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Sample records for abnormal semen samples

  1. 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.

  2. 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

  3. 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.

  4. Influence of genetic abnormalities on semen quality and male fertility: A four-year prospective study

    PubMed Central

    Elfateh, Fadlalla; Wang, Ruixue; Zhang, Zhihong; Jiang, Yuting; Chen, Shuang; Liu, Ruizhi

    2014-01-01

    Background: Wide range of disorders ranging from genetic disorders to coital difficulties can influence male fertility. In this regard, genetic factors are highlighted as the most frequent, contributed to 10-15%, of male infertility causes. Objective: To investigate the influence of genetic abnormalities on semen quality and reproductive hormone levels of infertile men from Northeast China. Materials and Methods: 2034 infertile men including 691 patients with abnormal sperm parameters were investigated retrospectively. Semen analysis was performed according to the World Health Organization guidelines. Y chromosome micro deletions were detected by polymerase chain reaction assays. Chromosome analysis was performed using G-banding. Results: The incidence of abnormal chromosomal karyotype in the patients with abnormal sperm parameters was 12.01% (83/691). The most frequent cause was Klinefelter's syndrome 37.35% (31/83). As the same as chromosomal abnormalities group, the volumes of testes (p=0.000 and 0.000, respectively) and the levels of testosterone (T) (p=0.000), and testosterone/ luteinizing hormone (T/LH) (p=0.000) of patients with Y chromosome micro deletions were significantly lower than those of fertile group. In addition, the levels of follicle-stimulating hormone (FSH) (p=0.000), and luteinizing hormone (LH) (p=0.000) were significantly higher in patients with Y chromosome micro deletions than those in the fertile group. Translocation abnormalities displayed slight effect on sperm motility. Conclusion: Y chromosome micro deletions and sex chromosome disorders particularly Klinefelter’s (47, XXY), have severe adverse influence on normal hormone levels, testicular volume and sperm count, whereas translocation abnormalities may inversely correlate with sperm motility. PMID:24799866

  5. 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.

  6. 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.

  7. 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

  8. Prevalence of chromosomal abnormalities and Y chromosome microdeletion among men with severe semen abnormalities and its correlation with successful sperm retrieval

    PubMed Central

    Mascarenhas, Mariano; Thomas, Sumi; Kamath, Mohan S.; Ramalingam, Ramya; Kongari, Ann Marie; Yuvarani, S; Srivastava, Vivi M.; George, Korula

    2016-01-01

    AIM: To estimate the prevalence of chromosomal abnormalities and Y chromosome microdeletion among men with azoospermia and severe oligozoospermia and its correlation with successful surgical sperm retrieval. SETTING AND DESIGN: A prospective study in a tertiary level infertility unit. MATERIALS AND METHODS: In a prospective observation study, men with azoospermia and severe oligozoospermia (concentration <5 million/ml) attending the infertility center underwent genetic screening. Peripheral blood karyotype was done by Giemsa banding. Y chromosome microdeletion study was performed by a multiplex polymerase chain reaction. RESULTS: The study group consisted of 220 men, 133 of whom had azoospermia and 87 had severe oligozoospermia. Overall, 21/220 (9.5%) men had chromosomal abnormalities and 13/220 (5.9%) men had Y chromosome microdeletions. Chromosomal abnormalities were seen in 14.3% (19/133) of azoospermic men and Y chromosome microdeletions in 8.3% (11/133). Of the 87 men with severe oligozoospermia, chromosomal abnormalities and Y chromosome microdeletions were each seen in 2.3% (2/87). Testicular sperm aspiration was done in 13 men and was successful in only one, who had a deletion of azoospermia factor c. CONCLUSIONS: Our study found a fairly high prevalence of genetic abnormality in men with severe semen abnormalities and a correlation of genetic abnormalities with surgical sperm retrieval outcomes. These findings support the need for genetic screening of these men prior to embarking on surgical sperm retrieval and assisted reproductive technology intracytoplasmic sperm injection. PMID:27803587

  9. Aneuploidy frequency in spermatozoa of Egyptian men with normal and abnormal semen parameters using fluorescence in situ hybridisation.

    PubMed

    Younan, D; Sorour, A; Genedy, R

    2015-03-01

    Chromosome anomalies were suggested to be more frequent in infertile males so our case-control study aimed at evaluating the incidence of spermatic aneuploidies in forty males with severe oligoasthenoteratozoospermia (OAT) and comparing it with that in another forty males having normal semen parameters. Semen samples were collected and analysed in the Clinical Pathology Department according to criteria of the World Health Organization (WHO laboratory manual for the examination and processing of human semen, 2010, WHO Press). Fluorescence in situ hybridisation (FISH) was performed on decondensed spermatozoa from fresh semen ejaculates, using dual coloured chromosome-specific DNA probes labelled with fluorochromes to study sperm aneuploidies in chromosomes 13, 21, X and Y. There was no statistical significant difference between cases and controls regarding disomy frequencies for chromosomes 13, 21 or both combined. However, 13, 21 diploidy frequency was significantly higher among OAT cases. Regarding chromosomes X and Y, both cases and controls showed similar results for disomy/diploidy frequency for both chromosomes; however, there was a statistical significant increase in YY disomy/diploidy frequency among OAT patients. X chromosome-bearing spermatozoa were found to be significantly higher among controls. Patients with severe OAT have a higher total sperm aneuploidy rate, regarding chromosomes 13, 21, X and Y but without a statistical significant difference.

  10. 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...

  11. Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men: a randomized, double-blind, placebo-controlled clinical trial.

    PubMed

    Kolahdooz, M; Nasri, S; Modarres, S Zadeh; Kianbakht, S; Huseini, H Fallah

    2014-05-15

    In recent years, wide utilization of herbal drugs has encouraged scientists to determine their impressive effects on health. Since Nigella sativa L. seed (N. sativa) has many uses including infertility in traditional medicine, the effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men with abnormal semen quality are of interest. This study was conducted on Iranian infertile men with inclusion criteria of abnormal sperm morphology less than 30% or sperm counts below 20×10(6)/ml or type A and B motility less than 25% and 50% respectively. The patients in N. sativa oil group (n=34) received 2.5mlN. sativa oil and placebo group (n=34) received 2.5ml liquid paraffin two times a day orally for 2 months. At baseline and after 2 months, the sperm count, motility and morphology and semen volume, pH and round cells as primary outcomes were determined in both groups. Results showed that sperm count, motility and morphology and semen volume, pH and round cells were improved significantly in N. sativa oil treated group compared with placebo group after 2 months. It is concluded that daily intake of 5ml N. sativa oil for two months improves abnormal semen quality in infertile men without any adverse effects.

  12. Decreased levels of genuine large free hCG alpha in men presenting with abnormal semen analysis

    PubMed Central

    2011-01-01

    Background The pregnancy hormone human chorionic gonadotropin (hCG) and its free subunits (hCG alpha, hCG beta) are produced in the male reproductive tract and found in high concentrations in seminal fluid, in particular hCG alpha. This study aimed to elucidate changes in peptide hormone profiles in patients showing abnormal semen analyses and to determine the genuineness of the highly abundant hCG alpha. Methods Seminal plasma was obtained from 45 male patients undergoing semen analysis during infertility workups. Comprehensive peptide hormone profiles were established by a panel of immunofluorometric assays for hCG, hCG alpha, hCG beta and its metabolite hCG beta core fragment, placental lactogen, growth hormone and prolactin in seminal plasma of patients with abnormal semen analysis results (n = 29) versus normozoospermic men (n = 16). The molecular identity of large hyperglycosylated hCG alpha was analyzed by mass-spectrometry and selective deglycosylation. Results hCG alpha levels were found to be significantly lower in men with impaired semen quality (1346 +/- 191 vs. 2753 +/- 533 ng/ml, P = 0.022). Moreover, patients with reduced sperm count had reduced intact hCG levels compared with normozoospermic men (0.097 +/- 0.022 vs. 0.203 +/- 0.040 ng/ml, P = 0.028). Using mass-spectrometry, the biochemical identity of hCG alpha purified from seminal plasma was verified. Under non-reducing conditions in SDS-PAGE, hCG alpha isolated from seminal plasma migrated in a manner comparable with large free hCG alpha with an apparent molecular mass (Mr, app) of 24 kDa, while hCG alpha dissociated from pregnancy-derived holo-hCG migrated at approximately 22 kDa. After deglycosylation with PNGase F under denaturing conditions, all hCG alpha variants showed an Mr, app of 15 kDa, indicating identical amino acid backbones. Conclusions The findings indicate a pathophysiological relevance of hCG, particularly its free alpha subunit, in spermatogenesis. The alternative glycosylation

  13. Exercise protects against obesity induced semen abnormalities via downregulating stem cell factor, upregulating Ghrelin and normalizing oxidative stress.

    PubMed

    Alhashem, Fahaid; Alkhateeb, Mahmoud; Sakr, Hussein; Alshahrani, Mesfer; Alsunaidi, Mohammad; Elrefaey, Hesham; Alessa, Riyad; Sarhan, Mohammad; Eleawa, Samy M; Khalil, Mohammad A

    2014-01-01

    Increased oxidative stress and hormonal imbalance have been hypothesized to underlie infertility in obese animals. However, recent evidence suggests that Ghrelin and Stem Cell Factor (SCF) play an important role in fertility, in lean individuals. Therefore, this study aimed at investigating whether changes in the levels of Ghrelin and SCF in rat testes underlie semen abnormal parameters observed in obese rats, and secondly, whether endurance exercise or Orlistat can protect against changes in Ghrelin, SCF, and/or semen parameters in diet induced obese rats. Obesity was modelled in male Wistar rats using High Fat Diet (HFD) 12-week protocol. Eight week-old rats (n=40) were divided into four groups, namely, Group I: fed with a standard diet (12 % of calories as fat); Group II: fed HFD (40 % of calories as fat); Group III: fed the HFD with a concomitant dose of Orlistat (200 mg/kg); and Group IV: fed the HFD and underwent 30 min daily swimming exercise. The model was validated by measuring the levels of testosterone, FSH, LH, estradiol, leptin, triglycerides, total, HDL, and LDL cholesterol, and final change in body weight. Levels were consistent with published obesity models (see Results). As predicted, the HFD group had a 76.8 % decrease in sperm count, 44.72 % decrease in sperm motility, as well as 47.09 % increase in abnormal sperm morphology. Unlike the control group, in the HFD group (i.e. obese rats) Ghrelin mRNA and protein were elevated, while SCF mRNA and protein were diminished in the testes. Furthermore, in the HFD group, SOD and GPx activities were significantly reduced, 48.5±5.8 % (P=0.0012) and 45.6±4.6 % (P=0.0019), respectively, while TBARS levels were significantly increased (112.7±8.9 %, P=0.0001). Finally, endurance exercise training and Orlistat administration individually and differentially protected semen parameters in obese rats. The mechanism includes, but is not limited to, normalizing the levels of Ghrelin, SCF, SOD, GPx and TBARS. In rat

  14. Characterization of the temporal persistence of chromosomal abnormalities in the semen of Hodkin`s disease patients after treatment with NOVP chemotherapy using multi-chromosome fluorescence in situ hybridization

    SciTech Connect

    Cassel, M.J.; Robbins, W.A.; Wyrobek, A.J.; Meistrich, M.L.

    1994-12-31

    Three-chromosome fluorescence in situ hybridization (FISH) was applied to sperm of men with Hodgkin`s disease to measure the persistence of chromosomally abnormal sperm within the time interval of 3 to 33 months after the end of treatment. NOVP chemotherapy includes the agents novantrone, oncovin, vinblastine, and prednisone, two of which are spindle poisons expected to induce aneuploidy. Semen samples were evaluated for the frequencies of fluorescence phenotypes representing hyperhaploidy, hypohaploidy, and genomic duplications using DNA probes specific for repetitive sequences on chromosomes X,Y, and 8. Using this procedure, NOVP was previously shown to induce chromosomally abnormal sperm in treated patients. In a longitudinal assessment of 11 semen samples from 2 men, frequencies of abnormal sperm appeared to return to pre-treatment levels at {approximately}6 months after the end of treatment and remained at these levels up to 33 months after the end of treatment. However, pre-treatment frequencies of chromosomally abnormal cells in Hodgkin`s patients were elevated above those found in normal healthy men. Additional patients are being evaluated to determine how long after therapy Hodgkin`s disease patients remain at increased risk for producing chromosomally abnormal sperm.

  15. 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.

  16. 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

  17. 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.

  18. Evidence for obtaining a second successive semen sample for intrauterine insemination in selected patients: results from 32 consecutive cases

    PubMed Central

    Ortiz, Alejandra; Ortiz, Rita; Soto, Evelyn; Hartmann, Jonathan; Manzur, Alejandro

    2016-01-01

    Objective The goal of this study was to compare the semen parameters of two successive samples obtained within an interval of less than 60 minutes from patients planning to undergo intrauterine insemination (IUI) whose first samples exhibited low semen quality. Methods Thirty-two consecutive patients were enrolled in the study. On the day of IUI, the semen analysis of the samples initially presented by all patients met at least two of the following criteria: sperm concentration <5×106/mL, total sperm count <10×106, progressive sperm motility (a+b) in the native sample <30%, and total motile sperm count (TMSC) <4×106. A successive semen sample was obtained no more than 60 minutes after the first sample. Results Compared to the first sample, the second exhibited significantly (p<0.05) improved sperm concentration, TMSC, progressive motility, and vitality. Regarding TMSC, the most critical parameter on the day of IUI, 23 patients (71.8%) improved it, while nine (28.2%) displayed poorer outcomes. Conclusion In defined cases, requesting a second successive ejaculate on the day of insemination may result in a high percentage of cases in an improvement of the quality of the sample. PMID:27358828

  19. Comparison of culture and a multiplex probe PCR for identifying Mycoplasma species in bovine milk, semen and swab samples

    PubMed Central

    Parker, Alysia M.; House, John K.; Hazelton, Mark S.; Bosward, Katrina L.; Sheehy, Paul A.

    2017-01-01

    Mycoplasma spp. are a major cause of mastitis, arthritis and pneumonia in cattle, and have been associated with reproductive disorders in cows. While culture is the traditional method of identification the use of PCR has become more common. Several investigators have developed PCR protocols to detect M. bovis in milk, yet few studies have evaluated other sample types or other important Mycoplasma species. Therefore the objective of this study was to develop a multiplex PCR assay to detect M. bovis, M. californicum and M. bovigenitalium, and evaluate its analytical performance against traditional culture of bovine milk, semen and swab samples. The PCR specificity was determined and the limit of detection evaluated in spiked milk, semen and swabs. The PCR was then compared to culture on 474 field samples from individual milk, bulk tank milk (BTM), semen and swab (vaginal, preputial, nose and eye) samples. Specificity analysis produced appropriate amplification for all M. bovis, M. californicum and M. bovigenitalium isolates. Amplification was not seen for any of the other Mollicutes or eubacterial isolates. The limit of detection of the PCR was best in milk, followed by semen and swabs. When all three Mycoplasma species were present in a sample, the limit of detection increased. When comparing culture and PCR, overall there was no significant difference in the proportion of culture and PCR positive samples. Culture could detect significantly more positive swab samples. No significant differences were identified for semen, individual milk or BTM samples. PCR identified five samples with two species present. Culture followed by 16S-23S rRNA sequencing did not enable identification of more than one species. Therefore, the superior method for identification of M. bovis, M. californicum and M. bovigenitalium may be dependent on the sample type being analysed, and whether the identification of multiple target species is required. PMID:28264012

  20. Normal sperm morphology and changes of semen characteristics and abnormal morphological spermatozoa among peri-mating seasons in captive japanese black bears (Ursus thibetanus japonicus).

    PubMed

    Okano, Tsukasa; Murase, Tetsuma; Nakamura, Sachiko; Komatsu, Takeshi; Tsubota, Toshio; Asano, Makoto

    2009-04-01

    The objectives of this study were to obtain morphological data for normal spermatozoa and to investigate seasonal changes (the early, mid- and post-mating seasons) in abnormal morphology of spermatozoa and the characteristics of semen in Japanese black bears. Semen was collected by electroejaculation from 34 captive male Japanese black bears a total of 74 times. Length of head, width of head, length of midpiece and total length of the spermatozoa were 6.3 +/- 0.4, 4.5 +/- 0.3, 10.4 +/- 0.7 and 69.6 +/- 3.1 mum (mean +/- SD; 20 semen, 200 spermatozoa), respectively. In the semen collected during the mid-mating season, ejaculate volume, ejaculate pH, sperm concentration, total sperm count, motility, viability and intact acrosomes were 0.46 +/- 0.36 ml, 7.3 +/- 0.4, 659 +/- 644 x 10(6)/ml, 214 +/- 208 x 10(6), 82.9 +/- 9.6%, 89.3 +/- 9.5% and 97.0 +/- 3.2% (mean +/- SD; n=21, in ejaculate pH n=8), respectively. Sperm motility and viability in the early (n=7) and mid-mating (n=21) seasons were significantly higher than in the post-mating (n=8) season. The rates of detached heads in the early and mid-mating season were significantly lower than in the post-mating season. The main abnormal morphologies observed (mean +/- SD%; n=23) were simply bent tail (19.9 +/- 22.6), distal droplets (13.5 +/- 11.7), proximal droplets (9.6 +/- 7.8), teratoid spermatozoa (6.7 +/- 10.7), knobbed acrosome (4.9 +/- 8.6), acrosome damage (3.7 +/- 2.8) and bent midpiece (3.7 +/- 5.1). The data will be useful for artificial breeding and further research on male reproductive physiology in this species.

  1. British Andrology Society guidelines for the assessment of post vasectomy semen samples (2002)

    PubMed Central

    Hancock, P; McLaughlin, E

    2002-01-01

    The British Andrology Society guidelines for the assessment of post vasectomy semen samples recommend that initial assessment is undertaken 16 weeks post vasectomy and after the patient has produced at least 24 ejaculates. The laboratory should examine a freshly produced seminal fluid specimen by direct microscopy and if no sperm are seen the centrifugate should be examined for the presence of motile and non-motile spermatozoa. It is recommended that the clinician should give clearance after the production of two consecutive sperm free ejaculates. In cases of persistent identification of non-motile spermatozoa the referring clinician should advise the patient regarding the cessation of other contraceptive precautions. Surgeons are responsible both preoperatively and postoperatively for the counselling of couples regarding complications and the possibility of late recanalisation after clearance. PMID:12401817

  2. 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.

  3. Y-STRs in forensic medicine: DNA analysis in semen samples of azoospermic individuals.

    PubMed

    Soares-Vieira, José Arnaldo; Billerbeck, Ana Elisa Correia; Iwamura, Edna Sadayo Miazato; Zampieri, Ricardo Andrade; Gattás, Gilka Jorge Fígaro; Munoz, Daniel Romero; Hallak, Jorge; Mendonca, Berenice Bilharinho; Lucon, Antonio Marmo

    2007-05-01

    The incidence of rape has increased, especially in metropolitan areas, such as the city of São Paulo. In Brazil, studies about it have shown that the majority of this type of crime is committed by the relatives and persons close to the victim. This has made the crime more difficult to be denounced, as only 10% of the cases are reported to competent police authorities. Usually, cytological exams are carried out in sex crime investigations. The difficulty in showing the presence of spermatozoa is frequent, but it does not exclude the presence of male DNA. The absence of spermatozoa in material collected from rape victims can be due to several factors, including the fact that the agressor suffers from azoospermia. This condition can be the result of a successful vasectomy. As the majority of DNA in the ejaculation sample is from spermatozoa, there is much less DNA to be analyzed. This study presents the application of Y-STRs (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, and DYS393) in DNA analysis of sperm samples from 105 vasectomized men. The study demonstrated a great variation in DNA concentration. DNA extraction and amplification was possible in all sperm samples even in the absence of spermatozoa. The same profile was observed, for each individual, from DNA extracted from blood, pre- and postvasectomy semen samples. The use of markers specific for Y chromosome in sex crime cases, especially in the absence of spermatozoa, is very important, mainly because in most situations there is a small quantity of the agressor's DNA in the medium and a large quantity of the victim's DNA.

  4. Seminal plasma anti-sperm antibodies and IVF: the effect of semen sample collection into 50% serum.

    PubMed

    Elder, K T; Wick, K L; Edwards, R G

    1990-02-01

    A retrospective analysis was carried out which studied IVF results for couples in whom the male partner had anti-sperm antibodies in seminal plasma, as judged by a positive MAR test. Of the 59 couples studied over a total of 113 cycles, 30 had IVF semen samples collected into sterile, dry pots. Thirty-eight of the couples had samples collected into medium containing 50% serum, and nine couples underwent separate treatment cycles with samples collected both dry and into 50% serum. The results demonstrate that the addition of serum to sample collection pots significantly improves the oocyte fertilization rate and is followed by a greater chance of conception for these couples.

  5. 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.

  6. Genital mycoplasmas in semen samples of males attending a tertiary care hospital in Nigeria: any role in sperm count reduction?

    PubMed

    Agbakoba, N R; Adetosoye, A I; Ikechebelu, J I

    2007-06-01

    Semen samples from 54 married men attending the outpatient clinics for problems of infertility and routine semen analysis were examined for the presence of genital mycoplasmas. The mean age of the men was 36.1 years with a range of 25 55 years. Majority of the men 57.4% (31 of 54) were in their fourth decade of life (30 39 years). This age group also had the highest percentage 57.2% (8 of 14) of positive isolates of genital mycoplasmas on semen culture. A total of 21 organisms obtained from 14 (26.0%) positive samples were isolated. Mycoplasma and Ureaplasma spp. separately isolated from the samples yielded frequencies of 1 (1.9%) and 6 (11.1%) respectively and the remaining 7 (13.0%) samples were infected with both organisms. A breakdown of the mycoplasma species include 5 (23.8%) M. hominis, 2 (9.5%) M. fermentans and 1 (4.8%) M. penetrans. Apart from one isolate of M. hominis other Mycoplasma species were found in association with Ureaplasma species. Fifteen (71.4%) of the 21 isolates [8 (53.3%) ureaplasmas and 7 (46.7%) mycoplasmas] were isolated from samples with sperm counts less than 20 million/ml while the remaining 6 (21.6%) isolates [5 (83.3%) ureaplasmas and 1 (16.7) mycoplasma] were from samples with counts greater than 20 million/ml. This finding could indicate a possible influence of genital mycoplasmas especially mycoplasmas species on sperm count.

  7. 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

  8. 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.

  9. 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

  10. In Vitro Antioxidant Treatment of Semen Samples in Assisted Reproductive Technology: Effects of Myo-Inositol on Nemaspermic Parameters

    PubMed Central

    Papale, Palma; Della Ragione, Antonietta; Quaranta, Giuseppa; Russo, Giovanni; Russo, Sabatino

    2016-01-01

    Male infertility and the poor quality of sperm seem to be influenced by oxidative stress. In particular, the reactive oxygen species (ROS) mainly produced by morphologically altered spermatozoa affect sperm motility, morphology, and integrity. The aim of this study was to evaluate the efficacy of Myo-Inositol (Myo-Ins) on a number of parameters such as viscosity and total and progressive motility of spermatozoa, in order to better validate its possible practical application in vitro, in order to improve the capacitation protocols commonly used in Assisted Reproductive Technology (ART). A total of 100 fresh and 25 thawed semen samples were analyzed in vitro prior to and after addition of Myo-Ins. Treatment of samples with Myo-Ins showed an increase in the sperm total and progressive motility in both fresh and thawed samples. Furthermore, Myo-Ins proved to be well tolerated by spermatozoa in vitro, demonstrating that it can be efficiently and safely used as antioxidant in the laboratory practice and for preparation of semen samples in ART. PMID:27672392

  11. 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.

  12. 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...

  13. Influence of sexual stimulation on sperm parameters in semen samples collected via masturbation from normozoospermic men or cryptozoospermic men participating in an assisted reproduction programme.

    PubMed

    Yamamoto, Y; Sofikitis, N; Mio, Y; Miyagawa, I

    2000-05-01

    To evaluate the influence of sexual stimulation via sexually stimulating videotaped visual images (VIM) on sperm function, two semen samples were collected from each of 19 normozoospermic men via masturbation with VIM. Two additional samples were collected from each man via masturbation without VIM. The volume of seminal plasma, total sperm count, sperm motility, percentage of morphologically normal spermatozoa, outcome of hypo-osmotic swelling test and zona-free hamster oocyte sperm penetration assay, and markers of the secretory function of prostate were significantly larger in semen samples collected via masturbation with VIM than masturbation without VIM. The improved sperm parameters in the samples collected via masturbation with VIM may reflect an enhanced prostatic secretory function and increased loading of the vas deferens at that time. In a similar protocol, two semen samples were collected via masturbation with VIM from each of 22 non-obstructed azoospermic men. Semen samples from these men had been occasionally positive in the past for a very small number of spermatozoa (cryptozoospermic men). Two additional samples were collected from each cryptozoospermic man via masturbation without VIM. The volume of seminal plasma, total sperm count, sperm motility, and a marker of the secretory function of prostate were significantly larger in semen samples collected via masturbation with VIM. Fourteen out of the 22 men were negative for spermatozoa in both samples collected via masturbation without VIM. These men demonstrated spermatozoa in both samples collected via masturbation with VIM. Six men with immotile spermatozoa in both samples collected via masturbation without VIM exposed motile spermatozoa in both samples collected via masturbation with VIM. High sexual stimulation during masturbation with VIM results in recovery of spermatozoa of greater fertilizing potential both in normozoospermic and cryptozoospermic men. The appearance of spermatozoa after

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. Sperm membrane integrity in fresh and frozen-thawed canine semen samples: a comparison of vital stains with the NucleoCounter SP-100.

    PubMed

    Daub, L; Geyer, A; Reese, S; Braun, J; Otzdorff, C

    2016-07-15

    The objective of this study was to assess sperm membrane integrity in canine semen samples using three different vital stains and the NucleoCounter SP-100 (NC). In addition, the occurrence of half-stained sperm heads, the influence of investigator, and storage-related artifacts using stained smears were examined. Forty fresh (30 dogs) and 40 frozen-thawed (28 dogs) canine semen samples were analyzed. The vital stains eosin (E), eosin-nigrosin (EN), and bromphenolblue-nigrosin (BN) were compared. Two smears per stain were prepared and a total of 200 sperm per slide were classified using bright field microscopy. Each slide was examined twice by three investigators. Spermatozoa with completely red (E, EN) or blue (BN) stained sperm heads were classified as "dead". Half-stained sperm heads were counted separately. Sperm concentration and viability were determined using the NC. The NC works with a built-in fluorescence microscope using propidium iodide as a fluorescence dye. Statistical analysis for comparison of results was made using mean values with standard deviation, Bland-Altman plot and coefficient of variation (CV). Staining with E led to a significant higher percentage of dead sperm compared with EN and BN (P < 0.05), which gave comparable results. Vital stains revealed higher CVs (range 8.8%-32.1%) than the NC (<6.5%). Interobserver viability ranged from 17.5% to 45.4% and was within the same range between stains. If only completely stained sperm heads were considered, best agreement was found between the E and the NC. In case of EN and BN, inclusion of half-stained sperm heads reduced the difference compared with NC. In general, the agreement between methods was better in samples with a low percentage of dead spermatozoa. In smears of fresh semen stored up to 3 months, no increase in the percentage of dead spermatozoa could be observed. In some smears of frozen-thawed samples stained with E (n = 12) or BN (n = 2), all previously unstained spermatozoa

  20. 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.

  1. The value of Bayes' theorem for interpreting abnormal test scores in cognitively healthy and clinical samples.

    PubMed

    Gavett, Brandon E

    2015-03-01

    The base rates of abnormal test scores in cognitively normal samples have been a focus of recent research. The goal of the current study is to illustrate how Bayes' theorem uses these base rates--along with the same base rates in cognitively impaired samples and prevalence rates of cognitive impairment--to yield probability values that are more useful for making judgments about the absence or presence of cognitive impairment. Correlation matrices, means, and standard deviations were obtained from the Wechsler Memory Scale--4th Edition (WMS-IV) Technical and Interpretive Manual and used in Monte Carlo simulations to estimate the base rates of abnormal test scores in the standardization and special groups (mixed clinical) samples. Bayes' theorem was applied to these estimates to identify probabilities of normal cognition based on the number of abnormal test scores observed. Abnormal scores were common in the standardization sample (65.4% scoring below a scaled score of 7 on at least one subtest) and more common in the mixed clinical sample (85.6% scoring below a scaled score of 7 on at least one subtest). Probabilities varied according to the number of abnormal test scores, base rates of normal cognition, and cutoff scores. The results suggest that interpretation of base rates obtained from cognitively healthy samples must also account for data from cognitively impaired samples. Bayes' theorem can help neuropsychologists answer questions about the probability that an individual examinee is cognitively healthy based on the number of abnormal test scores observed.

  2. Novel Y-STR typing strategies reveal the genetic profile of the semen donor in extended interval post-coital cervicovaginal samples.

    PubMed

    Hall, Ashley; Ballantyne, Jack

    2003-09-09

    For a variety of reasons, some victims of sexual assault provide vaginal samples more than 24-36 h after the incident. In these cases, the ability to obtain an autosomal STR profile of the semen donor from the living victim diminishes rapidly as the post-coital interval is extended. We have used a number of carefully selected Y-STR loci in a variety of multiplex or monoplex formats to extend the post-coital interval from which a genetic profile of the semen donor can be obtained. The proposed Y-STR typing strategies enable the routine detection of the male donor Y-STR haplotype in cervicovaginal samples recovered up to 4 days post-coitus. We attribute our success to a number of factors that significantly improve the sensitivity and specificity of the analysis. Firstly, we utilize a subset of Y-STR loci that have been carefully selected for their superior performance under stressed conditions in both multiplex and monoplex formats. Specifically these loci function with low copy number templates in the presence of a vast excess of potentially confounding female DNA. Secondly, sperm and non-sperm DNA is co-extracted without a differential extraction process to prevent the unnecessary loss of the small number of structurally fragile sperm remaining in the cervicovaginal tract several days after intercourse. Thirdly, low copy number detection is facilitated by increasing the cycle number to 34-35 cycles and by the ability to input up to 450 ng of co-extracted sperm/non-sperm DNA into the PCR reaction without the appearance of confounding female artifacts. Lastly, the proper collection of post-coital cervicovaginal samples, instead of the lower or mid-vaginal tract samples often taken, is required for optimal recovery of sperm for analysis. In this report we demonstrate that our previously described 19 Y-STR loci systems (MPI and MPII) permit a reliable high resolution haplotype determination of the semen donor in cervicovaginal samples taken up to 48 h after intercourse

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. Spectral Cytopathology of Cervical Samples: Detecting Cellular Abnormalities in Cytologically Normal Cells

    PubMed Central

    Schubert, Jennifer M.; Bird, Benjamin; Papamarkakis, Kostas; Miljković, Miloš; Bedrossian, Kristi; Laver, Nora; Diem, Max

    2010-01-01

    Aim Spectral Cytopathology (SCP) is a novel spectroscopic method for objective and unsupervised classification of individual exfoliated cells. The limitations of conventional cytopathology are well-recognized within the pathology community. In SCP, cellular differentiation is made by observing molecular changes in the nucleus and the cytoplasm, which may or may not produce morphological changes detectable by conventional cytopathology. This proof of concept study demonstrates SCP’s potential as an enhancing tool for cytopathologists by aiding in the accurate and reproducible diagnosis of cells in all states of disease. Method Infrared spectra are collected from cervical cells deposited onto reflectively coated glass slides. Each cell has a corresponding infrared spectrum that describes its unique biochemical composition. Spectral data are processed and analyzed by an unsupervised chemometric algorithm, Principal Component Analysis (PCA). Results In this blind study, cervical samples are classified by analyzing the spectra of morphologically normal looking squamous cells from normal samples and samples diagnosed by conventional cytopathology with low grade squamous intraepithelial lesions (LSIL). SCP discriminated cytopathological diagnoses amongst twelve different cervical samples with a high degree of specificity and sensitivity. SCP also correlated two samples with abnormal spectral changes: these samples had a normal cytopathological diagnosis but had a history of abnormal cervical cytology. The spectral changes observed in the morphologically normal looking cells are most likely due to an infection with human papillomavirus, HPV. HPV DNA testing was conducted on five additional samples, and SCP accurately differentiated these samples by their HPV status. Conclusions SCP tracks biochemical variations in cells that are consistent with the onset of disease. HPV has been implicated as the cause of these changes detected spectroscopically. SCP does not depend on

  8. Development and evaluation of a reverse transcription-insulated isothermal polymerase chain reaction (RT-iiPCR) assay for detection of equine arteritis virus in equine semen and tissue samples using the POCKIT™ system.

    PubMed

    Carossino, Mariano; Lee, Pei-Yu A; Nam, Bora; Skillman, Ashley; Shuck, Kathleen M; Timoney, Peter J; Tsai, Yun-Long; Ma, Li-Juan; Chang, Hsiao-Fen G; Wang, Hwa-Tang T; Balasuriya, Udeni B R

    2016-08-01

    Equine arteritis virus (EAV) is the causative agent of equine viral arteritis (EVA), a respiratory and reproductive disease of horses. Most importantly, EAV induces abortion in pregnant mares and can establish persistent infection in up to 10-70% of the infected stallions, which will continue to shed the virus in their semen. The objective of this study was to develop and evaluate a reverse transcription insulated isothermal polymerase chain reaction (RT-iiPCR) for the detection of EAV in semen and tissue samples. The newly developed assay had a limit of detection of 10 RNA copies and a 10-fold higher sensitivity than a previously described real-time RT-PCR (RT-qPCR). Evaluation of 125 semen samples revealed a sensitivity and specificity of 98.46% and 100.00%, respectively for the RT-qPCR assay, and 100.00% and 98.33%, respectively for the RT-iiPCR assay. Both assays had the same accuracy (99.2%, k=0.98) compared to virus isolation. Corresponding values derived from testing various tissue samples (n=122) collected from aborted fetuses, foals, and EAV carrier stallions are as follows: relative sensitivity, specificity, and accuracy of 88.14%, 96.83%, and 92.62% (k=0.85), respectively for the RT-qPCR assay, and 98.31%, 92.06%, and 95.08% (k=0.90), respectively for the RT-iiPCR assay. These results indicate that RT-iiPCR is a sensitive, specific, and a robust test enabling detection of EAV in semen and tissue samples with very considerable accuracy. Even though the RT-qPCR assay showed a sensitivity and specificity equal to virus isolation for semen samples, its diagnostic performance was somewhat limited for tissue samples. Thus, this new RT-iiPCR could be considered as an alternative tool in the implementation of EAV control and prevention strategies.

  9. Effect of semen collection by transrectal massage of accessory sexual glands or artificial vagina on the outcome of breeding soundness examinations of Italian yearling beef bulls.

    PubMed

    Sylla, Lakamy; Palombi, Claudio; Stradaioli, Giuseppe; Vagniluca, Antonio; Monaci, Maurizio

    2015-03-15

    Although semen quality is one of the major traits that influence breeding soundness examination outcomes in bulls, field conditions occasionally do not allow for the collection of semen samples by means of an artificial vagina. The aims of the present study were to report the results of a large number of semen collections that were performed via the transrectal massage (TRM) of the accessory sexual glands of Italian yearling beef bulls and compare this semen collection method to the artificial vagina (AV) method in term of breeding soundness examination outcomes; furthermore, we determined whether the breed affected the semen characteristics. In the TRM group (n = 475), the semen samples were collected via TRM of the accessory sexual glands, and in the AV group (n = 502), the AV method was used. In the TRM group, semen samples were obtained from 81.3% of the bulls and penile protrusion was observed in 87.6% of the animals during semen collection. The sperm concentrations (920.5 ± 439.0 vs. 281.0 ± 259.8 × 10(6)/mL) and the percentages of total abnormal spermatozoa (22.8 ± 15.0 vs. 18.8 ± 12.9) were significantly higher in the AV group than those in the TRM group. The percentage of bulls that did not meet the minimum requirement for normal cells (≥70%) was 6.2% higher in the AV group than that in the TRM group (P < 0.05). Moreover, the samples collected from Chianina bulls by TRM exhibited a lower percentage of motile sperm and a higher percentage of abnormal spermatozoa when compared with the other two breeds. The major drawbacks of the TRM technique were the inability to conduct complete evaluation of the libido and mating ability of the yearling bulls, a significant reduction of the number of spermatozoa collected, and an increase in the variability of the semen characteristics due to breed. In conclusion, despite the drawbacks, TRM guarantees that semen evaluation can be conducted in cases in which the semen samples cannot be collected

  10. Relationship of seminal reactive nitrogen and oxygen species and total antioxidant capacity with sperm DNA fragmentation in infertile couples with normal and abnormal sperm parameters.

    PubMed

    Khosravi, F; Valojerdi, M R; Amanlou, M; Karimian, L; Abolhassani, F

    2014-02-01

    The objective of this study was to investigate the association between the amount of superoxide anion, peroxynitrite as oxidative stress (OS) markers and total antioxidant capacity (TAC) with sperm DNA fragmentation in infertile men with abnormal semen parameters. Semen samples were obtained from 102 infertile couples and divided into groups with normal and abnormal semen parameters according to the World Health Organization (WHO). Peroxynitrite and superoxide anions were detected using spectrofluorometric assays combined with 2,7 dicholorofluorescein (DCF)-DA and 4-chloro-7-nitrobenzo-2-oxa -1, 3-diazole (NBD-CL). Colorimetric assay was used for evaluation of TAC, while DNA fragmentation was studied by using sperm chromatin dispersion test. Superoxide anion, peroxynitrite and DNA fragmentation were significantly higher in infertile couples with abnormal semen parameters as compared to infertile couples with normal semen (P < 0.01). TAC was significantly lower in infertile men with abnormal semen parameters (P < 0.01). There was also a significant positive correlation between OS markers with sperm DNA fragmentation (r = 0.59, P < 0.01 and r = 0.67, P < 0.01, respectively). We have found that imbalance between superoxide anion and peroxynitrite with antioxidant capacity in infertile men with abnormal sperm parameters is associated with higher sperm DNA fragmentation.

  11. Sperm shape abnormalities in carbaryl-exposed employees

    PubMed Central

    Wyrobek, A. J.; Watchmaker, G.; Gordon, L.; Wong, K.; Moore, D.; Whorton, D.

    1981-01-01

    Semen was collected from 50 men occupationally exposed to carbaryl (1-naphthyl methyl carbamate) in a produciton plant for durations of 1 to 18 years and compared to semen from a control group of 34 unexposed, newly-hired workers. Employment, fertility, health, personal data, and blood samples were collected for each individual. Semen samples were analyzed for changes in sperm count, morphology, and frequency of sperm carrying double flourescent bodies (YFF). As a group, the exposed workers showed a significantly higher proportion of sperm with abnormal head shapes than did the control group (p < 0.005). Age, smoking habits, and medical problems did not appear to affect this result. This finding appears to be limited to men working in the carbaryl production area at the time of sampling. Sperm count and YFF did not show similar differences, which may be because they are known to be statistically less sensitive to small changes. Formerly exposed workers (away from carbaryl for an average of 6.3 years) showed a marginally significant elevation in sperm abnormalities compared to controls (p < .05, one-tailed statistical analyses) suggesting that the increase in abnormal morphology may not be reversible. However, the question of reversibility is sensitive to confounding factors and small sample sizes and, therefore, requires further study. With these data a definitive link between carbaryl exposure and human seminal defects cannot be established. Although a distinct effect on sperm morphology was seen in the exposed group, the increases in sperm shape abnormalities were not related to exposure dose (estimated by number of years on the job or job classification during the year prior to semen collection). Inexplicably, the increases in sperm abnormalities were seen primarily in currently exposed men who had worked with carbaryl for less than approximately 6 years. These findings suggest the need for further study since other workplace-related factor(s) may be responsible

  12. 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.

  13. 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.

  14. [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.

  15. 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

  16. Abnormal gel flotation caused by contrast media during adrenal vein sampling

    PubMed Central

    Lima-Oliveira, Gabriel; Lippi, Giuseppe; Salvagno, Gian Luca; Gelati, Matteo; Bassi, Antonella; Contro, Alberto; Pizzolo, Francesca; Guidi, Gian Cesare

    2016-01-01

    Introduction During adrenal venous sampling (AVS) procedure, radiologists administer a contrast agent via the catheter to visualize the proper catheter position. Materials and methods A patient with primary aldosteronism diagnostic-hypothesis was admitted for AVS. A venogram was performed to
confirm the catheter’s position with 2mL of Iopamidol 300 mg/mL. Samples were collected with syringe connected to a hydrophilic coated catheter by low-pressure aspiration from each of the four collection sites: inferior vena cava in the suprarenal portion, inferior vena cava in the infrarenal portion, left adrenal vein, and right adrenal vein; then immediately transferred from syringe to tubes with gel separator. All tubes were centrifuged at 1200 x g for 10 minutes. Results At the end of centrifugation process, primary blood tubes containing blood from inferior vena cava and left adrenal vein exhibited the standard gel separator barrier, while tubes from right adrenal vein showed abnormal flotation of gel separator. The radiologist confirmed the usage of 2.6 mL instead of 2.0 mL of Iopamidol 300 mg/mL. This iodinated contrast media, with 1.33 g/cm3 of density, was used close to the right adrenal vein due to some difficulty to access it. Conclusion The abnormal flotation of gel separator in samples taken from right adrenal vein can be explained by the usage of the iodinated
contrast media. We suggest using plain-tubes (without gel separator) for AVS in order to avoid preanalytical nonconformities. Moreover, a blood volume equivalent to twice the catheter extension should be discarded to eliminate residual contrast media before collection of samples for laboratory assays. PMID:27812311

  17. 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

  18. 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).

  19. An efficient sampling algorithm for uncertain abnormal data detection in biomedical image processing and disease prediction.

    PubMed

    Liu, Fei; Zhang, Xi; Jia, Yan

    2015-01-01

    In this paper, we propose a computer information processing algorithm that can be used for biomedical image processing and disease prediction. A biomedical image is considered a data object in a multi-dimensional space. Each dimension is a feature that can be used for disease diagnosis. We introduce a new concept of the top (k1,k2) outlier. It can be used to detect abnormal data objects in the multi-dimensional space. This technique focuses on uncertain space, where each data object has several possible instances with distinct probabilities. We design an efficient sampling algorithm for the top (k1,k2) outlier in uncertain space. Some improvement techniques are used for acceleration. Experiments show our methods' high accuracy and high efficiency.

  20. 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

  1. 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.

  2. 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.

  3. Classification of raw and roasted Semen Cassiae samples with the use of Fourier transform infrared fingerprints and least squares support vector machines.

    PubMed

    Lai, Yanhua; Ni, Yongnian; Kokot, Serge

    2010-06-01

    Raw and roasted Semen Cassiae seeds, a complex traditional Chinese medicine (TCM), are used as examples to research and develop a method of classification analysis based on measurements of Fourier transform infrared (FT-IR) spectral fingerprints. Eighty samples of the TCM were measured in the mid-infrared range, 400-2000 cm(-1) (KBr pellets), and the complex overlapping spectra were submitted for interpretation to a principal component analysis least squares support vector machine (PC-LS-SVM), kernel principal component analysis least squares support vector machine (KPC-LS-SVM), and radial basis function artificial neural networks (RBF-ANN). The LS-SVM models were developed with an RBF kernel function and a grid search technique. Training models were constructed with the use of raw and first-derivative spectra and these were then verified by another data set containing both raw and roasted spectral objects. It was demonstrated that the first-derivative data set produced the best separation of the spectral objects. In general, satisfactory analytical performance was obtained with the PC-LS-SVM, KPC-LS-SVM, and RBF-ANN training models and with the classification of the verification spectral objects. With regard to chemometrics modeling, the performance of KPC-LS-SVM was somewhat more economical than that of the PC-LS-SVM model. It would appear that the latter relatively simple model would be sufficient for application to most small to medium sized FT-IR fingerprint data sets, but with larger matrices the more complex models, such as the RBF-ANN and KPC-LS-SVM, may be more advantageous on a computational basis.

  4. Spectrum of Cytogenomic Abnormalities Revealed by Array Comparative Genomic Hybridization on Products of Conception Culture Failure and Normal Karyotype Samples.

    PubMed

    Zhou, Qinghua; Wu, Shen-Yin; Amato, Katherine; DiAdamo, Autumn; Li, Peining

    2016-03-20

    Approximately 30% of pregnancies after implantation end up in spontaneous abortions, and 50% of them are caused by chromosomal abnormalities. However, the spectrum of genomic copy number variants (CNVs) in products of conception (POC) and the underlying gene-dosage-sensitive mechanisms causing spontaneous abortions remain largely unknown. In this study, array comparative genomic hybridization (aCGH) analysis was performed as a salvage procedure for 128 POC culture failure (POC-CF) samples and as a supplemental procedure for 106 POC normal karyotype (POC-NK) samples. Chromosomal abnormalities were detected in 10% of POC-CF and pathogenic CNVs were detected in 3.9% of POC-CF and 5.7% of POC-NK samples. Compiled results from this study and relevant case series through a literature review demonstrated an abnormality detection rate (ADR) of 35% for chromosomal abnormalities in POC-CF samples, 3.7% for pathogenic CNVs in POC-CF samples, and 4.6% for pathogenic CNVs in POC-NK samples. Ingenuity Pathway Analysis (IPA) was performed on the genes from pathogenic CNVs found in POC samples. The denoted primary gene networks suggested that apoptosis and cell proliferation pathways are involved in miscarriage. In summary, a similar spectrum of cytogenomic abnormalities was observed in POC culture success and POC-CF samples. A threshold effect correlating the number of dosage-sensitive genes in a chromosome with the observed frequency of autosomal trisomy is proposed. A rationalized approach using firstly fluorescence in situ hybridization (FISH) testing with probes of chromosomes X/Y/18, 13/21, and 15/16/22 for common aneuploidies and polyploidies and secondly aCGH for other cytogenomic abnormalities is recommended for POC-CF samples.

  5. 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

  6. Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms

    PubMed Central

    YAMANAKA, Masaya; TOMITA, Kazuhisa; HASHIMOTO, Shu; MATSUMOTO, Hiroshi; SATOH, Manabu; KATO, Hiromi; HOSOI, Yoshihiko; INOUE, Masayasu; NAKAOKA, Yoshiharu; MORIMOTO, Yoshiharu

    2016-01-01

    Density gradient centrifugation (DGC) and swim-up techniques have been reported for semen preparation in assisted reproductive techniques in humans. We investigated whether semen preparation using a combination of DGC and swim-up techniques could effectively decrease morphologically abnormal human sperms at the ultrastructural level. Semen samples were obtained from 16 infertile males and fractionated by swim-up following DGC. Ultrastructural abnormalities of sperms obtained from original semen, lower layer of swim-up following DGC, and upper layer of swim-up following DGC were analyzed by transmission electron microscopy. The correlation among ultrastructural head abnormality in sperms from the upper layer of swim-up, fertilization in in vitro fertilization, and pregnancy after embryo transfer was also investigated. Furthermore, sperms with DNA fragmentation in the samples processed via a combination of DGC and swim-up was assessed in a sperm chromatin structure assay. Ultrastructural abnormalities in sperm heads and tails in the upper layer after swim-up following DGC was the lowest among the three groups. Sperms with nuclear vacuoles were the most difficult to eliminate using a combination of DGC and swim-up in all types of head abnormalities. A negative correlation was confirmed between the fertilization rates of intracytoplasmic sperm injection and head abnormality of sperms obtained from the upper layer of the swim-up following DGC. Sperms with DNA fragmentation were effectively decreased using the combination of two techniques. In conclusion, the combination of DGC and swim-up effectively decreased the number of sperms with ultrastructural abnormalities both in the head and in the tail. However, sperms with ultrastructural abnormalities that cannot be completely decreased using a combination of DGC and swim-up may impair fertilization in some cases of intracytoplasmic sperm injection. PMID:27616283

  7. 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.

  8. 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.

  9. 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

  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. 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.

  12. 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.

  13. 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

  14. Increased sperm ubiquitination correlates with abnormal chromatin integrity.

    PubMed

    Hodjat, M; Akhondi, M A; Al-Hasani, S; Mobaraki, M; Sadeghi, M R

    2008-09-01

    Ubiquitin, a 8.5 kDa peptide that marks other proteins for proteasomal degradation, tags defective spermatozoa during epididymal passage and is proposed as a biomarker for sperm quality. The present study was designed to evaluate the relationships between sperm ubiquitination, sperm chromatin integrity and semen parameters. Semen samples from 63 couples were collected and analysed according to World Health Organization criteria. Each sample was evaluated for sperm ubiquitination by the direct immunofluorescence method, using anti-ubiquitin antibodies. Chromatin integrity of the same samples was analysed using acridine orange (AO) and toluidine blue (TB) tests. A positive correlation was found between ubiquitinated spermatozoa and the percentage of spermatozoa with abnormal chromatin (AO: r = 0.58, P < 0.001 and TB: r = 0.48, P < 0.001). Negative correlations were obtained between sperm ubiquitination and: sperm count (r = -0.2, P = 0.048), sperm morphology (r = -0.36, P = 0.003), rapidly progressive motility (r = -0.25, P = 0.044) and slow progressive motility (r = -0.28, P = 0.022). Sperm ubiquitination was positively correlated with the percentage of immotile spermatozoa. These results show that among semen parameters, chromatin abnormality is more closely associated with sperm ubiquitination and further validate sperm ubiquitination as a suitable marker for sperm quality.

  15. HPV is detectable in virtually all abnormal cervical cytology samples after reinvestigation of HPV negatives with multiple alternative PCR tests.

    PubMed

    Evans, Mark Francis; Adamson, Christine Stewart-Crawford; Schned, Laura Meredith; St John, Timothy Louis; Leiman, Gladwyn; Ashikaga, Takamaru; Cooper, Kumarasen

    2010-09-01

    The demonstration of human papillomavirus (HPV) in 99.7% of cervical carcinoma surgical specimens from around the world required investigations by multiple alternative polymerase chain reaction (PCR) assays. A similar approach may therefore be necessary to best characterize HPV prevalence and genotype distribution among cervical cytology samples. In an earlier study, 752 of 799 (94.1%) abnormal and 82 of 300 (27.3%) normal cytology specimens tested HPV positive after PCR using GP5+/6+primers. This study has reinvestigated the "HPV negative" abnormal samples (20 atypical squamous cells of undetermined significance, 5 low-grade squamous intraepithelial lesion, 14 atypical squamous cells, cannot exclude HSIL, 6 high-grade squamous intraepithelial lesion) and an age-matched cohort of "HPV negative" normal (negative for an intraepithelial lesion or malignancy) samples by PCR using PGMY09/11, FAP59/64, and LCR-E7 primers. PGMY09/11-GP5+/6+ nested PCR was performed on samples that were HPV negative by PGMY09/11 PCR. After the first 3 assays, HPV was detected in 41 of 45 (91.1%) abnormal and in 10 of 47 (21.3%) normal samples (P<0.0001). Eighteen HPV genotypes were detected and in some samples the genotype that was identified differed between the tests. The nondetection of common HPV genotypes (eg, HPVs 6, 11, 16, and 18) was notable. High-grade histopathology was found for 2 patients with HPV52-positive cytopathology. Combined with our earlier study, HPV (40 different genotypes) is shown in 99.5% of abnormal samples (99.8% inclusive of the nested PCR data). These findings show that HPV genotype and prevalence estimates are dependent on the method(s) of detection and indicate that suboptimal analytical sensitivity for one or more of the less common high-risk HPV genotypes could lead to impaired clinical sensitivity. HPV may be causal in almost every instance of abnormal cervical cytology; however, passenger HPV that is incidental to an abnormality may also have been

  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. Post-thaw viability of european bison (Bison bonasus) semen frozen with extenders containing egg yolk or lipids of plant origin and examined with a heterologous in vitro fertilization assay.

    PubMed

    Pérez-Garnelo, S S; Oter, M; Borque, C; Talavera, C; Delclaux, M; Martínez-Nevado, E; Palasz, A T; De la Fuente, J

    2006-06-01

    Basic characteristics of European bison (Bison bonasus) semen were described and the efficacies of two extenders-Triladyl, containing egg yolk, and a synthetic extender, containing soybean lipids-were tested for semen cryopreservation. Seven ejaculates were collected by electroejaculation from a 10-yr-old, European bison bull. Each ejaculate was diluted at 37 degrees C to a final concentration of 200 x 10(6) sperm/ml with Triladyl or the synthetic extender. Extended semen samples were frozen according to a standard bull semen freezing protocol. After 2 wk of storage, one straw from each extender and ejaculate was thawed, and postthaw quality was evaluated by individual sperm motility and movement rate, numbers of sperm morphologic abnormalities and intact acrosomes, functional integrity of the sperm membranes determined by hypoosmotic swelling test (HOST), viability (live-dead, eosin-nigrosin stain), and a heterologous in vitro sperm penetration assay (SPA). A total of 600 in vitro-matured bovine oocytes were inseminated with 1 X 10(6) spermatozoa of Holstein semen frozen-thawed in Triladyl (control) or of European bison semen frozen in Triladyl or the synthetic extender. Nuclear status of the oocytes was determined after 18 h of sperm-oocyte coincubation. Extender had no effect on any evaluated parameters of semen after dilution and cooling (4 hr at 5 degrees C) or in postthaw individual motility, quality of movement, and sperm morphology. However, significantly (P < 0.05) higher numbers of spermatozoa with intact acrosomes, intact membranes (HOST), and viable sperm (P < 0.01) were in semen frozen in Triladyl than in the synthetic extender. Mean values for heterologous SPA for bull (control) and for bison semen frozen in the synthetic extender were very much alike-63.3+/-10.6% and 63.1 +/- 15.9%, respectively; bison semen frozen in Triladyl was lower, 43.0+/-24.2% but not significantly different. Cumulative results from a variety of viability assays of diluted

  18. [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.

  19. 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

  20. 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.

  1. Effects of hypotaurine, cysteamine and aminoacids solution on post-thaw microscopic and oxidative stress parameters of Angora goat semen.

    PubMed

    Bucak, Mustafa Numan; Tuncer, Pürhan Barbaros; Sariözkan, Serpil; Ulutaş, Pinar Alkim; Coyan, Kenan; Başpinar, Nuri; Ozkalp, Birol

    2009-12-01

    This study was conducted to determine the effects of cysteamine, hypotaurine and aminoacids solution (BME) on standard semen parameters, lipid peroxidation and antioxidant activities of Angora goat semen after the freeze-thawing process. Ejaculates collected from four Angora goats were evaluated and pooled at 37 degrees C. Semen samples, which were diluted with a Tris-based extender containing the antioxidants hypotaurine (5 mM) and cysteamine (5 mM), and an aminoacid solution (13%), and an extender containing no antioxidants (control), were cooled to 5 degrees C and frozen in 0.25-ml French straws in liquid nitrogen. Frozen straws were thawed individually at 37 degrees C for 20s in a water bath for evaluation. Supplementation with cysteamine, hypotaurine and BME caused significant (P<0.05) increases in sperm motility, and significant (P<0.05) decreases in total abnormality rates in comparison to the control group. While all in vitro treatments did not affect the acrosomal abnormality rates, hypotaurine and BME but not cysteamine significantly (P<0.05) increased the HOST results as compared to the control group. Supplementation with antioxidants and BME did not significantly affect MDA levels and CAT activity in comparison to the control group (P>0.05). The antioxidants hypotaurine and cysteamine decreased SOD activity when compared to the BME group and controls (P<0.001).

  2. Cysteamine supplementation revealed detrimental effect on cryosurvival of buffalo sperm based on computer-assisted semen analysis and oxidative parameters.

    PubMed

    Swami, Dheer Singh; Kumar, Pradeep; Malik, R K; Saini, Monika; Kumar, Dharmendra; Jan, M H

    2017-02-01

    The aim of this study was to investigate the effect of addition of cysteamine to the semen extender on post-thaw semen quality. A total of 30 ejaculates were collected from six bulls. Each ejaculate was divided into five equal parts and diluted to final concentration of 80 million sperms/mL using Optixcell(®)(IMV, France) semen extender supplemented with different concentrations of cysteamine (0, 0.75, 1.25, 2.5 and 5mM) and cryopreserved. In the frozen-thawed samples, the VAP, VSL, VCL ALH and sperm motility of control samples was greater (P<0.05) than cysteamine treated samples. The sperm abnormality and malondialdehyde (MDA) concentration were found highest in 5mM cysteamine treated samples. The cysteamine treated samples travelled significantly less distance in cervical mucus as compared to control. Further, cysteamine decreased acrosomal integrity of sperm. In incubation test, control samples showed better sperm motility as compared to treatment groups. Further, cysteamine supplementation decreased the total antioxidants and increased the MDA concentration of sperm. From the study, we hypothesized that cysteamine cannot stimulate synthesis of glutathione (GSH) intracellularly in sperm to combat free radicals because during the maturation, sperm lost its cytoplasm which is necessary for biochemical reaction in which cysteamine reacts with cystine to form a mixed disulfide which taken up by cells and split into cysteine in the cytoplasm. Synthesis of GSH depends on the availability of cysteine. In conclusion, the results of our study strongly emphasize that cysteamine would not be a suitable additive in extender for freezing buffalo bull semen.

  3. 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

  4. 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.

  5. 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.

  6. 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

    concentration, motility, vitality, morphology or DNA fragmentation index. The percentage of men with abnormal volume, concentration and total sperm increased with increasing body size (P < 0.05). No relationship between physical activity and semen parameters was identified. LIMITATIONS, REASONS FOR CAUTION Our cohort was largely overweight and sedentary, which may result in limited external validity, i.e. generalizability. The lack of physical activity did preclude examination of exercise more frequently than once per week, thus our ability to examine more active individuals is limited. WIDER IMPLICATIONS OF THE FINDINGS Body size (as measured by BMI or WC) is negatively associated with semen parameters with little influence of physical activity. Our findings are the first showing a relationship between WC and semen parameters in a sample of men without known infertility. Given the worldwide obesity epidemic, further study of the role of weight loss to improve semen parameters is warranted. STUDY FUNDING/COMPETING INTEREST(S) Supported by the Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development (Contracts #N01-HD-3-3355, N01-HD-3-3356 and N01-HD-3-3358). There are no competing interests. PMID:24306102

  7. 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.

  8. 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

  9. 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.

  10. 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.

  11. 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

  12. Semen samples showing an increased rate of spermatozoa with imprinting errors have a negligible effect in the outcome of assisted reproduction techniques

    PubMed Central

    Camprubí, Cristina; Pladevall, Marta; Grossmann, Mark; Garrido, Nicolás; Pons, Maria; Blanco, Joan

    2012-01-01

    The topic of imprinting defects present in the sperm of infertile patients has been addressed by several reports in the last few years. However, whether methylation abnormalities at one or few CpGs within an imprinted locus are pathological is a matter of debate. Moreover, whether imprinting anomalies in sperm could interfere with fertility treatment outcomes is still unknown. In this report we analyze the sperm DNA methylation profile of H19-ICR, KvDMR, SNRPN-ICR, IG-DMR and MEG3-DMR by pyrosequencing in 107 infertile men series and a control population of 30 proven fertile males. DNA methylation was statistically evaluated from two points of view: first, the methylation of each CpG was analyzed in the control population and the mean, standard deviation and range were determined and compared with infertile population data; second, in order to define altered methylation patterns for each region, a hierarchical cluster analysis was performed by which individuals were grouped in different clusters according to the degree of similarity of their methylation pattern. Two pieces of data supported the results obtained in the multi-variate analysis: the classification of the vast majority of control individuals in clusters with normal methylation patterns and the significant differences in methylation levels found between individuals within the normal and abnormal clusters. Individuals included in normal and abnormal methylation clusters were compared according to seminal parameters as well as to the outcome of assisted reproduction. PMID:22885410

  13. Effect of single-layer centrifugation or washing on frozen-thawed donkey semen quality: Do they have the same effect regardless of the quality of the sample?

    PubMed

    Ortiz, I; Dorado, J; Morrell, J M; Crespo, F; Gosálvez, J; Gálvez, M J; Acha, D; Hidalgo, M

    2015-07-15

    The aims of this study were to determine the sperm quality of frozen-thawed donkey sperm samples after single-layer centrifugation (SLC) using Androcoll-E in comparison to sperm washing or no centrifugation and to determine if the effect on the sperm quality after SLC or sperm washing depends on the quality of the sample. Frozen-thawed sperm samples from Andalusian donkeys were divided into three aliquots, and they were processed using three different techniques after thawing: uncentrifuged diluted control (UDC), sperm washing (SW), and SLC. Afterward, sperm quality index was estimated by integrating all parameters (total and progressive sperm motility, membrane integrity, and DNA fragmentation) in a single value. The relationship between the sperm quality of thawed UDC samples and the effect on sperm parameters in SW and SLC-selected samples was assessed. Sperm quality index was significantly higher (P < 0.001) in SLC (0.8 ± 0.0) samples than that in UDC (0.6 ± 0.0) and SW (0.6 ± 0.0) samples, regardless of the sperm quality index after thawing of the sperm sample. In conclusion, SLC of frozen-thawed donkey spermatozoa using Androcoll-E-Small can be a suitable procedure for selecting frozen-thawed donkey sperm with better quality, in particular in those samples where an improvement in motility is needed.

  14. 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...

  15. 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

  16. 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

  17. Effects of In Vitro Zinc Sulphate Additive to The Semen Extender on Water Buffalo (Bubalusbubalis) Spermatozoa before and after Freezing

    PubMed Central

    Dorostkar, Kamran; Alavi Shoushtari, Sayed Mortaza; Khaki, Amir

    2014-01-01

    Background The objective of the study was to investigate the effects of in vitro zinc sulphate additive to semen extender on sperm parameters (progressive motility, viability, membrane integrity and DNA stability) after cryopreservation. Materials and Methods In this Prospective longitudinal laboratory study, semen samples of 5 buffalo bulls of 3-5 years old were collected at 5 different occasions from Iran, Urmia during summer and autumn 2011, 25 samples were used in each treatment. Sperm progressive motility, viability and abnormal morphology were measured before and at 0.5 (T0), 1(T1) and 2(T2) hours after diluting semen(1:10 v/v) in Tris-citric acid based extender (without egg yolk and glycerol) at 37˚C containing none (control group), 0.072, 0.144, 0.288, 0.576 and 1.152 mg/L zinc sulphate to investigate dose and time effects. Next, a Tris-citric acid-egg yolk-glycerol extender (20% egg yolk and 7% glycerol) containing the same amount of zinc sulphate was prepared, diluted semen (1:10 v/v) was cooled and kept into a refrigerated chamber (4˚C) for 4 hours to equilibrate. Sperm progressive motility, viability, abnormal morphology, membrane integrity and DNA damage were estimated.The equilibrated semen was loaded in 0.5 ml French straws and frozen in liquid nitrogen. Later, the frozen semen was thawed and the same parameters as well as total antioxidant capacity (TAC) of the frozen-thawed semen were determined. Results The results showed that zinc sulphate additive at the rate of 0.288 mg/L gave a higher protection of sperm progressive motility (53.7 ± 1.8% vs. 40.5 ± 1.7%), viability (70.8 ± 1.8% vs. 60.1 ± 1.5%), membrane integrity (67.3 ± 1.6% vs. 56.6 ± 1.7%), DNA stability (10.1 ± 0.47% vs. 11.8 ± 0.33% damaged DNA) through the process of dilution, equilibration and freeze-thawing and caused a higher TAC level (81 ± 3.3% vs. 63 ± 3.2 µmol/L) after freez-thawing compared to the control group. Adding 0.576 and 1.152 mg/L zinc sulphate, however

  18. Associations between urinary phthalate concentrations and semen quality parameters in a general population

    PubMed Central

    Bloom, M.S.; Whitcomb, B.W.; Chen, Z.; Ye, A.; Kannan, K.; Buck Louis, G.M.

    2015-01-01

    STUDY QUESTION Are urinary phthalate concentrations associated with altered semen quality parameters among males recruited from the general population? SUMMARY ANSWER Urinary levels of metabolites of phthalate diesters are associated with lower total sperm counts, larger sperm head sizes, and higher percentages of morphologically abnormal sperm. WHAT IS KNOWN ALREADY High dose experiments in rats implicate phthalates as anti-androgens. Studies involving infertile men seeking care suggest that phthalates influence measures of semen quality raising concern about the implications for men in the general population. STUDY DESIGN, SIZE, DURATION This prospective cohort study comprised 501 male partners in couples discontinuing contraception to become pregnant, who were recruited from 16 US counties using population-based sampling frameworks from 2005 to 2009. PARTICIPANTS/MATERIALS, SETTING, METHODS Urine and semen samples were obtained at baseline from 473 (94%) men, of whom 378 (80%) men provided a second sample the following month. Urine was analyzed for 14 monoester metabolites of phthalate diesters by high-performance liquid chromatography coupled to tandem mass spectrometry. Semen samples were analyzed for 34 quality parameters categorized as general, motility, morphology, sperm head and sperm chromatin structure. MAIN RESULTS AND THE ROLE OF CHANCE Urinary mono-[2-(carboxymethyl) hexyl] phthalate (MCMHP), mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono-benzyl phthalate (MBzP), and mono-isononyl phthalate (MNP) were significantly associated with lower total sperm counts and concentrations, larger sperm head sizes, higher proportions of megalo head sperm morphology, and/or other morphological changes. Urinary mono-methyl phthalate (MMP) and mono-cyclohexyl phthalate (MCPP) were significantly associated with lower sperm motility, and urine mono-2-ethylhexyl phthalate (MEHP) was significantly associated with higher sperm motility. LIMITATIONS, REASONS FOR

  19. 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).

  20. 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

  1. 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.

  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. 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

  4. Semen Analysis Test

    MedlinePlus

    ... indicate fewer sperm, which diminishes opportunities for successful fertilization and subsequent pregnancy. Excessive seminal fluid may dilute ... know? While abnormal results decrease the chances of fertilization, some couples with poor results on infertility tests ...

  5. [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.

  6. 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

  7. 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

  8. 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

  9. 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.

  10. Congenital Abnormalities

    MedlinePlus

    ... Listen Español Text Size Email Print Share Congenital Abnormalities Page Content Article Body About 3% to 4% ... of congenital abnormalities earlier. 5 Categories of Congenital Abnormalities Chromosome Abnormalities Chromosomes are structures that carry genetic ...

  11. 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.

  12. 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.

  13. Levels of adipocytokines and vitamin D in a biracial sample of young metabolically healthy obese and metabolically abnormal obese women

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purpose: Adipocytokines and vitamin D (vitD) concentrations may contribute to cardiometabolic risk profiles in obese populations. The purpose was to determine if levels of adipocytokines and vitD differ between young metabolically healthy obese (MHO) and metabolically abnormal obese (MAO) black and ...

  14. Chronic environmental exposure to lead affects semen quality in a Mexican men population

    PubMed Central

    Morán-Martínez, Javier; Carranza-Rosales, Pilar; Morales-Vallarta, Mario; A. Heredia-Rojas, José; Bassol-Mayagoitia, Susana; Denys Betancourt-Martínez, Nadia; M. Cerda-Flores, Ricardo

    2013-01-01

    Background: Male infertility is affected by several factors. Lead is one of the heavy metals more bioavailable than usually modifies the sperm quality in humans. Objective: The aim of this study was to establish the role of lead in semen quality in environmentally exposed men. Materials and Methods: Semen and blood samples were obtained from two groups: the exposed group (EG=20) and the non-exposed group (NEG=27). Two semen aliquots were used, one to evaluate spermatic quality and the other for lead determination. Blood (PbB) and semen lead (PbS) determination was performed by atomic absorption spectrophotometry. Results: The PbB concentration was significantly greater in the EG, 10.10±0.97 µgdL-1 than in the NEG, 6.42±0.38 µgdL-1 (p<0.01), as well as the PbS concentration, with 3.28±0.35 and 1.76±0.14µgdL-1 in the EG and NEG respectively (p=0.043). A significant correlation between PbS and PbB concentration in the EG was found (r=0.573, p=0.038). Overall, the spermatic quality was lower in the EG than in the NEG. Specifically, there were significant differences in the spermatic concentration [EG=43.98±6.26 and NEG=68.78±8.51X106 cellmL-1 (p<0.01)], motility [EG=49±7 and NEG=67±4% (p=0.029)], viability [EG=36.32±3.59 and NEG=72.12±1.91% (p<0.01)] and abnormal morphology [EG=67±18 and NEG=32±12% (p<0.01)]. In the immature germ cells (IGC) concentration differences were found only for A cells (EG=8.1±1.1x100 and NEG=3.2±1.9X100 spermatozoa) (p<0.01) and for Sab cells (EG=3.4±2.2x100 and NEG=1.1±1.0X100 spermatozoa) (p=0.041). Conclusion: These results suggest that chronic environmental exposure to low levels of lead adversely affect the spermatic quality. PMID:24639755

  15. 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.

  16. Effects of the seminal plasma zinc content and catalase activity on the semen quality of water buffalo (Bubalus bubalis) bulls.

    PubMed

    Alavi-Shoushtari, S M; Rezai, S Asri; Ansari, M H Kh; Khaki, A

    2009-01-15

    In order to determine zinc and catalase content of seminal plasma in the buffalo and to study their associations with the semen characteristics, 54 semen samples were collected from 10 buffalo bulls; semen volume and sperm concentration, gross and progressive motility and viability were evaluated, seminal plasma was then harvested by centrifugation and its zinc content was estimated by atomic absorption spectrophotometer and its catalase activity determined by using a commercial kit. The zinc content of the seminal plasma (Mean +/- SEM) was recorded as 154.40 +/- 1.74 mg L(-1), while, the mean catalase value was 32.00 +/- 0.42 U mL(-1). The mean zinc values was highly correlated with sperm progressive motility and viability and with catalase values (p = 0.000 for all) and also was associated with gross motility (p = 0.020) and negatively with abnormal morphology (p = 0.049). The catalase values were highly associated with sperm progressive motility, viability and zinc content (p = 0.000 for all) and was associated with sperm gross motility (p = 0.024). For further clarification of these correlations, the samples were categorized in three groups of excellent (Ex, >90% motile, n = 33), good (Go, 80-89% motile, n = 15) and moderate (Mo, <79% motile, n = 6) according to their percentage of sperm motility. The mean progressive motility in Ex group was 92.54 +/- 0.51%, in Go group was 81.66 +/- 0.62% and in Mo group was 71.66 +/- 1.05%. The mean zinc and catalase values were recorded as 161.07 +/- 1.63 mg L(-1) and 33.41 +/- 0.34 U mL(-1) in Ex, 146.70 +/- 1.91 mg L(-1) and 31.01 +/- 0.67 in Go and 136.42 +/- 4.97 mg L(-1) and 26.51 +/- 0.87 U mL(-1) in Mo groups. The mean zinc value in Ex group was highly associated with sperm motility, viability and catalase values, in Go group was associated with catalase values and highly associated with sperm abnormal morphology and in Mo group it was highly associations with catalase values only. The mean catalase value in Ex group

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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...

  5. 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.

  6. Sperm in poor quality semen from bulls during heat stress have a lower affinity for binding hydrogen-3 heparin

    SciTech Connect

    Ax, R.L.; Gilbert, G.R.; Shook, G.E.

    1987-01-01

    Binding assays with (/sup 3/H) heparin were performed using spermatozoa collected prior to, during, and following summer heat stress to dairy bulls. Ejaculates collected in August 1983 after a period of ambient temperatures exceeding 29.4/sup 0/C exhibited a high frequency of abnormal sperm, and motility was reduced in some samples. Sperm in samples collected during heat stress possessed dissociation constants for binding (/sup 3/H) heparin ranging from 134.5 to 163.2 nmol. In contrast, sperm in semen collected prior to and after heat stress had significantly lower dissociation constants (higher affinity) for (/sup 3/H)heparin, 12.9 to 56.4 nmol. The number of binding sites for (/sup 3/H) heparin on sperm did not change among collection periods. It was concluded that the binding affinity for (/sup 3/H) heparin may reflect membrane integrity of bull sperm.

  7. 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.

  8. 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.

  9. Scrotal insulation and its relationship to abnormal morphology, chromatin protamination and nuclear shape of spermatozoa in Holstein-Friesian and Belgian Blue bulls.

    PubMed

    Rahman, Mohammad Bozlur; Vandaele, Leen; Rijsselaere, Tom; Maes, Dominiek; Hoogewijs, Maarten; Frijters, Adrie; Noordman, Jakomien; Granados, Ana; Dernelle, Eric; Shamsuddin, Mohammed; Parrish, John J; Van Soom, Ann

    2011-10-15

    The objectives of this study were to identify the stages of spermatogenesis susceptible to elevated testicular temperature in terms of sperm motility, viability, morphology, chromatin protamination and nuclear shape. The latter two valuable parameters are not included in routine semen analysis. Scrotal insulation (SI) was applied for 48 h in 2 Holstein-Friesian (HF) and 2 Belgian Blue (BB) bulls and semen was collected at 7 d intervals along with semen collection of a non-insulated bull of each breed. Semen samples were frozen and assigned to 4 groups: period 1 (preinsulation) = -7 d and 0 d, where 0 d = initiation of SI after semen collection; period 2 = 7 d (sperm presumed in the epididymis during SI); period 3 = 14 d to 42 d (cells presumed at spermiogenesis and meiosis stages during SI); period 4 = 49 d to 63 d (cells presumed at spermatocytogenesis stage during SI). The percentages of progressively motile and viable spermatozoa as assessed by computer-assisted sperm analysis (CASA) and fluorescence microscopy, respectively were decreased whereas abnormal sperm heads, nuclear vacuoles and tail defects were increased at period 3 (P < 0.05) compared to period 1, 2 or 4 in SI bulls of both HF and BB breeds. Protamine deficient spermatozoa as observed by chromomycin A(3) (CMA(3)) staining were more present (P < 0.05) at period 2 and 3 in both breeds compared to period 1 or 4. Sperm nuclear shape as determined by Fourier harmonic amplitude (FHA) was most affected by heat stress during period 3 (P < 0.01) and a higher response was observed in BB bulls than HF bulls. In conclusion, sperm cells at the spermiogenic and meiotic stages of development are more susceptible to heat stress. The lack of chromatin protamination is the most pertinent result of heat stress, together with subtle changes in sperm head shape, which can be detected by FHA but not by conventional semen analysis.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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...

  15. 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

  16. 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.

  17. 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

  18. 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.

  19. 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%.

  20. 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).

  1. 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.

  2. The effect of BSO-induced oxidative stress on histologic feature of testis: testosterone secretion and semen parameters in mice

    PubMed Central

    Sajjadian, Fakhrosadat; Roshangar, Leila; Hemmati, Alireza; Nori, Mohammad; Soleimani-Rad, Sara; Soleimani-Rad, Jafar

    2014-01-01

    Objective(s): Buthionine sulfoximine (BSO) inhibits synthesis of glutathione as the main intracellular antioxidant. The aim of the present study is to investigate the effect of BSO-induced oxidative stress on histological structure of testis, testosterone secretion and semen parameters. Materials and Methods: Thirty male BALB/c mice were divided into 3 groups. In control group, the mice did not receive any chemical. In the experimental group, the mice received 2 mmol/kg BSO for 35 days. In the sham group, the mice received the solvent of BSO (0.9% saline). After the treatment, the mice were sacrificed. Their testes were fixed in Buein's fixative, embedded in paraffin and prepared for histological studies. To assess semen parameters, the sperms were collected from cauda epididymis. Blood samples were used for determination of super oxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPX), glutathione (GSH), catalase (CAT) and the serum testosterone level. The data analyzed using ANOVA and Dunnett's tests and SPSS software, version11.5. P- values at 0.05 level considered significant. Results: Data showed that in experimental group in comparison to control group; the concentration of CAT, GPX, SOD,GSH and the total level of testosterone is reduced while MDA level is increased significantly. The number of sperms with progressive motility were decreased (P<0.001) but sperms with abnormal morphology were increased (P<0.001). Histological studies revealed that the values for tubal differentiation index and spermatogenic index in experimental group were reduced (P<0.001). Conclusion: It is concluded that exposure to oxidative stress induced by BSO could affect testicular structure and semen parameters. PMID:25422755

  3. Induced lipid peroxidation in ram sperm: semen profile, DNA fragmentation and antioxidant status.

    PubMed

    Hamilton, Thais Rose dos Santos; de Castro, Letícia Signori; Delgado, Juliana de Carvalho; de Assis, Patrícia Monken; Siqueira, Adriano Felipe Perez; Mendes, Camilla Mota; Goissis, Marcelo Demarchi; Muiño-Blanco, Teresa; Cebrián-Pérez, José Álvaro; Nichi, Marcílio; Visintin, José Antonio; D'Ávila Assumpção, Mayra Elena Ortiz

    2016-04-01

    Action of reactive oxygen species, protamination failures and apoptosis are considered the most important etiologies of sperm DNA fragmentation. This study evaluated the effects of induced lipid peroxidation susceptibility on native semen profile and identified the mechanisms involved in sperm DNA fragmentation and testicular antioxidant defense on Santa Ines ram sperm samples. Semen was collected from 12 adult rams (Ovis aries) performed weekly over a 9-week period. Sperm analysis (motility, mass motility, abnormalities, membrane and acrosome status, mitochondrial potential, DNA fragmentation, lipid peroxidation and intracellular free radicals production); protamine deficiency; PRM1, TNP1 and TNP2 gene expression; and determination of glutathione peroxidase (GPx), glutathione reductase, catalase (CAT) and superoxide dismutase activity and immunodetection in seminal plasma were performed. Samples were distributed into four groups according to the sperm susceptibility to lipid peroxidation after induction with ascorbate and ferrous sulfate (low, medium, high and very high). The results were analyzed by GLM test and post hoc least significant difference. We observed an increase in native GPx activity and CAT immunodetection in groups with high susceptibility to induced lipid peroxidation. We also found an increase in total sperm defects, acrosome and membrane damages in the group with the highest susceptibility to induced lipid peroxidation. Additionally, the low mitochondrial membrane potential, susceptible to chromatin fragmentation and the PRM1 mRNA were increased in the group showing higher susceptibility to lipid peroxidation. Ram sperm susceptibility to lipid peroxidation may compromise sperm quality and interfere with the oxidative homeostasis by oxidative stress, which may be the main cause of chromatin damage in ram 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. 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.

  6. 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.

  7. Alveolar abnormalities

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/001093.htm Alveolar abnormalities To use the sharing features on this page, please enable JavaScript. Alveolar abnormalities are changes in the tiny air sacs in ...

  8. Nail abnormalities

    MedlinePlus

    Beau's lines; Fingernail abnormalities; Spoon nails; Onycholysis; Leukonychia; Koilonychia; Brittle nails ... 2012:chap 71. Zaiac MN, Walker A. Nail abnormalities associated with systemic pathologies. Clin Dermatol . 2013;31: ...

  9. 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.

  10. 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

  11. 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.

  12. 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

  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. 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.

  15. 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.

  16. 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

  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. 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.

  19. 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

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. Meiotic abnormalities

    SciTech Connect

    1993-12-31

    Chapter 19, describes meiotic abnormalities. These include nondisjunction of autosomes and sex chromosomes, genetic and environmental causes of nondisjunction, misdivision of the centromere, chromosomally abnormal human sperm, male infertility, parental age, and origin of diploid gametes. 57 refs., 2 figs., 1 tab.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  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. A short-term evaluation of semen and accessory sex gland function in phase III trial subjects receiving intravasal contraceptive RISUG.

    PubMed

    Chaki, S P; Das, H C; Misro, M M

    2003-01-01

    Following the intravasal injection of a new male contraceptive RISUG (reversible inhibition of sperm under guidance) in volunteers, routine semen analysis, semen biochemistry and germ cell morphology were evaluated in comparison with the corresponding preinjection samples for a maximum period of 6 months. Sperm counts in all 25 subjects before injection varied from 45 to 120 x 10(6)/ml. Out of 25 subjects, 6 became azoospermic after 1 month, 15 after 2 months, 3 after 3 months and 1 after 4 months of contraceptive injection. The mean volume of the ejaculates was found to be less as compared to preinjection samples. Occasional sperm or sperm heads and immature germ cells were identified in only a few postinjected subjects. However, no pregnancy was reported in these subjects during the study period. Abnormal morphology found in most of the sperm, but not in the accompanying immature germ cells, may be due to a charge-related effect on the former but not on the latter cells. Neutral alpha-glucosidase, the biochemical marker for epididymis, was estimated to be significantly lower in the seminal plasma of all the postinjected subjects. On the other hand, acid phosphatase activity and fructose levels in the seminal plasma were found to be in the normal range. Based on the above findings, it is concluded that at least for the present study period, RISUG, a new male contraceptive, is effective as a partially occluding agent in the vas deferens.

  20. Proteomic analysis of seminal plasma from asthenozoospermia patients reveals proteins that affect oxidative stress responses and semen quality.

    PubMed

    Wang, Jun; Wang, Jian; Zhang, Hua-Rong; Shi, Hui-Juan; Ma, Duan; Zhao, Hong-Xin; Lin, Biaoyang; Li, Run-Sheng

    2009-07-01

    Asthenozoospermia (AS) is a common cause of human male infertility. In one study, more than 80% of the samples from infertile men had reduced sperm motility. Seminal plasma is a mixture of secretions from the testis, epididymis and several male accessory glands, including the prostate, seminal vesicles and Cowper's gland. Studies have shown that seminal plasma contains proteins that are important for sperm motility. To further explore the pathophysiological character of AS, we separated the seminal plasma proteins from AS patients and healthy donors using sodium dodecyl sulfate polyacrylamide gel electrophoresis and in-gel digestion, and then subjected the proteins to liquid chromatography-mass spectrometry (LC-MS/MS) analysis. A total of 741 proteins were identified in the seminal plasma, with a false discovery rate of 3.3%. Using spectral counting, we found that 45 proteins were threefold upregulated and 56 proteins were threefold downregulated in the AS group when compared with the control. Most of these proteins originated from the epididymis and prostate. This study identified a rich source of biomarker candidates for male infertility and indicates that functional abnormalities of the epididymis and prostate can contribute to AS. We identified DJ-1-a protein that has been shown elsewhere to be involved in the control of oxidative stress (OS)-as a downregulated protein in AS seminal plasma. The levels of DJ-1 in AS seminal plasma were about half of those in the control samples. In addition, the levels of reactive oxygen species were 3.3-fold higher in the AS samples than in the controls. Taken together, these data suggest that downregulation of DJ-1 is involved in OS in semen, and therefore affects the quality of the semen.

  1. 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

  2. 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.

  3. 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.

  4. [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.

  5. 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.

  6. 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

  7. 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.

  8. 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%.

  9. 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

  10. 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.

  11. 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.

  12. Effect of Addition of Taurine on the Liquid Storage (5°C) of Mithun (Bos frontalis) Semen

    PubMed Central

    Perumal, P.; Vupru, Kezhavituo; Rajkhowa, C.

    2013-01-01

    The present study was undertaken to assess the effect of taurine on sperm motility, viability, total sperm abnormalities, acrosomal and plasma membrane integrity, enzymatic profiles such as reduced glutathione (GSH), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT), and biochemical profiles such as cholesterol efflux and malondialdehyde (MDA) production. A total of 50 ejaculates were collected twice a week from 8 mithun bulls, and semen was split into 4 equal aliquots and diluted with the TEYC extender. Group 1: semen was without additives (control); groups 2 to 4: semen was diluted with 25 mM, 50 mM, and 100 mM of taurine, respectively. Seminal parameters and enzymatic and biochemical profiles were assessed at 5°C. Inclusion of taurine into diluent resulted in significant (P < 0.05) decreases in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities after liquid storage compared with the control group. Additionally, taurine at 50 mM has significant improvement in quality of mithun semen than taurine at 25 or 100 mM stored in in vitro at 5°C. It was concluded that the possible protective effects of taurine on sperm parameters are from enhancing the function of antioxidant enzymes, preventing efflux of cholesterol from cell membranes and decreased MDA production. PMID:23853737

  13. 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

  14. [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.

  15. 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

  16. 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.

  17. Impact of seminal trace element and glutathione levels on semen quality of Tunisian infertile men

    PubMed Central

    2012-01-01

    Background Growing evidence indicates that oxidative stress can be a primary cause of male infertility. Non-enzymatic antioxidants play an important protective role against oxidative damages and lipid peroxidation. Human seminal plasma is a natural reservoir of antioxidants. The aim of this study was to determine glutathione (GSH) concentrations, trace element levels (zinc and selenium) and the lipid peroxidation end product, malondialdehyde (MDA), in the seminal plasma of men with different fertility potentials. Methods Semen samples from 60 fertile men (normozoospermics) and 190 infertile patients (74 asthenozoospermics, 56 oligozoospermics, and 60 teratozoospermics) were analyzed for physical and biochemical parameters. Zinc (Zn) and selenium (Se) levels were estimated by atomic absorption spectrophotometry. Total GSH (GSHt), oxidized GSH (GSSG), reduced GSH (GSHr) and MDA concentrations were measured spectrophotometrically. Results Zn and Se concentrations in seminal plasma of normozoospermics were more elevated than the three abnormal groups. Nevertheless, only the Zn showed significant differences. On the other hand, Zn showed positive and significant correlations with sperm motility (P = 0.03, r = 0.29) and count (P < 0.01, r = 0.49); however Se was significantly correlated only with sperm motility (P < 0.01, r = 0.36). GSHt, GSSG and GSHr were significantly higher in normozoospermics than in abnormal groups. We noted a significant association between seminal GSHt and sperm motility (P = 0.03). GSSG was highly correlated to sperm motility (P < 0.001) and negatively associated to abnormal morphology (P < 0.001). GSHr was significantly associated to total sperm motility (P < 0.001) and sperm count (P = 0.01). MDA levels were significantly higher in the three abnormal groups than in normozoospermics. Rates of seminal MDA were negatively associated to sperm motility (P < 0.01; r = -0.24) and sperm concentration (P = 0.003; r = -0.35) Meanwhile, there is a

  18. 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.

  19. 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

  20. Prospective Analysis on the Effect of Botanical Medicine (Tribulus terrestris) on Serum Testosterone Level and Semen Parameters in Males with Unexplained Infertility.

    PubMed

    Roaiah, Mohamed Farid; Elkhayat, Yasser Ibrahim; Saleh, Sameh Fayek GamalEl Din; Abd El Salam, Mohamed Ahmed

    2016-06-23

    We evaluated the role of Tribulus terrestris in males with unexplained infertility and its effect on serum testosterone and semen parameters. Thirty randomized male patients presenting to Andrology outpatient clinic complaining of idiopathic infertility were selected. They were given Tribulus terrestris (750 mg) in three divided doses for three months. The effect of Tribulus terrestris on serum testosterone (total and free) and luteinizing hormone (LH), as well as its impact on semen parameters in those patients, was studied. No statistically significant difference was observed in the levels of testosterone (total and free) and LH and semen parameters (sperm concentration or motility, or abnormal forms) before and after the treatment. In addition, no statistically significant correlations were observed between testosterone (free and total) and LH and semen parameters before and after the treatment. Tribulus terrestris was ineffective in the treatment of idiopathic infertility.

  1. Effect of vitamin E and selenium nanoparticles on post-thaw variables and oxidative status of rooster semen.

    PubMed

    Safa, Soroush; Moghaddam, Gholamali; Jozani, Raziallah Jafari; Daghigh Kia, Hossein; Janmohammadi, Hossein

    2016-11-01

    This study was conducted to determine the combined effects of adding vitamin E (VitE) and selenium nanoparticle (Nano-Se) as antioxidant supplements to rooster semen extender for freezing. Semen samples were collected from 12 White Leghorn roosters and pooled. Subsequently, the samples were divided into nine equal groups using modified Beltsville extender. Extenders were supplemented with either two amounts of VitE (5 and 10μg/mL) or two amounts of Nano-Se (1% and 2%) or a combination of both VitE and Nano-Se, and no antioxidants extender (control group). Using 5μg/mL VitE and 1% of Nano-Se improved (P<0.05) total sperm motility (79.28±3.86%), progressive sperm motility (18.03±1.02%), sperm viability (81.46±2.16%) and integrity of the sperm membrane (77.21±2.12%) after the freeze-thawing process compared with control group (54.08±3.86%, 10.96±1.02%, 60.53±2.16%, and 54.47±2.12%; respectively). Also, extenders supplemented with 5μg/mL Vit E or 5μg/mL VitE and 1% Nano-Se had a lesser malondialdehyde (MDA) concentration compared to control extender (1.15±0.32, and 1.29±0.32, respectively). Total abnormal morphology of sperm was decreased (P<0.05) by adding 5 or 10μg/mL VitE alone or in combined with 1% or 2% Nano-Se. Moreover, extenders containing Nano-Se demonstrated greater glutathione peroxidase (GPx) activity compared to other extenders. Catalase (CAT) activity was greater in extender supplemented with 10μg/mL VitE and 2% Nano-Se. Moreover, higher TAC was observed in extenders supplemented with VitE and Nano-Se. It can be concluded that addition of 5μg/mL vitamin E in combined with 1% Nano-Se improved the post-thawing quality and oxidative variables of rooster semen.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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%).

  9. 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.

  10. 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

  11. 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

  12. Leukocyte abnormalities.

    PubMed

    Gabig, T G

    1980-07-01

    Certain qualitative abnormalities in neutrophils and blood monocytes are associated with frequent, severe, and recurrent bacterial infections leading to fatal sepsis, while other qualitative defects demonstrated in vitro may have few or no clinical sequelae. These qualitative defects are discussed in terms of the specific functions of locomotion, phagocytosis, degranulation, and bacterial killing.

  13. 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.

  14. 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.

  15. 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.

  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. 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

  18. Large nuclear vacuoles are indicative of abnormal chromatin packaging in human spermatozoa.

    PubMed

    Franco, J G; Mauri, A L; Petersen, C G; Massaro, F C; Silva, L F I; Felipe, V; Cavagna, M; Pontes, A; Baruffi, R L R; Oliveira, J B A; Vagnini, L D

    2012-02-01

    The aim of this investigation was to determine the presence of abnormal sperm chromatin packaging in spermatozoa with large nuclear vacuoles (LNV) selected via high magnification by analysing the pattern of chromomycin A3 (CMA3) staining. A prospective observational study was designed to analyse semen samples obtained from 66 men undergoing infertility diagnosis and treatment. The numbers of cells with normal (dull yellow staining of the sperm head/CMA3-negative) and abnormal (bright yellow fluorescence of the sperm head/CMA3-positive) chromatin packaging were determined on slides with normal and LNV spermatozoa. The presence of bright yellow fluorescence (CMA3-positive) was significantly higher (p < 0.0001) in spermatozoa with LNV than in normal spermatozoa (719/1351; 53.2% vs. 337/835; 40.3%, respectively), reflecting a higher percentage of abnormal chromatin packaging in spermatozoa with large LNV. Our data support the hypothesis that the presence of LNV reflects the presence of abnormal chromatin packaging, which may facilitate sperm DNA damage. As sperm nuclear vacuoles are evaluated more precisely at high magnifications using motile sperm organelle morphology examination (MSOME), the present results support the use of high-magnification sperm selection for intracytoplasmic sperm injection (ICSI).

  19. Supplementation of soybean lecithin-based semen extender by antioxidants: complementary flowcytometric study on post-thawed ram spermatozoa.

    PubMed

    Sharafi, Mohsen; Zhandi, Mahdi; Akbari Sharif, Abbas

    2015-06-01

    The purpose of the current study was to evaluate the effects of cysteine (C) and glutathione (G) on the post-thawed ram sperm quality. Collected semen samples from four mature rams were diluted with five soybean lecithin (SL)-based extenders containing: no antioxidant (SL-0), 5 mM cysteine (SL-C5), 10 mM cysteine (SL-C10), 5 mM glutathione (SL-G5) and 10 mM glutathione (SL-G10). After freeze-thawing process, motion and velocity parameters, plasma membrane integrity and functionality, morphological abnormality, lipid peroxidation, acrosomal status, mitochondria activity, and apoptosis status of post-thawed ram spermatozoa were assessed. The results showed that SL-C10 increased the total motility and plasma membrane integrity (p < 0.05) of post-thawed ram spermatozoa (55.86 ± 1.37 and 60.57 ± 1.34 %) compared to other extenders. Progressive motility was significantly higher in SL-C10 (24.71 ± 1.13 %) compared to SL-0 (20 ± 1.13 %) and SL-G10 (15 ± 1.13 %). Mitochondrial activity was significantly higher in SL-C10 (56.83 ± 2.29 %) compared to SL-G10 (38.75 ± 2.29 %). Capacitation and acrosomal status, lipid peroxidation, and the percentage of dead spermatozoa were not affected by different extenders. The percentage of live spermatozoa was higher in SL-C10 (56.33 ± 1.35 %) compared to other extenders. Also, SL-C10 resulted in a lower percentage of apoptotic spermatozoa (14.17 ± 0.53 %) compared to other extenders. The results of this study showed that supplementation of SL-based ram semen extender with 10 mM cysteine resulted in an improved quality of post-thawed ram spermatozoa.

  20. l-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm.

    PubMed

    Fattah, A; Sharafi, M; Masoudi, R; Shahverdi, A; Esmaeili, V; Najafi, A

    2017-02-01

    Rooster semen cryopreservation is not efficient for artificial insemination in breeder flocks. l-Carnitine (LC) has been evaluated for effectiveness in cryopreservation media on the characteristics of rooster sperm after freeze-thawing. Motility characteristics, membrane functionality, abnormal morphology, apoptotic like changes, mitochondria activity and lipid peroxidation of rooster sperms were assessed after freeze-thawing with different concentrations of LC in Beltsville medium. Semen samples were collected from 12 roosters, twice a week, and diluted in the extenders that contained different concentrations of LC. Supplementation of Beltsevile with 1 and 2 mM LC was found to result in higher total motility (68.2± 1.7% and 69.1± 1.7%, respectively), progressive motility (28.4± 1.6%, 29.8± 1.6%), membrane functionality (76.2± 1.9% and 75.9± 1.9%), viability (58.2 ± 1.1%, 59.1 ± 1.1%) and lower significant of lipid peroxidation (2.53 ± 0.08 nmol/ml, 2.49 ± 0.08 nmol/ml) compared to control group containing no LC. Lower motility, progressive motility, and viability were observed in frozen-thawed sperm in extender containing 8 mM LC (35.8± 1.7%, 9.6± 1.2% and 27.1 ± 1.2%, respectively) compared to control. Morphology and mitochondrial activity were not affected by different concentrations of LC. Our results showed that supplementation of Beltsville extender with 1 and 2 mM LC significantly improved the quality of rooster sperm quality after freeze-thawing.

  1. 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.

  2. Effects of in vitro selenium addition to the semen extender on the spermatozoa characteristics before and after freezing in water buffaloes (Bubalus bubalis)

    PubMed Central

    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. PMID:25653769

  3. 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

  4. 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.

  5. 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.

  6. Heritabilities and genetic correlations for litter size and semen traits in Czech Large White and Landrace pigs.

    PubMed

    Wolf, J

    2010-09-01

    The objective of the present study was to estimate heritabilities and genetic correlations for semen and litter size traits using full animal model analyses. Litter size data were available from 28,485 Czech Large White and 10,410 Czech Landrace sows farrowing between 1995 and 2008. The number of litter records was 91,922 and 30,937, respectively. Records on semen traits (37,137 ejaculates from 778 Large White boars and 51,341 ejaculates from 841 Landrace boars) were available from 22 AI centers in the Czech Republic from 2000 to 2009. All calculations were carried out separately for each breed. Number of functional sperm was defined as total number of sperm times the fraction of motile sperm times (1 - the fraction of abnormal sperm). Among the semen traits, semen volume showed the greatest heritability (0.20 +/- 0.019 in Large White and 0.25 +/- 0.018 in Landrace), followed by sperm concentration with a heritability of 0.18 (SE = 0.012 and 0.014) in both breeds. The heritabilities of the remaining 4 traits (motility, percentage of abnormal sperm, total number of sperm, and number of functional sperm) were around 0.10 (SE = 0.016 to 0.031). Large negative genetic correlations were observed between semen volume and sperm concentration and between motility and percentage of abnormal sperm, especially in Large White. Positive and negative correlations among remaining semen traits were mostly of small magnitude. There was a tendency for increasing litter size to be associated with slight decreases in the total number of sperm and in the number of functional sperm, especially in the Large White breed (genetic correlations of -0.08 to -0.14 and -0.16 to -0.31, respectively, with SE between 0.100 and 0.114). Some of the correlations between semen and litter size traits (especially with the percentage of abnormal sperm) were breed-specific (positive up to 0.63 +/- 0.062 for Large White and negative until -0.41 +/- 0.106 in Landrace). Furthermore, parity

  7. 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.

  8. Evaluation of a container for collection and shipment of semen with potential uses in population-based, clinical, and occupational settings.

    PubMed

    Royster, M O; Lobdell, D T; Mendola, P; Perreault, S D; Selevan, S G; Rothmann, S A; Robbins, W A

    2000-01-01

    Large, population-based studies of semen quality are encumbered by the logistics and expense of obtaining semen samples from men who live in a variety of locations. A prototype semen collection and transportation kit, the TRANSEM100, can be distributed to study participants and then directly shipped to a central laboratory for analysis. This study was designed to evaluate the ability of male volunteers to correctly use the kit. Thirty volunteers aged 20 to 44 years with no history of diabetes, recent chemotherapy, fertility problems, or vasectomy were recruited through a newspaper advertisement, interviewed to obtain demographic information, and instructed on the use of the kit. Twenty-six of the initial subjects provided at least 1 semen specimen using the kit and returned the specimens by overnight delivery to the laboratory for analysis, 25 completed a follow-up interview on the use of the collection kit, and 20 submitted a second semen sample using the same method. The average volunteer was white, 27.8 years old, and held at least a college degree. Forty percent of the volunteers were married. In general, participants correctly followed the instructions for collecting, packaging, and shipping the semen samples. Volunteers were instructed to collect samples after at least 2, but no more than 7 days of abstinence. For the first and second samples submitted, participants collected semen samples after an average of 3.3 and 3.9 days of abstinence, respectively. Seventeen (65%) of the samples from the first sampling period and 16 (80%) of the samples from the second period were received in the laboratory the day after they had been collected. In summary, the TRANSEM100 may prove to be useful for collecting human semen in field studies. Further testing of this method is warranted to evaluate preservation of sample quality and use of the kit by men among diverse socioeconomic groups.

  9. 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

  10. Polychlorinated biphenyls and dibenzofurans increased abnormal sperm morphology without alterations in aneuploidy: The Yucheng study.

    PubMed

    Hsu, Ping-Chi; Li, Ming-Chieh; Lee, Yeu-Chin; Kuo, Pao-Lin; Guo, Yueliang Leon

    2016-12-01

    In 1979, more than 2000 persons ingested rice oil contaminated with polychlorinated biphenyls and polychlorinated dibenzofurans; this event was called the "Yucheng accident." An increased percentage of oligospermia, reduced ability of sperm to penetrate oocytes, and reduced percentage of male offspring were reported in Yucheng men. This study examined whether the sperm sex ratio and chromosome aneuploidy are responsible for our observed findings in Yucheng men. In 1999-2000, Yucheng men and their neighborhood referents aged 37-50 years were recruited for physical examination, followed by semen analysis. The semen samples were analyzed for chromosomal aneuploidy through fluorescent in situ hybridization according to an established procedure in our laboratory. A total of 50 Yucheng men and 34 neighborhood referents volunteered to participate in the study. Although abnormal morphology was mildly increased, no differences were observed in sperm percentages, with normal numbers of chromosomes X, Y, and 8 in the two groups. The percentage of sperm with aneuploidy of the sex chromosomes or chromosome 8 and of that with diploidy did not vary between both groups. The normal X/Y sperm ratio was not different between the groups. However, among Yucheng men, 8% had a normal X/Y sperm ratio of >1.4, and no neighborhood referent showed such an elevated X/Y ratio. Chromosomal aneuploidy was not elevated in Yucheng men. The mechanisms underlying the reduced sperm capability of oocyte penetration and changed offspring sex ratio in Yucheng men remain undetermined.

  11. 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.

  12. 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.

  13. 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

  14. 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

  15. 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.

  16. 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.

  17. Use of new field methods of semen analysis in the study of occupational hazards to reproduction: the example of ethylene dibromide

    SciTech Connect

    Schrader, S.M.; Ratcliffe, J.M.; Turner, T.W.; Hornung, R.W.

    1987-12-01

    Increasing attention has been paid to the use of semen analysis as an indicator of exposure to potential mutagenic and reproductive hazards. In the infertility clinic setting, semen evaluations include the measurement of sperm concentration, volume, pH, motility, velocity and morphology, the analysis of seminal plasma to evaluate accessory sex gland function and, in some cases, the in vitro evaluation of fertilization capacity and sperm-cervical mucus interaction. To date, however, the study of semen characteristics of occupationally exposed populations has been confined principally to the measurement of sperm concentration and sperm morphology. This has been largely due to the unavailability of portable equipment suitable for the measurement of other semen characteristics and the difficulty of obtaining fresh semen samples in the field setting. National Institute for Occupational Safety and Health researchers have developed mobile laboratory facilities which enable us to evaluate fresh samples, in the field, for semen characteristics in addition to concentration and morphology. This paper describes the application of these methods using the example of our recent cross-sectional study of workers occupationally exposed to ethylene dibromide in the papaya fumigation industry. We discuss our findings in the context of the usefulness of semen analysis as an indicator of occupational hazards to male reproduction.

  18. 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.

  19. 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...

  20. APPLICATION OF THE SPERM CHROMATIN STRUCTURE ASSAY TO THE TEPLICE PROGRAM SEMEN STUDIES: A NEW METHOD FOR EVALUATING SPERM NUCLEAR CHROMATIN DAMAGE

    EPA Science Inventory

    ABSTRACT
    A measure of sperm chromatin integrity was added to the routine semen end points evaluated in the Teplice Program male reproductive health studies. To address the hypothesis that exposure to periods of elevated air pollution may be associated with abnormalities in sp...

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. Blood serum and seminal plasma selenium, total antioxidant capacity and coenzyme q10 levels in relation to semen parameters in men with idiopathic infertility.

    PubMed

    Eroglu, Mustafa; Sahin, Sadik; Durukan, Birol; Ozakpinar, Ozlem Bingol; Erdinc, Nese; Turkgeldi, Lale; Sofuoglu, Kenan; Karateke, Ates

    2014-06-01

    In this case-control study, we aimed to evaluate the serum and seminal plasma levels of Selenium (Se), total antioxidant capacity (TAC), and Coenzyme Q10 (CoQ-10) and determine their relationship with sperm concentration, motility, and morphology in men with idiopathic infertility. A total of 59 subjects were enrolled in the study. Forty four patients were diagnosed with idiopathic male infertility and had abnormal sperm parameters, and 15 subjects had normal sperm parameters with proven fertility. Serum Se, semen Se, and semen TAC levels were significantly different in the fertile and infertile groups (p<0.01, p<0.001, and p<0.001, respectively). However, serum TAC, serum, and seminal plasma CoQ-10 levels did not differ between fertile and infertile groups. When the levels of the measured parameters were compared in serum and seminal plasma, serum levels of Se were found to be correlated positively with the semen levels in all subjects included into the study (N=59) (r=0.46, p<0.01). A relationship was found between neither serum and semen levels of TAC nor between serum and semen levels of CoQ-10. Correlations among measured serum and semen parameters with sperm parameters demonstrated that both the serum and semen levels of Se were correlated positively with spermatozoa concentration, motility, and morphology. Additionally, seminal plasma levels of TAC correlated positively with all these sperm parameters. On the other hand, seminal plasma levels of CoQ-10 correlated only with sperm morphology but not with concentration or motility. No relationship was observed between serum levels of TAC or serum levels of CoQ-10 and sperm parameters. In conclusion, serum and seminal plasma Se deficiency may be a prominent determinant of abnormal sperm parameters and idiopathic male infertility. Measurement of serum Se levels may help determine nutritional status and antioxidant capacity in infertile patients, which may help distinguish those patients who will benefit from

  6. 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

  7. 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

  8. 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.

  9. 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.

  10. The effect of season on spermatozoa motility, plasma membrane and acrosome integrity in fresh and frozen-thawed semen from Xinong Saanen bucks.

    PubMed

    Wang, W; Luo, J; Sun, S; Xi, L; Gao, Q; Haile, A B; Shi, H; Zhang, W; Shi, H

    2015-02-01

    The aim of this study was to evaluate whether the season of ejaculate collection influences seminal quality parameters of pre- and post-freeze-thawing in Xinong Saanen bucks. Ejaculates were collected from eight bucks throughout the four seasons (spring, summer, autumn and winter) in a 12 months' time period, identified in the Northern Hemisphere. Semen samples were evaluated by the combinations of conventional and Computer-Assisted Sperm Analysis (CASA) when fresh and after frozen-thawed, respectively. The results clearly demonstrated that season of ejaculate collection influenced (p < 0.05) fresh semen quality. Highest semen quality was observed during autumn. On the contrary, undesirable indices (significantly lower, p < 0.05) were observed in winter as compared with the other remaining seasons. CASA has clearly shown the influences of seasonal variations on semen motility parameters. Furthermore, season of ejaculate collection was also found to influence sperm freezability. Semen characteristics after frozen-thawed followed a similar pattern with that of fresh ejaculate except in spring. The results revealed that sperm quality was higher (p < 0.01) in summer and autumn than in spring and winter. In conclusion, seasonal variation influences semen quality in Xinong Saanen bucks. In addition to summer and autumn, fresh ejaculates in spring can also be successfully used for AI. Sperm from ejaculates collected during summer and autumn are more suitable for cryopreservation. Hence, it is possible to increase the efficiency of goat breeding by manipulating the seasonal variations of semen quality for immediate AI and/or cryopreservation.

  11. 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.

  12. 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

  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. Effect of single layer centrifugation using Androcoll-E-Large on the sperm quality parameters of cooled-stored donkey semen doses.

    PubMed

    Ortiz, I; Dorado, J; Ramírez, L; Morrell, J M; Acha, D; Urbano, M; Gálvez, M J; Carrasco, J J; Gómez-Arrones, V; Calero-Carretero, R; Hidalgo, M

    2014-02-01

    The aim of this study was to determine the effect of single layer centrifugation (SLC) using Androcoll-E-Large on donkey sperm quality parameters after 24 h of cool-storage. Ejaculates were collected from Andalusian donkeys and then cooled at 5°C. SLC was carried out after 24 h of cool-storage using Androcoll-E-Large. In the first experiment, all sperm parameters assessed (total and progressive sperm motility, viability, sperm morphology and sperm kinematics VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were statistically compared between semen samples processed or not with Androcoll-E-Large. Significant differences (P<0.05) were found between SLC-selected and unselected semen samples for all parameters assessed, obtaining better results after SLC. In the second experiment, semen samples were classified in two groups according to their sperm progressive motility (PM) before SLC. Then, the increments obtained in semen quality parameters after SLC were compared between groups. No significant differences were found between groups, indicating that SLC improved the sperm quality parameters of entire set of semen samples processed with independence to their original PM. In conclusion, SLC with Androcoll-E-Large can be used in donkeys, increasing the sperm quality of cooled-stored donkey semen doses after 24 h of cool storage.

  15. Electroejaculation and semen analysis and freezing in the giant panda (Ailuropoda melanoleuca).

    PubMed

    Platz, C C; Wildt, D E; Howard, J G; Bush, M

    1983-01-01

    Semen was collected by a standardized electroejaculation procedure from a giant panda on 4 occasions. Ejaculate volume, sperm count and % sperm motility were 2.3-3.6 ml, 62-562 x 10(6) spermatozoa/ml and 45-85%, respectively. The results, although limited to a single male, suggested a seasonal influence on ejaculate and gonadal parameters with improved ejaculate volume, sperm motility and increased testicular size in the season proximate to the female's oestrous period. Frozen-thawed spermatozoa were motile with no apparent abnormalities induced by the freezing procedure.

  16. 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...

  17. 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...

  18. 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

  19. 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.

  20. 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.

  1. 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

  2. 'They say Islam has a solution for everything, so why are there no guidelines for this?' Ethical dilemmas associated with the births and deaths of infants with fatal abnormalities from a small sample of Pakistani Muslim couples in Britain.

    PubMed

    Shaw, Alison

    2012-11-01

    This paper presents ethical dilemmas concerning the termination of pregnancy, the management of childbirth, and the withdrawal of life-support from infants in special care, for a small sample of British Pakistani Muslim parents of babies diagnosed with fatal abnormalities. Case studies illustrating these dilemmas are taken from a qualitative study of 66 families of Pakistani origin referred to a genetics clinic in Southern England. The paper shows how parents negotiated between the authoritative knowledge of their doctors, religious experts, and senior family members in response to the ethical dilemmas they faced. There was little knowledge or open discussion of the view that Islam permits the termination of pregnancy for serious or fatal abnormality within 120 days and there was considerable disquiet over the idea of ending a pregnancy. For some parents, whether their newborn baby would draw breath was a main worry, with implications for the baby's Muslim identity and for the recognition of loss the parents would receive from family and community. This concern sometimes conflicted with doctors' concerns to minimize risk to future pregnancies by not performing a Caesarean delivery if a baby is sure to die. The paper also identifies parents' concerns and feelings of wrong-doing regarding the withdrawal of artificial life-support from infants with multiple abnormalities. The conclusion considers some of the implications of these observations for the counselling and support of Muslim parents following the pre- or neo-natal diagnosis of fatal abnormalities in their children.

  3. 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.

  4. 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

  5. Effect of dietary Satureja khuzistanica powder on semen characteristics and thiobarbituric acid reactive substances concentration in testicular tissue of Iranian native breeder rooster

    PubMed Central

    Heydari, M. J.; Mohammadzadeh, S.; Kheradmand, A.; Alirezaei, M.

    2015-01-01

    Because of a paucity of information on the effect of Satureja khuzistanica in male chickens, this study was undertaken to determine the influence of dietary S. khuzistanica powder (SKP) on seminal characteristics and testes thiobarbituric acid reactive substances (TBARS) content in Iranian native breeder rooster. Thirty-six 40-week-old roosters were randomly allotted to 3 equal groups and received either a basal diet without SKP (T1 or control), or a diet containing 20 g/kg (T2) and 40 g/kg (T3) of SKP for 8-week-long experimental period. Semen samples were obtained weekly by abdominal massage to evaluate the seminal characteristics. At the end of the eighth week 18 birds (6 birds per each group) were randomly slaughtered, and sample was taken from right testes for TBARS evaluation. Administration of SKP improved all semen traits, except for sperm concentration. Likewise, TBARS content in SKP treatments did not significantly differ from the control (P>0.05). Seminal volume, live sperm percentage and plasma membrane integrity percentage in SKP-treated groups were higher than the control. Conversely, abnormal sperm percentages reduced in SKP-treated groups (P<0.05). Plasma membrane integrity in experimental treatments was significantly higher than the control in 2nd, 3rd and 7th weeks. However, at 6th and 8th weeks only T3 treatment was significantly different from the control. Notably, there was an increase in total sperm concentration in SKP-treated groups in compared to the control birds. In conclusion, this study indicated that addition of SKP in rooster diet improves sperm quality and also reduces their sperm membrane lipid peroxidation, which may lead to higher fertilization rate. PMID:27175185

  6. ETOPOSIDE INDUCES CHROMOSOMAL ABNORMALITIES IN SPERMATOCYTES AND SPERMATOGONIAL STEM CELLS

    SciTech Connect

    Marchetti, F; Pearson, F S; Bishop, J B; Wyrobek, A J

    2005-07-15

    Etoposide (ET) is a chemotherapeutic agent widely used in the treatment of leukemia, lymphomas and many solid tumors, such as testicular and ovarian cancers, that affect patients in their reproductive years. The purpose of the study was to use sperm FISH analyses to characterize the long-term effects of ET on male germ cells. We used a mouse model to characterize the induction of chromosomal aberrations (partial duplications and deletions) and whole chromosomal aneuploidies in sperm of mice treated with a clinical dose of ET. Semen samples were collected at 25 and 49 days after dosing to investigate the effects of ET on meiotic pachytene cells and spermatogonial stem-cells, respectively. ET treatment resulted in major increases in the frequencies of sperm carrying chromosomal aberrations in both meiotic pachytene (27- to 578-fold) and spermatogonial stem-cells (8- to 16-fold), but aneuploid sperm were induced only after treatment of meiotic cells (27-fold) with no persistent effects in stem cells. These results demonstrate that male meiotic germ cells are considerably more sensitive to ET than spermatogonial stem-cell and that increased frequencies of sperm with structural aberrations persist after spermatogonial stem-cell treatment. These findings predict that patients who undergo chemotherapy with ET may have transient elevations in the frequencies of aneuploid sperm, but more importantly, may have persistent elevations in the frequencies of sperm with chromosomal aberrations, placing them at higher risk for abnormal reproductive outcomes long after the end of their chemotherapy.

  7. 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

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. Effect of elemental nano-selenium on semen quality, glutathione peroxidase activity, and testis ultrastructure in male Boer goats.

    PubMed

    Shi, Li-guang; Yang, Ru-jie; Yue, Wen-bin; Xun, Wen-juan; Zhang, Chun-xiang; Ren, You-she; Shi, Lei; Lei, Fu-lin

    2010-04-01

    The objective of this experiment is to study the effects of novel elemental nano-selenium in the diet on testicular ultrastructure, semen quality and GSH-Px activity in male goats. Forty-two 2-month-old bucks were offered a total mixed ration which had been supplemented with nano-Se (0.3mg/kg Se) or unsupplemented (the control group only received 0.06mg/kg Se-background), for a period of 12 weeks (from weaning to sexual maturity). Results showed that the testicular Se level, semen glutathione peroxidase and ATPase activity increased significantly in the nano-Se supplementation group compared with control (P<0.05). The semen quality (volume, density, motility and pH) was not affected by added Se in diets, however, the sperm abnormality rate of control bucks was significantly higher than Se supplemented bucks (P<0.05). The testes of 5 goats in each group were examined by transmission electron microscopy (TEM), and showed that in Se-deficient bucks the membrane was damaged, and showed the occurrence of abnormalities in the mitochondria of the midpiece of spermatozoa. In conclusion, selenium deficiency resulted in abnormal spermatozoal mitochondria, and supplementation with nano-Se enhanced the testis Se content, testicular and semen GSH-Px activity, protected the membrane system integrity and the tight arrayment of the midpiece of the mitochondria. Further studies are required to research the novel elemental nano-Se with characterization of bioavailability and toxicity in small ruminants.

  13. 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.

  14. 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

  15. Liqui-Prep® versus conventional Papanicolaou smear to detect cervical cells abnormality by split-sample technique: a randomized double-blind controlled trial.

    PubMed

    Jesdapatarakul, Somnuek; Tangjitgamol, Siriwan; Nguansangiam, Sudarat; Manusirivithaya, Sumonmal

    2011-01-01

    To assess the diagnostic performances of LiquiPrep® (LP) to detect cervical cellular abnormality in comparison to Papanicolaou (Pap) smear in 194 women with abnormal cervical cytology who were scheduled for colposcopy at the institution between January 2008 and November 2008. The women were randomized to undergo a repeated cervical cytologic evaluation by Pap smear followed by LP, or the two methods in alternating order. The pathologist was blinded to previous cytologic diagnosis and the pair of slides assigned for each woman. Cytologic results from each method were compared to subsequent histopathology. Mean screening time for each LP and Pap slides were 4.3 ± 1.2 minutes and 5.4 ± 1.1 minutes, respectively (P < 0.001). From 194 cases, ASC or AGC were diagnosed in 72 cases (37.1%) from LP and 68 cases (35.1%) from Pap smear. After excluding the ASC/AGC group, the overall cytologic diagnostic agreement between the two tests were 69 of 87 cases (73.6%) while the agreements with histologic diagnoses were 39/87 cases from LP (44.8%) and 41 (47.1%) from Pap smear (P = 0.824). The accuracy of LP was not significantly different from Pap test, 43.4% (95% confidence interval [CI]: 34.8-52.1%) compared to 44.4% (95% CI: 35.7-53.1%). LP did not have superior performance over Pap test to detect high-grade lesions (≥ cervical intraepithelial neoplasia II) using ASC/AGC as the threshold with the sensitivity of 70.5% (95% CI: 64.0-76.9%) versus 77.3% (95% CI: 71.4-83.2%), respectively.

  16. 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

  17. 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.

  18. 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

  19. Effect of nutritional state on semen characteristics, testicular size and serum testosterone concentration in Sanjabi ram lambs during the natural breeding season.

    PubMed

    Ghorbankhani, F; Souri, M; Moeini, M M; Mirmahmoudi, R

    2015-02-01

    This experiment was conducted to investigate the effect of plane of nutrition on body weight, average daily gain, dry mater intake, semen characteristics, serum testosterone concentration and testicular circumference of Sanjabi ram lambs during the natural breeding season. Sanjabi ram lambs (n = 20) that were 8 months of age were penned under natural photoperiod at latitude 34°18'N for a period of 9 months. The control group (C, n = 10) received a diet consisting of 80% alfalfa and 20% concentrate, providing 2.18 Mcal metabolizable energy and 130.0 g /kg DM crude protein, whereas, treatment group (T, n = 10) was fed with 65% alfalfa and 35% concentrate, providing 2.34 Mcal metabolizable energy and 160.0 g/kg DM crude protein. Body weight, additive daily gain and feed intake in T group were significantly greater than those obtained in C group. Body weight and testicular circumference increased at a steady rate throughout the experiment. All semen variables (except percentage of abnormal sperm and semen pH), serum testosterone concentration and testicular circumference were positively influenced by nutritional state (P < 0.05). Interaction of nutritional state with season was found for semen volume, sperm concentration and abnormal sperm, but there was no interaction on the total sperm, progressive motility, live sperm, semen pH and semen index. It is concluded that the reproductive activity of growing Sanjabi ram lambs is affected by nutritional state. These results also demonstrated a monthly pattern in reproductive characteristics of Sanjabi ram lambs, independent of the nutritional state.

  20. Effect of sod (superoxide dismutase) protein supplementation in semen extenders on motility, viability, acrosome status and ERK (extracellular signal-regulated kinase) protein phosphorylation of chilled stallion spermatozoa.

    PubMed

    Cocchia, N; Pasolini, M P; Mancini, R; Petrazzuolo, O; Cristofaro, I; Rosapane, I; Sica, A; Tortora, G; Lorizio, R; Paraggio, G; Mancini, A

    2011-04-15

    New studies are underway to find new methods for supporting longer storage of cooled stallion semen. It is known that high concentrations of reactive oxygen species (ROS) cause sperm pathology. The metalloprotein superoxide dismutase (SOD) is responsible for H(2)O(2) and O(2) production, by dismutation of superoxide radicals. The aim of this study is to assess the quality of chilled stallion semen processed with extenders containing SOD at different concentrations as antioxidant additives. A total of 80 ejaculates collected from 5 standardbred stallions was divided into 5 aliquots treated as: native semen (control 1); native semen diluted 1:3 with Kenney semen extender (control 2); spermatozoa diluted after centrifugation in extender without (control 3) or with SOD at 25 IU/ml (experimental 1) or 50 IU/ml (experimental 2). Each sample was analyzed for motility, viability and acrosome status, immediately after semen preparation and again after storage at 5 °C for 24 h, 48 h and 7 2h. Acrosome integrity was evaluated by Chlortetracycline (CTC) and Fluorescent-labeled peanut lectin agglutinin (PNA-FITC conjugated staining). A proteomic approach of quantifying extracellular signal regulated kinase (ERK) was also evaluated as an indirect indicator of oxidative stress. In all samples sperm progressive motility and sperm acrosomal integrity showed a significant reduction between fresh and cooled spermatozoa at 24 h, 48 h and 72 h. Quality parameters of sperm were significantly higher (Progressive Motility P < 0.01; Viability P < 0.001) in aliquots supplemented with SOD. ERK phosphorylation was statistically higher (P < 0.01) in aliquots without SOD. The Authors concluded that addition of SOD to semen extenders improves the quality of chilled equine semen and reduces ERK activation.

  1. Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays.

    PubMed

    Gu, Xingnian; Davis, Rodney J; Walsh, Susan J; Melville, Lorna F; Kirkland, Peter D

    2014-01-01

    Infection with Bluetongue virus (BTV) is a significant impediment to the global movement of bovine semen. Repeat testing of blood from donor animals is specified in the World Organization for Animal Health (OIE) Manual for the export of semen from regions where BTV may be present. Screening of blood or semen samples has usually been carried out by virus isolation (VI) either by inoculation of chicken embryos followed by passage onto insect and mammalian cell cultures or in vivo inoculation of sheep followed by serology to detect seroconversion. Direct testing of semen for BTV would enable earlier release of semen samples and avoid repeat testing of the donor, as well as provide an option for releasing batches of semen that were collected without certification of the donor. Quantitative (real-time) reverse transcription polymerase chain reaction (qRT-PCR) assays overcome most of the limitations of other methods and have the potential to provide higher sensitivity. The present study compared 5 qRT-PCR assays, including 2 commercially available kits, for the detection of BTV in semen serially collected from 8 bulls over a period of 90 days after experimental infection. The results of the study show that at least one of the qRT-PCR assays is extremely reproducible and has both very high sensitivity and specificity to reliably detect all available serotypes. The preferred qRT-PCR gave consistently superior results to VI, sheep inoculation, and conventional RT-PCR. Therefore, the assay can be recommended for the screening of bovine semen for freedom from BTV.

  2. 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.

  3. Incidence, structure and morphological classification of abnormal sperm in the emu (Dromaius novaehollandiae).

    PubMed

    du Plessis, Lizette; Soley, John T

    2011-03-01

    Little detailed information is currently available on the incidence and morphological characteristics of abnormal sperm in the emu (Dromaius novaehollandiae) and of ratites in general. This situation is further compounded by the lack of a uniform system for the morphological classification of avian sperm defects. Considering the important role that sperm morphology plays in the assessment of semen quality, a detailed description of avian sperm defects is of paramount importance. Based on morphological data provided by light and electron microscopy, a mean of 17.3% abnormal sperm was recorded in semen samples collected from the distal deferent duct of four adult emus during the middle of the breeding season. Four categories of defects were identified. Head defects (57.2% of total defects) consisted of bent heads, macrocephalic heads, round heads and acephalic sperm. Zones of incomplete chromatin condensation and retained cytoplasmic droplets appeared to be implicated in head bending, while giant heads were often associated with multiple tails. Acephalic sperm revealed a complete tail devoid of a head which was replaced by a small spherical structure. Tail defects (22.6% of total defects) were subdivided into neck/midpiece defects and principal piece defects. In the neck/midpiece region disjointed sperm were the exclusive defect noted and were characterized by the complete separation of the head and midpiece in the neck region but within the confines of the plasmalemma. Defects observed in the principal piece were subdivided into short tails, coiled tails and multiple tails. No conclusive evidence was obtained that tail coiling represented the 'Dag' defect. Biflagellate sperm were the most common form of multiple tails, demonstrating two complete tails with all the normal structural elements. Cytoplasmic droplets (13.9% of total defects) were classified as a separate defect. The location and eccentric positioning of retained cytoplasmic droplets was similar to that

  4. 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...

  5. 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.

  6. Antioxidant effect of quercetin in an extender containing DMA or glycerol on freezing capacity of goat semen.

    PubMed

    Seifi-Jamadi, Afshin; Ahmad, Ejaz; Ansari, Mahdi; Kohram, Hamid

    2017-04-01

    This study was performed to evaluate the effectiveness of quercetin as a non-enzymatic antioxidant in combination with glycerol or Dimethylacetamide (DMA), on freezability of goat semen. Ejaculates from four healthy mature Mahabadi goats were collected using an artificial vagina. After primary processing, semen was pooled and extended by egg yolk based extender supplemented with different concentrations of quercetin (10 or 20 μM) along with 5% glycerol or DMA. The extended semen was frozen and sperm motility parameters, viability, abnormality, membrane integrity and lipid peroxidation were assessed after thawing. Results showed that sperm viability, total motility, progressive motility, straightness (STR) and linearity (LIN) were higher (P < 0.05), and abnormality percentage and MDA concentration were lower (P < 0.05) in extender containing DMA. Similarly, higher (P < 0.05) total motility, progressive motility, viability and membrane integrity along with lower (P < 0.05) MDA level were noted in Q10 group. The lowest (P < 0.05) MDA level was observed in DMA extender containing moderate level of quercetin (Q10D). Also the STR was higher (P < 0.05) in Q10D compared to Q10G and Q20G groups. In conclusion, supplementation of extender with 10 μM quercetin in combination with DMA improves the goat sperm motion kinetics and suppresses lipid peroxidation after freezing and thawing. Furthermore, DMA is more effective cryoprotectant for the freezing of goat sperm.

  7. Semen quality in papaya workers with long term exposure to ethylene dibromide.

    PubMed Central

    Ratcliffe, J M; Schrader, S M; Steenland, K; Clapp, D E; Turner, T; Hornung, R W

    1987-01-01

    To examine whether long term occupational exposure to ethylene dibromide (EDB) affects semen quality a cross sectional study of semen quality was conducted among 46 men employed in the papaya fumigation industry in Hawaii, with an average duration of exposure of five years and a geometric mean breathing zone exposure to airborne EDB of 88 ppb (eight hour time weighted average) and peak exposures of up to 262 ppb. The comparison group consisted of 43 unexposed men from a nearby sugar refinery. Statistically significant decreases in sperm count per ejaculate, the percentage of viable and motile sperm, and increases in the proportion of sperm with specific morphological abnormalities (tapered heads, absent heads, and abnormal tails) were observed among exposed men by comparison with controls after consideration of smoking, caffeine and alcohol consumption, subject's age, abstinence, history of urogenital disorders, and other potentially confounding variables. No effect of exposure to EDB on sperm velocity, the overall proportion of sperm with normal morphology, or YFF bodies was observed. These data strongly suggest that EDB may increase the risk of reproductive impairment in workers at exposure levels near the National Institute for Occupational Safety and Health recommended limit of 45 ppb (as an eight hour time weighted average) and far below the current standard of the Occupational Safety and Health Administration of 20 ppm. PMID:3297130

  8. [Effect of estradiol valerate on the male reproductive organs and various semen parameters in rats].

    PubMed

    Köhler-Samouilidis, G; Papaioannou, N; Kotsaki-Kovatsi, V P; Vadarakis, A

    1998-01-01

    The effect of the administration of estradiol valerate on the male reproductive organs, on their histological structure, and several semen parameters of Wistar rats was studied. In experiment A and B 140 micrograms estradiol valerate/kg b.w. were administered once a week for 4 weeks to 14 weeks old rats by s.c. injection. One week after the 4th injection the rats of experiment A were sacrificed, while the rats of experiment B lived 5 weeks without treatment for recovery. In both experiments, suppression of body weight, food consumption, decreases in absolute and relative weights of testes, epididymides, prostate and semen vesicles were observed along with testis, epididymis, seminal vesicle and prostate atrophy. The absolute and relative weights of adrenals and pituitary revealed a tendency for increase in both treated groups. The histopathological examination of the testes revealed degeneration of spermatozytes in experiment A, and degeneration of spermatozytes, spermatides, spermatozoa, Sertoli and Leydig cells in experiment B. In experiment A the motility, and number of sperms was significantly decreased, the sperm abnormalities were significantly increased. In experiment B the motility of sperms was slightly, the number significantly decreased and the abnormalities slightly increased.

  9. 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.

  10. 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.

  11. 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.

  12. Effect of semen collection method (artificial vagina vs. electroejaculation), extender and centrifugation on post-thaw sperm quality of Blanca-Celtibérica buck ejaculates.

    PubMed

    Jiménez-Rabadán, P; Ramón, M; García-Álvarez, O; Maroto-Morales, A; del Olmo, E; Pérez-Guzmán, M D; Bisbal, A; Fernández-Santos, M R; Garde, J J; Soler, A J

    2012-05-01

    The aim of this study was to evaluate the effect of semen collection method (artificial vagina compared to electroejaculation), season in which the semen was collected (breeding season compared to non-breeding season), freezing extender (Biladyl(®), Andromed(®) and skim milk based extender) and pre-treatment procedure (washing compared to non-washing) on post-thaw semen quality in buck. Ejaculates from seven bucks of the Blanca-Celtibérica breed were collected by artificial vagina and electroejaculation during the breeding (July to December) and non-breeding season (January to June). Samples were split in two aliquots and one of them was washed. Three freezing extenders were evaluated on washing and non-washing sperm samples. Ejaculates collected by artificial vagina had a greater sperm quality after thawing, with greater values (P≤0.05) for SM (sperm motility), NAR (acrosome intact), YO-PRO-1-/PI- (intact spermatozoa), and Mitotracker+/YO-PRO-1- (spermatozoa with active mitochondria) and lower % DFI (DNA fragmentation index). Thawed sperm samples which were collected during the breeding season had greater values (P≤0.05) for NAR, intact spermatozoa and spermatozoa with active mitochondria, than those semen samples obtained during the non-breeding season. Semen freezing with Biladyl(®) and Andromed(®) resulted in a greater sperm quality (P≤0.05) after thawing in relation to milk-based extender. Washing procedure had no effect on sperm parameters assessed at thawing. Results from the present study suggest that the success of semen cryopreservation in Blanca-Celtibérica goat depends on semen collection method and season, as well as on the extender used. Thus, the post-thaw sperm quality will be greater (P≤0.05) when samples are collected by artificial vagina during the breeding season and when Biladyl(®) or Andromed(®) are used as freezing extenders.

  13. 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

  14. 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.

  15. 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.

  16. 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...

  17. 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

  18. Cryopreservation of rabbit semen using non-permeable cryoprotectants: effectiveness of different concentrations of low-density lipoproteins (LDL) from egg yolk versus egg yolk or sucrose.

    PubMed

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

    2014-12-30

    This study was designed to identify the most effective non-permeable cryoprotectant (CPA) for the cryopreservation of rabbit semen by comparing the effects of different concentrations of low-density lipoproteins (LDL) on post-thaw sperm quality with those of whole egg yolk or sucrose. In a second experiment, the performance of the non-permeable CPAs identified as most effective was assessed in vivo by determining reproductive performances. Pooled semen samples were diluted to a ratio of 1:1 (v:v) in freezing extender (Tris-citrate-glucose and 16% dimethylsulfoxide as permeable CPA) containing as non-permeable CPAs 6, 8, 10 or 15% LDL from egg yolk, 0.1M sucrose, or 15% egg yolk. The semen was loaded in 0.25mL straws and frozen in liquid nitrogen vapor. After thawing, we determined sperm motility, viability, osmotic resistance, and acrosome and DNA integrity. Our results clearly revealed a significant effect of LDL concentration on semen quality. Also, at an optimal concentration of 10%, motility and acrosome integrity were improved over the values recorded for egg yolk (P<0.05). Based on the in vitro data, 3 groups of does (n=30 each) were inseminated with fresh semen or semen frozen using sucrose or 10% LDL. Sucrose led to a significantly higher conception rate than LDL and reproductive performance was similar to that observed for fresh semen. Our findings indicate the markedly better performance of sucrose in vivo as a non-permeable CPA for the cryopreservation of rabbit semen.

  19. 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

  20. HIV RNA and proviral HIV DNA can be detected in semen after 6 months of antiretroviral therapy although HIV RNA is undetectable in blood.

    PubMed

    Du, Peiwei; Liu, An; Jiao, Yanmei; Liu, Cuie; Jiang, Taiyi; Zhu, Weijun; Zhu, Yunxia; Wu, Hao; Sun, Lijun

    2016-03-01

    The risk of sexual transmission of HIV is strongly correlated with amounts of genital HIV RNA. Few studies have reported amounts of HIV RNA and HIV DNA in semen in HIV-infected Chinese patients undergoing antiviral treatment (ART). In this observational study, the amounts of HIV RNA and HIV DNA in semen were assessed after six months of ART in HIV-infected Chinese individuals, when HIV RNA was undetectable in blood . This study included 19 HIV-infected Chinese men undergoing ART for six months. Amounts of HIV in paired semen and blood samples were assessed using real-time PCR. The C2-V5 region of the HIV envelope (env) genes was cloned and sequenced and genotype and co-receptor usage predicted based on the sequence. It was found that HIV RNA was undetectable in the plasma of most patients (17/19), whereas HIV RNA could be detected in the semen of most patients (16/19). HIV DNA could be detected in both semen and blood. Genetic diversity of HIV between the seminal and blood compartments was identified. Thus, amounts of HIV RNA and HIV DNA remain high in semen of HIV-infected Chinese patients after six months of ART treatment, even when HIV RNA was undetectable in blood.

  1. Effects of monochromatic light sources on sex hormone levels in serum and on semen quality of ganders.

    PubMed

    Chang, Shen-Chang; Zhuang, Zi-Xuan; Lin, Min-Jung; Cheng, Chuen-Yu; Lin, Tsung-Yi; Jea, Yu-Shine; Huang, San-Yuan

    2016-04-01

    Light is an essential external factor influencing various physiological processes, including reproductive performance, in birds. Although several attempts have been made to understand the effect of light on poultry production, the effect of light of a particular wavelength (color) on the reproductive function in geese remains unclear. This study evaluated the effect of various monochromatic light sources on the levels of sex hormone and on semen quality of ganders. Of 30 male White Roman geese in their third reproductive season (average age=3 years), 27 were divided into three groups receiving monochromatic white or red or blue lights. The birds were kept in an environmentally controlled house with a lighting photoperiod of 7L:17D for six weeks as the adaptation period. The photoperiod was subsequently changed to 9L:15D and maintained for 24 weeks. Three ganders at the beginning of the study and three from each group at the end of the adjusting period and the 20th and 30th week of the study period were sacrificed, and their testes and blood samples were collected for determining the sex hormone levels. Semen samples were collected for determining semen quality parameters, including the semen collection index, sperm concentration, semen volume, sperm motility, sperm viability, sperm morphology, and semen quality factor. The results showed that the testosterone and estradiol levels remained unchanged in all three groups at all time points. The ratio of testosterone to estradiol of ganders exposed to white light was significantly higher than that of ganders exposed to red light at the 30th week (P<0.05). Semen collection index and sperm viability of ganders exposed to blue light were significantly the lowest (P<0.05). Moreover, sperm motility, sperm viability, and percentage of morphologically normal spermatozoa of ganders in white light were the highest (P<0.05). In conclusion, the results of this study suggested that artificial illumination with white light may

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. Bacterial contamination of ram semen, antibiotic sensitivities, and effects on sperm quality during storage at 15°C.

    PubMed

    Yániz, Jesús Luis; Marco-Aguado, María Angeles; Mateos, José Angel; Santolaria, Pilar

    2010-10-01

    This study was designed to determine the degree and type of bacterial contamination of ejaculated semen samples in fertile rams and its consequences on sperm quality during storage. In experiment 1, 68 ejaculates from 36 rams were divided into two aliquots, one of which was used for bacterial culture, while the other one was diluted, stored at 15°C and assessed for plasma membrane integrity and motility at 0, 24 and 48h after dilution. From the 68 ejaculates, 66 were positive for aerobic bacteria, including 20 species of bacteria from 14 genera. The most frequently isolated bacteria were Escherichia coli, Proteus mirabilis, Enterobacter cloacae, Staphylococcus epidermis, and Staphylococcus aureus species. These 5 bacteria were present in 97% of all contaminated samples. All contaminant bacteria were found to be sensitive to gentamicin and to ceftiofur, with variable percentages of resistance to the other antibiotics evaluated. In samples with total enterobacteria count lower than 100 colony-forming units (CFU)/ml, higher proportions of motile and progressive sperm and higher velocities of spermatozoa were observed at different times during storage. In experiment 2, pure cultures of the most frequently isolated bacteria were individually added to fresh semen samples of low contamination and tested for their effects on sperm quality during storage at 15°C. Semen with E. coli showed a drastic reduction in motility, velocity and viability during storage. This reduction was also significant, but less drastic, in semen with E. cloacae and P. mirabilis, whereas it was partial and less pronounced in the other groups (S. epidermidis and S. aureus). In conclusion, the contamination of ram semen with enterobacterial species reduced sperm quality during storage at 15°C, and the antibiotics gentamicin and ceftiofur showed the higher antimicrobial activities.

  7. Chromosomal abnormalities in human sperm

    SciTech Connect

    Martin, R.H.

    1985-01-01

    The ability to analyze human sperm chromosome complements after penetration of zona pellucida-free hamster eggs provides the first opportunity to study the frequency and type of chromosomal abnormalities in human gametes. Two large-scale studies have provided information on normal men. We have studied 1,426 sperm complements from 45 normal men and found an abnormality rate of 8.9%. Brandriff et al. (5) found 8.1% abnormal complements in 909 sperm from 4 men. The distribution of numerical and structural abnormalities was markedly dissimilar in the 2 studies. The frequency of aneuploidy was 5% in our sample and only 1.6% in Brandriff's, perhaps reflecting individual variability among donors. The frequency of 24,YY sperm was low: 0/1,426 and 1/909. This suggests that the estimates of nondisjunction based on fluorescent Y body data (1% to 5%) are not accurate. We have also studied men at increased risk of sperm chromosomal abnormalities. The frequency of chromosomally unbalanced sperm in 6 men heterozygous for structural abnormalities varied dramatically: 77% for t11;22, 32% for t6;14, 19% for t5;18, 13% for t14;21, and 0% for inv 3 and 7. We have also studied 13 cancer patients before and after radiotherapy and demonstrated a significant dose-dependent increase of sperm chromosome abnormalities (numerical and structural) 36 months after radiation treatment.

  8. Sperm oxidative stress and the effect of an oral vitamin E and selenium supplement on semen quality in infertile men.

    PubMed

    Keskes-Ammar, L; Feki-Chakroun, N; Rebai, T; Sahnoun, Z; Ghozzi, H; Hammami, S; Zghal, K; Fki, H; Damak, J; Bahloul, A

    2003-01-01

    Numerous studies have reported beneficial effects of antioxidant drugs on semen quality, but there is no well-defined therapeutical protocol in male infertility. This study aimed to test the effects of vitamin E and selenium supplementation on lipid peroxidation and on sperm parameters. The study included 54 voluntary and infertile men who produced semen samples for spermiogram and for spectrophotometric measurement of a lipid peroxidation marker, the malondialdehyde (MDA), and produced blood samples for high-performance liquid chromatography assessment of serum vitamin E level. The trial was randomized and open. Twenty-eight men were supplemented daily by vitamin E (400 mg) and selenium (225 microg), during 3 months. The remaining 26 patients received vitamin B (4,5 g/day) for the same duration. Only 20 patients achieved their treatment and returned for control analysis. MDA concentrations in sperm were much less than in seminal plasma and motility and viability were inversely correlated with semen MDA levels. In contrast to vitamin B supplementation, vitamin E and selenium supplementation produced a significant decrease in MDA concentrations and an improvement of sperm motility. The results confirm the protective and beneficial effects of vitamin E and selenium on semen quality and advocate their use in male infertility treatment.

  9. The acid phosphatase test two minute cut-off: an insufficient time to detect some semen stains.

    PubMed

    Redhead, Paul; Brown, Melanie K

    2013-06-01

    The ability to detect semen in sexual offence cases is a crucial first step to locating stains which may be suitable for DNA profiling. Since the development of the acid phosphatase test in the late 1950s by Stuart S. Kind, the process undertaken to perform the test has gone largely unchanged. The method currently accepted by operational forensic science laboratories allows 2 min for a reaction to be obtained, and until relatively recently, this has not been challenged. In this research, samples of semen were obtained from three donors and a range of dilutions for each sample were prepared. Each dilution was subjected to acid phosphatase testing using both direct testing and the 'press test' method. The results showed that semen could be detected in excess of 15 min in dilutions up to 1 in 400 using the press test method and in dilutions up to 1 in 1000 using the direct method. Of further significance was the observation that using the press test method, the two minute cut-off was insufficient to detect the majority of stains and in some cases, semen stains as strong as 1 in 20 dilutions. This research provides compelling evidence for protocols currently utilised in forensic practice to be reviewed in order that forensic scientists do not overlook potential evidential material that may prove suitable for body fluid identification such as DNA STR profiling.

  10. Effect of Long-Term Fish Oil Supplementation on Semen Quality and Serum Testosterone Concentrations in Male Dogs

    PubMed Central

    Risso, Analía; Pellegrino, Francisco Javier; Relling, Alejandro Enrique; Corrada, Yanina

    2016-01-01

    Background Manipulating the dietary fatty acid (FA) content can alter FA profiles of reproductive tissues. Numerous researchers have evaluated the effect of fish oil (FO) supplementation on reproductive characteristics in domestic animals, but reliable information concerning dietary FO effects on semen quality and testosterone concentrations in dogs has not been reported. Therefore, this study evaluated the effects of dietary FO on semen quality and serum testosterone concentrations in dogs. Materials and Methods In this cross-over experimental study, 5 male dogs consumed either a control diet or the same diet supplemented with 54 mg FO/kg metabolic body weight (BW) for 120 days. After the 120-day wash-out period, control (C) dogs received FO and FO-fed dogs consumed the control diet. In the first period, 2 dogs were allocated to the FO group and 3 to the C group. In the second period, 3 dogs were allocated to the FO group and 2 to the C group. Semen samples collected on days 0, 60, 90 and 120 were evaluated by standard methods. Day 120 semen samples were analyzed for FA profiles. Blood samples were collected on days 0, 30, 60, 90 and 120 to measure serum testosterone concentrations. Data were analyzed by analysis of variance with repeated measures using the Mixed Models procedure of SAS (version 9.0, SAS Institute Inc., Cary, NC, USA). Animals and period of time (first or second 120 days) were random variables; and treatment, time, and the treatment by time interaction were considered fixed effects. Results FO supplementation increased the percentage of motile sperm (P=0.02), total sperm count (P<0.01), total sperm viability (P<0.01), and total morphologically normal sperm (P<0.01). Supplementation decreased the percentage of viable sperm (P=0.03) and serum testosterone concentration (P<0.01). FO supplementation also increased the percentage of arachidonic acid, eicosapentaenoic acid, (EPA) and total n-3 in semen samples (P≤0.05). Conclusion These results are

  11. 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.

  12. The Association between Dietary Patterns and Semen Quality in a General Asian Population of 7282 Males

    PubMed Central

    Liu, Chin-Yu; Chou, Yu-Ching; Chao, Jane C. -J.; Hsu, Chien-Yeh; Cha, Tai-Lung; Tsao, Chih-Wei

    2015-01-01

    Objective To explore the associations between different dietary patterns and semen quality in a general Asian male population. Methods Cross-sectional study. Healthy Taiwanese men aged 18 years or older who participated in a standard medical screening program from 2008-2013 run by a private firm were included in this study. Semen parameters including sperm concentration (SC), total sperm motility (TSM), progressive motility (PRM) and normal sperm morphology (NSM) were recorded. A dietary questionnaire was used to categorize the participants into 5 groups: “Healthy diet”, “Western diet”, “High-carbohydrate diet”, “High sweet snacks & sugar-sweetened drinks” and “High-sodium diet”. Results A total of 7282 men completed the questionnaire regarding dietary pattern, and examination of anthropometric indexes was performed and laboratory data were obtained. A high intake of a “Western diet” resulted in statistically linear declines of SC and NSM (P < 0.001 and P < 0.001). Similarly, a greater intake of “High sweet snacks & sugar-sweetened drinks” was associated with a lower SC (P = 0.001). Increased intake of a “High-carbohydrate diet” was related to higher prevalences of abnormal TSM and PRM (P = 0.012 and P = 0.025). Similarly, a greater intake of a “High-sodium diet” was correlated with an elevated prevalence of abnormal NSM (P = 0.035). Conclusions This study showed that a greater intake of a “Western diet” is associated with poorer SC and NSM, a “High sweet snacks and sugar-sweetened drinks” intake is correlated with a lower SC, and high-carbohydrate food is related to elevated prevalences of abnormal TSM and PRM. PMID:26218796

  13. 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

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. Characterization and cryopreservation of semen from endangered markhor goats (Capra falconeri heptneri) with evaluation of reproductive seasonality.

    PubMed

    Bezjian, Marisa; Abou-Madi, Noha; Kollias, George V; Parks, John E; Cheong, Soon Hon; Beltaire, Katherine A

    2013-09-01

    The aims of this study were to determine the reproductive seasonality of four captive markhor goats (Capra falconeri heptneri), to characterize semen collected by electroejaculation, and to compare extenders and processing techniques for semen cryopreservation. Over the course of 1 yr, mean monthly scrotal circumference, serum testosterone, and fecal testosterone were measured and found to be inversely associated with day length. Maximum scrotal circumference (25.2 +/- 0.9 cm), serum testosterone (521.0 +/- 103.4 ng/dl), and fecal testosterone (382.5 +/- 90.3 ng/g) occurred in November, when day length was short (9.7 +/- 0.1 hr). Once a month for 3 mo (December, January, and February), bucks were anesthetized for electroejaculation and semen evaluation. Semen samples were divided into six aliquots for extension and cryopreservation in soy-based Bioxcell or Tris-based extender with 5 or 15% egg yolk, with and without centrifugation. Samples were then thawed for repeat evaluation 1-3 mo later. Postthaw evaluation revealed no significant differences between centrifuged and noncentrifuged samples. Sperm in Tris 5% and 15% egg yolk displayed higher total motility at 0, 3, and 6 hr postthaw and higher progressive motility postthaw compared with sperm in Bioxcell (P < 0.05). Sperm in Bioxcell displayed higher viability than sperm in both Tris-egg yolk extenders (P < 0.01), more intact acrosomes than sperm in Tris-15% egg yolk (P < 0.05), and a tendency for more intact acrosomes than sperm in Tris-5% egg yolk (P < 0.10). Sperm in Tris-5% egg yolk tended to have a higher percentage of morphologically normal sperm compared with Bioxcell (P < 0.10). This study provides evidence that markhor goats exhibit seasonality in scrotal circumference and testosterone levels and that centrifugation may be eliminated from the processing of markhor semen.

  19. Effect of age and environmental factors on semen quality, glutathione peroxidase activity and oxidative parameters in Simmental bulls.

    PubMed

    Balić, I Majić; Milinković-Tur, S; Samardžija, M; Vince, S

    2012-07-15

    Taking into account that semen quality depends on animal age and climate conditions and that oxidative stress has been reported to be a common cause of infertility, the objective of this study was to monitor indicators of oxidative stress and antioxidant protection during four seasonal periods in service bulls of various age to get better insight into the significance of these factors upon evaluating service bull semen. The research was conducted over a year on 19 Simmental service bulls. Animals were divided into two groups according to age; Group I consisted of younger bulls aged two to four yrs (n=9), and Group II was comprised of older bulls aged five to ten yrs (n=10). Semen samples were obtained once in the middle of every seasonal period and blood samples for biochemical analysis were collected by jugular venipuncture immediately after ejaculate collection. The activity of total glutathione peroxidase (T-GSH-Px), selenium-dependent glutathione peroxidase (Se-GSH-Px) and selenium-independent glutathione peroxidase (non-Se-GSH-Px), together with the intensity of lipid peroxidation (thiobarbituric acid reactive substances; TBARS) and oxidative protein damage (protein carbonyl content (PCC)) were measured in seminal plasma. In samples of spermatozoa and blood serum, the activity of Se-GSH-Px and TBARS and PCC concentrations were determined. Older service bulls had significantly higher ejaculate volume in summer in comparison with younger bulls, whereas the number of spermatozoa and progressive motility percentage did not significantly vary with age. Younger animals had lower progressive motility percentage during summer than in spring, with more intensive oxidative processes observed in seminal plasma (TBARS) and spermatozoa (TBARS and PCC). Based on the results presented here, it can be concluded that younger bulls are more sensitive to elevated ambient temperatures during the summer, when intensified prooxidative processes in semen plasma and spermatozoa

  20. 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.

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. Differences in the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques.

    PubMed

    Parrilla, Inma; del Olmo, David; Sijses, Laurien; Martinez-Alborcia, María J; Cuello, Cristina; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2012-05-01

    The present study aimed to evaluate the ability of spermatozoa from individual boar ejaculates to withstand different semen-processing techniques. Eighteen sperm-rich ejaculate samples from six boars (three per boar) were diluted in Beltsville Thawing Solution and split into three aliquots. The aliquots were (1) further diluted to 3×10(7) sperm/mL and stored as a liquid at 17°C for 72 h, (2) frozen-thawed (FT) at 1×10(9) sperm/mL using standard 0.5-mL straw protocols, or (3) sex-sorted with subsequent liquid storage (at 17°C for 6 h) or FT (2×10(7) sperm/mL using a standard 0.25-mL straw protocol). The sperm quality was evaluated based on total sperm motility (the CASA system), viability (plasma membrane integrity assessed using flow cytometry and the LIVE/DEAD Sperm Viability Kit), lipid peroxidation (assessed via indirect measurement of the generation of malondialdehyde (MDA) using the BIOXYTECH MDA-586 Assay Kit) and DNA fragmentation (sperm chromatin dispersion assessed using the Sperm-Sus-Halomax(®) test). Data were normalized to the values assessed for the fresh (for liquid-stored and FT samples) or the sorted semen samples (for liquid stored and the FT sorted spermatozoa). All of the four sperm-processing techniques affected sperm quality (P<0.01), regardless of the semen donor, with reduced percentages of motile and viable sperm and increased MDA generation and percentages of sperm with fragmented DNA. Significant (P<0.05) inter-boar (effect of boars within each semen-processing technique) and intra-boar (effect of semen-processing techniques within each boar) differences were evident for all of the sperm quality parameters assessed, indicating differences in the ability of spermatozoa from individual boars to withstand the semen-processing techniques. These results are the first evidence that ejaculate spermatozoa from individual boars can respond in a boar-dependent manner to different semen-processing techniques.

  9. 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.

  10. 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

  11. 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.

  12. Association of soybean-based extenders with field fertility of stored ram (Ovis aries) semen: a randomized double-blind parallel group design.

    PubMed

    Khalifa, Tarek; Lymberopoulos, Aristotelis; Theodosiadou, Ekaterini

    2013-02-01

    Two consecutive randomized double-blind field fertility experiments were conducted over a 4-month period and aimed at evaluating the association of two commercial soybean lecithin-based extenders (AndroMed [Minitub, Tiefenbach, Germany] and BioXcell [IMV Technologies, L'Aigle, France]) with pregnancy rates of chilled-stored (CS) and frozen-thawed (FT) ram semen. Semen samples with more than 2 × 10(9) sperm per mL and 70% progressive motile spermatozoa were collected via an artificial vagina from twelve proven fertile Chios rams, split-diluted with the above mentioned extenders, packaged in 0.25 mL straws and either stored at 5 ± 1 °C for 30 to 36 hours or frozen and thawed. Non-lactating multiparous ewes were inseminated in progestagen-synchronized estrus either with CS (AndroMed: N = 212 and BioXcell: N = 206; intracervical AI) or with FT (AndroMed: N = 114 and BioXcell: N = 92; laparoscopic intrauterine AI) semen. Ovulation was confirmed in all ewes based on determination of blood plasma progesterone (>1 ng/mL) 8 days post AI. Ewes were screened for pregnancy diagnosis by transabdominal ultrasonography 65 days post AI. BioXcell was superior to AndroMed in preserving the fertilizing potential of CS (P < 0.05) and FT (P < 0.005) semen. In AndroMed-stored semen, young rams (1.5-2.5 years old, N = 8) had a pregnancy rate (59.1%; 124/210) lower than that (72.4%; 84/116) of mature rams (4.5 to 5.5 years, N = 4; P < 0.025). Compared with AndroMed extender, processing of young ram semen in BioXcell extender improved pregnancy rates of CS (66.7%; 88/132 vs. 83.9%; 94/112; P < 0.005) and FT (46.2%; 36/78 vs. 71.0%; 44/62; P < 0.01) spermatozoa. Both extenders were similarly effective in preserving pregnancy rates of mature ram semen (P > 0.05). Ram-by-extender interactions were significant for pregnancy rates of CS and FT semen. Irrespective of extenders, overall pregnancy rates after intracervical and intrauterine AI were 75.1% and 62.2%, respectively (P < 0.001). In

  13. Quantification of leptin in seminal plasma of buffalo bulls and its correlation with antioxidant status, conventional and computer-assisted sperm analysis (CASA) semen variables.

    PubMed

    Kumar, Pradeep; Saini, Monika; Kumar, Dharmendra; Jan, M H; Swami, Dheer Singh; Sharma, R K

    2016-03-01

    The present study is the first to quantify leptin in seminal plasma of buffalo and investigate its relationship with seminal attributes. Ten ejaculates each from 10 Murrah buffalo bulls were collected. Semen quality variables such as semen volume, sperm concentration, sperm abnormalities, membrane integrity, antioxidant enzyme activities (superoxide dismutase, catalase and total antioxidant capacity), malondialdehyde (MDA) concentration, as well as sperm kinetics and motility variables were evaluated. The leptin concentration in serum and seminal plasma were estimated by the ELISA method. Bulls were classified in two groups on the basis of sperm concentration with Group I having >800 million sperm/mL and Group II <500 million sperm/mL. Greater (P<0.05) mean sperm abnormalities, seminal leptin concentrations and MDA concentrations were recorded in Group II than Group I. The seminal leptin was positively correlated with sperm abnormalities and MDA concentration while being negatively correlated with sperm concentration, but there was no correlation with sperm kinetic and motility variables, sperm membrane integrity and seminal plasma antioxidant enzyme activity. Thus, the data suggest that seminal leptin has a role in spermatogenesis and can be used as a marker for spermatogenesis to predict the capacity of buffalo bulls for semen production.

  14. [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.

  15. 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

  16. Improved semen collection method for wild felids: urethral catheterization yields high sperm quality in African lions (Panthera leo).

    PubMed

    Lueders, I; Luther, I; Scheepers, G; van der Horst, G

    2012-08-01

    For wild and domestic felids, electroejaculation (EE) is the most common semen collection method. However, the equipment is expensive, there is a risk of urine contamination and animals usually show strong muscular contraction despite general anesthesia. Accordingly, we tested the feasibility of a different approach using urethral catheterization (UC) in seven African lions, previously described for domestic cats only. After general anesthesia with the α2-agonist medetomidine (which also stimulates semen release into the urethra) and ketamine, a transrectal ultrasound was performed to locate the prostate. A commercial dog urinary catheter (2.6 or 3.3 mm in diameter) was advanced approximately 30 cm into the urethra to allow semen collection into the lumen of the catheter by capillary forces. After retraction, sperm volumes between of 422.86 ± 296.07 μl yielded motility of 88.83 ± 13.27% (mean ± SD) with a mean sperm concentration of 1.94 × 10(9)/ml. Here we describe a simple, field friendly and effective method to attain highly concentrated semen samples with excellent motility in lions and potentially other wild felid species as an alternative to electroejaculation.

  17. Development of a SYBR Green I based duplex real-time PCR for detection of bovine herpesvirus-1 in semen.

    PubMed

    Pawar, Sachin S; Meshram, Chetan D; Singh, Niraj K; Saini, Mohini; Mishra, B P; Gupta, Praveen K

    2014-11-01

    Bovine herpesvirus-1 (BoHV-1) is a viral pathogen found in infected bull semen, which is transmitted to inseminated cows by artificial insemination. BoHV-1 infection can cause reproductive disorders leading to significant economic loss to cattle industry. To detect BoHV-1 in semen, in this study, a SYBR Green I based duplex real-time PCR was developed. The assay included primers from BoHV-1 glycoprotein C (gC) and bovine growth hormone (bGH) genes for simultaneous detection in single tube. The result was interpreted by analysing melting temperature (Tm) peaks obtained after melt curve analysis of the amplified products at the end of reaction. The Tm peaks for BoHV-1-gC indicated presence of BoHV-1 while the bGH peak indicated reaction without inhibition. The sensitivity of the assay was to detect ten BoHV-1 genome copies per reaction. The analytical sensitivity was to detect 0.21 TCID50 infectious BoHV-1 in spiked semen. The assay was validated with 80 semen samples collected from breeding bulls. The diagnostic sensitivity and specificity of the assay was 100% with OIE recommended TaqMan probe based real-time PCR.

  18. Liquid storage of equine semen: Assessing the effect of d-penicillamine on longevity of ejaculated and epididymal stallion sperm.

    PubMed

    Brogan, P T; Beitsma, M; Henning, H; Gadella, B M; Stout, T A E

    2015-08-01

    Short-term storage of equine sperm at 5°C in an extender containing milk and/or egg yolk components is common practice in the equine breeding industry. Sperm motility, viability, DNA integrity and, consequently, fertilizing ability decline over time, partly due to reactive oxygen species (ROS) generation. We investigated whether adding the anti-oxidant d-penicillamine to a commercial milk/egg yolk extender delayed the decrease in semen quality. Semen was recovered on four consecutive days from eight 3-year old Warmblood stallions. On day 5, seven of the stallions were castrated and sperm recovered from the caudae epididymides. Ejaculated samples were split, and one portion was centrifuged and re-suspended to reduce seminal plasma content. All samples were diluted to 50millionsperm/ml and divided into two portions, one of which was supplemented with 0.5mM d-penicillamine. After 48h, 96h, 144h and 192h storage, sperm motility was assessed by computer-assisted semen analysis (CASA), viability by SYBR14/PI staining, and DNA integrity using the sperm chromatin structure assay (SCSA). d-Penicillamine had no effect on motility of ejaculated sperm (P>0.05) but reduced total and progressive motility of epididymal sperm. Sperm chromatin integrity was not influenced by storage time, seminal plasma or d-penicillamine. In short, adding d-penicillamine to a commercial semen extender was neither beneficial nor detrimental to the maintenance of quality in ejaculated semen stored at 5°C. The negative effect on motility of epididymal sperm may reflect differences in (membrane) physiology of spermatozoa that have not been exposed to seminal plasma.

  19. Elemental composition of human semen is associated with motility and genomic sperm defects among older men

    PubMed Central

    Schmid, Thomas E.; Grant, Patrick G.; Marchetti, Francesco; Weldon, Rosana H.; Eskenazi, Brenda; Wyrobek, Andrew J.

    2013-01-01

    BACKGROUND Older men tend to have poorer semen quality and are generally at higher risks for infertility and abnormal reproductive outcomes. METHODS We employed proton-induced X-ray emission (PIXE, 3 MeV proton beam) to investigate the concentrations of zinc, copper, calcium, sulfur, chlorine, potassium, titanium, iron and nickel in washed sperm and seminal plasma from non-smoking groups of 10 older men (65–80 years old) and 10 younger men (22–28 years old) who were concurrently assayed for sperm function and genomicly defective sperm. RESULTS The older group showed elevated zinc, copper and calcium in sperm and elevated sulfur in seminal plasma compared with the younger men. The older group also showed reduced motility as well as increased sperm DNA fragmentation, achondroplasia mutations, DNA strand breaks and chromosomal aberrations. Sperm calcium and copper were positively associated with sperm DNA fragmentation (P < 0.03). Seminal sulfur was positively associated with sperm DNA fragmentation and chromosomal aberrations (P < 0.04), and negatively associated with sperm motility (P < 0.05). Sperm calcium was negatively associated with sperm motility, independent of male age (P = 0.01). CONCLUSIONS We identified major differences in elemental concentrations between sperm and seminal plasma and that higher sperm copper, sulfur and calcium are quantitatively associated with poorer semen quality and increased frequencies of genomic sperm defects. PMID:23042799

  20. Occupational exposure to pesticides and consequences on male semen and fertility: a review.

    PubMed

    Mehrpour, Omid; Karrari, Parissa; Zamani, Nasim; Tsatsakis, Aristides M; Abdollahi, Mohammad

    2014-10-15

    Exposure to pesticides affects many body organs including reproductive system. Disorder of the reproductive system leads to infertility and therefore has been in the center of attention within the recent decades. Pesticides are one of the compounds that might reduce the semen quality in the exposed workers according to current knowledge. Although many underlying mechanisms have been proposed, the mechanisms of action are not clarified yet. The object of the present review was to criticize all the results of studies which evaluated the pesticide effects on male reproductive system. Results indicate that semen changes are multifactorial in the workers exposed to pesticides as there are numerous factors affecting sperm quality in occupational exposures. Majority of pesticides including organophosphoruses affect the male reproductive system by mechanisms such as reduction of sperm density and motility, inhibition of spermatogenesis, reduction of testis weights, reduction of sperm counts, motility, viability and density, and inducing sperm DNA damage, and increasing abnormal sperm morphology. Reduced weight of testes, epididymis, seminal vesicle, and ventral prostate, seminiferous tubule degeneration, change in plasma levels of testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH), decreased level and activity of the antioxidant enzymes in testes, and inhibited testicular steroidogenesis are other possible mechanisms. Moreover, DDT and its metabolites have estrogenic effects on males. Although effect of pesticides on sperm quality is undeniable, well-designed long-term studies are needed to elucidate all the possible affecting variables such as socioeconomic, cultural, nutritional, occupational, physical, and clinical characteristics alongside pesticides.

  1. Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa

    PubMed Central

    Bolanca, Ivan; Obhodas, Jasmina; Ljiljak, Dejan; Matjacic, Lidija; Kuna, Krunoslav

    2016-01-01

    We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower “linearity” and “straightness”, and as showed by this research “elongation”, were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20), asthenozoospermia (n = 54) and oligoasthenozoospermia (n = 23). The findings indicate that sperm quality of patients with normal spermiogram diagnosed according to WHO criteria, may be compromised by showing premature spontaneous hyperactivation which can decrease the chances of natural conception. We assessed synergistic effects of multiple chemical elements in ejaculated semen to find if premature spontaneous hyperactivation of spermatozoa can be a sign of imbalanced semen composition especially of elements K, Ca, Cu and Zn. Human semen samples showing low or high baseline status of chemical elements concentrations were found in samples from all three diagnostic groups. However, correlation of K/Ca and Cu/Zn ratios, taking into account samples from all three groups of men, were negative at statistical significance level p = 0.01. We tested if the negative correlation between K/Ca and Cu/Zn ratio works for greater number of semen samples. We found the negative correlation to be valid for 175 semen samples at statistical significance of p = 0.00002. The ratio of K/Ca and Cu/Zn, i.e. increased concentrations of K and Zn in comparison to concentrations of Ca and Cu, were associated with a decrease of “straightness” in the group of men with normal spermiogram and pronounced spontaneous hyperactivation of spermatozoa, implying that these elements act in synergy and that the balance of elements and not their absolute concentrations plays the major role in premature spermatozoa hyperactivation in ejaculated semen. PMID:27031102

  2. Synergetic Effects of K, Ca, Cu and Zn in Human Semen in Relation to Parameters Indicative of Spontaneous Hyperactivation of Spermatozoa.

    PubMed

    Bolanca, Ivan; Obhodas, Jasmina; Ljiljak, Dejan; Matjacic, Lidija; Kuna, Krunoslav

    2016-01-01

    We have observed that sperm quality parameters indicative of spermatozoa hyperactivation such are lower "linearity" and "straightness", and as showed by this research "elongation", were more pronounced in patients with normal spermiogram compared to the group of men with reduced sperm motility who were undergoing routine in vitro fertilisation. The research encompassed 97 men diagnosed with normozoospermia (n = 20), asthenozoospermia (n = 54) and oligoasthenozoospermia (n = 23). The findings indicate that sperm quality of patients with normal spermiogram diagnosed according to WHO criteria, may be compromised by showing premature spontaneous hyperactivation which can decrease the chances of natural conception. We assessed synergistic effects of multiple chemical elements in ejaculated semen to find if premature spontaneous hyperactivation of spermatozoa can be a sign of imbalanced semen composition especially of elements K, Ca, Cu and Zn. Human semen samples showing low or high baseline status of chemical elements concentrations were found in samples from all three diagnostic groups. However, correlation of K/Ca and Cu/Zn ratios, taking into account samples from all three groups of men, were negative at statistical significance level p = 0.01. We tested if the negative correlation between K/Ca and Cu/Zn ratio works for greater number of semen samples. We found the negative correlation to be valid for 175 semen samples at statistical significance of p = 0.00002. The ratio of K/Ca and Cu/Zn, i.e. increased concentrations of K and Zn in comparison to concentrations of Ca and Cu, were associated with a decrease of "straightness" in the group of men with normal spermiogram and pronounced spontaneous hyperactivation of spermatozoa, implying that these elements act in synergy and that the balance of elements and not their absolute concentrations plays the major role in premature spermatozoa hyperactivation in ejaculated semen.

  3. Processed meat intake is unfavorably and fish intake favorably associated with semen quality indicators among men attending a fertility clinic.

    PubMed

    Afeiche, Myriam C; Gaskins, Audrey J; Williams, Paige L; Toth, Thomas L; Wright, Diane L; Tanrikut, Cigdem; Hauser, Russ; Chavarro, Jorge E

    2014-07-01

    Emerging literature suggests that men's diets may affect spermatogenesis as reflected in semen quality indicators, but literature on the relation between meat intake and semen quality is limited. Our objective was to prospectively examine the relation between meat intake and indicators of semen quality. Men in subfertile couples presenting for evaluation at the Massachusetts General Hospital Fertility Center were invited to participate in an ongoing study of environmental factors and fertility. A total of 155 men completed a validated food-frequency questionnaire and subsequently provided 338 semen samples over an 18-mo period from 2007-2012. We used linear mixed regression models to examine the relation between meat intake and semen quality indicators (total sperm count, sperm concentration, progressive motility, morphology, and semen volume) while adjusting for potential confounders and accounting for within-person variability across repeat semen samples. Among the 155 men (median age: 36.1 y; 83% white, non-Hispanic), processed meat intake was inversely related to sperm morphology. Men in the highest quartile of processed meat intake had, on average, 1.7 percentage units (95% CI: -3.3, -0.04) fewer morphologically normal sperm than men in the lowest quartile of intake (P-trend = 0.02). Fish intake was related to higher sperm count and percentage of morphologically normal sperm. The adjusted mean total sperm count increased from 102 million (95% CI: 80, 131) in the lowest quartile to 168 million (95% CI: 136, 207) sperm in the highest quartile of fish intake (P-trend = 0.005). Similarly, the adjusted mean percentages of morphologically normal sperm for men in increasing quartiles of fish intake were 5.9 (95% CI: 5.0, 6.8), 5.3 (95% CI: 4.4, 6.3), 6.3 (95% CI: 5.2, 7.4), and 7.5 (95% CI: 6.5, 8.5) (P-trend = 0.01). Consuming fish may have a positive impact on sperm counts and morphology, particularly when consumed instead of processed red meats.

  4. The breed and season effects on scrotal circumference and semen characteristics of hair sheep rams under tropical conditions.

    PubMed

    Cárdenas-Gallegos, M A; Aké-López, J R; Centurión-Castro, F; Magaña-Monforte, J G

    2012-12-01

    The aim of the present work was to evaluate the breed and season effects on scrotal circumference (SC) and semen characteristics of 28 mature hair sheep rams kept under tropical conditions. SCs, sperm concentration (SPC) and abnormal sperm were significantly affected by breed effect (p < 0.001). The season effect was significant in SPC (p < 0.0001) while ejaculate volume, mass motility and SPC were affected by breed × season interaction effect (p < 0.001). It can be concluded that the magnitude of the breed and season effects were not sufficient to affect the reproductive capacity of hair sheep rams throughout the year.

  5. Sperm fertility and viability following 48h of refrigeration: evaluation of different extenders for the preservation of bull semen in liquid state.

    PubMed

    Crespilho, A M; Nichi, M; Guasti, P N; Freitas-Dell'Aqua, C P; Sá Filho, M F; Maziero, R R; Dell'aqua, J A; Papa, F O

    2014-05-01

    Two experiments were conducted to compare the effectiveness of different extenders conventionally used for semen cryopreservation to maintain the viability and fertility of cooled bull semen. In Experiment 1, sperm samples obtained from 20 Nellore bulls were preserved at 5°C for 48h using two extenders containing 20% of egg yolk [Tris (TRIS-R) and Botu-Bov(®) (BB)] and another composed of 1% soy lecithin [Botu-Bov(®)-Lecithin (BB-L)] as substitutes for animal origin products. The samples were evaluated at 6, 24 and 48h for plasma and acrosomal membrane integrity, quantification of thiobarbituric acid reactive substances (ng of TBARS/10(8) cells) and sperm motility parameters by computer-assisted semen analysis (CASA). In Experiment 2, pregnancy rate (P/AI) of 973 fixed-time artificially inseminated Nellore cows were compared when cows were inseminated with conventionally cryopreserved semen in TRIS-egg yolk glycerol (TRIS-C Control, n=253) or semen cooled for 48h in TRIS-R (n=233), BB (n=247) or BB-L (n=240). Although none of the extenders used was effective on maintaining total progressive motility and cellular integrity throughout the 48-h of the refrigeration period (P<0.01), BB-L conferred greater protection against oxidative stress (P<0.05) than egg yolk-based medias. The P/AI for semen samples preserved in TRIS-C, TRIS-R, BB and BB-L were 39.92(a), 25.32(b), 26.32(b) and 33.33(ab), respectively. These results demonstrate that the three conventional extenders used for semen cryopreservation do not provide the protection required to maintain bull semen fertility under refrigeration for a 48-h period, resulting in reduced pregnancy rates. However, the use of lecithin-based medium instead of egg yolk results in greater protection against lipid peroxidation, producing P/AI results comparable to those obtained using frozen semen.

  6. Role of amino acids as additives on sperm motility, plasma membrane integrity and lipid peroxidation levels at pre-freeze and post-thawed ram semen.

    PubMed

    Sangeeta, Sharon; Arangasamy, A; Kulkarni, S; Selvaraju, S

    2015-10-01

    The possibility of including amino acids for cryopreservation of ram semen to improve the quality of frozen semen was explored in this study in sheep model. 24 samples were collected in triplicate from 8 rams of 2-3 year old Bannur cross bred rams maintained at the Institute Experimental Livestock Unit. Semen was diluted in tris-egg yolk glycerol diluent and made into 7 aliquots as follows: aliquot 1 served as control, "l-alanine" was added at 100 and 135mM in the aliquots 2 and 3, "l-glutamine" was added at 20 and 25mM in the aliquots 4 and 5 and "l-proline" was added at 25 and 50mM in the aliquots 6 and 7, respectively. Diluted semen was filled in 0.25ml French straws and frozen in LN2. Inclusion of "l-proline" and "l-glutamine" in the diluent increased the percent live sperm (P<0.001), total motility (P<0.05) and maintained higher functional membrane and acrosomal integrity (P<0.001) by decreasing lipid peroxidation (P<0.001) compared to the control group. In contrast, "l-alanine" decreased the percentage of total motility, fast progressive spermatozoa and increased (P<0.01) the percentage of immotile spermatozoa. It can be concluded that 20mM "l-glutamine" and 25mM "l-proline" can be used as semen additive to freeze ram semen as they prevented cryoinjuries to sperm and improved the pre-freeze and post-thaw semen characteristics.

  7. Potential effect of smoking on semen quality through DNA damage and the downregulation of Chk1 in sperm

    PubMed Central

    CUI, XIANGRONG; JING, XUAN; WU, XUEQING; WANG, ZHENQIANG; LI, QIANG

    2016-01-01

    Previous studies have found that smoking is associated with decreased male fertility via altering the quality of semen. However, the mechanism by which cigarette smoking affects semen quality remains to be fully elucidated. Heavy smoking-induced DNA damage has been reported to correlate with abnormal spermatozoa and male infertility. It has been reported that, in response to DNA damage, activation of the checkpoint kinase 1 (Chk1) facilitates S and G2 checkpoint arrest. The aim of the present study was to investigate the expression levels of Chk1 in sperm cells of smoking and non-smoking men, and to further examine the correlation between DNA fragmentation rates and the expression levels of Chk1 with smoking. The present study was performed on a cohort of 841 smoking men and 287 non-smoking men. In the investigation, sperm concentration, motility, viability, seminal plasma zinc concentration, acrosin activity and sperm DNA fragmentation were examined. The gene and protein expression levels of Chk1 were detected using reverse transcription quantitative-polymerase chain reaction and western blot analyses, respectively. It was observed that the progressive motility of the sperm was significantly decreased in the moderate and heavy smoking groups, whereas no significant changes were observed in the mild smoking group. The sperm in the medium-term smoking group had significantly decreased progressive motility, and the semen concentration, sperm count and progressive motility vitality were markedly decreased in the long-term smoking group. Compared with the non-smoking group, the abnormal head rates in the heavy smoking group and long-term smoking group were significantly increased. The sperm viability and seminal plasma zinc concentration were markedly increased in the smoking group. Increased DNA fragmentation rates were found in the smoking group. The expression of Chk1 was significantly decreased in the smoking group, compared with the non-smoking group. Progressive

  8. Urine - abnormal color

    MedlinePlus

    ... medlineplus.gov/ency/article/003139.htm Urine - abnormal color To use the sharing features on this page, please enable JavaScript. The usual color of urine is straw-yellow. Abnormally colored urine ...

  9. Tooth - abnormal colors

    MedlinePlus

    ... medlineplus.gov/ency/article/003065.htm Tooth - abnormal colors To use the sharing features on this page, please enable JavaScript. Abnormal tooth color is any color other than white to yellowish- ...

  10. Abnormal Head Position

    MedlinePlus

    ... cause. Can a longstanding head turn lead to any permanent problems? Yes, a significant abnormal head posture could cause permanent ... occipitocervical synostosis and unilateral hearing loss. Are there any ... postures? Yes. Abnormal head postures can usually be improved depending ...

  11. Skeletal limb abnormalities

    MedlinePlus

    ... medlineplus.gov/ency/article/003170.htm Skeletal limb abnormalities To use the sharing features on this page, please enable JavaScript. Skeletal limb abnormalities refers to a variety of bone structure problems ...

  12. Influence of seminal plasma on cryopreservation of human spermatozoa in a biological material-free medium: study of normal and low-quality semen.

    PubMed

    Grizard, G; Chevalier, V; Griveau, J F; Le Lannou, D; Boucher, D

    1999-06-01

    The objective was to evaluate the efficiency of a biological material-free medium and the role of seminal plasma (SP) in the cryopreservation of human spermatozoa. Normal semen samples and low-quality semen samples were used for this study. After centrifugation of 300 microL fractions of whole semen, pellets were resuspended either in autologous SP or in a chemically defined medium (BM) supplemented or not with 3% bovine serum albumin (BSA); after 15 min at 37 degrees C, the samples were diluted (V/V) with cryoprotective medium (30 mM NaCl; 22 mM sodium citrate, 19.4 mM fructose; 80 mM glutamine; 14%, V/V, glycerol) and maintained for 15 min at room temperature before freezing. Assessment of viability and motility was performed using fresh semen (T0), after centrifugation and resuspension prior to adding the cryoprotectant (T15), after adding the cryoprotectant (T30) and after freezing and thawing (Tpost). In all three resuspending media used, sperm viability and motility (forward and total) decreased (p < 0.05) during both the equilibration period especially before addition of the cryoprotective medium (between T0 and T15) and during the freeze-thaw process comparison between T30 and Tpost. The recovery of viable and motile spermatozoa (post-thaw values/values of fresh samples) was higher (p < 0.05) in normal semen than in low-quality semen. In both groups, the recovery was slightly, but significantly, higher with SP than with BM and the presence of BSA has no beneficial effect. To conclude, these data suggest that SP may reduce the deleterious effects of cryopreservation. Nevertheless cryopreservation of spermatozoa in a medium containing neither SP nor biological substances could offer an acceptable cryoprotection of spermatozoa to be used in assisted fertilization procedures, especially for intracytoplasmic sperm injection.

  13. Abnormal Uterine Bleeding FAQ

    MedlinePlus

    ... PROBLEMS Abnormal Uterine Bleeding • What is a normal menstrual cycle? • When is bleeding abnormal? • At what ages is ... treat abnormal bleeding? •Glossary What is a normal menstrual cycle? The normal length of the menstrual cycle is ...

  14. 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.

  15. Lower levels of HIV RNA in semen in HIV-2 compared with HIV-1 infection: implications for differences in transmission

    PubMed Central

    Gottlieb, Geoffrey S.; Hawes, Stephen E.; Agne, Habibatou D.; Stern, Joshua E.; Critchlow, Cathy W.; Kiviat, Nancy B.; Sow, Papa Salif

    2013-01-01

    Background and objectives HIV-2 infection, in comparison with HIV-1, is characterized by lower plasma viral loads, slower CD4 cell count decline, decreased AIDS-related mortality, and lower rates of mother-to-child and sexual transmission. To gain further insight into why HIV-1 is more readily transmitted as compared with HIV-2, we analyzed semen and plasma HIV RNA levels in HIV-1 and HIV-2-positive men from Senegal. Design and methods Twenty-two HIV-1 and 10 HIV-2-infected subjects from the University of Dakar donated semen and blood samples for this analysis. HIV-1 and HIV-2 viral loads in semen and plasma were quantified using type-specific polymerase chain reaction assays. Results The mean age of the subjects was 37 and 40 years; mean CD4 cell count was 222 and 276 cells/µl and the mean plasma viral load was 4.7 and 3.0 log10 copies/ml for HIV-1 and HIV-2, respectively (P = 0.002). HIV RNA was detected in semen in 21 of 22 (95%) of HIV-1 and seven of 10 (70%) of HIV-2-infected subjects; P = 0.07). However, the levels of HIV RNA present in semen were markedly different between those with HIV-1 and HIV-2, with a mean of 4.4 log10 copies/ml among those with HIV-1 and a mean of 2.6 log10 copies/ml among those with HIV-2 (P < 0.001). In multivariate analysis, plasma viral load and HIV type, but not CD4 cell count, were independently predictive of semen viral load (P = 0.03, 0.05, 0.48, respectively) Conclusions These data suggest that differences in semen viral load between HIV-1 and HIV-2 may be in part responsible for the markedly different transmission rates of these two viruses. In addition, risk of male genital tract shedding strongly correlates with plasma viral loads. Interventions that decrease viral load may help decrease transmission of both HIV-1 and HIV-2. PMID:16549974

  16. Detection of Leptospira spp. in semen and vaginal fluids of goats and sheep by polymerase chain reaction.

    PubMed

    Lilenbaum, W; Varges, R; Brandão, F Z; Cortez, A; de Souza, S O; Brandão, P E; Richtzenhain, L J; Vasconcellos, S A

    2008-04-15

    Thirteen goat herds and seven sheep flocks in the state of Rio de Janeiro, Brazil were screened for leptospirosis. From the three herds and three flocks with greatest seroreactivity, 19 goats (16 females and three bucks) and 40 sheep (26 ewes and 14 rams) that were seropositive (specific anti-Leptospira titres > or =400, based on a microscopic agglutination test), were selected for more detailed studies. From those animals, samples of vaginal fluids or semen were collected for bacteriological and molecular assays. For both species of animals, the most prevalent reactions were to serovars Hardjo, Shermani, and Grippotyphosa. Although leptospires were detected by darkfield microscopy in three vaginal fluid samples (from two goats and one ewe), pure isolates were not obtained by bacteriological culture of vaginal fluids or semen. However, seven vaginal fluid samples (from four goats and three ewes) and six semen samples (all from rams) were positive on polymerase chain reaction (PCR). Based on these findings, in addition to analogous findings in cattle, we inferred that there is potential for venereal transmission of leptospirosis in small ruminants.

  17. 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

  18. 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...

  19. Effects of caffeine supplementation in post-thaw human semen over different incubation periods.

    PubMed

    Pariz, J R; Hallak, J

    2016-11-01

    This study aimed to evaluate the effects of caffeine supplementation in post-cryopreservation human semen over different incubation periods. After collection by masturbation, 17 semen samples were analysed according to World Health Organization criteria, processed and cryopreserved with TEST-yolk buffer (1 : 1) in liquid nitrogen. After a thawing protocol, samples were incubated with 2 mm of caffeine for 0, 5, 15, 30 or 60 min, followed by analysis of motility and mitochondrial activity using 3,3'-diaminobenzidine (DAB). Mean variance analysis was performed, and P < 0.05 was the adopted significance threshold. Samples incubated for 15 min showed increased progressive motility compared to other periods of incubation, as well as a reduced percentage of immotile spermatozoa (P < 0.05). In samples incubated for 5 min, increased mitochondrial activity above 50% was observed (DABI and DABII). Although cryosurvival rates were low after the cryopreservation process, incubation with caffeine was associated with an increase in sperm motility, particularly 15-min incubation, suggesting that incubation with caffeine can be an important tool in patients with worsening seminal quality undergoing infertility treatment.

  20. 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

  1. 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.

  2. Reversible meiotic abnormalities in azoospermic men with bilateral varicocele after microsurgical correction.

    PubMed

    North, M O; Lellei, I; Rives, N; Erdei, E; Dittmar, A; Barbet, J P; Tritto, G

    2004-05-01

    Because of a possible relationship between microenvironmental disturbances and meiotic abnormalities and of a straight relationship between lower-quality semen in patient carrying a varicocele and first meiotic non-disjunction, bilateral bipolar testicular biopsies are realized according the thermic differential gradient described in varicocele. Systematic meiotic studies of multiple testicular biopsies from 65 azoospermic men with bilateral varicocele were done in a multi-centric study on microsurgical correction of bilateral varicocele with microthermic intra-operative evaluation using minimally invasive thermal microsensors (Betatherm 10K3MCD2). In the present study abnormal temperature raising, histomorphometric abnormalities (spermatocyte arrest) and meiotic abnormalities (class IIC) are strongly correlated. In the ten patients submitted to another testicular biopsy procedure six months after surgery for TESE, normal thermal differential is registered and no meiotic abnormalities recurrences are found.

  3. Fruit and vegetable intake and their pesticide residues in relation to semen quality among men from a fertility clinic

    PubMed Central

    Chiu, Y.H.; Afeiche, M.C.; Gaskins, A.J.; Williams, P.L.; Petrozza, J.C.; Tanrikut, C.; Hauser, R.; Chavarro, J.E.

    2015-01-01

    STUDY QUESTION Is consumption of fruits and vegetables with high levels of pesticide residues associated with lower semen quality? SUMMARY ANSWER Consumption of fruits and vegetables with high levels of pesticide residues was associated with a lower total sperm count and a lower percentage of morphologically normal sperm among men presenting to a fertility clinic. WHAT IS KNOWN ALREADY Occupational and environmental exposure to pesticides is associated with lower semen quality. Whether the same is true for exposure through diet is unknown. STUDY DESIGN, SIZE, DURATION Men enrolled in the Environment and Reproductive Health (EARTH) Study, an ongoing prospective cohort at an academic medical fertility center. Male partners (n = 155) in subfertile couples provided 338 semen samples during 2007–2012. PARTICIPANTS/MATERIALS, SETTING, METHODS Semen samples were collected over an 18-month period following diet assessment. Sperm concentration and motility were evaluated by computer-aided semen analysis (CASA). Fruits and vegetables were categorized as containing high or low-to-moderate pesticide residues based on data from the annual United States Department of Agriculture Pesticide Data Program. Linear mixed models were used to analyze the association of fruit and vegetable intake with sperm parameters accounting for within-person correlations across repeat samples while adjusting for potential confounders. MAIN RESULTS AND THE ROLE OF CHANCE Total fruit and vegetable intake was unrelated to semen quality parameters. High pesticide residue fruit and vegetable intake, however, was associated with poorer semen quality. On average, men in highest quartile of high pesticide residue fruit and vegetable intake (≥1.5 servings/day) had 49% (95% confidence interval (CI): 31%, 63%) lower total sperm count and 32% (95% CI: 7%, 58%) lower percentage of morphologically normal sperm than men in the lowest quartile of intake (<0.5 servings/day) (P, trend = 0.003 and 0

  4. 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.

  5. 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).

  6. 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

  7. 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.

  8. 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.

  9. 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...

  10. Semen Quality, Hormonal Levels, and Androgen Receptor Gene Polymorphisms in a Population of Young Male Volunteers from Two Different Regions of Poland

    PubMed Central

    Kamieniczna, Marzena; Fraczek, Monika; Malcher, Agnieszka; Rozwadowska, Natalia; Czernikiewicz, Anna; Jedrzejczak, Piotr; Semczuk, Marian; Kurpisz, Maciej

    2015-01-01

    Background The population of healthy Polish men has not been frequently and systematically investigated for fertility status. The aim of this study was to assess the quality of semen in a randomly recruited population of young males. The most important task was to find a relationship between semen parameters, sex hormones, and AR gene polymorphism. Material/Methods Semen and blood samples from young men from the Poznan (n=113) and Lublin regions (n=89) were collected for semen analysis, assessment of hormonal concentrations, and calculation of the CAG and GGN repeats of the AR gene. Results Statistical comparisons of the hormones and circulating proteins and the seminological parameters revealed significant differences between the regional groups of males studied. Among the correlations found, we emphasize the positive relationship between inhibin B levels and both the number of spermatozoa per ml (R=0.37; p=0.0001) and the total sperm concentration (R=0.40; p=0.00003). Positive correlations between IGF1 and sperm morphology was also found (R=0.40; p=0.000004). The mean number of CAG repeats in our tested groups was 21.93±2.79, in a range from 16 to 31. The mean number of GGN repeats was 23.2±1.66 and ranged from 16 to 29. Numerous significant correlations were found between CAG or GGN repeats and blood hormones or circulating proteins and semen parameters; however, Spearman’s rank correlations revealed rather weak coefficients. Conclusions This report attempted to determine the quality of semen samples and sex hormones in a population of Polish young men. The results were found to be similar to data obtained in Scandinavia. The calculated means and range of CAG or GGN repeats of the AR gene in Polish males were similar to West European epidemiological data. PMID:26299772

  11. Influence of Selected Extenders for Liquid Storage at 4 degrees C of Breeding Chinchilla (Chinchilla lanigera) Semen on Sperm DNA Integrity.

    PubMed

    Niedbała, Piotr; Szeleszczuk, Olga; Kuchta-Gładysz, Marta; Joneczek, Maria; Dobrzyńska, Małgorzata; Maj, Dorota

    2015-01-01

    The influence of two commercial and two laboratory oriented extenders on survival rate and DNA integrity of chinchilla (Chinchilla lanigera) sperm was determined during liquid storage. Semen was collected using an electroejaculator from 6 adult male chinchillas. Ejaculates (n = 16) were diluted with extenders to obtain a concentration of 40 x 10 (3) sperm/5 μl. After dilution the semen samples were stored at 4"C. The percent motility, progressive motility, and morphology were assessed conventionally, whereas DNA integrity was evaluated by Single Cell Gel Electrophoresis (comet) assay at 0 (just after dilution), 24, 48 and 72 h. Conventional assessment of sperm quality showed that commercial extenders are characterized by the lowest sperm survival parameters out of the investigated extenders. In commercial extenders spermatozoa lost their capacity for progressive motility compared to laboratory extenders. After 24 h storage, from 21.67% to 30% of motile sperms were observed in commercial extender whereas total sperm motility was 63.33% (41.67% with progressive motility) in samples in which stallion semen extender was used. After 72 h storage, 10% of sperm were motile in stallion semen extender while no sperm movement was observed in tubes containing the commercial extender. Furthermore, a lower percentage of damaged spermatozoa in laboratory oriented extenders was demonstrated. It was also stated that along with the extended time of semen storage at 4 degrees C, commercial extenders lost their protective action. An analysis of DNA content in the heads of sperm cells and tail moment (TM) showed that the most useful extender for liquid preservation of chinchilla semen was the extender for stallions.

  12. Effect of ketoprofen treatment on the uterine inflammatory response after AI of jennies with frozen semen.

    PubMed

    Vilés, K; Rabanal, R; Rodríguez-Prado, M; Miró, J

    2013-04-15

    Artificial insemination (AI) involving the placing of frozen-thawed semen directly into the jenny uterine body is associated with very low pregnancy rates. This might be because of an exacerbation of the acute response of the endometrium to sperm, as seen in mares with persistent induced mating endometritis. Pregnancy rates can be increased in such mares, however, by including anti-inflammatory treatments in the insemination protocol (Bucca S, Carli A, Buckley T, Dolci G, Fogarty U. The use of dexamethasone administered to mares at breeding time in the modulation of persistent mating induced endometritis. Theriogenology 2008;70:1093-100; Rojer H, Aurich C. Treatment of persistent mating-induced endometritis in mares with the non-steroid anti-inflammatory drug vedaprofen. Reprod Domest Anim 2010;45:e458-60). To investigate the endometritis caused by the use of frozen-thawed semen in jennies, and to assess the response to ketoprofen treatment, endometrial cytological samples and biopsies from six healthy jennies were examined in a crossover design experiment. Samples were taken from jennies in estrus (E; control) and at 6 hours after AI with or without ketoprofen (+K and -K, respectively). Ketoprofen was administered iv 24 hours before and for 4 days after insemination (total = 2.2 mg/kg/24 hours for 5 days). All animals showed a severe inflammatory response to semen deposition. Polymorphonuclear neutrophil numbers in the cytological smears and biopsies differed significantly between the +K and E animals. No significant differences were recorded, however, between the +K and -K treatments. Eosinophils were observed in all sample types from all groups; these cells appear to be a feature of the normal jenny endometrium. Slight fibrosis was observed in some biopsies, but no significant relationship with inflammation was found. Intense cyclooxygenase-2 (COX-2) immunohistochemical labeling was detected in the -K biopsies. Less intense labeling was seen in those of the +K

  13. Different concentrations of cysteamine and ergothioneine improve microscopic and oxidative parameters in ram semen frozen with a soybean lecithin extender.

    PubMed

    Najafi, Abozar; Kia, Hossein Daghigh; Mohammadi, Hossein; Najafi, Mir Hossein; Zanganeh, Zaynab; Sharafi, Mohsen; Martinez-Pastor, Felipe; Adeldust, Hamideh

    2014-08-01

    The aim of this study was to evaluate the effects of ergothioneine and cysteamine as antioxidant supplements in a soybean lecithin extender for freezing ram semen. Twenty-four ejaculates were collected from four rams and diluted with extenders (1.5% soybean lecithin, 7% glycerol) containing no supplements (control) and cysteamine or ergothioneine (2, 4, 6 or 8mM). Motility by CASA, viability, plasma membrane functionality (HOS test), total abnormality, lipid peroxidation, glutathione peroxidase (GPx) activity and capacitation status (CTC staining) were assessed after thawing. Using 6mM of either antioxidant improved total motility. Cysteamine at 6mM and ergothioneine at 4 and 6mM improved viability and reduced lipid peroxidation (malondialdehyde concentration). Both antioxidants improved membrane functionality significantly, except at 8mM. Progressive motility, kinematic parameters, GPx activity, capacitation status and sperm abnormalities were not influenced by the antioxidant supplements. In conclusion, cysteamine at 6mM and ergothioneine at 4 or 6mM seem to improve the post-thawing quality of ram semen cryopreserved in a soybean lecithin extender.

  14. Effect of different levels and sources of zinc supplementation on quantitative and qualitative semen attributes and serum testosterone level in crossbred cattle (Bos indicus x Bos taurus) bulls.

    PubMed

    Kumar, Nishant; Verma, Ramesh Prashad; Singh, Lallan Prasad; Varshney, Vijay Prakash; Dass, Ram Sharan

    2006-01-01

    An experiment was conducted on 16 crossbred bulls (about 2 years of age, 316.2+/-0.77 kg average body weight), divided into groups I, II, III and IV to study the effect of different levels of Zn supplementation from inorganic and organic sources on semen quality. The animals in the first 3 groups were supplemented with 0, 35 and 70 ppm Zn from Zn sulfate, respectively and the animals in-group IV were supplemented with 35 ppm Zn as Zn propionate. Semen collection and evaluation was done in the first month (to assess semen quality at the start of the experiment) and 7th, 8th and 9th month of experimental feeding to evaluate the effect of supplemental Zn on semen attributes. We gave 6 months for Zn feeding, so that 3 sperm cycles of spermatogenesis had passed and the collected semen reflected the complete effect of Zn supplementation. Six ejaculates from each bull were collected and evaluated for semen quantitative (ejaculate volume, sperm concentration and sperm number per ejaculate) and qualitative characteristics (semen pH, mass motility, individual motility, sperm livability percent and abnormal sperm percent, percent intact acrosome, bovine cervical mucus penetration test, hypo-osmotic sperm swelling test) and activity of seminal plasma enzymes i.e., alkaline phosphatase, acid phosphatase, GOT and GPT. Testosterone level in the blood serum of crossbred bulls was also estimated. Mean values of semen quantitative and qualitative characteristics at the start of the experiment were statistically non significant (P > 0.05) in all the crossbred cattle bulls, however, there were statistically significant differences among the bulls of different groups after 6 months of zinc supplementation. Mean ejaculate volume (mL) was 2.37, 4.70, 5.86 and 6.38, respectively in groups I to IV, indicating a statistically significant (P < 0.05) higher semen volume in Zn-supplemented groups as compared to the control group of bulls. Similarly, sperm concentration (million.mL(-1)), live

  15. 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.

  16. [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.

  17. 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

  18. Effects of presumptive test reagents on the ability to obtain restriction fragment length polymorphism (RFLP) patterns from human blood and semen stains.

    PubMed

    Hochmeister, M N; Budowle, B; Baechtel, F S

    1991-05-01

    Some of the commonly used presumptive test reagents for identification of blood and semen could potentially affect the recovery of intact high-molecular-weight deoxyribonucleic acid (DNA) from evidentiary samples. Thus, the capability of performing restriction fragment length polymorphism (RFLP) analysis on evidentiary samples could be compromised. In order to investigate the potential effects of presumptive test reagents on the DNA present in these samples, bloodstains on cotton and glass were exposed directly to luminol, benzidine, phenolphthalein, o-tolidine, and leucomalachite green, while semen stains and vaginal swabs containing semen were exposed directly to bromochloroindolyl phosphate (BCIP) and sodium thymolphthalein monophosphate (STMP) reagents. The yield gels for DNA quality and quantity and RFLP results indicated that bloodstains exposed to luminol, benzidine dissolved in ethanol, and phenolphthalein, as well as semen stains and vaginal swabs exposed to BCIP and STMP yield RFLP patterns consistent with that of the uncontaminated control. Except for the phenolphthalein treatment, the quantity of extractable, high-molecular-weight DNA obtained was comparable with that of untreated stains. Therefore, evidentiary material purposely or inadvertently contaminated with these reagents can be successfully typed. However, stains exposed to benzidine dissolved in glacial acetic acid, leucomalachite green, and o-tolidine failed to yield high-molecular-weight DNA or to produce any RFLP patterns.

  19. Structurally abnormal human autosomes

    SciTech Connect

    1993-12-31

    Chapter 25, discusses structurally abnormal human autosomes. This discussion includes: structurally abnormal chromosomes, chromosomal polymorphisms, pericentric inversions, paracentric inversions, deletions or partial monosomies, cri du chat (cat cry) syndrome, ring chromosomes, insertions, duplication or pure partial trisomy and mosaicism. 71 refs., 8 figs.

  20. 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.

  1. 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.

  2. The Effect of Chronic Prostatitis/Chronic Pelvic Pain Syndrome (CP/CPPS) on Semen Parameters in Human Males: A Systematic Review and Meta-Analysis

    PubMed Central

    Fu, Weihua; Zhou, Zhansong; Liu, Shijian; Li, Qianwei; Yao, Jiwei; Li, Weibing; Yan, Junan

    2014-01-01

    Background Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is one of the risk factors of impaired male fertility potential. Studies have investigated the effect of CP/CPPS on several semen parameters but have shown inconsistent results. Hence, we performed a systematic literature review and meta-analysis to assess the association between CP/CPPS and basic semen parameters in adult men. Methods Systematic literature searches were conducted with PubMed, EMBASE and the Cochrane Library up to August 2013 for case-control studies that involved the impact of CP/CPSS on semen parameters. Meta-analysis was performed with Review Manager and Stata software. Standard mean differences (SMD) of semen parameters were identified with 95% confidence intervals (95% CI) in a random effects model. Results Twelve studies were identified, including 999 cases of CP/CPPS and 455 controls. Our results illustrated that the sperm concentration and the percentage of progressively motile sperm and morphologically normal sperm from patients with CP/CPPS were significantly lower than controls (SMD (95% CI) −14.12 (−21.69, −6.63), −5.94 (−8.63, −3.25) and −8.26 (−11.83, −4.66), respectively). However, semen volume in the CP/CPPS group was higher than in the control group (SMD (95% CI) 0.50 (0.11, 0.89)). There was no significant effect of CP/CPPS on the total sperm count, sperm total motility, and sperm vitality. Conclusions The present study illustrates that there was a significant negative effect of CP/CPPS on sperm concentration, sperm progressive motility, and normal sperm morphology. Further studies with larger sample sizes are needed to better illuminate the negative impact of CP/CPPS on semen parameters. PMID:24743301

  3. Fertilization rates and in vitro embryo production using sexed or non-sexed semen selected with a silane-coated silica colloid or Percoll.

    PubMed

    Rodríguez Villamil, P; Wei, H; Moreira, G; Caccia, M; Fernandez Taranco, M; Bó, G A

    2012-07-01

    The aim of this study was to evaluate sperm fertilization rates and in vitro embryo development rates for sexed and non-sexed semen selected using a silane-coated silica colloid method (Isolate) or Percoll. Frozen/thawed, sexed and unsexed semen samples from four Holstein bulls were randomly allocated to one of two different density gradient selection methods. Sperm quality (motility, concentration, morphology and membrane integrity) were evaluated and compared before and after sperm selection. Sperm motility and morphology improved (P < 0.005) after the sperm selection process with no differences between the two methods. For non-sexed semen, Percoll gradient increased the mean (± SEM) percentage of sperm recovered (57.3 ± 2.8) compared to Isolate (46.0 ± 1.8; P < 0.01). However, membrane integrity was higher after Isolate than Percoll (sexed semen: 41.0 ± 0.6 vs. 38.8 ± 0.8 and non-sexed semen 60.8 ± 1.6 vs. 58.8 ± 0.5; P < 0.05). The percentage of blastocysts produced was higher when either sexed or non-sexed semen was selected by Isolate (14.0 ± 1.0; 22.0 ± 1.1) than by Percoll (10.5 ± 1.5; 17.0 ± 2.1, respectively; P < 0.05). In summary, Isolate was a more effective method for the recovery of high quality sperm for in vitro fertilization embryo production.

  4. HIV Migration Between Blood and Cerebrospinal Fluid or Semen Over Time

    PubMed Central

    Chaillon, Antoine; Gianella, Sara; Wertheim, Joel O.; Richman, Douglas D.; Mehta, Sanjay R.; Smith, David M.

    2014-01-01

    Previous studies reported associations between neuropathogenesis and human immunodeficiency virus (HIV) compartmentalization in cerebrospinal fluid (CSF) and between sexual transmission and human immunodeficiency virus type 1 (HIV) compartmentalization in semen. It remains unclear, however, how compartmentalization dynamics change over time. To address this, we used statistical methods and Bayesian phylogenetic approaches to reconstruct temporal dynamics of HIV migration between blood and CSF and between blood and the male genital tract. We investigated 11 HIV-infected individuals with paired semen and blood samples and 4 individuals with paired CSF and blood samples. Aligned partial HIV env sequences were analyzed by (1) phylogenetic reconstruction, using a Bayesian Markov-chain Monte Carlo approach; (2) evaluation of viral compartmentalization, using tree-based and distance-based methods; and (3) analysis of migration events, using a discrete Bayesian asymmetric phylogeographic approach of diffusion with Markov jump counts estimation. Finally, we evaluated potential correlates of viral gene flow across anatomical compartments. We observed bidirectional replenishment of viral compartments and asynchronous peaks of viral migration from and to blood over time, suggesting that disruption of viral compartment is transient and directionally selected. These findings imply that viral subpopulations in anatomical sites are an active part of the whole viral population and that compartmental reservoirs could have implications in future eradication studies. PMID:24302756

  5. Morphological abnormalities among lampreys

    USGS Publications Warehouse

    Manion, Patrick J.

    1967-01-01

    The experimental control of the sea lamprey (Petromyzon marinus) in the Great Lakes has required the collection of thousands of lampreys. Representatives of each life stage of the four species of the Lake Superior basin were examined for structural abnormalities. The most common aberration was the presence of additional tails. The accessory tails were always postanal and smaller than the normal tail. The point of origin varied; the extra tails occurred on dorsal, ventral, or lateral surfaces. Some of the extra tails were misshaped and curled, but others were normal in shape and pigment pattern. Other abnormalities in larval sea lampreys were malformed or twisted tails and bodies. The cause of the structural abnormalities is unknown. The presence of extra caudal fins could be genetically controlled, or be due to partial amputation or injury followed by abnormal regeneration. Few if any lampreys with structural abnormalities live to sexual maturity.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. Investigation of polycyclic aromatic hydrocarbon level in blood and semen quality for residents in Pearl River Delta Region in China.

    PubMed

    Song, Xiao Fei; Chen, Zhi Yuan; Zang, Zhi Jun; Zhang, Ya Nan; Zeng, Feng; Peng, Yen Ping; Yang, Chen

    2013-10-01

    This study is the first one investigating the correlation between the concentration of polycyclic aromatic hydrocarbon (PAHs) in blood and semen qualities for residents in the Pearl River Delta (PRD) region in China. Blood samples from 53 infertile volunteers were studied for measures of semen quality and 16 PAHs. Information on the study subjects' living habits (such as smoking, drinking and preference of consumption for food) and general information (age, body-mass-index (BMI) and educational background) were also collected. Statistical results showed that age and BMI were significantly and negatively related to semen motilities. The total concentrations of PAHs (∑16 PAHs) in the blood were 12,010, 7493, 9105 and 8647ng/g for factory workers, office workers, technicians and salespersons, respectively. In addition, ∑16 PAHs in the blood of smokers, drinkers and heavy-taste food consumers were 11,950, 11,266 and 12,141ng/g, which were higher than those observed in nonsmokers (10,457ng/g), nondrinkers (10,920ng/g) and light-taste food consumers (9202ng/g), individually. Furthermore, the Pearson correlation analysis results showed significant positive correlations between BMI and ∑16 PAHs in the blood. Statistically significant correlations were observed between semen motilities and ∑16 PAHs in the blood as well. Logistic regression results showed that for each 1ng/g increase in ∑16 PAHs in blood samples, the log odds of experiencing a pregnancy decrease by 0.039 on average. However, more evidences are needed to clarify the impact of PAHs in the blood to male infertility.

  12. Effects of dilution and centrifugation on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey semen.

    PubMed

    Miró, J; Taberner, E; Rivera, M; Peña, A; Medrano, A; Rigau, T; Peñalba, A

    2009-11-01

    The aim of this work was to study the effects of dilution and centrifugation (i.e., two methods of reducing the influence of the seminal plasma) on the survival of spermatozoa and the structure of motile sperm cell subpopulations in refrigerated Catalonian donkey (Equus asinus) semen. Fifty ejaculates from nine Catalonian jackasses were collected. Gel-free semen was diluted 1:1, 1:5 or 1:10 with Kenney extender. Another sample of semen was diluted 1:5, centrifuged, and then resuspended with Kenney extender until a final dilution of 25x10(6) sperm/ml was achieved (C). After 24 h, 48 h or 72 h of refrigerated storage at 5 degrees C, aliquots of these semen samples were incubated at 37 degrees C for 5 min. The percentage of viable sperm was determined by staining with eosin-nigrosin. The motility characteristics of the spermatozoa were examined using the CASA system (Microptic, Barcelona, Spain). At 24h, more surviving spermatozoa were seen in the more diluted and in the centrifuged semen samples (1:1 48.71%; 1:5 56.58%, 1:10 62.65%; C 72.40%). These differences were maintained at 48 h (1:1 34.31%, 1:5 40.56%, 1:10 48.52%, C 66.30%). After 72 h, only the C samples showed a survival rate of above 25%. The four known donkey motile sperm subpopulations were maintained by refrigeration. However, the percentage of motile sperms in each subpopulation changed with dilution. Only the centrifuged samples, and only at 24h, showed exactly the same motile sperm subpopulation proportions as recorded for fresh sperm. However, the 1:10 dilutions at 24 and 48 h, and the centrifuged semen at 48 h, showed few variations compared to fresh sperm. These results show that the elimination of seminal plasma increases the survival of spermatozoa and the maintenance of motility patterns. The initial sperm concentration had a significant (P<0.05) influence on centrifugation efficacy, but did not influence the number of spermatozoa damaged by centrifugation. In contrast, the percentage of live

  13. An Optimized Centrifugal Method for Separation of Semen from Superabsorbent Polymers for Forensic Analysis.

    PubMed

    Camarena, Lucy R; Glasscock, Bailey K; Daniels, Demi; Ackley, Nicolle; Sciarretta, Marybeth; Seashols-Williams, Sarah J

    2017-03-01

    Connection of a perpetrator to a sexual assault is best performed through the confirmed presence of semen, thereby proving sexual contact. Evidentiary items can include sanitary napkins or diapers containing superabsorbent polymers (SAPs), complicating spermatozoa visualization and DNA analysis. In this report, we evaluated the impact of SAPS on the current forensic DNA workflow, developing an efficient centrifugal protocol for separating spermatozoa from SAP material. The optimized filtration method was compared to common practices of excising the top layer only, resulting in significantly higher sperm yields when a core sample of the substrate was taken. Direct isolation of the SAP-containing materials without filtering resulted in 20% sample failure; additionally, SAP material was observed in the final eluted DNA samples, causing physical interference. Thus, use of the described centrifugal-filtering method is a simple preliminary step that improves spermatozoa visualization and enables more consistent DNA yields, while also avoiding SAP interference.

  14. 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...

  15. 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.

  16. 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.

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. 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...

  6. 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...

  7. 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

  8. 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.

  9. 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...

  10. "Jeopardy" in Abnormal Psychology.

    ERIC Educational Resources Information Center

    Keutzer, Carolin S.

    1993-01-01

    Describes the use of the board game, Jeopardy, in a college level abnormal psychology course. Finds increased student interaction and improved application of information. Reports generally favorable student evaluation of the technique. (CFR)

  11. Results of the 2003-2004 GEP-ISFG collaborative study on mitochondrial DNA: focus on the mtDNA profile of a mixed semen-saliva stain.

    PubMed

    Crespillo, Manuel; Paredes, Miguel R; Prieto, Lourdes; Montesino, Marta; Salas, Antonio; Albarran, Cristina; Alvarez-Iglesias, V; Amorin, Antonio; Berniell-Lee, Gemma; Brehm, Antonio; Carril, Juan C; Corach, Daniel; Cuevas, Nerea; Di Lonardo, Ana M; Doutremepuich, Christian; Espinheira, Rosa M; Espinoza, Marta; Gómez, Felix; González, Alberto; Hernández, Alexis; Hidalgo, M; Jimenez, Magda; Leite, Fabio P N; López, Ana M; López-Soto, Manuel; Lorente, Jose A; Pagano, Shintia; Palacio, Ana M; Pestano, José J; Pinheiro, Maria F; Raimondi, Eduardo; Ramón, M M; Tovar, Florangel; Vidal-Rioja, Lidia; Vide, Maria C; Whittle, Martín R; Yunis, Juan J; Garcia-Hirschfel, Julia

    2006-07-13

    We report here a review of the seventh mitochondrial DNA (mtDNA) exercise undertaken by the Spanish and Portuguese working group (GEP) of the International Society for Forensic Genetics (ISFG) corresponding to the period 2003-2004. Five reference bloodstains from five donors (M1-M5), a mixed stain of saliva and semen (M6), and a hair sample (M7) were submitted to each participating laboratory for nuclear DNA (nDNA; autosomal STR and Y-STR) and mtDNA analysis. Laboratories were asked to investigate the contributors of samples M6 and M7 among the reference donors (M1-M5). A total of 34 laboratories reported total or partial mtDNA sequence data from both, the reference bloodstains (M1-M5) and the hair sample (M7) concluding a match between mtDNA profiles of M5 and M7. Autosomal STR and Y-STR profiling was the preferred strategy to investigate the contributors of the semen/saliva mixture (M6). Nuclear DNA profiles were consistent with a mixture of saliva from the donor (female) of M4 and semen from donor M5, being the semen (XY) profile the dominant component of the mixture. Strikingly, and in contradiction to the nuclear DNA analysis, mtDNA sequencing results yield a more simple result: only the saliva contribution (M4) was detected, either after preferential lysis or after complete DNA digestion. Some labs provided with several explanations for this finding and carried out additional experiments to explain this apparent contradictory result. The results pointed to the existence of different relative amounts of nuclear and mtDNAs in saliva and semen. We conclude that this circumstance could strongly influence the interpretation of the mtDNA evidence in unbalanced mixtures and in consequence lead to false exclusions. During the GEP-ISFG annual conference a validation study was planned to progress in the interpretation of mtDNA from different mixtures.

  12. 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

  13. The Stability of Prostate-Specific Antigen in Semen Under Various Temperatures.

    PubMed

    Srettabunjong, Supawon; Betset, Parimol; Limawongpranee, Suvit; Ekpo, Pattama

    2015-11-01

    Prostate-specific antigen (PSA) is most commonly used for identifying semen, especially in the absence of sperm. However, PSA concentration varies according to storage temperature and duration, and little is known about its stability in semen. This study was therefore aimed to determine the stability under five different temperatures: -80, -20, 4, 25, and 37°C; and nine different durations: 1, 2, 3, 5, 7, 14, 30, 90, and 180 days. All samples were stored at -80°C after being secreted from the volunteers' body until analyzed. Results showed that the PSA concentration declined significantly over time under all temperatures studied except -80°C. At -20 and 4°C, PSA was still detectable on Day 180 with 50% and 70% decrease from its original concentration, respectively. At 25 and 37°C, PSA was detected up to Day 7 and 3, respectively. This information might assist forensic scientists understand more about PSA nature and integrate it into their works.

  14. Concentration of trace elements in human semen and relation to spermatozoa quality.

    PubMed

    Slivkova, Jana; Popelkova, Miroslava; Massanyi, Peter; Toporcerova, Silvia; Stawarz, Robert; Formicki, Grzegorz; Lukac, Norbert; Putała, Aldona; Guzik, Marek

    2009-03-01

    In this study the concentrations of trace elements such as lead, cadmium, iron, nickel, copper and zinc in the human semen (n = 47), occurrence of pathological spermatozoa, and correlations of these elements to pathological forms were investigated. For each sample of human spermatozoa at least 500 spermatozoa were evaluated. Metal contents were determined by the voltametric method and flame absorption spectrophotometry method. The concentrations of trace elements in human semen were: lead 1.49 +/- 0.40 mg x kg(- 1), cadmium 0.13 +/- 0.15 mg x kg(- 1), iron 2.59 +/- 0.21 mg x kg(- 1), nickel 0.40 +/- 0.07 mg x kg(- 1), copper 0.28 +/- 0.06 mg x kg(- 1), and zinc 153.93 +/- 67.08 mg x kg(- 1), respectively. The total percentage of pathological spermatozoa was 41.61 +/- 9.80% with predominancy of broken flagellum, flagellum torso and separated flagellum. In relation to trace elements the analysis showed correlation between copper and lead (r = -0.47), nickel and iron (r = 0.36), lead and flagellum ball (r = -0.39), cadmium and large heads (r = 0.37) and between iron and other forms of pathological spermatozoa (r = -0.32). Results of this study describe possible effects of trace elements on the spermatozoa quality in normal human sperm.

  15. P-Nitrophenol-alpha-D-glucopyranoside as substrate for measurement of maltase activity in human semen.

    PubMed

    Chapdelaine, P; Tremblay, R R; Dubé, J Y

    1978-02-01

    Hitherto, seminal plasma maltase has been measured with maltose as substrate; this method is time consuming and lacks specificity. The use of a synthetic substrate, p-nitrophenol-alpha-D-glucopyranoside, allows accurate and rapid determination of this activity. When maltase is added to the incubation medium (the substrate and reduced glutathione in potassium phosphate buffer, pH 6.8), maintained at 37 degrees C, hydrolysis of the original substrate to p-nitrophenol goes at a constant rate during 4 h. Under optimal conditions of incubation, the Michaelis constant of the reaction, calculated by the Hanes method, was 2.92 +/- 0.84 (SD) X 10(-3) for six different semen samples. Isomaltase appeared to be absent from seminal plasma. The enzyme is stable to freezing and slow thawing and can be stored for at least 26 days at -80 degrees C. Its molecular weight is 259 000. Tris(hydroxymethyl)aminomethane (pH 6.8) exerts a noncompetitive inhibition on the enzyme activity. In 68 men 23 to 45 years old, whose semen analyses were normal, the seminal plasma maltase activity was 467 +/- 135 (SD) mU/g of protein. It was generally decreased in patients with infertility disorders.

  16. [Determination of sinigrin in semen Thlaspi from Sichuan and Tibet using near infrared diffuse reflectance spectroscopy].

    PubMed

    Wang, Lei-Lei; Chen, Cong; Zhou, Min; Wang, Jian-Zhong; Luo, Xia; Huang, Guo; Ye, Li-Ming

    2009-10-01

    The objective of the present study was to develop a method for the determination of sinigrin in semen Thlaspi from Sichuan using near infrared diffuse reflectance spectroscopy. Near infrared spectra (NIR) in the region of 7,502.1-5,446.2 cm(-1) were recorded for the 246 semen Thlaspi samples containing sinigrin in the content of 1.962%-3.917%. Calibration models were established using the PLS (partial least squares). Different spectra pretreatment methods were compared. The study showed that spectral information can be extracted thoroughly by minimum and maximum normalization pretreatment methods. In this calibration model, the correlation coefficient (R2) was 0.9280, the SEC (standard deviation of the calibration sets) was 0.314 and the SEP (standard deviation of the prediction sets) was 0.388. Results indicated that near infrared diffuse reflectance spectroscopy method can be used to rapidly analyze the valid component in traditional Chinese medicine, and also can be used in the quality control of traditional Chinese medicine.

  17. Concentration of copper, iron, zinc, cadmium, lead, and nickel in boar semen and relation to the spermatozoa quality.

    PubMed

    Massányi, Peter; Trandzík, Jozef; Nad, Pavol; Koréneková, Beáta; Skalická, Magdaléna; Toman, Robert; Lukác, Norbert; Strapák, Peter; Halo, Marko; Turcan, Ján

    2003-01-01

    The concentration of copper, iron, zinc, cadmium, lead, and nickel as well as its relation to spermatozoa quality was investigated. The semen samples were analyzed by atomic absorption spectrophotometry (AAS). The concentration of copper in boar semen was 1.64 +/- 0.28 mg kg(-1) and of iron 16.14 +/- 10.35 mg kg(-1). The concentration of zinc in boar semen reached an average value of 171.74 +/- 64.72 mg kg(-1) and the level of cadmium reached 0.01-0.16 mg kg(-1) with the average value of 0.05 mg kg(-1). The analysis of lead showed that the concentration of this element in boar semen was 0.02 +/- 0.03 mg kg(-1) and the average level of nickel was 0.06 +/- 0.08 mg kg(-1). The total percentage of pathological spermatozoa was 9.82 +/- 1.47%. Detail analysis determined 3.18% of separated flagellum, 2.26% knob twisted flagellum, 0.88% flagellum torso, 0.85% flagellum ball, 0.42% broken flagellum, 0.23% retention of the cytoplasmic drop, 0.14% small heads, 0.03% large heads, and 1.83% forms other of pathological changes. Correlation analysis showed significant (p < 0.05) positive correlation between copper and lead (r = 0.52). High correlation between small head and knob twisted tail (r = 0.67), small head and broken flagellum (r = 0.88) as well as between small head and total number of pathological spermatozoa (r = 0.73) was determined.

  18. PLCz functional haplotypes modulating promoter transcriptional activity are associated with semen quality traits in Chinese Holstein bulls.

    PubMed

    Pan, Qing; Ju, Zhihua; Huang, Jinming; Zhang, Yan; Qi, Chao; Gao, Qin; Zhou, Lei; Li, Qiuling; Wang, Lingling; Zhong, Jifeng; Liu, Mei; Wang, Changfa

    2013-01-01

    The sperm-specific phospholipase C zeta (PLCz) is a candidate sperm-borne oocyte-activating factor that triggers a characteristic series of physiological stimuli via cytoplasmic Ca(2+) oscillations during fertilization. The molecular mechanisms involved in the regulation of PLCz gene expression remain largely unknown. To explore the genetic variations in the 5'-flanking region of the PLCz gene and their common haplotypes in Chinese Holstein bulls, as well as to determine whether these variations affect bovine semen quality traits and transcriptional activity, DNA samples were collected from Chinese Holstein bulls and sequenced for the identification of genetic variants in the 5'-flanking region of PLCz. Two genetic variants were identified, and their haplotypic profiles were constructed. The two novel genetic variations (g. -456 G>A and g. +65 T>C) were genotyped in 424 normal Chinese Holstein bulls. Bioinformatics analysis revealed that both loci are in transcription factor binding sites of the core promoter region. The association studies revealed that the two genetic variations and their haplotype combinations significantly affected semen quality traits. Using serially truncated constructs of the bovine PLCz promoters and the luciferase reporter, we found that a 726 bp (-641 nt to +112 nt) fragment constitutes the core promoter region. Furthermore, four haplotypes, H1H1 (GTGT), H2H2 (GCGC), H3H3 (ATAT), and H4H4 (ACAC), were significantly associated with semen quality traits and successfully transfected into MLTC-1 cell lines. The luciferase reporter assay showed that the different haplotypes exhibited distinct promoter activities. Maximal promoter activity was demonstrated by the H2H2 haplotypes, as compared with the other haplotypes. To the best of our knowledge, this study is the first report on genetic variants and their respective haplotypes in the 5'-flanking region of PLCz gene that can influence the semen quality of Chinese Holstein bulls as well as

  19. Effect of argan oil on liquid storage of ram semen in Tris or skim milk based extenders.

    PubMed

    Allai, Larbi; Druart, Xavier; Contell, Jesus; Louanjli, Noureddine; Moula, Anass Ben; Badi, Abdelmoughit; Essamadi, Abdelkhalid; Nasser, Boubker; El Amiri, Bouchra

    2015-09-01

    Due to its high antioxidant content, the argan oil could play a beneficial role in liquid storage of ram semen. The aim of this study was to investigate effects of different concentration of argan oil (ARO) on spermatologic parameters, lipid peroxidation and DNA fragmentation during liquid storage of ram semen until 48 h. Also effects of extenders and temperature on same parameters were assessed. For these aims, semen samples were collected from Boujaâd rams, extended with Tris egg yolk or skim milk extenders without (control) or supplemented with different concentrations of ARO (1%, 2%, 5% and 10% v/v) at a final concentration of 0.8 × 10(9) sperm/mL and stored until 48 h at 5 °C or 15 °C. The sperm quality assessments were performed at different intervals during storage (0, 8, 24 and 48 h). Sperm progressive motility started to decrease after 8h of storage in all temperatures--extenders combinations and dropped steadily during the 8-48 h interval. However, sperm viability, progressive motility and membrane integrity were markedly higher in ARO groups (especially in 1% in Tris and 5% in skim milk) until 24h and 48 h storage at both temperatures compared to controls. The argan oil also decreased the level of spontaneous and induced malondialdehyde (MDA) and the sperm DNA fragmentation until 48 h storage. In conclusion, it was determined that addition of argan oil to conventional extenders may improve the quality of ram semen during liquid storage in different temperatures.

  20. Donkey jack (Equus asinus) semen cryopreservation: studies of seminal parameters, post breeding inflammatory response, and fertility in donkey jennies.

    PubMed

    Rota, A; Panzani, D; Sabatini, C; Camillo, F

    2012-11-01

    The aims of this study were (1) to evaluate motility parameters of donkey jack (jack; Equus asinus) semen cryopreserved in INRA-96 (INRA; IMV Technologies, France, 2% egg-yolk enriched) using either glycerol (GLY) or ethylene glycol (EG) as a cryoprotector; (2) to compare in vitro the postthaw re-extension with homologous seminal plasma (SPL) or INRA; (3) to compare fertility in donkey jennies (jennies; Equus asinus) timed artificially inseminated with jack semen cryopreserved using GLY or EG, re-extended with INRA; (4) to compare fertility in jennies timed artificially inseminated with jack semen cryopreserved using GLY re-extended with SPL, INRA, or not re-extended (NN); and (5) to describe some preliminary results of the inflammatory uterine response postbreeding. Semen from two jacks was collected and frozen in an INRA-2% egg yolk extender added of either 2.2% GLY or 1.4% EG. Postthaw motility was evaluated by a computer-assisted motility analyzer. Uterine inflammatory response and fertility were evaluated after artificial insemination (AI) of 13 jennies with frozen-thawed semen, either further extended with INRA (Group GLY-INRA, 13 cycles, and EG-INRA, 8 cycles), or with SPL (Group GLY-SPL, 13 cycles), or not re-extended (GLY-NN, 5 cycles). In each cycle, jennies were bred twice with 500 × 10(6) sperm cells (250 × 10(6) from each jack), at fixed times after induction of ovulation, and uterus was flushed at 6 and 10 h after first and second breeding, respectively. Cells in the recovered fluid were counted and distinguished as polymorphonuclear neutrophils (PMN) or other cell types. Total and progressive motility did not differ between cryoprotectants, but were higher when semen samples were re-extended in INRA, compared with SPL (P < 0.05). Pregnancy was diagnosed by transrectal palpation and ultrasonography examinations at 14 and 16 days postovulation. In 7/13 (53.8%) jennies and 12/39 (30.4%) cycles postbreeding intrauterine fluid accumulation was observed

  1. 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.

  2. 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.

  3. 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

  4. Comparison of Oxidative Stress/DNA Damage in Semen and Blood of Fertile and Infertile Men

    PubMed Central

    Guz, Jolanta; Gackowski, Daniel; Foksinski, Marek; Rozalski, Rafal; Zarakowska, Ewelina; Siomek, Agnieszka; Szpila, Anna; Kotzbach, Marcin; Kotzbach, Roman; Olinski, Ryszard

    2013-01-01

    Abnormal spermatozoa frequently display typical features of oxidative stress, i.e. excessive level of reactive oxygen species (ROS) and depleted antioxidant capacity. Moreover, it has been found that a high level of oxidatively damaged DNA is associated with abnormal spermatozoa and male infertility. Therefore, the aim of our study was the comparison of oxidative stress/DNA damage in semen and blood of fertile and infertile men. The broad range of parameters which describe oxidative stress and oxidatively damaged DNA and repair were analyzed in the blood plasma and seminal plasma of groups of fertile and infertile subjects. These parameters include: (i) 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) and 8-oxo-7,8-dihydroguanine (8-oxoGua) levels in urine; (ii) 8-oxodG level in DNA isolated from leukocytes and spermatozoa; (iii) antioxidant vitamins (A, C and E) and uric acid. Urinary excretion of 8-oxodG and 8-oxoGua and the level of oxidatively damaged DNA in leukocytes as well as the level of antioxidant vitamins were analyzed using HPLC and HPLC/GC/MS methods. The results of our study demonstrate that 8-oxodG level significantly correlated with every parameter which describe sperm quality: sperm count, motility and morphology. Moreover, the data indicate a higher level of 8-oxodG in sperm DNA compared with DNA of surrogate tissue (leukocytes) in infertile men as well as in healthy control group. For the whole study population the median values of 8-oxodG/106 dG were respectively 7.85 and 5.87 (p = 0.000000002). Since 8-oxodG level in sperm DNA is inversely correlated with urinary excretion rate of 8-oxoGua, which is the product of OGG1 activity, we hypothesize that integrity of spermatozoa DNA may be highly dependent on OGG1 activity. No relationship between the whole body oxidative stress and that of sperm plasma was found, which suggests that the redox status of semen may be rather independent on this characteristic for other tissues. PMID:23874641

  5. 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

  6. Minimizing microbial contamination of sperm samples

    USGS Publications Warehouse

    Jenkins, Jill A.; Tiersch, Terrence R.; Green, Christopher C.

    2011-01-01

    Taken from the Methods section: With the collection and translocation of gametes from aquatic species, a potential hazard exists for microbial transfer. Contamination of semen can occur during collection, processing, storage, and transport. Some preventative measures are described below for limiting the spread and amplification of microorganisms such as bacteria, viruses, fungi, mycoplasmas, and parasites. Generally, sanitation during collection is essential. Materials and equipment used to freeze semen should be sterile. Following good practice guidelines for handling and processing samples collected for freezing is especially important for non-domestic animals where disease-free status cannot be guaranteed and unsophisticated technology is used (Russell et al. 1977).

  7. Semen characteristics and biochemical composition of cloacal foam of male Japanese quails (Coturnix coturnix Japonica) fed diet incorporated with selenium.

    PubMed

    Biswas, A; Mohan, J; Mandal, A B; Lal, N

    2017-04-01

    An attempt was made to investigate the effect of dietary selenium (Se) on physical and cloacal gland size, foam production, biochemical composition of foam and semen biochemical characteristics of male Japanese quail (Coturnix coturnix Japonica). Two hundred twenty-five (225)-day-old male Japanese quail were randomly distributed to three dietary treatment groups for a period of 20 weeks. Each treatment comprised of three replicates, each containing 25 chicks. Three experimental diets were supplemented with 0, 0.5 and 1.0 mg Se/kg (T1 , T2 and T3, respectively), and diet T1 was considered as control. Sodium selenite was used as the source of selenium. All the birds were provided with feed and water ad libitum. Cloacal foam characteristics, that is cloacal gland index and foam weight, were significantly higher in T2 group. However, body weight, frequency of foam discharge and testes weight (left and right) did not differ significantly (p > 0.05). Physical characteristics of semen, that is semen volume and sperm concentration, did not differ (p > 0.05) among the Se-treated groups. The sperm motility, live-dead count and abnormality improved significantly (p < 0.05) in 0.5 mg/Se-supplemented group compared to 0 or 1.0 mg/Se-supplemented groups. Similarly, fertility and hatchability percentages were higher (p < 0.05) in 0.5 mg/Se-supplemented group than in control or 1.0 mg/Se-supplemented counterparts. The biochemical characteristics of foam in terms of total protein, acid phosphatase (ACP) and nitric oxide did not differ (p > 0.05), while the concentration of glucose was higher (p < 0.05) in 0.5 mg/Se-supplemented diet. On the other hand, alkaline phosphatase (ALP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were lower (p < 0.05) in 0.5 mg/Se-supplemented group compared to control or 1.0 mg/Se-supplemented groups. From this study, it was concluded that supplementation of 0.5 mg Se/kg diet was beneficial for foam

  8. 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.

  9. 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.

  10. Cholesterol concentration in seminal plasma as a predictive tool for quality semen evaluation.

    PubMed

    Beer-Ljubić, B; Aladrović, J; Marenjak, T S; Laskaj, R; Majić-Balić, I; Milinković-Tur, S

    2009-11-01

    The aim of this study was to investigate the relationship between lipid composition of bovine serum and seminal plasma, seasonality, and semen quality. The experiment was carried out in two groups of Simmental breeding bulls: Group I (ages 2 to 4 yr) and Group II (ages 5 to 10 yr). Blood samples were collected from jugular vein, and bovine semen was sampled with an artificial vagina once per season. Serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triacylglycerols, nonesterified fatty acids (NEFAs), and lipoprotein electrophoretic patterns were determined. Seminal plasma concentrations of total cholesterol, HDL-C, and LDL-C were assayed. Serum concentration of triacylglycerols in young bulls was significantly higher in winter compared with that in autumn, whereas serum NEFA concentration was significantly higher in autumn compared with that in other seasons. Serum concentration of total cholesterol, LDL-C, and LDL lipoproteins in older bulls was significantly higher in winter than in spring. Seminal plasma concentration of total cholesterol in young bulls was significantly higher in spring compared with that in summer, whereas in older bulls it was significantly higher in winter compared with that in autumn samples. Sperm volume of both groups was significantly higher in summer compared with that in autumn and winter. Sperm motility in young bulls was lowest in summer and differed significantly from the values recorded in other seasons. The changes observed in seminal plasma cholesterol concentration were associated with extracellular lipid use and appeared to be applicable as a biochemical marker of sperm quality.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. A retrospective clinical study of endoscopic-assisted transcervical insemination in the bitch with frozen-thawed dog semen.

    PubMed

    Mason, S J

    2016-11-24

    Since the conclusion of data collation from previously published work, a further 352 inseminations using frozen-thawed dog semen by endoscopic-assisted transcervical insemination (EIU) have been performed by the author. Insemination was performed on the second day in which crenulation of the anterior vagina was detected in conjunction with a progesterone concentration of >10 ng/ml. All semen samples were analysed for total number of sperm, total motility and progressive motility using computer-assisted semen analysis (CASA). The insemination dose was based on the progressively motile normal spermatozoa (PMNS). Insemination was performed on all bitches as previously described using a ureterorenoscope. Additional extender was inseminated subsequent to the semen to expand and fill the uterus. The semen and additional extender were inseminated slowly over a period of 15-20 min. Pregnancy was determined by B-mode ultrasound equipped with a 7.5-MHz probe whilst standing and/or via the whelping rate. The number of sperm inseminated ranged from 9 × 10(6) PMNS to 519 × 10(6) PMNS, with progressive motility values ranging between 20% and 80%. The overall pregnancy rate was 68% (238/352). When stratified by PMNS, pregnancy rates were as follows: >150 × 10(6) PMNS - 76% (110/145), 100-150 × 10(6) - 68% (87/128) and <100 × 10(6) PMNS - 52% (41/79). Pregnancy rate was significantly higher when >150 × 10(6) PMNS (p = .003) or 100-150 ×10(6) PMNS (p = .027) were inseminated compared to <100 × 10(6) PMNS. These data are concordant with previous reports recommending the insemination of >150 × 10(6) PMNS to maximize pregnancy rate. These results indicate that one optimally timed EIU insemination results in similar pregnancy rates to previous publications of one optimally timed, or two or more non-optimally timed inseminations using the Norwegian catheter.

  16. [The relativity of abnormity].

    PubMed

    Nilson, Annika

    2006-01-01

    In the late 19th century and in the beginning of the 20th century, mental diseases and abnormal behavior was considered to be a great danger to culture and society. "Degeneration" was the buzzword of the time, used and misused by artists and scientists alike. At the same time, some scientists saw abnormity as the key to unlock the mysteries of the ordinary mind. Naturalistic curiosity left Pandoras box open when religion declined in Darwins wake. Two swedish scientists, the physician Bror Gadelius (1862-1938) and his friend the philosopher Axel Herrlin (1870-1937), inspired by the French psychologist Theodule Ribots (1839-1916) "psychology without a soul", denied all fixed demarcation lines between abnormity and normality. All humans are natures creatures ruled by physiological laws, not ruled by God or convention. Even ordinary morality was considered to be an utterly backward explanation and guideline for complex human behavior. Different forms of therapy, not various kinds of penalties for wicked and disturbing behavior, are the now the solution for lots of people, "normal" as well as "abnormal". Psychiatry is expanding.

  17. Abnormalities of gonadal differentiation.

    PubMed

    Berkovitz, G D; Seeherunvong, T

    1998-04-01

    Gonadal differentiation involves a complex interplay of developmental pathways. The sex determining region Y (SRY) gene plays a key role in testis determination, but its interaction with other genes is less well understood. Abnormalities of gonadal differentiation result in a range of clinical problems. 46,XY complete gonadal dysgenesis is de