Se in both the high-Se and placebo groups. Moreover, sperm straight-line velocity and percent normal morphology increased in Se-treated and placebo-treated participants. The lack of an increase in sperm Se suggests that testicular Se stores were unaffected, even though the participants' dietary Se intake was tripled and their total body Se approximately doubled by supplementation. These results are consistent with animal studies showing the Se status of testes to be unresponsive to dietary Se intake.

  14. Decline of semen quality during IVF is not associated with subjective male stress.

    PubMed

    Nouri, Kazem; Litschauer, Brigitte; Sator, Michael; Tiringer, Denise; Ott, Johannes; Walch, Katherina; Hefler, Lukas A; Tempfer, Clemens B

    2014-01-01

    The aim of the present study was to assess if semen quality declines during in vitro fertilization (IVF) and whether or not this phenomenon is triggered by chronic male stress. In order to test this hypothesis, we first investigated a retrospective cohort of 155 male IVF patients (testing cohort). Subsequently, we started a prospective cohort study in men undergoing their first IVF and assessed semen quality and subjective male chronic stress using a validated tool, i.e. the Fertility Problem Inventory (FPI) questionnaire. The association between stress and sperm quality decline measured 4-6 weeks before the start of IVF (T1) and at the day of oocyte retrieval (T2) was the primary outcome. Live birth rate, first trimester abortion and rate of poor responders were secondary outcomes. In the testing cohort, mean progressive motility, but not mean sperm density significantly declined. There were 78/154 (51%) men who showed a decline in semen density and 50/154 (32%) men who showed a decline in progressive motility. In the validation cohort, progressive motility declined, whereas, sperm density increased from T1 to T2. Of 78 men, 27 men had increased stress (FPI-score > 146). Sperm density and progressive motility were not significantly different in men with and without stress. However, in the presence of male stress, couples had a higher rate of poor responders, miscarriages and a lower rate of live births. Subjective stress is not associated with a decline in semen quality observed during IVF but may be associated with adverse pregnancy outcome.

  15. Physically active men show better semen parameters and hormone values than sedentary men.

    PubMed

    Vaamonde, Diana; Da Silva-Grigoletto, Marzo Edir; García-Manso, Juan Manuel; Barrera, Natalibeth; Vaamonde-Lemos, Ricardo

    2012-09-01

    Physical exercise promotes many health benefits. The present study was undertaken to assess possible semen and hormone differences among physically active (PA) subjects and sedentary subjects (SE). The analyzed qualitative sperm parameters were: volume, sperm count, motility, and morphology; where needed, additional testing was performed. The measured hormones were: follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), cortisol (C), and the ratio between T and C (T/C). Maximum oxygen consumption was also assessed to check for differences in fitness level. Statistically significant differences were found for several semen parameters such as total progressive motility (PA: 60.94 ± 5.03; SE: 56.07 ± 4.55) and morphology (PA: 15.54 ± 1.38, SE: 14.40 ± 1.15). The seminological values observed were supported by differences in hormones, with FSH, LH, and T being higher in PA than in SE (5.68 ± 2.51 vs. 3.14 ± 1.84; 5.95 ± 1.11 vs. 5.08 ± 0.98; 7.68 ± 0.77 vs. 6.49 ± 0.80, respectively). Likewise, the T/C ratio, index of anabolic versus catabolic status, was also higher in PA (0.46 ± 0.11 vs. 0.32 ± 0.07), which further supports the possibility of an improved hormonal environment. The present study shows that there are differences in semen and hormone values of physically active subjects and sedentary subjects. Physically active subjects seem to have a more anabolic hormonal environment and a healthier semen production.

  16. Clinical relevance of polymorphonuclear (PMN-) elastase determination in semen and serum during infertility investigation.

    PubMed

    Eggert-Kruse, W; Zimmermann, K; Geissler, W; Ehrmann, A; Boit, R; Strowitzki, T

    2009-08-01

    The polymorphonuclear (PMN) elastase is secreted by activated granulocytes and is widely used as a marker of male accessory gland infection. However, the clinical value of routine determination of seminal plasma (SP) PMN elastase in asymptomatic patients during infertility investigation has not clearly been established and not much is known about the significance of PMN-elastase levels in serum as a potential biochemical determinant associated with infection/inflammation of the male genital tract. This prospective study included a total of 221 asymptomatic males from unselected subfertile couples, to evaluate the relationship of (i) serum and (ii) same-day SP PMN elastase concentrations with established semen quality parameters, including sperm functional capacity, local antisperm antibodies (ASA), seminal leucocytes, and the outcome of semen cultures including typical sexually transmitted disease pathogens, and a potential association with patients' medical history and results of clinical andrological examination. Furthermore, couples were followed up for subsequent fertility (controlled for female infertility factors). The concentrations of PMN elastase in serum and in SP were not significantly related to semen quality [with regard to microscopic (e.g. count, motility, morphology) as well as biochemical parameters, and also to local ASA of the IgG- or IgA-class]. There was no strong relationship with sperm functional capacity. No significant relationship with the outcome of the microbial screening was found. PMN-elastase levels in serum and SP were not significantly correlated and there was no association with subsequent fertility. Therefore, the value of routine determination of PMN elastase in semen and/or serum samples, particularly when used as a single parameter to screen for subclinical infection/inflammation in males under infertility investigation is limited.

  17. Technology Readiness Assessments (TRAs)

    DTIC Science & Technology

    2006-05-11

    Pictures are of: Left side: Edmond Halley (above), Michael Faraday (below) Right side: Leonardo da Vinci (above), Alfred Nobel (below) Agencies need to...Propulsion Š AV Sys Software Š Comm/ID Š Central Computer Š Fire Control Š Auto Flight Control Š Weapons Delivery Air Vehicle (AV) System T...Configuration) • Does the design incorporate a configuration that has not been used in flight ? • How similar is the configuration to that of

  18. Quality and developmental rate of embryos produced with sex-sorted and conventional semen from superovulated dairy cattle.

    PubMed

    Mikkola, M; Taponen, J

    2017-01-01

    This study investigated the effect of sex-sorted semen compared with conventional semen on the outcome of embryo recovery, placing special emphasis on the quality, and developmental stage of embryos. Data were analyzed for 443 embryo collections with sex-sorted semen (SEX group) and 1528 with conventional semen (CONV group) in superovulated dairy heifers and cows. The insemination protocol for conventional semen included two inseminations, comprising a total dose of 30 million sperm passing into the uterine body. For sex-sorted semen, two (30%) to three (70%) deep uterine inseminations were performed, the total dose ranging from eight to 12 million sperm. The data were analyzed separately for heifers and cows. The total number of recovered structures was similar among the groups. The number of viable embryos decreased in the SEX groups compared with the CONV (with 1.4 and 3.2 fewer embryos in heifers and cows, correspondingly, P < 0.001), and correspondingly the proportions of unfertilized ova and degenerated embryos increased in the SEX groups (P < 0.001). The proportion of unsuccessful collections, yielding no transferable embryos, increased in the SEX groups for both heifers (from 7.2% to 11.2%, P = 0.025) and cows (from 9.0% to 20.7%, P < 0.001). Regarding the quality of viable embryos, the quality grades were superior in the CONV group compared with the SEX group for heifers (P < 0.001) and cows (P < 0.001). The proportion of grade 1 embryos decreased by 6.5 percentage points in heifers and 11.9 percentage points in cows when sex-sorted semen was used. Correspondingly, the proportions of grade 2 and 3 embryos increased in heifers and cows when sexed semen was used. The mean developmental stages of embryo collections were numerically slightly lower in the SEX group. In heifers, the delay in developmental stage was statistically significant (P = 0.001), but in cows, there was only a tendency toward that (P = 0.067). In conclusion, sex-sorted sperm

  19. Constitution of semen samples from XYY and XXY males as analysed by in-situ hybridization.

    PubMed

    Martini, E; Geraedts, J P; Liebaers, I; Land, J A; Capitanio, G L; Ramaekers, F C; Hopman, A H

    1996-08-01

    A brightfield microscopical in-situ hybridization (ISH) technique was applied to semen samples of two 47,XYY males, one 46,XY/47,XXY/XXY male with fertility problems, and two normal 46,XY men, who served as controls. The use of a standardized nuclear DNA decondensation method, together with double-target ISH and morphological staining, allowed an accurate study of the sex chromosomal content and morphology of spermatozoa. In the males carrying an extra sex chromosome, we detected X- and Y-bearing spermatozoa in a ratio which did not differ significantly from the 1:1 ratio found in normal males. Aneuploidy for the sex chromosomes was found in approximately 15% of the spermatozoa of both XYY males and in 3% of the XXY male. The most striking finding was the relatively low percentage of spermatozoa in these patients, with an average of 65% in the XYY males and 84% in the XXY male. The other cells represented immature germ cells (IGC), including spermatogonia and spermatocytes arrested at various stages of spermatogenesis. Apparently, in XYY or XXY men, these IGC are shed into the semen to an increased extent as compared to normal, fertile men. The sex chromosome constitution of these IGC was heterogeneous. However, the finding that the majority of spermatozoa in semen of 47,XYY and 47,XXY males carried a single sex chromosome strengthens the hypothesis that a 46,XY germ cell line must be present, apparently with a proliferative advantage over the 47,XYY or 47,XXY cells.

  20. [Dimensions of the personality of semen donors and their social behavior].

    PubMed

    Taus, L; Gerzová, J

    1991-02-22

    The authors examined using three generally accepted methods, the personality structure of 80 semen donors (Cattell's 16-factor questionnaire, 16PF, Eysenck's personality questionnaire EOD and Leary's method of interpersonal diagnosis of personality). The donors were selected by means of the Questionnaire of semen donors. The group is subdivided into four subgroups by the grade of education, i.e. university graduates, men with secondary and elementary education and university students. All are 20-40 years old. The authors describe the assembled results in different sub-groups and in the group as a whole and compare them mutually and with the standardized norm. With regard to the specificity of individual methods and their application the findings are summarized. The donors are balanced personalities, slightly extrovert, emotionally well developed with a realistic outlook. They have positive, sensitive relations with their environment an behaviour towards other people, they are considerate, careful and disciplined. They respect social norms as regards preservation of originality of personality. They have a slight tendency of sheltering behaviour, they wish to be somewhat more aggressive. No pathological phenomena were observed in the donors. Their intelligence is above average. They make a favourable impression with regard to the demand of mental health and transmission of genetic information. The authors evaluate favourably the Questionnaire for semen donors as the method for selection of donors.

  1. Sericin supplementation improves semen freezability of buffalo bulls by minimizing oxidative stress during cryopreservation.

    PubMed

    Kumar, Pradeep; Kumar, Dharmendra; Sikka, P; Singh, P

    2015-01-01

    The variety of mammalian cells has been successfully cryopreserved by use of the silk protein sericin due to its strong free-radical-scavenging and potent antioxidant activity. The present study was conducted to examine the protective role of sericin on buffalo spermatozoa during cryopreservation. Semen of four breeding bulls was collected twice a week using artificial vagina technique. The ejaculates of four bulls were pooled, divided into five equal fractions, diluted with the extender supplemented with different concentrations of sericin (0, 0.25, 0.5, 1.5 and 2%) and then cryopreserved. Post-thawed motility was objectively assessed by computer assisted sperm analyzer. Sperm plasma membrane integrity was assessed by hypo-osmotic swelling test (HOST). Malondialdehyde (MDA) concentration, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in frozen-thawed extended seminal plasma by spectrophotometry. The extender supplemented with 0.25, 0.5 and 1% sericin resulted in the higher sperm motility and GPx acivity. Furthermore, plasma membrane integrity and SOD activity were found to be higher (P<0.05) in group supplemented with 0.25 and 0.5% sericin (P<0.05). The MDA concentration was found to be significantly lower (P<0.05) in 0.25 and 0.5% sericin treated groups than control and other treated groups. In conclusion, the supplementation of 0.25-0.5% sericin in semen extender improves frozen-thawed semen quality through protecting sperm from oxidative stress.

  2. Sperm Cell Population Dynamics in Ram Semen during the Cryopreservation Process

    PubMed Central

    Ramón, Manuel; Pérez-Guzmán, M. Dolores; Jiménez-Rabadán, Pilar; Esteso, Milagros C.; García-Álvarez, Olga; Maroto-Morales, Alejandro; Anel-López, Luis; Soler, Ana J.; Fernández-Santos, M. Rocío; Garde, J. Julián

    2013-01-01

    Background Sperm cryopreservation has become an indispensable tool in biology. Initially, studies were aimed towards the development of efficient freezing protocols in different species that would allow for an efficient storage of semen samples for long periods of time, ensuring its viability. Nowadays, it is widely known that an important individual component exists in the cryoresistance of semen, and efforts are aimed at identifying those sperm characteristics that may allow us to predict this cryoresistance. This knowledge would lead, ultimately, to the design of optimized freezing protocols for the sperm characteristics of each male. Methodology/Principal Findings We have evaluated the changes that occur in the sperm head dimensions throughout the cryopreservation process. We have found three different patterns of response, each of one related to a different sperm quality at thawing. We have been able to characterize males based on these patterns. For each male, its pattern remained constant among different ejaculates. This latter would imply that males always respond in the same way to freezing, giving even more importance to this sperm feature. Conclusions/Significance Changes in the sperm head during cryopreservation process have resulted useful to identify the ability of semen of males for freezing. We suggest that analyses of these response patterns would represent an important tool to characterize the cryoresistance of males when implemented within breeding programs. We also propose follow-up experiments to examine the outcomes of the use of different freezing protocols depending on the pattern of response of males. PMID:23544054

  3. Epididymis microlithiasis and semen abnormalities in young adult kidney transplant recipients.

    PubMed

    Bozzini, G; Lunelli, L; Berlingheri, M; Groppali, E; Carmignani, L

    2013-10-01

    Microlithiasis of the epididymis is a rare ultrasound finding in the general population, but the incidence of calcifications in various organs of patients with end-stage renal disease (ESRD) is extremely high. The aim of this study was to describe epididymal microlithiasis in 22 previously dialysed patients who received kidney transplantations at a median age of 19 years (range 9-30). The patients underwent scrotum ultrasonography, semen analysis and laboratory tests (renal function, sexual hormones, Ca, P and PTH) and were administered the International Index of Erectile Function questionnaire. Seventeen presented calcifications of the epididymis, two of whom had concomitant testicular calcifications; a further three patients had isolated testicular calcifications without epididymis involvement. It was not possible to investigate the fertility of all of the patients but 12 of the 13 whose semen was analysed showed abnormalities: five were azoospermic and seven oligospermic with various degrees of morphological anomalies. To the best of our knowledge, these are the first published data concerning the prevalence of epididymal calcifications in young dialysed patients undergoing renal transplantation. Epididymal microlithiasis and infertility were common findings and so performing a spermiogram and preserving semen before ESRD for future paternity may be good advice in this selected population.

  4. Microbial contamination of embryos and semen during long term banking in liquid nitrogen.

    PubMed

    Bielanski, A; Bergeron, H; Lau, P C K; Devenish, J

    2003-04-01

    We report on microbial contamination of embryos and semen cryopreserved in sealed plastic straws and stored for 6-35 years in liquid nitrogen. There were 32 bacterial and 1 fungal species identified from randomly drawn liquid nitrogen, frozen semen, and embryos samples stored in 8 commercial and 8 research facility liquid nitrogen (LN) tanks. The identified bacteria represented commensal or environmental microorganisms and some, such as Escherichia coli, were potential or opportunistic pathogens for humans and animals. Stenotrophomonas maltophilia was the most common contaminant identified from the samples and was further shown to significantly suppress fertilization and embryonic development in vitro. Analysis of the strains by pulsed field gel electrophoresis revealed restriction patterns with no relatedness indicating that there was no apparent cross-contamination of S. maltophilia strains between the germplasm and liquid nitrogen samples. In addition, no transmission of bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1) from infected semen and embryos straws to clean germplasm stored in the same LN tanks or LN was detected.

  5. Semen preparation methods and sperm telomere length: density gradient centrifugation versus the swim up procedure

    PubMed Central

    Zhao, Feifei; Yang, Qingling; Shi, Senlin; Luo, Xiaoyan; Sun, Yingpu

    2016-01-01

    Previous studies have shown that both density gradient centrifugation (DGC) and swim up (SU) procedures can select spermatozoa with longer telomeres for assisted reproduction techniques (ART). However, it is unknown which approach is more effective. The aim of the present study was to compare the effects of these two methods on sperm telomere length (STL). A total of 150 normozoospermic subjects were recruited. STL, DNA fragmentation index (DFI), reactive oxygen species (ROS) content and progressive motility of semen samples were detected before and after the procedures of DGC and SU. When compared to raw semen, the average length of sperm telomeres was significantly longer after the two sperm preparation methods. However, no significant difference was found between the DGC and SU procedures. We also found that semen prepared by the two methods had lower DNA fragmentation, ROS content and sperm progressive motility. However, no significant difference was found in those parameters between the two procedures. This is the first study that compares the effects of the DGC and SU procedures on STL, and the results show that both methods can recover a sperm population with longer STL and better DNA integrity for ART. PMID:27958357

  6. Semen collection, characterisation and artificial insemination in the beluga (Delphinapterus leucas) using liquid-stored spermatozoa.

    PubMed

    O'Brien, J K; Steinman, K J; Schmitt, T; Robeck, T R

    2008-01-01

    Ejaculates were collected from a beluga (Delphinapterus leucas) to gain an understanding of sperm biology and develop a short-term sperm preservation method for use in artificial insemination (AI). Ejaculate parameters and biochemistry, semen production and serum testosterone concentrations of an adult male were characterised for 21 months. Sperm viability, acrosome integrity and morphology did not change (P > 0.05) but ejaculate volume, sperm concentration and total spermatozoa per ejaculate were higher (P < 0.05) from January to June than from July to December. Peak testosterone concentrations (P < 0.05) were observed from October to April (8.0 +/- 1.6 ng mL(-1)). The effects of hyaluronic acid (HA), antioxidants, storage temperature and time on in vitro sperm characteristics were examined. Motility parameters and viability were improved (P < 0.05) when semen was stored at 5 degrees C compared with 21 degrees C. During the first 24 h of storage sperm agglutination was absent only at 5 degrees C in the presence of HA. A nulliparous 28-year-old female was inseminated endoscopically with liquid-stored semen. A pregnancy and birth of a calf was achieved following AI for the first time in this species, thereby validating both the AI technique and the fertility of beluga spermatozoa after chilled storage in a specialised diluent.

  7. Objective assessment of the cryoprotective effects of dimethylformamide for freezing goat semen.

    PubMed

    Bezerra, Francisco Silvestre B; Castelo, Thibério S; Alves, Heron M; Oliveira, Isabelle R S; Lima, Gabriela L; Peixoto, Gislayne C X; Bezerra, Ana Carla S D; Silva, Alexandre R

    2011-12-01

    The aim of this work was to assess the cryoprotective effects of dimethylformamide (DMF) for freezing goat semen, using an objective analysis by computer-assisted sperm analysis (CASA). Twenty-one ejaculates (seven per animal) were collected from three stud bucks with the aid of an artificial vagina and immediately evaluated for gross and microscopic characteristics. The semen was diluted in two steps with a Tris-egg yolk extender containing 6% glycerol or 6% DMF, frozen in 0.50-mL straws, and stored in liquid nitrogen. Samples were accessed for sperm morphology, sperm membrane structural and functional integrity, and by CASA, immediately after thawing. There were differences (P<0.05) between glycerol and DMF with regard to subjective progressive motility (23.9±2.2% vs. 16.6±2.0%), objective progressive motility (3.5±0.4% vs. 1.8±0.3%), linearity (53.9±1.6% vs. 48.1±1.4%) and amplitude of lateral head (2.3±0.1 vs. 2.9±0.1 mm), which confirmed the efficiency of glycerol. In conclusion, dimethylformamide could be used as an alternative cryoprotectant for goat semen freezing. However it was showed that no benefits were derived by using dimethylformamide to replace glycerol at an equal 6% concentration.

  8. Blood and semen paraoxonase-arylesterase activities in normozoospermic and azoospermic men.

    PubMed

    Gulum, M; Gumus, K; Yeni, E; Dogantekin, E; Ciftci, H; Akin, Y; Savas, M; Altunkol, A

    2016-12-21

    Paraoxonase and arylesterase enzymes are corner stones of antioxidant defence. We aimed to compare azoospermic infertile men and normozoospermic individuals with respect to total antioxidant status (TAS), total oxidant status (TOS), oxidative stress index (OSI), paraoxonase and arylesterase levels in the blood and seminal plasma. Two-hundred consecutive infertility patients and voluntarily participated were included. In the normozoospermic group, TAS, PON, arylesterase values were statistically significantly higher when compared with those in the azoospermic group, while lower TOS and OSI levels were observed in the blood and seminal plasma of azoospermic group. In the semen analyses of normozoospermic group, the correlation between semen volume, sperm concentration, sperm motility and morphology and TAS, TOS, OSI, PON and arylesterase values was examined. A negative correlation was determined between semen volume and OSI. Levels of serum oxidative parameters were higher in the azoospermic group relative to normozoospermic group, but antioxidant parameters were lower than those of the normozoospermic group. Oxidative stress performs an essential role in the aetiology of male infertility by negatively influencing sperm quality and function. Assessment of blood and seminal plasma oxidative profiles might be an important tool to better evaluation of sperm reproductive capacity and functional competence.

  9. Organic selenium supplementation increases PHGPx but does not improve viability in chilled boar semen.

    PubMed

    Martins, S M M K; De Andrade, A F C; Zaffalon, F G; Bressan, F F; Pugine, S M P; Melo, M P; Chiaratti, M R; Marino, C T; Moretti, A S; Arruda, R P

    2015-02-01

    This study evaluated the effects of dietary organic selenium (Se) on viability of chilled boar semen. Twelve boars were divided into three groups: control (CON), 0.3 mg kg(-1) sodium selenite; inorganic (INO), 0.5 mg kg(-1) sodium selenite and organic (ORG), 0.5 mg kg(-1) Se yeast. The experiment was conducted within 10 weeks, and analysis was performed fortnightly, in storage semen by 72 h. No effect was observed on motility; however, straightness and linearity percentages were higher (P < 0.05) in the animals receiving CON diet compared with INO group. Percentages of cells with both plasma and acrosomal intact membranes, lipidic membrane peroxidation and mitochondrial membrane potential were similar on all treatments. Animals receiving CON diet presented higher (P < 0.05) values of ATP when compared with INO group. The PHGPx was higher (P < 0.05) in animals that received ORG in comparison with INO group. In conclusion, organic selenium supplementation increases PHGPx but does not improve chilled semen viability in 72 h.

  10. Effect of pasteurized egg and Rosmarinus officinalis supplementation on quality of cryopreserved ram semen.

    PubMed

    Mascaro, F; Gil, L; Malo, C; Gonzales, N; Martinez, F; de Blas, I

    2013-01-01

    The aim was to assess the in vitro effect of pasteurized egg (PE) and rosemary (Rosmarinus officinalis) on frozen-thawed ram semen. Ejaculates from three mature rams of the Rasa Aragonesa breed were cryopreserved using a 2-step dilution method (Fraction 1: F1; Fraction 2: F2). In Experiment 1, semen was frozen in egg yolk (EY) or PE extenders. After thawing, similar results were obtained in terms of total and progressive motility, viability, hypo-osmotic swelling test (HOST) and acrosome integrity after 2 h incubation. In Experiment 2, addition of rosemary to F1, F2 or both fractions to EY extenders was evaluated. Rosemary in F1 decreased progressive motility (p = 0.013) after 2 h incubation. Finally, PE can be used as a substitute for EY to reduce hygienic risks in extenders and is easier to standardize. Supplementation of EY extender with rosemary in F1 reduced progressive motility. Rosemary supplementation in F2 does not affect semen quality.

  11. Dimethylformamide as a cryoprotectant for canine semen diluted and frozen in ACP-106C.

    PubMed

    Mota Filho, A C; Teles, C H A; Jucá, R P; Cardoso, J F S; Uchoa, D C; Campello, C C; Silva, A R; Silva, L D M

    2011-10-15

    The objective was to assess the effect of adding various concentrations of dimethylformamide on characteristics of canine semen diluted in powdered coconut water (ACP-106C; ACP Biotecnologia, Fortaleza, CE, Brazil) and frozen at -196°C. Fifteen ejaculates were collected by manual stimulation from five adult Boxer dogs. The sperm-rich fraction was diluted in ACP-106C (ACP Biotecnologia) containing 10% egg yolk and divided into four aliquots. The cryoprotectants used for each aliquot were 6% glycerol (control group; CG) or 2%, 4%, or 6% dimethylformamide (DF2, DF4, and DF6, respectively). After thawing, total motility (mean ± SEM) for CG (58.4 ± 24.6) was higher (P < 0.05) than that of the other groups (2% dimethylformamide, 24.4 ± 12.3; 4% dimethylformamide, 26.5 ± 16.1; and 6% dimethylformamide, 21.7 ± 17.9). Furthermore, there was a greater percentage of fast, average, and slow moving sperm (assessed with computer-aided semen analysis; CASA) in CG in comparison with the other three groups. Therefore, based on concentrations tested in this study, dimethylformamide, together with ACP-106C (ACP Biotecnologia) and 10% egg yolk as a diluent, yielded unsatisfactory in vitro results for freezing canine semen.

  12. Human semen refrigeration at + 4 degrees C: bio-kinetic characteristics.

    PubMed

    Dondero, Franco; Rossi, Tiziana; Delfino, Michele; Imbrogno, Norina; Cannistrà, Stefania; Mazzilli, Fernando

    2006-01-01

    The aim of our study was to evaluate the bio-kinetic characteristics of human semen refrigerated for different periods and to compare the effects of refrigeration at +4 degrees C against cryopreservation of human sperm at -196 degrees C. Semen was obtained from 30 male partners of infertile couples (infertile subjects) with the following semen profile: sperm count >or=10 x 10(6)/ml; progressive motility >or=20%; atypical forms <70% and white blood cells <1.0 x 10(6)/ml. Fifteen normospermic subjects were also selected as controls (control subjects). The following tests were carried out on basal, refrigerated and cryopreserved sperm: a) sperm kinetic properties (by Superimposed Image Analysis System); b) the Hypoosmotic Viability Test (HVT) (combined Hypoosmotic Swelling and Viability Test). The results of the study showed that the percentage recovery of kinetic properties and of HVT were optimum for up to 48 h. After refrigeration for 72 h, a drastic decrease in straight motility recovery was observed. No significant differences were observed between cryopreservation and refrigeration at +4 degrees C for 48 h for motility or HVT recoveries in samples from control subjects. However, in infertile subjects, a significant decrease in straight progressive motility and HVT recoveries was observed in cryopreserved samples compared to those refrigerated for 48 h. Neither refrigeration nor cryopreservation led to the growth of pathogenic bacteria in any of the cases studied. Based on the above results, refrigeration could represent a useful alternative to the cryopreservation method.

  13. High-resolution melt analysis of DNA methylation to discriminate semen in biological stains.

    PubMed

    Antunes, Joana; Silva, Deborah S B S; Balamurugan, Kuppareddi; Duncan, George; Alho, Clarice S; McCord, Bruce

    2016-02-01

    The goal of this study was to develop a method for the detection of semen in biological stains using high-resolution melt (HRM) analysis and DNA methylation. To perform this task, we used an epigenetic locus that targets a tissue-specific differentially methylated region for semen. This specific locus, ZC3H12D, contains methylated CpG sites that are hypomethylated in semen and hypermethylated in blood and saliva. Using this procedure, DNA from forensic stains can be isolated, processed using bisulfite-modified polymerase chain reaction (PCR), and detected by real-time PCR with HRM capability. The method described in this article is robust; we were able to obtain results from samples with as little as 1 ng of genomic DNA. Samples inhibited by humic acid still produced reliable results. Furthermore, the procedure is specific and will not amplify non-bisulfite-modified DNA. Because this process can be performed using real-time PCR and is quantitative, it fits nicely within the workflow of current forensic DNA laboratories. As a result, it should prove to be a useful technique for processing trace evidence samples for serological analysis.

  14. Mucuna pruriens Reduces Stress and Improves the Quality of Semen in Infertile Men

    PubMed Central

    Shukla, Kamla Kant; Ahmad, Mohammad Kaleem; Jaiswar, Shyam Pyari; Shankwar, Satya Narain; Tiwari, Sarvada Chandra

    2010-01-01

    The present investigation was undertaken to assess the role of Mucuna pruriens in infertile men who were under psychological stress. Study included 60 subjects who were undergoing infertility screening and were found to be suffering from psychological stress, assessed on the basis of a questionnaire and elevated serum cortisol levels. Age-matched 60 healthy men having normal semen parameters and who had previously initiated at least one pregnancy were included as controls. Infertile subjects were administered with M. pruriens seed powder (5 g day−1) orally. For carrying out morphological and biochemical analysis, semen samples were collected twice, first before starting treatment and second after 3 months of treatment. The results demonstrated decreased sperm count and motility in subjects who were under psychological stress. Moreover, serum cortisol and seminal plasma lipid peroxide levels were also found elevated along with decreased seminal plasma glutathione (GSH) and ascorbic acid contents and reduced superoxide dismutase (SOD) and catalase activity. Treatment with M. pruriens significantly ameliorated psychological stress and seminal plasma lipid peroxide levels along with improved sperm count and motility. Treatment also restored the levels of SOD, catalase, GSH and ascorbic acid in seminal plasma of infertile men. On the basis of results of the present study, it may be concluded that M. pruriens not only reactivates the anti-oxidant defense system of infertile men but it also helps in the management of stress and improves semen quality. PMID:18955292

  15. Impact of male obesity on semen quality and serum sex hormones.

    PubMed

    Eskandar, Mamdoh; Al-Asmari, Manal; Babu Chaduvula, Suresh; Al-Shahrani, Mesfer; Al-Sunaidi, Mohammed; Almushait, Mona; Donia, Osman; Al-Fifi, Suliman

    2012-01-01

    Introduction. To investigate the association of high Body Mass Index (BMI) with semen parameters and reproductive hormones in men of reproductive age. Setting. The Saudi Center for Assisted Reproduction. Method. This study was conducted during the period from February 2009 to February 2011. Subjects were exposed through medical history evaluation as well as physical examination. BMI was calculated. Two semen samples about 1 week apart were taken from each participant by masturbation after 2-5 days of abstinence. The samples were assessed according to the WHO Criteria. Blood samples (5 ml) were withdrawn; centrifuged and the resulting sera were preserved at -4 degrees Centigrade. Serum FSH, LH, PRL, and Testosterone levels were estimated by the ELISA method. Results. There was no significant correlation between BMI and any of semen and hormonal parameters. There was significant negative correlation between age and total motility. Only the advanced paternal age has shown significant association with low motility (P = 0.007). Conclusion. Our study showed a significant effect of aging on sperm motility and concentration.

  16. The male reproductive system in classic galactosemia: cryptorchidism and low semen volume.

    PubMed

    Gubbels, Cynthia S; Welt, Corrine K; Dumoulin, John C M; Robben, Simon G F; Gordon, Catherine M; Dunselman, Gerard A J; Rubio-Gozalbo, M Estela; Berry, Gerard T

    2013-09-01

    Previous studies examining reproductive parameters in men with galactosemia have inconsistently demonstrated abnormalities. We hypothesized that men with galactosemia may demonstrate evidence of reproductive dysfunction. Pubertal history, physical examination, hormone levels and semen analyses were examined in 26 males with galactosemia and compared to those in 46 controls. The prevalence of cryptorchidism was higher in men with galactosemia than in the general population [11.6% vs. 1.0% (95%CI: 0.75-1.26; p <0.001)]. Testosterone (461±125 vs. 532± 33 ng%; p=0.04), inhibin B (144±66 vs. 183±52 pg/mL; p=0.002) and sperm concentration (46±36 vs. 112±75×10(6) spermatozoa/mL; p=0.01) were lower and SHBG was higher (40.7±21.5 vs 26.7±14.6; p=0.002) in men with galactosemia compared to controls. Semen volume was below normal in seven out of 12 men with galactosemia. Men with galactosemia have a higher than expected prevalence of cryptorchidism and low semen volumes. The subtle decrease in testosterone and inhibin B levels and sperm count may indicate mild defects in Sertoli and Leydig cell function, but does not point towards severe infertility causing reproductive impairment. Follow-up studies are needed to further determine the clinical consequences of these abnormalities.

  17. Enrichment of high-quality spermatozoa in bovine semen: relative effectiveness of three filtration matrixes.

    PubMed

    Cisale, H O; Fischman, M L; Blasi, C D; Fernandez, H A; Gledhill, B L

    2001-05-01

    To be practical, any method for improving bull semen must yield a large quantity of motile spermatozoa. Some separation methods based on physical properties, e.g. filtration, chromatography, centrifugation, washing and pooling, have been reported as satisfactory, but generally are not repeatable. Nevertheless, filtration methods appear to allow the attainment of an acceptable number of spermatozoa, thus allowing such a technique to be introduced in the production of standard bovine semen doses for artificial insemination. The aim of this work was to evaluate systematically the relative effects of three filtration matrixes (silica oxide, glass beads or Sephadexätrade mark) on the improvement of whole ejaculate quality. Analysis of the type of matrix and the volume and height of the filtration column was performed. The only characteristic of the columns that appears to influence ejaculate quality after filtering is the matrix volume. While all matrixes produced improvement of semen quality, SephadexTM was better than the other matrixes tested. An explanation for the mechanism of column filtration is proposed.

  18. Atropine-induced inhibition of sperm and semen transport impairs fertility in male rats.

    PubMed

    Sato, Takahiro; Ban, Yoshiki; Uchida, Miki; Gondo, Eri; Yamamoto, Masakatsu; Sekiguchi, Yoshiko; Sakaue, Akiko; Kemi, Masayuki; Nakatsuka, Toshio

    2005-08-01

    Previous studies revealed that atropine reduced male fertility in rats without any effects on mating performance, sperm production and motility, and testicular morphology. The present study was conducted to investigate whether the impairment of male fertility induced by atropine was related to the inhibition of sperm and semen transports from the vas deferens and seminal vesicle to the urethra during the process of emission. Male rats were treated with atropine at 125 mg/kg/day for 10-17 days prior to mating with untreated females. After confirmation of mating, male rats were euthanized and sperm number in the vas deferens and weights of the seminal vesicle and copulatory plug were determined as indicators of inhibition of sperm and semen transports, respectively. Reproductive status of mated females was determined on gestation days 15-17. A low pregnancy rate associated with a decreased number of implants was observed in females that mated with the atropine-treated males. The average number of sperm in the vas deferens was increased in the atropine-treated males. The average seminal vesicle weight in the atropine-treated males was greater than that of controls. The copulatory plug weights were decreased in the atropine-treated males. These results suggest that inhibitions of sperm and semen transports from the vas deferens and seminal vesicle to the urethra during the process of emission result in reduced male fertility in rats.

  19. Comparison of differents methods of sperm selection of llama raw semen.

    PubMed

    Santa Cruz, R; Giuliano, S M; Gambarotta, M C; Morrell, J M; Abraham, M C; Miragaya, M H; Carretero, M I

    2016-10-01

    The objective of this study was to compare the efficiency of different sperm selection methods applied to the same llama ejaculate. Four treatments were compared: two variants of the swim up technique (with and without seminal plasma), and two different colloids, Androcoll-E-Large and Percoll(®). Using electroejaculation, 21 semen samples were obtained from 7 llama males (n=7, r=3). The ejaculates were incubated in a solution of 0.1% collagenase, to decrease thread formation, and then split into 4 aliquots: one aliquot was layered over a column of Androcoll-E-Large (SLC) and the second over a column of Percoll (45%). The third aliquot was deposited in a tube with culture medium and was incubated at a 45° angle for 30min at 37°C (SU1). The last aliquot was centrifuged to separate the spermatozoa and seminal plasma. The sperm pellet obtained was resuspended, and transferred to a tube with culture medium which was incubated at an angle of 45° for 30min at 37°C (SU2). Both aliquots SLC and P showed higher proportions of progressive motility and plasma membrane functionality (p≤0.05) than raw semen. There were no significant differences (p>0.05) in sperm viability and in normal spermatozoa between raw semen and treatments. Nevertheless, only SLC did not have a significant increase of bent tails. In conclusion SLC centrifugation would be the method of choice for selecting llama spermatozoa.

  20. Male facial attractiveness and masculinity may provide sex- and culture-independent cues to semen quality.

    PubMed

    Soler, C; Kekäläinen, J; Núñez, M; Sancho, M; Álvarez, J G; Núñez, J; Yaber, I; Gutiérrez, R

    2014-09-01

    Phenotype-linked fertility hypothesis (PLFH) predicts that male secondary sexual traits reveal honest information about male fertilization ability. However, PLFH has rarely been studied in humans. The aim of the present study was to test PLFH in humans and to investigate whether potential ability to select fertile partners is independent of sex or cultural background. We found that on the contrary to the hypothesis, facial masculinity was negatively associated with semen quality. As increased levels of testosterone have been demonstrated to impair sperm production, this finding may indicate a trade-off between investments in secondary sexual signalling (i.e. facial masculinity) and fertility or status-dependent differences in investments in semen quality. In both sexes and nationalities (Spanish and Colombian), ranked male facial attractiveness predicted male semen quality. However, Spanish males and females estimated facial images generally more attractive (gave higher ranks) than Colombian raters, and in both nationalities, males gave higher ranks than females. This suggests that male facial cues may provide culture- and sex-independent information about male fertility. However, our results also indicate that humans may be more sensitive to facial attractiveness cues within their own populations and also that males may generally overestimate the attractiveness of other men to females.

  1. The impact of male overweight on semen quality and outcome of assisted reproduction

    PubMed Central

    Thomsen, Lise; Humaidan, Peter; Bungum, Leif; Bungum, Mona

    2014-01-01

    It is well-documented that male overweight and obesity causes endocrine disorders that might diminish the male reproductive capacity; however, reports have been conflicting regarding the influence of male body mass index (BMI) on semen quality and the outcome of assisted reproductive technology (ART). The aim of this study was to investigate whether increased male BMI affects sperm quality and the outcome of assisted reproduction in couples with an overweight or obese man and a non-obese partner. Data was prospectively collected from 612 infertile couples undergoing ART at a Danish fertility center. Self-reported information on paternal height and weight were recorded and BMI was calculated. The men were divided into four BMI categories: underweight BMI < 20 kg m−2, normal BMI 20–24.9 kg m−2, overweight BMI 25–29.9 kg m−2 and obese BMI > 30 kg m−2. Conventional semen analysis was performed according to the World Health Organization guideline and sperm DNA integrity was analyzed by the Sperm Chromatin Structure Assay (SCSA). No statistically significant effect of male BMI was seen on conventional semen parameters (sperm concentration, total sperm count, seminal volume and motility) or on SCSA-results. Furthermore, the outcome of ART regarding fertilization rate, number of good quality embryos (GQE), implantation and pregnancy outcome was not influenced by the increasing male BMI. PMID:24759576

  2. Comparative fertility of freshly collected vs frozen-thawed semen with laparoscopic oviductal artificial insemination in domestic cats.

    PubMed

    Lambo, C A; Grahn, R A; Lyons, L A; Bateman, H l; Newsom, J; Swanson, W F

    2012-12-01

    Artificial insemination (AI) is potentially invaluable as an adjunct to natural breeding for the conservation management of non-domestic felid populations. The efficacy of AI, however, must be substantially improved for applied use, especially when using frozen semen. Our recent advances in using laparoscopic oviductal AI (LO-AI) with low sperm numbers and freezing of cat semen in a soy lecithin-based cryoprotectant medium suggest that combining these two approaches might improve pregnancy outcomes with frozen-thawed spermatozoa. In this study, our objectives were to (i) assess the effect of two gonadotropin dosages (100 vs 150 IU eCG) on ovarian response in domestic cats and (ii) compare the relative fertility of frozen-thawed and fresh semen in vivo following LO-AI. All 16 females ovulated after gonadotropin treatment and were inseminated with fresh semen from one male and frozen-thawed semen from a second male. There were no differences between gonadotropin dosages in CL number, pregnancy percentage or litter size. Half (8/16) of the females conceived, with seven females giving birth to a total of 36 offspring. Paternity analysis showed that more kittens resulted from LO-AI with fresh (28/36, 78%) than frozen-thawed (8/36, 22%) semen, possibly due to impaired motility and longevity of thawed sperm. These results demonstrated that viable offspring can be produced by AI using semen frozen in a soy lecithin-based medium. Insemination with greater numbers of frozen-thawed spermatozoa, combined with further refinement of cat sperm cryopreservation methods, may be necessary to optimize pregnancy success with LO-AI in domestic and nondomestic cats.

  3. Use of cholesterol-loaded cyclodextrin in donkey semen cryopreservation improves sperm viability but results in low fertility in mares.

    PubMed

    Oliveira, R R; Rates, D M; Pugliesi, G; Ker, P G; Arruda, R P; Moraes, E A; Carvalho, G R

    2014-10-01

    The use of cholesterol-loaded cyclodextrin (CLC) on semen cryopreservation has been related with better sperm viability in several species; however, the effect on fertility is not known in donkey semen. Ejaculates (n = 25) from five donkeys were diluted in S-MEDIUM with 0, 1, 2 or 3 mg of CLC/120 × 10(6) spermatozoa. Semen was frozen, and thawed samples were evaluated by computer-assisted sperm analyser system (CASA), supravital test, hyposmotic swelling test and fluorescent dyes to assess the integrity of sperm membranes. Mares (n = 60) were inseminated with frozen-thawed semen treated with the doses of 0 or 1 mg CLC. Percentages of sperm with progressive motility and with functional plasma membrane were greater (p < 0.05) in the CLC-treated groups than in the control. Percentages of intact plasma membrane and intact plasma membrane and acrosome detected by fluorescent dyes were also greater (p < 0.05) in CLC-treated groups. Although no difference (p > 0.05) in conception rates was detected between groups (control, 3/30, 10%; CLC-treated, 1/30, 3.3%), fertility was low for artificial insemination programs in mares. Therefore, we firstly demonstrated that frozen semen treated with CLC in S-MEDIA extender before freezing improves the in vitro sperm viability, but semen treated or not with CLC in S-MEDIUM extender results in a very low conception rate in mares inseminated with thawed donkey semen.

  4. Semen collection and polymerase chain reaction-based sex determination of black-headed and straw-necked ibis.

    PubMed

    Kaneko, K; Uematsu, E; Takahashi, Y; Tong, B; Takino, S; Wajiki, Y; Kimura, T; Yamashiro, H; Kaneko, Y; Iwaisaki, H; Sugiyama, T; Yamada, T; Yamagishi, S

    2013-12-01

    This study aimed to develop a polymerase chain reaction (PCR)-based sexing and effective semen collection methods for black-headed and straw-necked ibis species. However, most birds are not sexually dimorphic, that is, the sexes appear similar. Therefore, the gender should be determined before semen collection. DNA was extracted from the blood samples of 11 black-headed and 4 straw-necked ibis. The sex was determined after PCR amplification of the EE0.6 region of W-chromosome. The PCR products were separated using gel electrophoresis. A single band indicated the presence of the EE0.6 region and that the individual was a female, while no band indicated that the individual was a male. Further, the single bands from seven specimens were amplified. Semen collection was performed by massage or a combination of massage with electro-ejaculation and was attempted during all four seasons. The semen was successfully collected in March from male straw-necked ibis using the massage method. Limited motility, viability and concentration of straw-necked ibis sperm were observed. The sperm length was 180 μm and that of the nucleus was 30 μm with acrosome located at the tip of the nucleus. Thus, the PCR-based sexing proved to be an accurate molecular sexing method for black-headed and straw-necked ibis. Furthermore, we successfully collected semen and observed the stained sperm nucleus and acrosome of the straw-necked ibis sperm. We propose that the use of this PCR methodology can be applied as a routine method for sex determination and semen collection in ibis species for future ecological research. However, considering our limited success, further studies on semen collection method are required.

  5. Comparison of the effect of semen from HIV-infected and uninfected men on CD4+ T-cell infection

    PubMed Central

    Camus, Céline; Matusali, Giulia; Bourry, Olivier; Mahe, Dominique; Aubry, Florence; Bujan, Louis; Pasquier, Christophe; Massip, Patrice; Ravel, Célia; Zirafi, Onofrio; Munch, Jan; Roan, Nadia R.; Pineau, Charles; Dejucq-Rainsford, Nathalie

    2016-01-01

    Objectives: Semen composition is influenced by HIV-1 infection, yet the impact of semen components on HIV infection of primary target cells has only been studied in samples from HIV-uninfected donors. Design: We compared the effect of seminal plasma (SP) from chronically HIV-infected (SP+) versus uninfected donors (SP–) on HIV-1 infection of peripheral blood mononuclear cells (PBMCs) and CD4+ T cells. Methods: Primary cells were infected with HIV-1 in the presence of SP+ or SP– and analyzed for infection level, metabolic activity, HIV receptor expression, proliferation and activation. SP+ and SP– were compared for infection-enhancing peptides, cytokines and prostaglandin E2 levels. Results: SP– efficiently enhanced HIV-1 R5 infection of CD4+ T cells, whereas SP+ enhancing activity was significantly reduced. RANTES (CCL5) concentrations were elevated in SP+ relative to SP–, whereas the concentrations of infectivity-enhancing peptides [semen-derived enhancer of viral infection (SEVI), SEM1, SEM2] were similar. CCR5 membrane expression levels were reduced on CD4+ T cells shortly postexposure to SP+ compared with SP– and correlated to R5-tropic HIV-1 infection levels, and CCR5 ligands’ concentrations in semen. SP+ and SP– displayed similar enhancing activity on PBMC infection by X4-tropic HIV-1. Addition/depletion of RANTES (regulated on activation, normal T-cell expressed and secreted) from SPs modulated their effect on PBMC infection by R5-tropic HIV-1. Conclusion: Semen from HIV-infected donors exhibits a significantly reduced enhancing potential on CD4+ T-cell infection by R5-tropic HIV-1 when compared with semen from uninfected donors. Our data indicate that elevated seminal concentrations of RANTES in HIV-infected men can influence the ability of semen to enhance infection. PMID:26854806

  6. Effect of vitamin E on lipid peroxidation and fertility after artificial insemination with liquid-stored turkey semen.

    PubMed

    Long, J A; Kramer, M

    2003-11-01

    Turkey sperm plasma membranes contain high levels of polyunsaturated fatty acids that are susceptible to lipid peroxidation during in vitro storage at 4 degrees C. Herein we assessed the degree of lipid peroxidation and fertility potential of semen liquid-stored for 24 h with the antioxidant vitamin E. Semen was collected weekly from 44 males and pooled as pairs (total = 22); the individuals in paired samples exhibited similar semen quality parameters. After initial semen evaluation, pooled samples were extended with Beltsville Poultry Semen Extender containing no supplement (control) or 10 or 40 microg/mL vitamin E and then stored at 4 degrees C with constant aeration for 24 h. Lipid peroxidation was determined by measuring malonaldehyde (MDA) in aliquots (50 x 10(6) sperm) of fresh (0 h) and stored (24 h) semen. Sperm mobility was also evaluated. A total of 176 hens (8 hens/tom pair; 4 hens/0 h, 4 hens/24 h) were inseminated (150 x 10(6) sperm) weekly for 6 wk, and fertility was determined after 7 d of incubation. Initial MDA values of the 22 tom pairs ranged from 0.928 to 1.36 uM. Males varied in production of MDA during in vitro storage, with most pairs exhibiting a threefold increase. Results indicated that supplemental vitamin E did not reduce lipid peroxidation during liquid storage. Not surprisingly, artificial insemination with stored semen (with much higher MDA values) yielded lower fertility rates than control regardless of the presence of vitamin E. These results demonstrate that lipid peroxidation is a significant factor affecting the fertility of stored turkey sperm and that methods to prevent or reduce lipid peroxidation remain to be elucidated.

  7. RNA/DNA co-analysis from human saliva and semen stains--results of a third collaborative EDNAP exercise.

    PubMed

    Haas, C; Hanson, E; Anjos, M J; Banemann, R; Berti, A; Borges, E; Carracedo, A; Carvalho, M; Courts, C; De Cock, G; Dötsch, M; Flynn, S; Gomes, I; Hollard, C; Hjort, B; Hoff-Olsen, P; Hríbiková, K; Lindenbergh, A; Ludes, B; Maroñas, O; McCallum, N; Moore, D; Morling, N; Niederstätter, H; Noel, F; Parson, W; Popielarz, C; Rapone, C; Roeder, A D; Ruiz, Y; Sauer, E; Schneider, P M; Sijen, T; Court, D Syndercombe; Sviežená, B; Turanská, M; Vidaki, A; Zatkalíková, L; Ballantyne, J

    2013-02-01

    A third collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling was organized by the European DNA Profiling Group (EDNAP). Twenty saliva and semen stains, four dilution series (10-0.01 μl saliva, 5-0.01 μl semen) and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by 20 participating laboratories using an RNA extraction or RNA/DNA co-extraction method. Two novel mRNA multiplexes were used: a saliva triplex (HTN3, STATH and MUC7) and a semen pentaplex (PRM1, PRM2, PSA, SEMG1 and TGM4). The laboratories used different chemistries and instrumentation and a majority (16/20) were able to successfully isolate and detect mRNA in dried stains. The simultaneous extraction of RNA and DNA from individual stains not only permitted a confirmation of the presence of saliva/semen (i.e. tissue/fluid source of origin), but allowed an STR profile of the stain donor to be obtained as well. The method proved to be reproducible and sensitive, with as little as 0.05 μl saliva or semen, using different analysis strategies. Additionally, we demonstrated the ability to positively identify the presence of saliva and semen, as well as obtain high quality DNA profiles, from old and compromised casework samples. The results of this collaborative exercise involving an RNA/DNA co-extraction strategy support the potential use of an mRNA based system for the identification of saliva and semen in forensic casework that is compatible with current DNA analysis methodologies.

  8. Conception rates in ewes after AI with ram semen preserved in milk-egg yolk extenders supplemented with glycerol.

    PubMed

    Olivera-Muzante, J; Fierro, S; Gil, J

    2011-06-01

    This study aimed at comparing the effect of ram semen preserved at 5 °C on two milk-based extenders (UHT skim milk or INRA-96(®) , 5% egg yolk) supplemented with 2% glycerol, and the preservation time (24 and 48 h) on conception rates after cervical AI of ewes. In two field trials, 1198 Merino ewes were cervical AI in spontaneous oestrus. In Experiment 1, pooled semen (6 rams) was extended in UHT-base (fresh, control) or chilled for 24 h in UHT5Y (UHT-base 5% egg yolk), INRA5Y (INRA-96(®) 5% egg yolk), UHT5Y2G (UHT5Y 2% glycerol) or INRA5Y2G (INRA5Y 2% glycerol). In Experiment 2, AI was performed with pooled semen (7 rams) used fresh (extended in UHT-base or UHT5Y2G, control groups) or chilled (extended in UHT5Y2G) for 24 or 48 h. Conception rate was determined by ultrasound 40 days after AI. INRA-96(®) - had similar conception as UHT-preserved semen (56.7 vs 55.4%, p>0.05). Addition of 2% glycerol did not modify the results (56.8 vs 55.2%, p>0.05). Fresh semen extended in UHT-base, and UHT5Y2G yielded similar conception rates (60 vs 64%, p>0.05). Preservation for 24 or 48 h in UHT5Y2G gave similar results (49 vs 47%; p>0.05). In conclusion, ram semen chilled for 24 h in UHT- or INRA-96(®) -based extenders yielded similar results, and glycerol addition did not have a detrimental effect. UHT5Y2G might be used to extend ram semen for fresh AI, or to preserve it for 24 or 48 h with acceptable results.

  9. Cryopreservation of collared peccary (Pecari tajacu) semen using different freezing curves, straw sizes, and thawing rates.

    PubMed

    Silva, M A; Peixoto, G C X; Castelo, T S; Lima, G L; Silva, A M; Oliveira, M F; Silva, A R

    2013-08-01

    The objective of this study was to verify the effect of different freezing curves, straw sizes, and thawing rates on the cryopreservation of collared peccary semen. Twelve ejaculates were obtained from captive adult males by electroejaculation, and evaluated for sperm motility, kinetic rating, viability, morphology, and functional membrane integrity. The ejaculates were diluted in a coconut water extender (ACP-116c) with egg yolk and glycerol, packaged into 0.25 mL or 0.50 mL plastic straws and cryopreserved in liquid nitrogen following a slow (-10 °C/min) or a fast (-40 °C/min) freezing curve. After one week, samples were thawed at 37 °C/1 min or 70 °C/8s and evaluated as reported for fresh semen, and also for kinematic parameters (computerized analysis). A significant decrease in sperm motility and kinetic rating was observed after glycerol addition at 5 °C and also after thawing for all the treatments (P<0.05). Regarding post-thaw semen variables, no differences were verified between freezing curves when the same straw size and thawing rate were taken as reference (P>0.05). In general, values for sperm characteristics found after thawing at 37 °C were better preserved than at 70 °C (P<0.05), both in the use of 0.25 mL or 0.50 mL straws, which were similar for semen packaging (P>0.05). The evaluation of the kinematic parameters of sperm motility confirmed these results at values varying from 20% to 30% motile sperm for the samples tha wed at 37 °C, and values fewer than 12% motile sperm for samples thawed at 70 °C (P<0.05). In conclusion, we recommend the use of a fast freezing curve that reduces the time spent on the cryopreservation of collared peccary semen, which could be packaged both in 0.25 mL or 0.50 mL straws, but the thawing should be conducted at 37 °C/1 min.

  10. Endocrine alterations around the time of abortion in mares impregnated with donkey or horse semen.

    PubMed

    Boeta, M; Zarco, L

    2010-08-01

    The objective of this study was to monitor and compare the concentrations of equine chorionic gonadotropin (eCG), progesterone and estrone sulphate during normal and failed pregnancies of mares impregnated with donkey or horse semen, relating their individual endocrine profiles to the time of pregnancy loss, and to the histopathologic findings in the aborted fetuses and placenta. Mares (n=54) were used, 32 of them impregnated with donkey semen and 22 impregnated with horse semen. Blood samples were taken twice a week from Day 35 to 120 of pregnancy. Ultrasonographic observations of the fetus were carried out twice a week. The incidence of abortion in mares impregnated with donkey semen (30%) was greater (P<0.05) than the 5% observed in mares impregnated with horse semen. From Week 8 to the end of the sampling period, the mean progesterone concentrations of mares with normal mule pregnancies were less (P<0.05) than those of mares with normal pregnancies with equine fetuses. The concentrations of eCG were less (P<0.05) in mule pregnancies from Week 6. Estrone sulphate concentrations were only different (P<0.05) between types of pregnancy on Weeks 13 and 14, being in this case greater with the mule pregnancies. Most of the abortions of mule fetuses were associated with lesser progesterone concentrations than the average for mares with successful mule pregnancies. Four of the abortions of mule fetuses and the only abortion of horse fetus occurred in mares with lesser progesterone and very low eCG concentrations, and were classified as caused by luteal impairment secondary to eCG deficiency; estrone sulphate concentrations were less than normal or absent before these abortions. Two mares aborted after several weeks of low progesterone concentrations in the presence of eCG concentrations that were normal for mule pregnancies, suggesting primary luteal deficiency. In three mares carrying a mule fetus, the concentrations of progesterone and estrone sulphate decreased

  11. Risks of transmitting ruminant spongiform encephalopathies (prion diseases) by semen and embryo transfer techniques.

    PubMed

    Wrathall, A E; Holyoak, G R; Parsonson, I M; Simmons, H A

    2008-09-15

    Early experiments suggested that scrapie transmission via sheep embryos was a possibility, and gave rise to much controversy. However, when account is taken of the complex genetic effects on ovine susceptibility to scrapie, and of the several different scrapie strains with different clinical and pathological effects, the overall conclusion now is that transmission of classical scrapie by embryo transfer is very unlikely if appropriate precautions are taken. Recent embryo transfer studies have confirmed this. Other studies in sheep have shown that from about the middle of pregnancy the placental trophoblast is liable to scrapie infection in genetically susceptible ewes if the fetus is also susceptible. Since the contrary is also true, use of resistant ewes as embryo recipients could add to the safety of the embryo transfer, at least for classical scrapie. There has been little recent research on scrapie transmission via semen in sheep, and, with hindsight, the early studies, though negative, were inadequate. There is scant information on scrapie transfer via goat semen or embryos, although one study did find that bovine spongiform encephalopathy (BSE) was not transmitted via goat embryos. In cattle it has been shown that, if appropriate precautions are taken, the risks of transmitting BSE via semen and in vivo-derived embryos are negligible, and this conclusion has gained worldwide acceptance. Research on TSE transmission via reproductive technologies in deer has not yet been done, but information on the pathogenesis and epidemiology of chronic wasting disease (CWD) of deer, and on transmission risks in other species, provides optimism that transmission of CWD via semen and embryos of deer is unlikely. The presence of TSE infectivity in blood and various other tissues of infected animals, particularly sheep, gives rise to concerns that certain biological products currently used in reproductive technologies, e.g. pituitary gonadotrophins for superovulation, and

  12. The role of international transport of equine semen on disease transmission.

    PubMed

    Metcalf, E S

    2001-12-03

    Despite the numerous benefits of having the capability to transport semen internationally, there are serious potential ramifications if that semen is contaminated with a communicable disease. Bacteria: Many commensal bacteria colonize the exterior of the stallion penis and are not regarded as pathogenic. They may be cultured from an ejaculate. Alterations of the normal bacterial flora on the exterior genitalia may cause the growth of opportunistic bacteria such as Klebsiella pneumonia, Pseudomonas aeruginosa, Streptococcus zooepidemicus, which, if inseminated, may cause infertility in susceptible mares. Contagious equine metritis (CEM), a highly transmissible, true venereal disease of horses, is caused by the gram-negative coccobacillis, Taylorella equigenitalis. Even with the use of rigorous testing protocols, the current techniques used may not ensure accuracy of results. Viruses: Equine coital exanthema (equine herpes virus type 3; EHV-3) is a highly contagious virus that causes painful lesions on the stallion's penis and mare's vulva. Although it is primarily transmitted through coitus, infected fomites have also been implicated in its spread. Therefore, it is possible that the virus can potentially be transmitted to the ejaculate through penile contact with an artificial vagina or sleeve. Equine arteritis virus appears to be becoming more prevalent in recent years. The most common method of transmission is through respiratory disease, but the organism can also be shed in the semen of asymptomatic stallions. Equine infectious anemia virus has also been found to be present in the semen of an infected stallion, although no evidence exists at this time that there is venereal transmission of this disease. Protozoa: Dourine, caused by Trympanosoma equiperidum, is a venereal disease found only in Africa, South and Central America and the Middle East. Serological testing using complement fixation is recommended for diagnosis. Piroplasmosis, a disease caused by Babesia

  13. Communication requested: Boar semen transport through the uterus and possible consequences for insemination.

    PubMed

    Rath, D; Knorr, C; Taylor, U

    2016-01-01

    Recent insemination techniques bypass the interactions between sperm and the uterine wall because the semen is deposited deep into the tip of uterine horn or directly into the oviduct. Such techniques allow high dilution of the ejaculates. After normal mating, semen entering the uterus communicates with the uterine milieu. Intact sperm of high mitochondrial membrane potential bind to uterine epithelial cells, whereas most of the unbound sperm in the uterine lumen have damaged membranes. Lectins are the most likely factors to mediate these sperm-uterine interactions. The lectin wheat germ agglutinin is known to induce the strongest binding of sperm, whereas binding is impaired when sialic acid receptors are blocked by wheat germ agglutinin. This suggests that sialic acid is involved in porcine sperm-endometrium interactions, and it is hypothesized that the use of a semen extender supplemented with sialidase would allow insemination with reduced sperm numbers. A lack of contact of sperm and seminal plasma with the uterine wall, as a result of deep insemination, may adversely affect (1) events during ovulation, (2) induction of immunologic tolerance against paternal antigens, (3) preparation of the endometrium for implantation and placentation, and (4) immunologic support required for the fetus during pregnancy. Seminal plasma is known to signal post-insemination changes in the uterine endometrium involving the redistribution of leukocytes. This may involve migration of leukocytes from the uterine wall to the ovary, as seminal plasma particularly increases the appearance of the major histocompatibility complex class II-positive cells. Uterine epithelial cells respond to sperm binding by the production of pro- or anti-inflammatory cytokines. These cytokines may include synchronizing substances, transferred through a counter-current pathway to the ipsilateral ovary, thereby accelerating the final maturation of preovulatory follicles and advancing time of ovulation. In

  14. 107 RESEARCH PRIORITIES FOR SAFE SANITARY TRADE OF EMBRYO AND SEMEN.

    PubMed

    Fieni, F; Grant, C; Gard-Schnuelle, J; Perry, G; Wrenzycki, C; Blondin, P

    2016-01-01

    Embryo transfer and artificial insemination are utilised nationally and internationally for the introduction, improvement, and preservation of livestock genetics. Embryos present a lower risk of infectious disease transmission than do live animals. In order to maintain the sanitary security and to facilitate the trade of embryos and semen worldwide, the Health and Scientific Advisory Committee of the International Embryo Technology Society designed, developed, and conducted a survey to determine research priority. The survey questionnaire was sent to 32 government representatives and 76 embryo transfer and artificial insemination industry representatives around the world to countries where artificial breeding industries are active and well developed. A total of 16 answers were received, 9/32 (28%) from government representatives and 7/76 (9%) from industry representatives. The global feedback was 15%. The survey indicated that, in terms of research priority, embryos and semen were equally important. With regards to embryo research priorities, the survey results ranked in vitro-produced embryos research as the most important, followed by in vitro-derived embryos, and then oocytes. Apart from scrapie for embryos and Campylobacteriosis for semen, research priorities were similar for the major pathogens of embryos and semen, in particular, bovine viral diarrhoea and paratuberculosis (Johne's disease; Table 1). Emerging or less common diseases were not forgotten. Other diseases suggested but not listed in Table 1 included bluetongue, foot and mouth disease, lentivirus, arbovirus, bovine tuberculosis, porcine epidemic diarrhoea, porcine reproductive and respiratory syndrome, African swine fever, and ovine pulmonary adenocarcinoma (Jaagsiekte). The survey highlighted the need to focus research largely on ruminant species (Table 1). Other issues identified by the survey included (i) alternative or indirect processes for determining the sanitary quality of in vitro

  15. Factors affecting sperm recovery rates and survival after centrifugation of equine semen.

    PubMed

    Ferrer, M S; Lyle, S K; Eilts, B E; Eljarrah, A H; Paccamonti, D L

    2012-11-01

    Conventional centrifugation protocols result in important sperm losses during removal of the supernatant. In this study, the effect of centrifugation force (400 or 900 × g), duration (5 or 10 min), and column height (20 or 40 mL; Experiment 1); sperm concentration (25, 50, and 100 × 10(6)/mL; Experiment 2), and centrifugation medium (EZ-Mixin CST [Animal Reproduction Systems, Chino, CA, USA], INRA96 [IMV Technologies, Maple Grove, MN, USA], or VMDZ [Partnar Animal Health, Port Huron, MI, USA]; Experiment 3) on sperm recovery and survival after centrifugation and cooling and storage were evaluated. Overall, sperm survival was not affected by the combination of centrifugation protocol and cooling. Total sperm yield was highest after centrifugation for 10 min at 400 × g in 20-mL columns (95.6 ± 5%, mean ± SD) or 900 × g in 20-mL (99.2 ± 0.8%) or 40-mL (91.4 ± 4.5%) columns, and at 900 × g for 5 min in 20-mL columns (93.8 ± 8.9%; P < 0.0001). Total (TMY) and progressively motile sperm yield followed a similar pattern (P < 0.0001). Sperm yields were not significantly different among samples centrifuged at various sperm concentrations. However, centrifugation at 100 × 10(6)/mL resulted in significantly lower total sperm yield (83.8 ± 10.7%) and TMY (81.7 ± 6.8%) compared with noncentrifuged semen. Centrifugation in VMDZ resulted in significantly lower TMY (69.3 ± 22.6%), progressively motile sperm yield (63.5 ± 18.2%), viable yield (60.9 ± 36.5%), and survival of progressively motile sperm after cooling (21 ± 10.8%) compared with noncentrifuged semen. In conclusion, centrifuging volumes of ≤ 20 mL minimized sperm losses with conventional protocols. With 40-mL columns, it may be recommended to increase the centrifugal force to 900 × g for 10 min and dilute the semen to a sperm concentration of 25 to 50 × 10(6)/mL in a milk- or fractionated milk-based medium. The semen extender VMDZ did not seem well suited for centrifugation of equine semen.

  16. Urinary Bisphenol A Levels in Young Men: Association with Reproductive Hormones and Semen Quality

    PubMed Central

    Frederiksen, Hanne; Jensen, Tina Kold; Petersen, Jørgen Holm; Joensen, Ulla N.; Main, Katharina M.; Skakkebaek, Niels E.; Juul, Anders; Jørgensen, Niels; Andersson, Anna-Maria

    2014-01-01

    Background: Few human studies have examined bisphenol A (BPA) exposure in relation to semen quality and reproductive hormones in men, and results are divergent. Objectives: We examined associations between urinary BPA concentration and reproductive hormones, as well as semen quality, in young men from the general population. Methods: Our study population consisted of 308 young men from the general population. Urinary BPA concentration was measured by isotope dilution TurboFlow-liquid chromatography–tandem mass spectrometry. We used multiple linear regression analysis to estimate associations between BPA concentration and reproductive hormones and semen quality, adjusting for confounding factors. Results: We found that 98% of the men had detectable urinary levels of BPA. Median (5th–95th percentiles) BPA concentration was 3.25 ng/mL (0.59–14.89 ng/mL). Men with BPA concentrations above the lowest quartile had higher concentrations of serum testosterone, luteinizing hormone (LH), estradiol, and free testosterone compared with the lowest quartile (ptrend ≤ 0.02). Men in the highest quartile of BPA excretion had on average 18% higher total testosterone (95% CI: 8, 28%), 22% higher LH (95% CI: 6, 39%), and 13% higher estradiol (95% CI: 4, 24%) compared with lowest quartile. Men in the highest quartile of BPA also had significantly lower percentage progressive motile spermatozoa compared with men in the lowest quartile (–6.7 percentage points, 95% CI: –11.76, –1.63). BPA was not associated with other semen parameters. Adjusting for dietary patterns did not influence the results. Conclusions: The pattern of associations between BPA and reproductive hormones could indicate an antiandrogenic or antiestrogenic effect, or both, of BPA on the hypothalamic–pituitary–gonadal hormone feedback system, possibly through a competitive inhibition at the receptor level. However, additional research is needed to confirm our findings and to further test the suggested

  17. Effect of age and season on semen quality parameters in Sahiwal bulls.

    PubMed

    Bhakat, Mukesh; Mohanty, T K; Raina, V S; Gupta, A K; Khan, H M; Mahapatra, R K; Sarkar, M

    2011-08-01

    The objective of the study was to determine the effect of season, period, age, bull, and ejaculate on semen quality in Sahiwal bulls. Semen production records from 1996 to 2006 of 5,483 ejaculates from 46 Sahiwal bulls maintained at Artificial Breeding Complex, NDRI, Karnal, India were analyzed using least square analysis of variance by LSML software package. The overall least squares means of ejaculate volume (VOL), total volume per day (VOLD), mass activity (MA), initial motility (IM), sperm concentration per ml (SPC), and sperm concentration per ejaculate (SPCE) were 3.79 ± 0.02 ml, 5.81 ± 0.06 ml, 2.32 ± 0.01, 55.47 ± 0.001%, 766.69 ± 5.50 × 10(6)/ml and 3023.25 ± 30.15 × 10(6), respectively. All semen traits (VOL, VOLD, MA, IM and SPCE) were significantly (P < 0.01) affected by age groups, season and period, whereas season had significant effect on VOL at 5% level. During hot-humid season, highest value of VOL, VOLD, MA, IM, SPC, and SPCE were observed followed by summer and cold season. Highest value of VOL, VOLD, IM, and SPCE were observed during period-3 (2004-2006), whereas highest value of MA and SPC were observed during period-1 (1996-1999). However, lowest magnitude of MA, IM, SPC, and SPCE during period-2 (2000-2003) was observed. Ejaculate characteristics like VOL, VOLD, and SPCE increased with the increasing age of bull up to 5 years and then decreased. Significant (P < 0.01) bull to bull variation was found in VOL, VOLD, MA, IM, SPC, and SPCE traits. First ejaculate had significantly (P < 0.01) higher MA, IM, SPC, and SPCE. Hence, it could be concluded that during rainy season and period-1 and period-3 the quality of semen is quantitatively and qualitatively good. Better quality semen was obtained up to 5 years of age in Sahiwal bulls.

  18. Evaluation of the presence of equine viral herpesvirus 1 (EHV-1) and equine viral herpesvirus 4 (EHV-4) DNA in stallion semen using polymerase chain reaction (PCR).

    PubMed

    Hebia-Fellah, Imen; Léauté, Anne; Fiéni, Francis; Zientara, Stéphan; Imbert-Marcille, Berthe-Marie; Besse, Bernard; Fortier, Guillaume; Pronost, Stephane; Miszczak, Fabien; Ferry, Bénédicte; Thorin, Chantal; Pellerin, Jean-Louis; Bruyas, Jean-François

    2009-06-01

    In the horse, the risk of excretion of two major equine pathogens (equine herpesvirus types 1 (EHV-1) and 4 (EHV-4)) in semen is unknown. The objective of our study was to assess the possible risks for the horizontal transmission of equine rhinopneumonitis herpesviruses via the semen and the effect of the viruses on stallion fertility. Samples of stallion semen (n=390) were gathered from several different sources. Examination of the semen involved the detection of viral DNA using specific PCR. The mean fertility of the stallions whose sperm tested positive for viral DNA and the mean fertility of stallions whose sperm did not contain viral DNA, were compared using the Student's t-test. EHV-4 viral DNA was not detected in any of the semen samples. EHV-1 DNA was identified in 51 of the 390 samples, (13%). One hundred and eighty-two samples came from 6 studs and there was significant difference (p<0.05) among the proportion of stallions whose semen tested positive for viral DNA from 0 to 55% between the studs. There was a significant difference (p<0.014) between the fertility of stallions whose semen tested positive for viral DNA and those whose semen was free from viral DNA. The stallions that excreted the EHV-1 virus in their semen appeared to be more fertile than the non-excretors, but this difference was in fact related to the breeding technique since higher proportion of excretors were found among those whose semen was used fresh rather than preserved by cooling or freezing. In conclusion, this study suggests that the EHV-1 virus may be transmitted via the semen at mating or by artificial insemination as demonstrated with other herpes viruses in other species.

  19. Seminal traits, suitability for semen preservation and fertility in the native Portuguese horse breeds Puro Sangue Lusitano and Sorraia: Implications for stallion classification and assisted reproduction.

    PubMed

    Gamboa, Sandra; Machado-Faria, Manuel; Ramalho-Santos, João

    2009-07-01

    The Puro Sangue Lusitano (PSL) is the major national breed of horse in Portugal, but no studies exist on its seminal characteristics, or on the possibility of conserving semen for future use. The aim of this study was to evaluate semen parameters, fertility and the aptness to semen preservation in Lusitano Stallions. In order to compare characteristics defined by a single or by multiple semen collections per stallion 152 ejaculates obtained from 152 Lusitano stallions presented at an annual breeding soundness examination as well as data related to 371 ejaculates obtained from 9 PSL were analyzed. These latter samples were also evaluated in terms of their possible use in assisted reproduction and were compared with 113 ejaculates obtained from 4 Sorraia horses, a rare and endangered Portuguese breed. The percentage of motile spermatozoa (PMS) was assessed after collection (AC), after semen dilution (AD) and at 24h of cool-storage. Mean values obtained for sperm motility and morphology and semen pH observed after semen collection differ significantly (P<0.05) between single collection/multiple stallions and multiple collections/limited stallions, and no age related effects were detected. Overall, Lusitano semen quality was comparable to that of related breeds, while Sorraia stallions had very poor semen quality. The response to cool-storage of diluted semen samples differed among stallions and breeds, and the best results for progressive motile sperm cells at 24h were in a range of 35-53% for PSL stallions and were lower for Sorraia stallions. Fertility rates obtained with artificial insemination (AI) averaged at 85% for PSL. With the exception of PMS AC, sperm vitality and semen pH no other seminal trait seemed to influence fertility rates in the Lusitano breed.

  20. Post-Prostatic Massage Fluid/Urine (post-PMF/U) as an Alternative to Semen for Studying Male Genitourinary HIV-1 Shedding

    PubMed Central

    Graham, Susan M.; Krieger, John N.; Githua, Peter L. M.; Wamuyu, Lorraine W.; Wale, Steven; Ramko, Kelly M.; Dragavon, Joan A.; Muller, Charles H.; Holte, Sarah E.; Mandaliya, Kishor N.; McClelland, R. Scott; Peshu, Norbert M.; Sanders, Eduard J.; Coombs, Robert W.

    2011-01-01

    Objectives Genitourinary tract samples are required to investigate male HIV-1 infectivity. Because semen collection is often impractical, we evaluated the acceptability, feasibility, and validity of post-prostatic massage fluid/urine (post-PMF/U) for studying male genitourinary HIV-1 shedding. Methods HIV-1-seropositive men were evaluated after 48 hours of sexual abstinence. At each visit, a clinician performed prostatic massage, then post-PMF/U and blood were collected. Participants provided semen specimens one week later. An audio computer-assisted self-interview (ACASI) administered after each specimen collection evaluated acceptability, adherence to instructions, and recent genitourinary symptoms. HIV-1 RNA was quantified using a real-time PCR assay. Detection and quantitation of HIV-1 RNA and stability over visits were compared for semen, post-PMF/U, and blood. Results Post-PMF/U was successfully obtained at 106 visits (64%) and semen at 136 visits (81%, p<0.001). In ACASI, discomfort was rated higher for post-PMF/U collection (p=0.003), but there was no significant difference in acceptability. Detection of HIV-1 RNA in post-PMF/U was associated with detection in semen (p=0.02). Semen and post-PMF/U HIV-1 RNA levels were correlated (ρ=0.657, p<0.001). Concordance of results at repeat visits was 78.9% for post-PMF/U (κ = 0.519, p = 0.02), and 89.5% for both blood and semen (κ = 0.774, p = 0.001). Conclusions Although semen collections were more successful, both post-PMF/U and semen collections were acceptable to many participants. HIV-1 RNA detection and levels were closely associated in semen and post-PMF/U, and results were relatively stable across visits. To assess male HIV-1 infectivity, post-PMF/U may represent a valid alternative when semen cannot be obtained. PMID:21278399

  1. A Longitudinal Study of Peripubertal Serum Organochlorine Concentrations and Semen Parameters in Young Men: The Russian Children’s Study

    PubMed Central

    Mínguez-Alarcón, Lidia; Sergeyev, Oleg; Burns, Jane S.; Williams, Paige L.; Lee, Mary M.; Korrick, Susan A.; Smigulina, Luidmila; Revich, Boris; Hauser, Russ

    2016-01-01

    Background: Exposures to endocrine-disrupting chemicals during critical phases of testicular development may be related to poorer semen parameters. However, few studies have assessed the association between childhood organochlorine (OC) exposure and adult semen parameters. Objective: We examined whether peripubertal serum OC concentrations are associated with semen parameters among young Russian men. Methods: From 2003 through 2005, 516 boys were enrolled at age 8–9 years and followed for up to 10 years. Serum OCs were measured in the enrollment samples using high-resolution mass spectrometry. At 18–19 years, 133 young men provided 1 or 2 semen samples (256 samples) collected approximately 1 week apart, which were analyzed for volume, sperm concentration, and motility. Unadjusted and adjusted linear mixed models were used to examine the associations of quartiles of lipid-standardized concentrations of dioxins [2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), polychlorinated dibenzo-p-dioxins (PCDDs)], furans, polychlorinated biphenyls (PCBs), and corresponding toxic equivalents (TEQs) with semen parameters. Results: The median (range) for TCDD was 2.9 (0.4–12.1) pg/g lipid and PCDD TEQ was 8.7 (1.0–36.0) pg TEQ/g lipid. Higher quartiles of TCDD and PCDD TEQs were associated with lower sperm concentration, total sperm count, and total motile sperm count (p-trends ≤ 0.05). The highest quartile of peripubertal serum TCDD concentrations was associated with a decrease (95% CI) of 40% (18, 66%), 29% (3, 64%), and 30% (2, 70%) in sperm concentration, total sperm count, and total motile sperm count, respectively, compared with the lowest quartile. Similar associations were observed for serum PCDD TEQs with semen parameters. Serum PCBs, furans, and total TEQs were not associated with semen parameters. Conclusion: Higher peripubertal serum TCDD concentrations and PCDD TEQs were associated with poorer semen parameters. Citation: Mínguez-Alarcón L, Sergeyev O, Burns JS

  2. Detection of Simian Immunodeficiency Virus in Semen, Urethra, and Male Reproductive Organs during Efficient Highly Active Antiretroviral Therapy

    PubMed Central

    Matusali, G.; Dereuddre-Bosquet, N.; Le Tortorec, A.; Moreau, M.; Satie, A.-P.; Mahé, D.; Roumaud, P.; Bourry, O.; Sylla, N.; Bernard-Stoecklin, S.; Pruvost, A.; Le Grand, R.

    2015-01-01

    ABSTRACT A number of men receiving prolonged suppressive highly active antiretroviral therapy (HAART) still shed human immunodeficiency virus (HIV) in semen. To investigate whether this seminal shedding may be due to poor drug penetration and/or viral production by long-lived cells within male genital tissues, we analyzed semen and reproductive tissues from macaques chronically infected with simian immunodeficiency virus mac251 (SIVmac251) who were treated for 4 months with HAART, which was intensified over the last 7 weeks with an integrase inhibitor. We showed that a subset of treated animals continued shedding SIV in semen despite efficient HAART. This shedding was not associated with low antiretroviral drug concentrations in semen or in testis, epididymis, seminal vesicles, and prostate. HAART had no significant impact on SIV RNA in the urethra, whereas it drastically reduced SIV RNA levels in the prostate and vas deferens and to a lesser extent in the epididymis and seminal vesicle. The only detectable SIV RNA-positive cells within the male genital tract after HAART were urethral macrophages. SIV DNA levels in genital tissues were not decreased by HAART, suggesting the presence throughout the male genital tract of nonproductively infected cells. In conclusion, our results demonstrate that 4 months of HAART induced variable and limited control of viral infection in the male reproductive organs, particularly in the urethra, and suggest that infected long-lived cells in the male genital tract may be involved in persistent seminal shedding during HAART. These results pave the way for further investigations of male genital organ infection in long-term-treated infected individuals. IMPORTANCE A substantial subset of men receiving prolonged HAART suppressing viral loads in the blood still harbor HIV in semen, and cases of sexual transmission have been reported. To understand the origin of this persistence, we analyzed the semen and male reproductive tissues from SIV

  3. Urinary Phytoestrogens Are Associated with Subtle Indicators of Semen Quality among Male Partners of Couples Desiring Pregnancy123

    PubMed Central

    Mumford, Sunni L; Kim, Sungduk; Chen, Zhen; Barr, Dana Boyd; Louis, Germaine M Buck

    2015-01-01

    Background: Phytoestrogens have been associated with subtle hormonal changes, although effects on male fecundity are largely unknown. Objective: We evaluated associations between male urinary phytoestrogen (isoflavone and lignan) concentrations and semen quality. Methods: This study was a prospective cohort study of 501 male partners of couples desiring pregnancy and discontinuing contraception. Each participant provided up to 2 semen samples that were analyzed for 35 semen quality endpoints the following day. Linear mixed-effects models were used to estimate associations between baseline urinary phytoestrogen concentrations and semen quality parameters, adjusted for age, body mass index (BMI), research site, and serum lipid and cotinine concentrations. Results: Most associations between urinary phytoestrogens and semen quality parameters were null. However, select individual phytoestrogens were associated with semen quality parameters, with associations dependent on the class of phytoestrogens and modified by BMI. Specifically, genistein and daidzein were associated with a lower percentage of normal sperm and increased abnormalities in semen morphology, with reduced associations observed as BMI increased (P < 0.05) [percentages (95% CIs) of normal morphology by WHO traditional criteria: genistein, main effect: −5.61% (−9.42%, −1.79%); interaction: 0.19% (0.06%, 0.31%) per log unit increase; daidzein, main effect: −5.35% (−9.36%, −1.34%); interaction: 0.18% (0.05%, 0.32%) per log unit increase]. Enterolactone was associated with fewer abnormalities in semen morphometry and morphology and decreased DNA fragmentation, with reduced associations observed as BMI increased (P < 0.05) [percentages (95% CIs) of abnormalities in the neck and midpiece: enterolactone, main effect: −3.35% (−6.51%, −0.19%); interaction: 0.11% (0.01%, 0.21%) per log unit increase]. Conclusions: These results suggest that male urinary phytoestrogen concentrations characteristic

  4. HIV-1 RNA May Decline More Slowly in Semen than in Blood following Initiation of Efavirenz-Based Antiretroviral Therapy

    PubMed Central

    Graham, Susan M.; Holte, Sarah E.; Dragavon, Joan A.; Ramko, Kelly M.; Mandaliya, Kishor N.; McClelland, R. Scott; Peshu, Norbert M.; Sanders, Eduard J.; Krieger, John N.; Coombs, Robert W.

    2012-01-01

    Objectives Antiretroviral therapy (ART) decreases HIV-1 RNA levels in semen and reduces sexual transmission from HIV-1-infected men. Our objective was to study the time course and magnitude of seminal HIV-1 RNA decay after initiation of efavirenz-based ART among 13 antiretroviral-naïve Kenyan men. Methods HIV-1 RNA was quantified (lower limit of detection, 120 copies/mL) in blood and semen at baseline and over the first month of ART. Median log10 HIV-1 RNA was compared at each time-point using Wilcoxon Signed Rank tests. Perelson’s two-phase viral decay model and nonlinear random effects were used to compare decay rates in blood and semen. Results Median baseline HIV-1 RNA was 4.40 log10 copies/mL in blood (range, 3.20–5.08 log10 copies/mL) and 3.69 log10 copies/mL in semen (range, <2.08–4.90 log10 copies/mL). The median reduction in HIV-1 RNA by day 28 was 1.90 log10 copies/mL in blood (range, 0.56–2.68 log10 copies/mL) and 1.36 log10 copies/mL in semen (range, 0–2.66 log10 copies/mL). ART led to a decrease from baseline by day 7 in blood and day 14 in semen (p = 0.005 and p = 0.006, respectively). The initial modeled decay rate was slower in semen than in blood (p = 0.06). There was no difference in second-phase decay rates between blood and semen. Conclusions Efavirenz-based ART reduced HIV-1 RNA levels more slowly in semen than in blood. Although this difference was of borderline significance in this small study, our observations suggest that there is suboptimal suppression of seminal HIV-1 RNA for some men in the early weeks of treatment. PMID:22912795

  5. Comparison of fertility of liquid or frozen semen when varying the interval from CIDR removal to insemination.

    PubMed

    Richardson, Brittany N; Larimore, Erin L; Walker, Julie A; Utt, Matthew D; DeJarnette, J Mel; Perry, George A

    2017-03-01

    Cryopreservation allows for long-term storage of semen; however, it leads to damage of sperm that may result in complete loss of viability or changes that possibly decrease sperm functionality. Liquid semen is not exposed to these stressors and may result in a longer lifespan in the female reproductive tract, thus increasing the range in timing of insemination without affecting fertility. The objective of this study was to compare fertility of liquid and frozen semen when varying the interval from CIDR removal to AI using the 7-day CO-Synch+CIDR protocol for synchronization of time of estrus. Within age group, crossbred cows (n=389) were randomly assigned to insemination at 36 or 60h after CIDR removal with either liquid or frozen semen (36L, 60L, 36F, and 60F) from one of two Angus bulls. Cows were monitored for estrous activity from CIDR removal until 60h thereafter. Cows that failed to exhibit estrus received GnRH (100μg, i.m.), and a blood sample was collected for analysis of estradiol concentration. There was no difference in pregnancy rates when liquid or frozen semen (53% and 52%) was used, but cows inseminated at 60h had a greater (P<0.01) pregnancy rate than those inseminated at 36h (72% and 31%). There was no time of AI by semen type interaction (P=0.57). Estrus was detected in 63%, 61%, 56%, and 62% of 36F, 36L, 60F, and 60L, respectively (only 5% and 1% of 36F and 36L were detected in estrus before insemination). Overall cows that exhibited estrus had a greater pregnancy rate compared with cows that did not (P<0.01; 79% compared with 24%). Among cows that did not exhibit estrus, those inseminated with liquid semen tended to have greater pregnancy rates than those inseminated with frozen semen (P=0.06). Cows that became pregnant had greater (P<0.01) concentrations of estradiol at 60h than those that did not (10.7±0.55 compared with 7.9±0.26pg/mL). In summary, there was no difference in pregnancy success between liquid and frozen semen. However, cows

  6. [Effects of occupational exposure to pesticides on semen quality of workers in an agricultural community of Merida state, Venezuela].

    PubMed

    Miranda-Contreras, Leticia; Cruz, Ibis; Osuna, Jesús A; Gómez-Pérez, Roald; Berrueta, Lisbeth; Salmen, Siham; Colmenares, Melisa; Barreto, Silvio; Balza, Alirio; Morales, Yasmin; Zavala, Leisalba; Labarca, Emilitza; García, Nelly; Sanchez, Beluardi; Contreras, Carlos A; Andrade, Henry

    2015-06-01

    Numerous studies report adverse effects of pesticides on male reproductive health. The objectives of this study were to investigate whether there is a relationship between occupational exposure to pesticides and semen quality, and to determine whether chronic exposure to pesticides differentially affects semen quality in men of different ages. A comparative study of 64 farmers and 64 control men was performed. The farmers were interviewed to determine their occupational history and particularly, activities that may involve exposure to pesticides. Semen parameters were evaluated and a comparative analysis of semen variables between exposed and control groups, as well as between age groups: 18-29, 30-37 and 38-60 years was done. Significant alterations of some semen parameters in the exposed group were found, such as: decreases in sperm concentration, slow progressive motility and sperm membrane integrity; at the same time, increases in eosin Y positive and sperm DNA fragmentation index. The results obtained by age groups showed significant differences between exposed and control groups for the parameters of membrane integrity, eosin Y positive and sperm DNA fragmentation index, being the exposed group between 18-29 years that showed the highest altered cases of these parameters. Our results prove that occupational pesticide exposure is associated with alterations in sperm quality, creating a risk to farm workers in their reproductive capacity.

  7. Long-term preservation of chilled canine semen using vitamin C in combination with green tea polyphenol.

    PubMed

    Wittayarat, Manita; Kimura, Taichi; Kodama, Risa; Namula, Zhao; Chatdarong, Kaywalee; Techakumphu, Mongkol; Sato, Yoko; Taniguchi, Masayasu; Otoi, Takeshige

    2012-01-01

    Vitamin C and green tea polyphenol are known to have antioxidant effects. The aim of this study was to evaluate the quality of canine semen after preservation with diluents containing vitamin C and polyphenol at 5 degree C for 4 weeks. In experiment 1, we investigated the effects of vitamin C combined with polyphenol supplementation on chilled semen quality. The addition of vitamin C (0.5 or 1 mM) with 0.75 mg per mL polyphenol to semen extender provided significantly higher percentages of sperm motility and viability during cold storage compared to unsupplemented semen. In experiment 2, we determined the optimal working concentration of vitamin C in the semen extender by comparison of a range of concentrations between 0.1 and 20 mM. Supplementation of 0.5 mM vitamin C plus polyphenol yielded the highest percentages of sperm motility and viability; however, there was no beneficial effect on the plasma membrane and acrosomal integrity of the spermatozoa.

  8. Cryopreservation of canine semen using a coconut water extender with egg yolk and three different glycerol concentrations.

    PubMed

    Cardoso, Rita de Cássia Soares; Silva, Alexandre Rodrigues; Uchoa, Daniel Couto; da Silva, Lúcia Daniel Machado

    2003-02-01

    Semen was collected by digital manipulation from six adult dogs. The second fraction of the ejaculate was used in this study. The semen was assessed by macroscopic and microscopic criteria 1 min after collection, first dilution, cooling, glycerol addition and thawing. Experiments were conducted to compare the effect of three different concentrations of glycerol in coconut water extender. The freezing method employed was that one described for caprine semen with slight changes. Semen was thawed at 37 degrees C for 1 min. Spermatozoal motility after thawing was 49.2+/-26, 44.2+/-18.3 and 35.8+/-26.8% for groups with 4, 6 and 8% glycerol, respectively. The vigor after thawing was 2.6+/-1.1, 2.7+/-1.0 and 2.1+/-1.2 for these groups, respectively. There was no difference among groups in motility and vigor. However, a smaller percentage of total and secondary abnormalities was observed using 6% glycerol in coconut water extender. In conclusion, the three glycerol concentrations (4, 6 or 8%) can be used successfully in cryopreservation of canine semen using a coconut water extender.

  9. Glycol ethers and semen quality: a cross‐sectional study among male workers in the Paris Municipality

    PubMed Central

    Multigner, L; Brik, E Ben; Arnaud, I; Haguenoer, J M; Jouannet, P; Auger, J; Eustache, F

    2007-01-01

    Objectives Apparent increases in human male reproductive disorders, including low sperm production, may have occurred because of increased chemical exposure. Various glycol ether‐based solvents have pronounced adverse effects on sperm production and male fertility in laboratory animals. The authors investigated the effects of past and current exposure to glycol ether‐containing products on semen quality and reproductive hormones among men employed by the Paris Municipality. Methods Between 2000 and 2001 the authors recruited 109 men who gave semen, blood and urine samples and underwent an andrological examination. Information on lifestyle, occupation, exposure and medical history was obtained by interview. According to their job and chemical products used during the period 1990–2000, men were classified as either occupationally exposed or non‐exposed. Current exposure levels to glycol ethers at the time of the study were evaluated by biological monitoring of six urinary metabolites. Results Previous exposure to glycol ethers was associated with an increased risk for sperm concentration, for rapid progressive motility and for morphologically normal sperm below the World Health Organization semen reference values. No effect of previous glycol ether exposure on hormones levels was observed. By contrast, current glycol ether exposure levels were low and not correlated with either seminal quality or hormone levels. Conclusions This study suggests that most glycol ethers currently used do not impact on human semen characteristics. Those that were more prevalent from the 1960s until recently may have long lasting negative effects on human semen quality. PMID:17332140

  10. Prevention of urethral blockage following semen collection in two species of lemur, Varecia variegata variegata and Lemur catta.

    PubMed

    Chatfield, Jenifer; Penfold, Linda

    2007-06-01

    Lemurs are a diverse group of primates comprised of five families, all of which are found only on Madagascar and the Comoro Islands. Of the 60 known species, 17 are endangered and 5 of these are considered critically endangered. The effects of inbreeding on population health and viability have been well described; though negative inbreeding effects can be ameliorated through the introduction of new genetic material. Introduction of new individuals into a population can be extremely challenging because of the highly social nature of lemurs. Semen collection in lemur species is notoriously challenging, as the ejaculate forms a coagulum. During normal breeding, the coagulum forms a copulatory plug in the female. However, this coagulum can present a life-threatening situation when retained in the urethra abnormally following electroejaculation. This study investigates the use of ascorbic acid in preventing urethral blockage in two lemur species during semen collection, demonstrates successful collection of semen by electroejaculation from two species of lemur during the breeding season, and discusses removal of urethral plugs subsequent to semen collection. Semen was collected successfully from all animals. Urethral plugs formed during each collection and were abnormally retained in 2/11 collections. Both plugs were successfully and immediately removed with the use of retropulsion through a urethral catheter. Although the results of this study are encouraging, more investigation is required to establish whether or not this procedure can be safely performed in the field.

  11. Implications of the pH and temperature of diluted, cooled boar semen on fresh and frozen-thawed sperm motility characteristics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boar semen is typically collected, diluted and cooled for AI use over numerous days, or frozen immediately after shipping to capable laboratories. The storage temperature and pH of the diluted, cooled boar semen could potentially influence the fertility of boar sperm. Therefore, the purpose of thi...

  12. Effect of non-sperm cells removal with single-layer colloidal centrifugation on myeloperoxidase concentration in post-thaw equine semen.

    PubMed

    Ponthier, Jérôme; Teague, Sheila R; Franck, Thierry Y; de la Rebière, Geoffroy; Serteyn, Didier D; Brinsko, Steven P; Love, Charles C; Blanchard, Terry L; Varner, Dickson D; Deleuze, Stéfan C

    2013-12-01

    Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in and released by neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with decreased sperm motility. Recently, MPO concentration in post-thaw semen was shown to be associated with the presence of non-sperm cells (NSC). The objective of this study was to evaluate the effect of a single-layer colloidal centrifugation before cryopreservation on NSC and MPO concentrations in equine semen. The experimental design consisted of freezing semen with or without previous centrifugation through two concentrations of single-layer colloid media. Non-sperm cells and MPO concentrations were assessed in pellet and upper layer at each step of the procedure and MPO was detected in cells by immunocytochemistry. Single-layer colloid centrifugation decreased NSC and MPO concentrations in post-thaw semen. The MPO concentration was correlated with concentration of NSC in the upper layer of the supernatant. In post-thaw semen, with or without previous single-layer colloid centrifugation, MPO concentration was correlated with concentration of NSC. Overall, neutrophils were rarely observed and NSC were mainly epithelial cells or cellular debris, as demonstrated by MPO immunocytochemistry. At all steps of the semen processing and cryopreservation, MPO immunostaining was clearly identified only on NSC. In conclusion, our study shows that NSC present in fresh semen release MPO during freezing.

  13. PCV2 Vaccination Prevented Clinical PCVAD and Reduced PCV2 Viremia and Semen Shedding in Boars Concurrently Infected with PCV2b and Mycoplasma hyopneumoiae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction. It has been determined that porcine circovirus type 2 (PCV2) DNA is shed in semen of naturally and experimentally infected boars (1). Recently, it also has been shown that PCV2 DNA present in semen is infectious in a swine bioassay model (2). However, under experimental conditions the ...

  14. Effect of separating bull semen into X and Y chromosome-bearing fractions on the sex ratio of resulting embryos.

    PubMed Central

    Hagele, W C; Hare, W C; Singh, E L; Grylls, J L; Abt, D A

    1984-01-01

    Seventy-six, day 12 to day 15 bovine embryos, collected from 14 donors which had been inseminated with either X or Y chromosome-bearing spermatozoa fractions of semen separated by a thermal convection counterstreaming sedimentation and forced convection galvanization process, were processed for sexing by chromosomal analysis. Fifty-seven embryos were sexed; 20 from Y chromosome-bearing and 37 from X chromosome-bearing fractions of semen. Statistical analysis of the sexing data indicated that there was no significant difference in the male: female ratio for donors receiving male fractions compared to those receiving female fractions. The Y chromosome-bearing fractions produced a male: female ratio that was indistinguishable from the expected 1:1 ratio. However, the X chromosome-bearing fractions of semen produced a highly significant deviation from the expected 1:1 ratio towards the male. PMID:6478299

  15. The Effects of Frequent Electroejaculation on the Semen Characteristics of a Captive Siberian Tiger (Panthera tigris altaica)

    PubMed Central

    FUKUI, Daisuke; NAGANO, Masashi; NAKAMURA, Ryohei; BANDO, Gen; NAKATA, Shinichi; KOSUGE, Masao; SAKAMOTO, Hideyuki; MATSUI, Motozumi; YANAGAWA, Yojiro; TAKAHASHI, Yoshiyuki

    2013-01-01

    Artificial insemination (AI) can help to avoid inbreeding and genetic degeneration for sustaining genetically healthy populations of endangered species in captivity. Collection of a sufficient quantity of viable sperm is an essential first step in the AI process. In the present study, we examined the effects of frequent electroejaculation on semen characteristics in a Siberian tiger. We collected semen in all 17 trials during 6 breeding seasons (6 years). The mean number of sperm and the percentage of motile sperm were 294.3 ± 250.2×106/ejaculate and 82.4 ± 11.4%, respectively. The number of motile sperm tended to increase during frequent electroejaculation in the same breeding season. Semen collection by electroejaculation can be performed effectively up to the fourth sequential ejaculate, which contained the most sperm in the study. In conclusion, frequent collection of sperm by electroejaculation from tigers may be effective for collection of a large number of motile sperm. PMID:23774799

  16. Exosomes in human semen restrict HIV-1 transmission by vaginal cells and block intravaginal replication of LP-BM5 murine AIDS virus complex

    PubMed Central

    Madison, Marisa N.; Jones, Philip H.; Okeoma, Chioma M.

    2015-01-01

    Exosomes are membranous extracellular nanovesicles secreted by diverse cell types. Exosomes from healthy human semen have been shown to inhibit HIV-1 replication and to impair progeny virus infectivity. In this study, we examined the ability of healthy human semen exosomes to restrict HIV-1 and LP-BM5 murine AIDS virus transmission in three different model systems. We show that vaginal cells internalize exosomes with concomitant transfer of functional mRNA. Semen exosomes blocked the spread of HIV-1 from vaginal epithelial cells to target cells in our cell-to-cell infection model and suppressed transmission of HIV-1 across the vaginal epithelial barrier in our trans-well model. Our in vivo model shows that human semen exosomes restrict intravaginal transmission and propagation of murine AIDS virus. Our study highlights an antiretroviral role for semen exosomes that may be harnessed for the development of novel therapeutic strategies to combat HIV-1 transmission. PMID:25880110

  17. Effect of density gradient centrifugation on reactive oxygen species in human semen.

    PubMed

    Takeshima, Teppei; Yumura, Yasushi; Kuroda, Shinnosuke; Kawahara, Takashi; Uemura, Hiroji; Iwasaki, Akira

    2017-03-23

    Density gradient centrifugation can separate motile sperm from immotile sperm and other cells for assisted reproduction, but may also remove antioxidants from seminal plasma, resulting in oxidative stress. Therefore, we investigated reactive oxygen species (ROS) concentrations and distribution in semen before and after density gradient centrifugation. We assessed semen volume, sperm concentration, sperm motility, and ROS levels before and after density gradient centrifugation (300 x g for 20 minutes) in 143 semen samples from 118 patients. The ROS removal rate was evaluated in ROS-positive samples and ROS formation rate in ROS-negative samples. Thirty-eight of 143 untreated samples (26.6%) were ROS-positive; sperm motility was significantly lower in these samples than in ROS-negative samples (p < 0.05). After density gradient centrifugation, only seven of the 38 ROS-positive samples (18.42%) exhibited a ROS-positive lower layer (containing motile sperm) with a ROS removal rate of 81.58%, whereas the upper layer was ROS-positive in 24 samples (63.16%). In the ROS-negative group (n = 105), ROS was detected in 19 samples after centrifugation (18.10%, ROS generation rate), of which 18 were ROS-positive only in the upper layer or interface and the other was ROS-positive in both layers. Density gradient centrifugation can separate motile sperm from immotile sperm as well as remove ROS (including newly generated ROS). This data supports the view that density gradient centrifugation can select motile spermatozoa without enhancing oxidative stress.

  18. The effect of superoxide dismutase mimetic and catalase on the quality of postthawed goat semen.

    PubMed

    Shafiei, Mojtaba; Forouzanfar, Mohsen; Hosseini, Sayyed Morteza; Esfahani, Mohammad Hossein Nasr

    2015-05-01

    Manganese(III) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin chloride (MnTE) is a cell-permeable superoxide dismutase mimetic agent which can convert superoxide to hydrogen peroxide (H2O2). Supplementation of MnTE to a commercial semen extender can protect sperm from superoxide but not H2O2. Therefore, we proposed that addition of catalase (0.0, 200, or 400 IU/mL) in combination with MnTE (0.1 μM) may further improve the cryopreservation efficiency of goat semen in commercially optimized freezing media such as Andromed. Therefore, ejaculates were obtained from three adult bucks twice a week during the breeding season and diluted with Andromed supplemented with or without MnTE and catalase and were frozen in liquid nitrogen. Sperm parameters and reactive oxygen species contents were evaluated 2 hours after dilution (before freezing) and after freezing/thawing. The results revealed that all the treatments significantly (P ≤ 0.05) improved sperm motility, viability, and membrane integrity after freezing and reduced reactive oxygen species content compared with the control group, but maximum improvement was obtained in MnTE + 400 IU/mL catalase. In addition, supplementation with these antioxidants significantly (P ≤ 0.05) increases the cleavage rate after IVF. In conclusion, the results of present study suggest that addition of antioxidant MnTE or catalase to commercial optimized media, such as Andromed, improves total motility, membrane integrity, and viability of goat semen samples after thawing. But the degree of improvement for these parameters significantly (P ≤ 0.05) higher when MnTE and catalase were simultaneously added to the cryopreservation media.

  19. The effects of storing and transporting cryopreserved semen samples on dry ice

    PubMed Central

    Til, David; Amaral, Vera L L; Salvador, Rafael A; Senn, Alfred; de Paula, Thais S

    2016-01-01

    Objective This study aimed to test the effects on sperm viability of transporting cryopreserved semen samples on dry ice. Methods Twenty normozoospermic semen samples were cryopreserved and divided into five groups. The samples in Group 1 were immersed in liquid nitrogen throughout the experiment in cryogenic storage tanks; the cryopreserved straws in Group 2 were placed in a Styrofoam box containing dry ice and kept under these conditions for 48 hours; the samples in Group 3 were kept for 48 hours on dry ice under the same conditions as the Group 2 samples, and were then moved to a storage tank filled with liquid nitrogen; Group 4 samples were also kept for 48 hours in dry ice storage, and the Styrofoam box containing the samples was shipped by plane to assess the effects of shipping; the samples in Group 5 were shipped together with the Group 4 samples and were placed in a storage tank with liquid nitrogen after spending 48 hours stored on dry ice. After thawing, sperm parameters were analyzed for viability, vitality, and motility; spermatozoa were also tested for mitochondrial activity. Results Significant decreases in motility recovery rates (P=0.01) and vitality (P=0.001) were observed in all groups when compared to the control group. Mitochondrial activity was significantly decreased only in Group 5 (P=0.04), as evidenced by greater numbers of sperm cells not stained by reagent 3,3'-diaminobenzidine. Conclusions Transportation did not affect the quality of cryopreserved semen samples, but dry ice as a means to preserve the samples during transportation had detrimental effects upon the sperm parameters assessed in this study. PMID:28050956

  20. Shorter Anogenital Distance Predicts Poorer Semen Quality in Young Men in Rochester, New York

    PubMed Central

    Mendiola, Jaime; Stahlhut, Richard W.; Jørgensen, Niels; Liu, Fan

    2011-01-01

    Background: In male rodents, anogenital distance (AGD) provides a sensitive and continuous correlate of androgen exposure in the intrauterine environment and predicts later reproductive success. Some endocrine-disrupting chemicals can alter male reproductive tract development, including shortening AGD, in both rodents and humans. Whether AGD is related to semen quality in human is unknown. Objective: We examined associations between AGD and semen parameters in adult males. Methods: We used multiple regression analyses to model the relationships between sperm parameters and two alternative measures of AGD [from the anus to the posterior base of the scrotum (AGDAS) and to the cephalad insertion of the penis (AGDAP)] in 126 volunteers in Rochester, New York. Results: AGDAS, but not AGDAP, was associated with sperm concentration, motility, morphology, total sperm count, and total motile count (p-values, 0.002–0.048). Men with AGDAS below (vs. above) the median were 7.3 times more likely (95% confidence interval, 2.5–21.6) to have a low sperm concentration (< 20 × 106/mL). For a typical study participant, sperm concentrations were 34.7 × 106/mL and 51.6 × 106/mL at the 25th and 75th percentiles of (adjusted) AGDAS. Conclusions: In our population, AGDAS was a strong correlate of all semen parameters and a predictor of low sperm concentration. In animals, male AGD at birth reflects androgen levels during the masculinization programming window and predicts adult AGD and reproductive function. Our results suggest, therefore, that the androgenic environment during early fetal life exerts a fundamental influence on both AGD and adult sperm counts in humans, as demonstrated in rodents. PMID:21377950

  1. Association between mobile phone use and semen quality: a systemic review and meta-analysis.

    PubMed

    Liu, K; Li, Y; Zhang, G; Liu, J; Cao, J; Ao, L; Zhang, S

    2014-07-01

    Possible hazardous health effects of radiofrequency electromagnetic radiations emitted from mobile phone on the reproductive system have raised public concern in recent years. This systemic review and meta-analysis was prepared following standard procedures of the Cochrane Collaboration and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement and checklist. Relevant studies published up to May 2013 were identified from five major international and Chinese literature databases: Medline/PubMed, EMBASE, CNKI, the VIP database and the Cochrane Central Register of Controlled Trials in the Cochrane Library. Eighteen studies with 3947 men and 186 rats were included in the systemic review, of which 12 studies (four human studies, four in vitro studies and four animal studies) with 1533 men and 97 rats were used in the meta-analyses. Systemic review showed that results of most of the human studies and in vitro laboratory studies indicated mobile phone use or radiofrequency exposure had negative effects on the various semen parameters studied. However, meta-analysis indicated that mobile phone use had no adverse effects on semen parameters in human studies. In the in vitro studies, meta-analysis indicated that radiofrequency radiation had detrimental effect on sperm motility and viability in vitro [pooled mean difference (MDs) (95% CI): -4.11 (-8.08, -0.13), -3.82 (-7.00, -0.65) for sperm motility and viability respectively]. As for animal studies, radiofrequency exposure had harmful effects on sperm concentration and motility [pooled MDs (95% CI): -8.75 (-17.37, -0.12), -17.72 (-32.79, -2.65) for sperm concentration and motility respectively]. Evidence from current studies suggests potential harmful effects of mobile phone use on semen parameters. A further multicentred and standardized study is needed to assess the risk of mobile phone use on the reproductive system.

  2. Lactotransferrin in Asian elephant (Elephas maximus) seminal plasma correlates with semen quality.

    PubMed

    Kiso, Wendy K; Selvaraj, Vimal; Nagashima, Jennifer; Asano, Atsushi; Brown, Janine L; Schmitt, Dennis L; Leszyk, John; Travis, Alexander J; Pukazhenthi, Budhan S

    2013-01-01

    Asian elephants (Elephas maximus) have highly variable ejaculate quality within individuals, greatly reducing the efficacy of artificial insemination and making it difficult to devise a sperm cryopreservation protocol for this endangered species. Because seminal plasma influences sperm function and physiology, including sperm motility, the objectives of this study were to characterize the chemistry and protein profiles of Asian elephant seminal plasma and to determine the relationships between seminal plasma components and semen quality. Ejaculates exhibiting good sperm motility (≥65%) expressed higher percentages of spermatozoa with normal morphology (80.3±13.0 vs. 44.9±30.8%) and positive Spermac staining (51.9±14.5 vs. 7.5±14.4%), in addition to higher total volume (135.1±89.6 vs. 88.8±73.1 ml) and lower sperm concentration (473.0±511.2 vs. 1313.8±764.7×10⁶ cells ml⁻¹) compared to ejaculates exhibiting poor sperm motility (≤10%; P<0.05). Comparison of seminal plasma from ejaculates with good versus poor sperm motility revealed significant differences in concentrations of creatine phosphokinase, alanine aminotransferase, phosphorus, sodium, chloride, magnesium, and glucose. These observations suggest seminal plasma influences semen quality in elephants. One- and two-dimensional (2D) gel electrophoresis revealed largely similar compositional profiles of seminal plasma proteins between good and poor motility ejaculates. However, a protein of ∼80 kDa was abundant in 85% of ejaculates with good motility, and was absent in 90% of poor motility ejaculates (P<0.05). We used mass spectrometry to identify this protein as lactotransferrin, and immunoblot analysis to confirm this identification. Together, these findings lay a functional foundation for understanding the contributions of seminal plasma in the regulation of Asian elephant sperm motility, and for improving semen collection and storage in this endangered species.

  3. The bitch uterine response to semen deposition and its modification by male accessory gland secretions.

    PubMed

    England, G C W; Russo, M; Freeman, S L

    2013-02-01

    Little is known about the response of the bitch's reproductive tract to semen deposition. In this study, an influx of polymorphonuclear neutrophils (PMNs) into the uterus was detected after artificial insemination, but there was normal fertility. Doppler ultrasonography showed that insemination induced an increase in uterine artery blood velocity and a decrease in the resistance index of short duration, indicating vasodilation. Semen that was extended in fluid from the sperm rich fraction of the ejaculate (seminal plasma, SP), or third fraction of the ejaculate (prostatic fluid, PF), produced a similar magnitude of effect but of longer duration. It was hypothesised that vasodilation following insemination was largely induced by SP and PF which, together with PMN influx, was part of a normal uterine response. Physiological concentrations of PMNs in vitro reduced the ability of spermatozoa to attach to uterine epithelium, most likely as a result of spermatozoa becoming attached to PMNs. However, both SP and PF increased attachment of spermatozoa to the uterine epithelium by reducing sperm attachment to PMNs, and potentially by an additional mechanism that did not involve inhibition of sperm binding to PMNs. These are the first canine studies to document an apparent physiological response by the uterus to semen, associated with uterine artery vasodilation and PMN influx. Moreover, these investigations are the first to demonstrate that canine SF and PF are part of the mechanism for increasing uterine perfusion and that both fluids have a modulatory effect on PMN-induced inhibition of spermatozoal attachment to uterine epithelium, most likely mediated by reduced sperm attachment to PMNs.

  4. A Preliminary Study: N-acetyl-L-cysteine Improves Semen Quality following Varicocelectomy

    PubMed Central

    Barekat, Foroogh; Tavalaee, Marziyeh; Deemeh, Mohammad Reza; Bahreinian, Mahsa; Azadi, Leila; Abbasi, Homayoun; Rozbahani, Shahla; Nasr-Esfahani, Mohammad Hossein

    2016-01-01

    Background Surgery is considered the primary treatment for male infertility from clinical varicocele. One of the main events associated with varicocele is excessive production of reactive oxygen species (ROS). N-acetyl-L-cysteine (NAC), an antioxidant that scavenges free radicals, is considered a supplement to alleviate glutathione (GSH) depletion during oxidative stress. Despite beneficial effects of NAC in other pathological events, there is no report on the effect of NAC in individuals with varicocele. Therefore, the aim of this study is to evaluate the outcome of NAC on semen quality, protamine content, DNA damage, oxidative stress and fertility following varicocelectomy. Materials and Methods This prospective clinical trial included 35 infertile men with varicocele randomly divided into control (n=20) and NAC (n=15) groups. We assessed semen parameters, protamine content [chromomycin A3 (CMA3)], DNA integrity [terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL)] and oxidative stress [2', 7'-dichlorodihydrofluorescein-diacetate (DCFH-DA)] before and three months after varicocelectomy. Results Percentage of abnormal semen parameters, protamine deficiency, DNA fragmentation and oxidative stress were significantly decreased in both groups compared to before surgery. We calculated the percentage of improvement in these parameters compared to before surgery for each group, then compared the results between the groups. Only percentage of protamine deficiency and DNA fragmentation significantly differed between the NAC and control groups. Conclusion The results of this study, for the first time, revealed that NAC improved chromatin integrity and pregnancy rate when administered as adjunct therapy post-varico- celectomy (Registeration Number: IRCT201508177223N5). PMID:27123209

  5. Lactotransferrin in Asian Elephant (Elephas maximus) Seminal Plasma Correlates with Semen Quality

    PubMed Central

    Kiso, Wendy K.; Selvaraj, Vimal; Nagashima, Jennifer; Asano, Atsushi; Brown, Janine L.; Schmitt, Dennis L.; Leszyk, John; Travis, Alexander J.; Pukazhenthi, Budhan S.

    2013-01-01

    Asian elephants (Elephas maximus) have highly variable ejaculate quality within individuals, greatly reducing the efficacy of artificial insemination and making it difficult to devise a sperm cryopreservation protocol for this endangered species. Because seminal plasma influences sperm function and physiology, including sperm motility, the objectives of this study were to characterize the chemistry and protein profiles of Asian elephant seminal plasma and to determine the relationships between seminal plasma components and semen quality. Ejaculates exhibiting good sperm motility (≥65%) expressed higher percentages of spermatozoa with normal morphology (80.3±13.0 vs. 44.9±30.8%) and positive Spermac staining (51.9±14.5 vs. 7.5±14.4%), in addition to higher total volume (135.1±89.6 vs. 88.8±73.1 ml) and lower sperm concentration (473.0±511.2 vs. 1313.8±764.7×106 cells ml−1) compared to ejaculates exhibiting poor sperm motility (≤10%; P<0.05). Comparison of seminal plasma from ejaculates with good versus poor sperm motility revealed significant differences in concentrations of creatine phosphokinase, alanine aminotransferase, phosphorus, sodium, chloride, magnesium, and glucose. These observations suggest seminal plasma influences semen quality in elephants. One- and two-dimensional (2D) gel electrophoresis revealed largely similar compositional profiles of seminal plasma proteins between good and poor motility ejaculates. However, a protein of ∼80 kDa was abundant in 85% of ejaculates with good motility, and was absent in 90% of poor motility ejaculates (P<0.05). We used mass spectrometry to identify this protein as lactotransferrin, and immunoblot analysis to confirm this identification. Together, these findings lay a functional foundation for understanding the contributions of seminal plasma in the regulation of Asian elephant sperm motility, and for improving semen collection and storage in this endangered species. PMID:23976974

  6. A study of the effect of perchloroethylene exposure on semen quality in dry cleaning workers

    SciTech Connect

    Eskenazi, B.; Wyrobek, A.J.; Fenster, L.; Katz, D.F.; Sadler, M.; Lee, J.; Hudes, M.; Rempel, D.M. )

    1991-01-01

    The purpose of this investigation was to determine the effects of perchloroethylene (PCE) exposure on human semen quality. We compared the semen quality of 34 dry cleaners with that of 48 laundry workers. We examined the relationships of 17 semen parameters to expired air levels of PCE and to an index of exposure based on job tasks in the last three months. The average sperm concentration was over 80 million for both dry cleaners and laundry workers, but approximately one-quarter of each group was oligospermic. The overall percentage of abnormal forms was similar for the two groups; however, sperm of dry cleaners were significantly more likely to be round (t = -3.29, p = 0.002) and less likely to be narrow (t = 2.35, p = 0.02) than the sperm of laundry workers. These effects were dose-related to expired air levels and to the exposure index after controlling for potential confounders (e.g., heat exposure). The average percent motile sperm for both groups was slightly over 60%; however, sperm of dry cleaners tended to swim with greater amplitude of lateral head displacement (ALH) than those of laundry workers (t = -1.73, p = 0.09), and level of PCE in expired air was a significant predictor of ALH in the multiple regression model (t = 2.00, p = 0.05). In addition, exposure index was a significant negative predictor of the sperm linearity parameter (t = -2.57, p = 0.01). These results suggest that occupational exposures to PCE can have subtle effects on sperm quality. Additional analyses are required to determine whether these effects are associated with changes in fertility.

  7. Development of an artificial insemination protocol in llamas using cooled semen.

    PubMed

    Giuliano, S M; Chaves, M G; Trasorras, V L; Gambarotta, M; Neild, D; Director, A; Pinto, M; Miragaya, M H

    2012-04-01

    The objective of this study was to design an AI protocol using cooled semen to obtain pregnancies in the llama. Each raw ejaculate was subdivided into four aliquots which were extended 1:1 with: (1) 11% lactose-egg yolk (L-EY), (2) Tris-citrate-fructose-egg yolk (T-F-EY), (3) PBS-llama serum (S-PBS) and (4) skim milk-glucose (K). Each sample reached 5°C in 2.5 h and remained at that temperature during 24 h. Percentages of the semen variables (motility, live spermatozoa) in ejaculates and samples cooled with L-EY were significantly greater than those obtained when cooling with the other extenders; therefore this extender was used (1:1) for all inseminations. Females were randomly divided into four groups (A-D) according to insemination protocol. Group A: females were inseminated with a fixed dose of 12 × 10(6) live spermatozoa kept at 37°C. Group B: females were inseminated with a fixed dose of 12 × 10(6) live spermatozoa, cooled to 5°C and kept for 24 h. Group C: females were inseminated with the whole ejaculate (variable doses), cooled to 5°C and kept for 24 h. These groups (A-C) were inseminated between 22 and 24 h after induction of ovulation. Group D: females were inseminated with the whole ejaculate (variable doses), cooled to 5°C, kept for 24 h and AI was carried out within 2 h after ovulation. Pregnancy rates were 75%, 0%, 0% and 23% for groups A, B, C and D respectively. These results indicate that it is possible to obtain llama pregnancies with AI using cooled semen and that the success of the technique would depend on the proximity to ovulation.

  8. Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen.

    PubMed

    Anzar, Muhammad; Kroetsch, Tom; Buhr, Mary M

    2009-01-01

    Assessing semen quality is crucially important for the exploitation of genetically superior sires in an artificial insemination (AI) program. In this study, we compare modern and conventional techniques to estimate bovine sperm concentration and membrane integrity. First, the NucleoCounter SP-100 was validated for sperm concentration and provided statistically reliable and repeatable estimates among aliquots and replicates of 25 fresh ejaculates. Sperm concentrations in 78 ejaculates were then determined with hemacytometer, flow cytometer, and NucleoCounter SP-100 and were significantly correlated (P < .001), with regression coefficients among these 3 techniques close to 1 (P < .01). However, the sperm concentration determined by hemacytometer was lower (P < .01) than by flow cytometer and NucleoCounter SP-100. Forty frozen-thawed semen samples were then assessed for sperm concentration and membrane integrity with hemacytometer, flow cytometer and NucleoCounter SP-100. Significant relationships were found for sperm concentration determined by hemacytometer and NucleoCounter SP-100 and for sperm membrane integrity determined by flow cytometer and NucleoCounter SP-100 (P < .01). Finally, the standard curves of sperm concentrations in 6 spectrophotometers, comparing optical density against counts drawn by hemacytometer and NucleoCounter SP-100 (n = 94 fresh ejaculates) showed different (P < .01) intercepts and regression coefficients (linear, quadratic, cubic). It was calculated that a breeding station can improve its production potential by 13% with the use of NucleoCounter SP-100 instead of hemacytometer for calibration of spectrophotometers. Flow cytometer and NucleoCounter SP-100 can be used with equal confidence to estimate sperm concentration and membrane integrity in domestic animals and human semen.

  9. Separation of motile spermatozoa from frozen-thawed buffalo semen: swim-up vs filtration procedures.

    PubMed

    Mustafa, G; Anzar, M; Arslan, M

    1998-07-15

    Three experiments were conducted to maximize the recovery rate of motile spermatozoa from frozen-thawed buffalo semen. In Experiment 1, the swim-up of motile spermatozoa was performed in the presence or absence of HEPES in TALP medium and CO2 in the environment. The recovery rate of motile spermatozoa in TALP medium (control), TALP + HEPES + CO2, TALP + HEPES and TALP + CO2 was 15, 18, 12 and 10%, respectively (P > 0.05), with sperm motility at 87, 89, 90 and 90%, respectively (P > 0.05). In Experiment 2, the pH of TALP medium was adjusted to 7.0, 7.5, 8.0, 8.5 and 9.0, and swim-up procedure was performed in the presence of HEPES and CO2. The recovery rate of motile spermatozoa at different pH was 14, 20, 24, 27 and 16%, respectively (P < 0.05). Motility of separated spermatozoa was 88, 91, 90, 89 and 90%, respectively (P > 0.05). In Experiment 3, the efficiency of ion-exchange filtration and Swim-up procedure in separating motile spermatozoa from frozen-thawed buffalo semen was compared. The recovery rate of motile spermatozoa was 95% in filtration procedure and 33% in swim-up procedure (P < 0.005). In all experiments, normal acrosomes did not vary due to treatments (P > 0.05). In conclusion, HEPES and CO2 had no significant effect on swim-up of buffalo spermatozoa. The pH 8.5 of TALP improved the recovery rate of motile spermatozoa in swim-up procedure. The ion-exchange filtration was found superior to swim-up procedure in harvesting maximum number of motile spermatozoa from frozen-thawed buffalo semen (95 vs 33%; P < 0.001).

  10. Comparison of semen variables, sperm DNA damage and sperm membrane proteins in two male layer breeder lines.

    PubMed

    M, Shanmugam; T R, Kannaki; A, Vinoth

    2016-09-01

    Semen variables are affected by the breed and strain of chicken. The present study was undertaken to compare the semen quality in two lines of adult chickens with particular reference to sperm chromatin condensation, sperm DNA damage and sperm membrane proteins. Semen from a PD3 and White Leghorn control line was collected at 46 and 47 weeks and 55 weeks of age. The semen was evaluated for gross variables and sperm chromatin condensation by aniline blue staining. Sperm DNA damage was assessed by using the comet assay at 47 weeks of age and sperm membrane proteins were assessed at 55 weeks of age. The duration of fertility was studied by inseminating 100 million sperm once into the hens of the same line as well as another line. The eggs were collected after insemination for 15days and incubated. The eggs were candled on 18th day of incubation for observing embryonic development. The White Leghorn control line had a greater sperm concentration and lesser percentage of morphologically abnormal sperm at the different ages where assessments occurred. There was no difference in sperm chromatin condensation, DNA damage and membrane proteins between the lines. Only low molecular weight protein bands of less than 95kDa were observed in samples of both lines. The line from which semen was used had no effect on the duration over which fertility was sustained after insemination either when used in the same line or another line. Thus, from the results of the present study it may be concluded that there was a difference in gross semen variables between the lines that were studied, however, the sperm chromatin condensation, DNA damage, membrane proteins and duration over which fertility was sustained after insemination did not differ between the lines.

  11. Semen quality and insulin-like factor 3: Associations with urinary and seminal levels of phthalate metabolites in adult males.

    PubMed

    Chang, Wei-Hsiang; Wu, Meng-Hsing; Pan, Hsien-An; Guo, Pao-Lin; Lee, Ching-Chang

    2017-04-01

    Certain phthalates have adverse effects on male reproductive functions in animals, and potentially affect human testicular function and spermatogenesis, but little is known about the active mechanisms. We measured the urinary and seminal phthalate metabolites and explored their associations on insulin-like factor 3 (INSL3) and semen quality. Urine, blood, and semen samples were collected from the male partners of subfertile (n = 253) and fertile (n = 37) couples in a reproductive center in southern Taiwan. INSL3, reproductive hormones, semen-quality, and 11 phthalate metabolites in urine and semen were measured. There were significant correlations in the distribution pattern of metabolites, such as the relative contribution of low or high molecular weight phthalate metabolites. The significantly monotonic trends in semen volume, sperm concentration and motility were associated with increasing quartiles of INSL3 (all p-trend < 0.001). In adjusted regression models, increases in urinary phthalate metabolites levels were adversely associated with sperm concentration (monobenzyl phthalate [MBzP], mono-2-ethylhexyl phthalate [MEHP] and MEHP%), motility (MBzP and MEHP) and INSL3 (MBzP, MEHP and MEHP%) (all p < 0.01). Higher seminal phthalate metabolite levels were associated with decreases in sperm concentration (MEHP and mono-2-ethyl-5-hydroxyhexyl phthalate), motility (mono-ethyl phthalate [MEP] and di-(2-ethylhexyl) phthalate [DEHP] metabolites), normal morphology (MEP), and INSL3 (monomethyl phthalate and MEP) (all p < 0.05). Our data suggest that INSL3 secretion, reproductive hormone balance, and sperm production and quality might be simultaneously adversely affected for individuals excreting increasing levels of phthalates metabolites (especially di-ethyl phthalate, butylbenzyl phthalate, and DEHP) in urine and semen samples.

  12. Bivalent response to long-term storage in liquid-preserved boar semen: a flow cytometric analysis.

    PubMed

    Henning, Heiko; Petrunkina, Anna M; Harrison, Robin A P; Waberski, Dagmar

    2012-07-01

    The fertility of liquid-preserved boar semen declines during storage at 17°C, insemination trials even indicating early losses in fertilizing ability within the first 24-48 h of storage. Standard semen parameters barely reflect these changes in semen quality, and new approaches for assessment of functional changes in stored spermatozoa are needed. Capacitation, the essential prefertilization step for spermatozoa in the female genital tract, is specifically induced in vitro by bicarbonate. Therefore, we have investigated changes in responsiveness of boar spermatozoa to bicarbonate during storage. Ejaculates of 14 boars were diluted in Beltsville thawing solution, cooled to 17°C and stored for 12, 24, 72, 120, and 168 h before investigation. At each time, basic semen quality was characterized by sperm motility and viability. Subsequently, washed subsamples were incubated in variants of an in vitro fertilization (IVF) medium and assessed for kinetic changes of viability (plasma membrane integrity) and intracellular calcium concentration using flow cytometry in combination with propidium iodide and Fluo-3. By this means, it was possible to determine specific effects of bicarbonate and calcium on sperm subpopulations over incubation time. During storage, standard semen parameters remained on a high level. However, flow cytometric analysis of sperm responses to capacitating and control media revealed two opposing effects of storage. There was a loss of response to bicarbonate in part of the live sperm population but an increasing degree of instability in the rest. Assessment of response to capacitating media by flow cytometry appears a markedly more sensitive way of monitoring sperm functionality during storage than the standard semen parameters of motility and viability.

  13. Mathematical prediction of freezing times of bovine semen in straws placed in static vapor over liquid nitrogen.

    PubMed

    Santos, M V; Sansinena, M; Zaritzky, N; Chirife, J

    2013-02-01

    A widespread practice in cryopreservation is to freeze spermatozoa by suspending the straws in stagnant nitrogen vapor over liquid nitrogen (N(2)V/LN(2)) for variable periods of time before plunging into liquid nitrogen (-196°C) for indefinite storage. A mathematical heat transfer model was developed to predict freezing times (phase change was considered) required for bull semen and extender packaged in 0.5ml plastic straws and suspended in static liquid nitrogen vapor. Thermophysical properties (i.e. thermal conductivity, specific heat, density, initial freezing temperature) of bovine semen and extender as a function of temperature were determined considering the water change of phase. The non-stationary heat transfer partial differential equations with variable properties (nonlinear mathematical problem) were numerically solved considering in series thermal resistances (semen suspension-straw) and the temperature profiles were obtained for both semen suspension and plastic straw. It was observed both the external heat transfer coefficient in stagnant nitrogen vapor and its temperature (controlled by the distance from the surface of liquid nitrogen to the straw) affected freezing times. The accuracy of the model to estimate freezing times of the straws was further confirmed by comparing with experimental literature data. Results of this study will be useful to select "safe" holding times of bull semen in plastic straws placed N(2)V/LN(2) to ensure that complete freezing of the sample has occurred in the nitrogen vapor and avoid cryodamage when plunging in LN(2). Freezing times predicted by the numerical model can be applied to optimize freezing protocols of bull semen in straws.

  14. Estimation of genetic parameters and effects of cytoplasmic line on scrotal circumference and semen quality traits in Angus bulls.

    PubMed

    Garmyn, A J; Moser, D W; Christmas, R A; Minick Bormann, J

    2011-03-01

    The purpose of this study was to estimate the heritability of scrotal circumference (SC) and semen traits, genetic correlations between SC and semen quality traits, and the effect of cytoplasmic line on SC and semen traits. Breeding soundness exam (BSE) data were collected on registered Angus bulls at 4 ranches over 7 yr. The American Angus Association provided historical pedigree information to estimate the effect of cytoplasmic line on SC and semen quality traits. After editing, the evaluated data set contained 1,281 bulls with breeding soundness exam data that traced back to 100 founder dams. Data were analyzed using a 2-trait animal model to obtain heritability, genetic correlation between SC and semen quality traits, as well as the effect of cytoplasmic line as a random effect for SC, percent motility (MOT), percent primary abnormalities (PRIM), percent secondary abnormalities (SEC), and percent total abnormalities (TOT) using multiple-trait derivative-free REML. Fixed effects included source ranch and collection year, and test age was used as a covariate. Estimates of heritability for SC, MOT, PRIM, SEC, and TOT were 0.46, 0.05, 0.27, 0.23, and 0.25, respectively. Genetic correlations between SC and MOT, PRIM, SEC, and TOT were 0.36, -0.19, -0.11, and -0.23, respectively. The proportions of phenotypic variance accounted for by cytoplasmic line for SC, MOT, PRIM, SEC, and TOT were <0.001, 0.013, 0.023, 0.002, and <0.001, respectively. Genetic correlations between SC and semen quality traits were low to moderate and favorable. Cytoplasmic line may have a marginal effect on MOT and PRIM, but is likely not a significant source of variation for SC, SEC, or TOT.

  15. The use of beef bull semen reduced the risk of abortion in Neospora-seropositive dairy cows.

    PubMed

    López-Gatius, F; Santolaria, P; Yániz, J L; Garbayo, J M; Almería, S

    2005-03-01

    There is an evidence that the epidemiology of neosporosis differs in dairy and beef cattle, such that beef cattle carry a lower risk of abortion. The aim of the present study was to establish whether artificial insemination using semen from beef bulls could reduce the risk of abortion in dairy cows seropositive for the Neospora caninum parasite. Our study was based on yearly serological screening for neosporosis and on the confirmation of Neospora infection in aborted fetuses in two high-producing dairy herds with a mean 28% seroprevalence of N. caninum antibodies. The study population comprised of 273 pregnancies in seropositive animals: 156 pregnancies monitored after insemination using Holstein-Friesian semen and 117 after insemination using beef bull semen. Abortion rates for these animals were 28.2% (77 of 273), 34.6% (54 of 156) and 19.7% (23 of 117). Logistic regression analysis indicated no significant effects of lactation number and previous abortion on the abortion rate. Based on the odds ratio, a 1-unit increase in the Neospora antibody titre yielded a 1.01-fold increase in the abortion rate. The likelihood of abortion was two times higher for cows in one of the two herds and 2.8 times lower (one of 0.36) for pregnant cows inseminated with beef bull semen rather than Holstein-Friesian semen. Our results indicate that the use of beef bull semen can reduce the risk of abortion in dairy cows, and suggest that annual screening for neosporosis, specifically the antibody titre to the protozoan, could be an useful predictor of abortion risk in reproductive health programmes.

  16. Localization and functional modification of L-type voltage-gated calcium channels in equine spermatozoa from fresh and frozen semen.

    PubMed

    Albrizio, M; Moramarco, A M; Nicassio, M; Micera, E; Zarrilli, A; Lacalandra, G M

    2015-02-01

    It is well known that insemination of cryopreserved semen always results in lower fertility when compared with fresh semen, but there is an increased interest and demand for frozen equine semen by the major breeder associations because of the utility arising from semen already "on hand" at breeding time. In this article, we report that equine sperm cells express L-type voltage-gated calcium channels; their localization is restricted to sperm neck and to the principal piece of the tail in both fresh and frozen-thawed spermatozoa. We also studied the causes of cryoinjury at the membrane level focusing on the function of L-type calcium channels. We report that in cryopreserved spermatozoa the mean basal value of [Ca(2+)]i is higher than that of spermatozoa from fresh semen (447.130 vs. 288.3 nM; P < 0.001) and L-type channels function differently in response to their agonist and antagonist in relation to semen condition (fresh or frozen-thawed). We found that on addition of agonist to the culture medium, the increase in intracellular calcium concentrations ([Ca(2+)]i) was greater in frozen semen than in fresh semen (Δ[Ca(2+)]i = 124.59 vs. 16.04 nM; P < 0.001), whereas after the addition of antagonist the decrease in [Ca(2+)]i was lower in frozen semen than in fresh semen (Δ[Ca(2+)]i = 32.5 vs. 82.5 nM; P < 0.001). In this article, we also discuss the impact of cryopreservation on sperm physiology.

  17. Effect of varicocele on semen characteristics according to the new 2010 World Health Organization criteria: a systematic review and meta-analysis

    PubMed Central

    Agarwal, Ashok; Sharma, Reecha; Harlev, Avi; Esteves, Sandro C

    2016-01-01

    This study investigated the effects of varicocele on semen parameters in infertile men based on the new 2010 World Health Organization laboratory manual for the examination of human semen. Semen analysis results (volume, sperm count, motility, and morphology) were the primary outcomes. An electronic search to collect the data was conducted using the Medline/PubMed, SJU discover, and Google Scholar databases. We searched articles published from 2010 to August 2015, i.e., after the publication of the 2010 WHO manual. We included only those studies that reported the actual semen parameters of adult infertile men diagnosed with clinical varicocele and contained a control group of either fertile men or normozoospermic men who were not diagnosed with varicocele. Ten studies were included in the meta-analysis, involving 1232 men. Varicocele was associated with reduced sperm count (mean difference: −44.48 × 106 ml−1; 95% CI: −61.45, −27.51 × 106 ml−1; P < 0.001), motility (mean difference: −26.67%; 95% CI: −34.27, −19.08; P < 0.001), and morphology (mean difference: −19.68%; 95% CI: −29.28, −10.07; P < 0.001) but not semen volume (mean difference: −0.23 ml; 95% CI: −0.64, 0.17). Subgroup analyses indicated that the magnitude of effect was influenced by control subtype but not WHO laboratory manual edition used for semen assessment. We conclude that varicocele is a significant risk factor that negatively affects semen quality, but the observed pooled effect size on semen parameters does not seem to be affected by the WHO laboratory manual edition. Given most of the studies published after 2010 still utilized the 1999 manual for semen analysis, further research is required to fully understand the clinical implication of the 2010 WHO laboratory manual on the association between varicocele and semen parameters. PMID:26780872

  18. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis).

    PubMed

    Dorostkar, Kamran; Alavi-Shoushtari, Sayed Mortaza; Mokarizadeh, Aram

    2012-01-01

    The aim of the present study was to investigate the effect of in vitro supplementation of selenium on fresh and frozen spermatozoa quality of buffalo (Bubalus bubalis) bulls. Five healthy buffalo bulls (5 ejaculates from each bull) were used. Each ejaculate was diluted at 37 ˚C with tris-based extender containing 0 (control), 0.5, 1, 2, 4 and 8 µg mL(-1) sodium selenite and the sperm motility and viability were evaluated at 0 (T0) (immediately after dilution), 60 (T1) and 120 (T2) min after diluting semen. In the second step, semen samples were diluted with tris-egg yolk-glycerol extender containing the same amounts of sodium selenite, cooled to 4 ˚C, equilibrated and semen parameters (motility, viability, membrane integrity and DNA damage) were estimated. Then, the semen was packed in 0.5 mL French straws and frozen in liquid nitrogen. Later, the semen was thawed and analyzed for the same parameters, as well as total antioxidant capacity. Results showed that addition of 1 and 2 µgmL(-1) selenium to the semen extender significantly increased the sperm motility of fresh and equilibrated semen compared to the control without affecting other parameters. However, in frozen-thawed semen, extenders containing 1 and 2 µg mL(-1) selenium significantly improved sperm motility, viability, membrane integrity and semen total antioxidant capacity and also resulted in lower DNA damaged sperms. In this study selenium supplementation of semen extender of 4 and 8 µg mL(-1) had deleterious effects on sperm parameters as early as the samples were prepared for freezing.

  19. Effectiveness of semen washing to prevent HIV transmission and assist pregnancy in HIV-discordant couples: a systematic review and meta-analysis

    PubMed Central

    Zafer, Maryam; Horvath, Hacsi; Mmeje, Okeoma; van der Poel, Sheryl; Semprini, Augusto; Rutherford, George; Brown, Joelle

    2015-01-01

    Objective To evaluate the effectiveness of semen washing in HIV-discordant couples in which the male partner is infected Design Systematic review and meta-analysis Setting All countries Patient(s) Forty single-arm, open label studies among HIV-discordant couples that underwent intrauterine insemination (IUI) or in vitro fertilization (IVF) with or without intracytoplasmic sperm injection (ICSI) using washed semen Intervention(s) Semen washing followed by IUI, IVF, or IVF/ICSI Main outcome measure(s) Primary outcome: HIV transmission to HIV-uninfected women; secondary outcomes: HIV transmission to newborns and proportion of couples achieving a clinical pregnancy Result(s) No HIV transmission occurred in 11,585 cycles of assisted reproduction us