Early and late effects of Ibuprofen on mouse sperm parameters, chromatin condensation, and DNA integrity in mice

Background: There are few studies indicating the detrimental effects of ibuprofen on sperm fertility potential and DNA integrity. Objective: To determine the effects of Ibuprofen on sperm parameters, chromatin condensation and DNA integrity of mice. Materials and Methods: In this experimental study, 36 adult male mice with average weight 37 gr were divided into three groups, including control (group I, n=12), normal dosage of ibuprofen (group II, n=12) and high dosage (group III, n=12). Ibuprofen with different doses was dissolved in daily water of animals. After 35, 70 and 105 days, the cauda epididymis of mice were cut and incubated in Ham’s F10 media. Sperm samples were analyzed for parameters (motility, morphology and count), DNA integrity (SCD test) and chromatin condensation (chromomycin A3 and Aniline blue staining). Results: After 35 days, in addition to above mentioned sperm parameters, all of the treated mice showed statistically significant increase in spermatozoa with immature chromatin (P<0.05). However, after 70 days, the rate of sperm DNA fragmentation assessed by SCD was increased in group II (66.5±0.7) and the percentage of immature spermatozoa (AB+ and CMA3+) was higher in group III (77.5±0.7 and 49.5±6.3 respectively) than other groups. After 105 days, the AB+ spermatozoa were increased in both normal dose and high dose groups. Conclusion: Ibuprofen may cause a significant reduction in sperm parameters and sperm chromatin/DNA integrity in mice. It should be noted that these deleterious effects are dose-dependent and can be seen in early and late stage of drug treatments.

Evaluation of sperm retrieval rate with bilateral testicular sperm extraction in infertile patients with azoospermia

Background: About 10% to 15% of infertile men have azoospermia, which could be obstructive or non-obstructive. Diagnostic biopsy from the testis and recently testicular sperm extraction (TESE) are the most precise investigations in these patients. Testicular biopsy can be done unilaterally or bilaterally. The worth of unilateral or bilateral testicular biopsy in men with azoospermia is controversial. Objective: To evaluate the necessity of bilateral diagnostic biopsy from the testis in new era of diagnosis and treatment of male infertility. Materials and Methods: In this retrospective study, we reviewed the results of testis biopsy in 419 azoospermic men, referred to Yazd Research and Clinical Center for Infertility from 2009-2013. Patients with known obstructive azoospermia were excluded from the study. Results: In totally, 254 infertile men (60.6%) were underwent unilateral TESE, which in 175 patients (88.4%) sperm were extracted from their testes successfully. Bilateral testis biopsy was done in 165 patients (39.4%) which in 37 patients (22.4%), sperm were found in their testes tissues. Conclusion: Due to the low probability of positive bilateral TESE results especially when we can’t found sperm in the first side, we recommend that physicians re-evaluate the risk and benefit of this procedure in era of newer and more precise technique of sperm retrieval like micro TESE.

Serum levels of lycopene, beta-carotene, and retinol and their correlation with sperm DNA damage in normospermic and infertile men

Background: Oxidative stress in reproductive system leads to sperm DNA damage and sperm membrane lipid peroxidation and may play an important role in the pathogenesis of male infertility, especially in idiopathic cases. Antioxidants such as carotenoids function against free radical damages. Objective: The aim of this study was to determine the levels of lycopene, beta-carotene and retinol in serum and their relationship with sperm DNA damage and lipid peroxidation in infertile and normospermic males. Materials and Methods: Sixty two infertile men and 71 normospermic men participated in this study. Blood and semen samples were collected from all subjects. Sperm DNA damage was measured using TUNEL method. Carotenoids, retinol, and malonedildehyde in serum were also determined. Results: DNA fragmentation was higher in infertile group comparing to control group. Serum levels of lycopene, beta-carotene and, vitamin A in infertile men were significantly lower than normospermic men (p< 0.001, =0.005, and =0.003 respectively). While serum MDA was not significantly different between two groups, MDA in seminal plasma of infertile men was significantly higher than control group (p< 0.001). Conclusion: We concluded that lycopene, beta-carotene, and retinol can reduce sperm DNA fragmentation and lipid peroxidation through their antioxidant effect. Therefore the DNA fragmentation assay and determination of antioxidants factors such as lycopene, beta-carotene and retinol, along with sperm analysis can be useful in diagnosis and treatment of men with idiopathic infertility.

Artificial fertilization of oocytes and sperm activation in pacu: effects of the spermatozoa:oocyte ratio, water volume, and in natura semen preservation

Objetivou-se foi avaliar a fertilização artificial e a duração da motilidade espermática em pacus com diferentes doses inseminantes, volumes de água e preservação do sêmen in natura. Foram realizados quatro experimentos para avaliação do efeito de doses inseminantes (7x10³, 7x10(4), 7x10(5), 7x10(6) e 7x10(7) espermatozoides ovócito-1) sobre a fertilização artificial dos ovócitos; do efeito do volume de água (0,5; 15,0; 30,0; 45,0 e 60,0 mL de água mL-1 de ovócitos) com doses inseminantes de 105.481 e 210.963 espermatozoides ovócito-1; do efeito de diluição do sêmen (0,005; 0,05; 0,5 e 5,0 µL de sêmen mL-1 de água) sobre a duração da motilidade espermática; e do efeito do armazenamento a 15 ºC por 9 h sobre a duração da motilidade espermática e o índice de sobrevivência espermática. Os maiores resultados obtidos foram: doses inseminantes entre 7x10³ e 7x10(7) espermatozoides ovócito-1; 15 a 60 mL de água mL-1 de ovócitos; diluição de 0.005 µL sêmen mL-1 de água e 98,65% de sobrevivência espermática até o tempo de preservação de 2h45min36s. A preservação a 15ºC por 9 horas não influencia a duração da motilidade espermática. As maiores taxas de fertilização podem ser observadas no emprego de 0,27 a 270 µL de sêmen mL-1 de ovócitos, com 15 a 60 mL de água para ativação.

Effects of the spermatozoa: oocyte ratio, water volume and water temperature on artificial fertilization and sperm activation of cascudo-preto

The objective of this study was to evaluate the effects of water volume and water temperature on the sperm motility duration and the number of spermatozoa, and the water volume on the fertilization rates of oocytes of Rhinelepis aspera. Experiments were carried out to evaluate the effect of semen dilutions (1.74×10-5, 1.74×10-4, 1.74×10-3, 1.74×10-2, 1.74×10-1 and 1.00 mL of sperm.mL-1 of water) and water temperature (5, 10, 15, 20, 25, 30, 35, 40, 45, and 50 ºC) on spermatozoa motility duration. In addition, the effects of insemination dose (7×10³, 7×10(4), 7×10(5), 7×10(6) and 7×10(7) spermatozoa.oocyte-1) and water volume (1.0, 30.0, 60.0, 90.0 and 120.0 mL water.2.0 mL-1 oocytes) on the artificial fertilization rates of oocytes were evaluated. The longest sperm motility duration were observed for the semen dilution of 1.74×10-5 mL semen.mL-1 water and in water at 5 ºC. The highest fertilization rates were obtained for insemination doses between 7.00×10³ and 1.23×10(7) spermatozoa. oocyte-1 and water volume of 28.11 mL water.2.0 mL-1 oocytes.

Measuring sperm backflow following female orgasm: a new method

Background: Human female orgasm is a vexed question in the field while there is credible evidence of cryptic female choice that has many hallmarks of orgasm in other species. Our initial goal was to produce a proof of concept for allowing females to study an aspect of infertility in a home setting, specifically by aligning the study of human infertility and increased fertility with the study of other mammalian fertility. In the latter case - the realm of oxytocin-mediated sperm retention mechanisms seems to be at work in terms of ultimate function (differential sperm retention) while the proximate function (rapid transport or cervical tenting) remains unresolved. Method: A repeated measures design using an easily taught technique in a natural setting was used. Participants were a small (n=6), non-representative sample of females. The introduction of a sperm-simulant combined with an orgasm-producing technique using a vibrator/home massager and other easily supplied materials. Results: The sperm flowback (simulated) was measured using a technique that can be used in a home setting. There was a significant difference in simulant retention between the orgasm (M=4.08, SD=0.17) and non-orgasm (M=3.30, SD=0.22) conditions; t (5)=7.02, p=0.001. Cohen’s d=3.97, effect size r=0.89. This indicates a medium to small effect size. Conclusions: This method could allow females to test an aspect of sexual response that has been linked to lowered fertility in a home setting with minimal training. It needs to be replicated with a larger sample size.

Influence of mini-Percoll techniques in sperm capacitation and plasma membrane integrity of ram frozen-thawed sperm.

2016

Effect of ß-Mercaptoetanol and cysteine on post-thawing quality and oxidative activity of ram sperm and on the viability of vitrified sheep embryos.

2016

Effect of different melatonin concentration in integrity of acrosomal membrane in swine sperm cryopreserved.

2016

Viabilility of Rams Epididymal Sperm after Preservation in Low Temperature (5oc)

The purpose of this research was to evaluate the viability of ram epididymal sperm collected from fresh caudal epididymis (H-0) or after storage in low temperature (5oC, in refrigerator) for one (H-1), two (H-2), and three (H-3) days.  Collected sperm were diluted in modified Tris extender and they were preserved in refrigerator up to four days.  The viability of diluted sperm was evaluated daily base on motility and sperm live.  Results indicated that mean sperm concentration after sperm diluted with 0.05 ml Tris extender of caudal epididymis was 2745 million/ml. Sperm motility and percentage of live for H-0 (71.25% and 82.83%) and H-1 (70.00% and 79.17%) were significantly higher (P<0.05) than H-2 (61.25% and 69.83%) and H-3 (51.67% and 66.17%).  Percentages of sperm motility and live of diluted sperm and preserved in refrigerator for H-0 were significantly higher (P<0.05) than H-1, H-2, and H-3.  These results showed that epididymal sperm collected from caudal epididymis up to three days of preservation (without further storage of the diluted sperm) could be used for artificial insemination or in vitro fertilization programs.  Diluted sperm of H-0 and H-1 could be preserved in refrigerator for two days and H-2 for one day. (Animal Production 6(1): 30-36 (2004) Key Words: Epididymal Sperm, Viability, Rams

The effect of Physiological NaCl Dilution levels and Storage Duration to Sperm Motility and Abnormality in Muscovy Duck

The aim of this research are to know the effects of physiological NaCl dilution levels and storage duration on sperm motility and abnormality of  Muscovy duck. Materials which used in this research were semen that collected of 1  - 1,5 years of four Muscovy duck. Semen were treated by physiological NaCl dilution levels nol , three, six and nine and storage duration ( t0 = 0 minute t1 =45minutes and t2=90 minute). Replication was three tapping periode . The method of this research was laboratory experimental with Randomized Complete Blok Design (RBD) in Split Plot Design (SPD)  pattern ,as main plot is Physiological NaCl dilation levels and as sub plot is storage duration. Result of the experiment showed that physiological NaCl dilution levels was very significantly influences on sperm motility (p<0,01) and significantly influences and sperm abnormality (p<0,05) . The storage duration was very significantly influences and sperm motility and abnormality (p<0,01) .Orthogonal duration polynomial assayed showed that the effect of physiological NaCl dilution levels to sperm motility  had a linier regression line with the a equation as Y =69.94 - 2.65 X,  r=0,49 R2=0.24 with Y presented to sperm motility and X represented to physiological NaCl dilution levels  sperm and effect of storage duration to sperm motility had a linier regression line with the equation as Y =74.24-0.35 X, r =0.73 R2=0.53 with Y represented to sperm motility and X represented to storage duration . The effect of physiological NaCl dilution levels to sperm abnormality had a linier regression line with the equation as Y =5.61+0.34X, r=0.52 R2=0.27 with Y represented to sperm to abnormality and X represented to regression line with the equation as Y =6.74+0.01X, r=0.24R2=0.060 with Y represented to sperm abnormality and X represented to storage duration. It is can be concluded that a higher physiological NaCl dilation levels and storage duration caused sperm to motility decreased and sperm abnormality increased. (Animal Production 3(2): 45-52 (2001)Key Word: sperm Muscovy duck.

Quality Of Pe Buck Sperm Stored In Different Packaging

AI for cattle has been develop in Indonesia in contrast, AI for small ruminants (sheep and goat) are less developed. Its due to the lack of facilities, processing and packaging during storage. This research aims to compare the effectivity and efficiency of two different packaging technique (test tube and straw). Test tube and 0.25 ml mini straw were tested . Semen was obtained from adult PE buck (3 yr) after collection by using  artificial vagina. The fresh semen was then evaluated and diluted 5 folds with 2.9 percent Na-citrate. Diluted semen then packed in test tube and mini straw, and stored in refrigerator (100C) for 7 days. Observations were done everyday on sperm motility, abnormality and percentage of  live sperm. Observation were made at 370C. Observation on fresh ejaculate showed that semen has 6x 109/ml concentration, 90 percent motility, 8 percent abnormality and  95 percent  live sperm. Five folds dilution reduced sperm concentration to 1.2 x 109/ml, but did not change sperm motility, abnormality and percent of live sperm. Sperm was then packed according  to the treatments. Storage in both packaging did significantly reduce  (P<0.01) sperm motility and percent live sperm, but not for sperm abnormality. All sperm stored in test tube were classified death at day 5 storage. However, sperm in the straw were remain live eventhough the percentages was low. It can be concluded that straw has a better result than test tube. Eventhough sperm motility was extremely low, it remains valuable for cervical insemination. (Animal Production 1(1) : 24-29 (1999).  Key Words : Sperm, PE Buck, Straw, Test Tube

The Effect of Level Testosteron e Addition in Diluents and Level Dilution on Speed Movement and Abnormality of Kedu Chicken Sperm

The research entitled “The Effect of Level Testosterone Addition in Diluents and Level Dilution on Speed Movement and Abnormality of Kedu Chicken Sperm” was conducted in Laboratory of Physiology and Reproduction, Faculty of Animal Science, UNSOED, started at August 15th to September 15th , 2001. The aims of  this research were to obtain influences of level testosterone and dilution on speed movement and secondary abnormality kedu chicken sperm and obtain interaction between the treatments. The tapped sperm from nine kedu chickens were used in this research. This Experiment was performed 4 x 3 Factorials with Randomized  Completely Block Design (RCBD) as the basic design. The treatment combinations were level testosterone 0, 300, 600 and 900 μg (t0, t1, t2 and t3) and level of dilution 4, 6 and 8 time (in a row p1, p2 and p3). The tapping period was replicated four times as a group (replicated) with two days interval. The result of this research showed that the interaction between level of testosterone addition and level of dilution gave a non-significant effect to speed movement sperm but significant to abnormality of kedu chicken sperm. The group gave a significant influence (P<0.05) to speed movement sperm and non significant to sperm abnormality. The interaction of level testosterone addition and level dilution of kedu chicken semen (T x P) has a quadratic regression to sperm abnormality with regression comparison is Y = 24.418 – 0.014 X +1.187E – 05 X², with peak point is (543.76: 20.23) of correlation coefficient (r) 0.55 and determination coefficient (R²) as 30.34%. The addition of 600 μg testosterone level with 6 level dilutions was the best to defend sperm abnormality. (Animal Production 4(2): 60-70 (2002) Key Words : Spermatozoa, Kedu Chicken, Testosterone, Dilution, Speed Movement and Abnormality

Listening to late discovery adoption and donor offspring stories : adoption, ethics and implications for contemporary donor insemination practices

For most of the 20th Century a ‘closed’ system of adoption was practised throughout Australia and other modern Western societies. This ‘closed’ system was characterised by sealed records; amended birth certificates to conceal the adoption, and prohibited contact with all biological family. Despite claims that these measures protected these children from the taint of illegitimacy the central motivations were far more complex, involving a desire to protect couples from the stigma of infertility and to provide a socially acceptable family structure (Triseliotis, Feast, & Kyle, 2005; Marshall & McDonald, 2001). From the 1960s significant evidence began to emerge that many adopted children and adults were experiencing higher incidences of psychological difficulties, characterised by problems with psychological adjustment, building self-esteem and forming a secure personal identity. These difficulties became grouped under the term ‘genealogical bewilderment’. As a result, new policies and practices were introduced to try to place the best interests of the child at the forefront. These changes reflected new understandings of adoption; as not only an individual process but also as a social and relational process that continues throughout life. Secrecy and the withholding of birth information are now prohibited in the overwhelming majority of all domestic adoptions processed in Australia (Marshall & McDonald, 2001). One little known consequence of this ‘closed’ system of adoption was the significant number of children who were never told of their adoptive status. As a consequence, some have discovered or had this information disclosed to them, as adults. The first study that looked at the late discovery of genetic origins experiences was conducted by the Post Adoption Resource Centre in New South Wales in 1999. This report found that the participants in their study expressed feelings of disbelief, confusion, anger, sorrow and loss. Further, the majority of participants continued to struggle with issues arising from this intentional concealment of their genetic origins (Perl & Markham, 1999). A second and more recent study (Passmore, Feeney & Foulstone, 2007) looked at the issue of secrecy in adoptive families as part of a broader study of 144 adult adoptees. This study found that secrecy and/or lies or misinformation on the part of adoptive parents had negative effects on both personal identity and relationships with others. The authors noted that those adoptees who found out about their adoption as adults were ‘especially likely to feel a sense of betrayal’ (p.4). Over recent years, stories of secrecy and late discovery have also started to emerge from sperm donor conceived adults (Spencer, 2007; Turner & Coyle, 2000). Current research evidence shows that although a majority of couples during the donor assisted conception process indicate that they intend to tell the offspring about their origins, as many as two-thirds or more of couples continue to withhold this information from their children (Akker, 2006; Gottlieb, A. McWhinnie, 2001; Salter-Ling, Hunter, & Glover, 2001). Why do they keep this secret? Infertility involves a range of complex factors that are often left unresolved or poorly understood by those choosing insemination by donor as a form of family building (Schaffer, J. A., & Diamond, R., 1993). These factors may only impact after the child is born, when resemblance talk becomes most pronounced. Resemblance talk is an accepted form of public discourse and a social convention that legitimises the child as part of the family and is part of the process of constructing the child’s identity within the family. Couples tend to become focused on resemblance as this is where they feel most vulnerable, and the lack of resemblance to the parenting father may trigger his sense of loss (Becker, Butler, & Nachtigall, 2005).

Ureaplasma parvum and ureaplasma urealyticum detected in semen after washing prior to assisted reproductive technology procedures

Objective: This study investigated: (i) the prevalence of ureaplasmas in semen and washed semen and (ii) the effect of ureaplasmas on semen andrology parameters. Design: Prospective study. Setting: IVF unit -private hospital, Brisbane, Australia. Patient(s): Three hundred and forty three men participating in an assisted reproductive technology (ART) treatment cycle. Intervention(s): Semen and washed semen tested by culture, PCR assays and indirect immunofluorescent antibody assays. Statistical differences were determined by a t-test, Wilcoxon or Pearson’s Chi- square test where appropriate. Main Outcome Measure(s): The prevalence of ureaplasmas in semen and washed semen and the effect of these microorganisms on semen andrology parameters. Result(s): Ureaplasmas were detected in 73/343 (22%) semen samples and 29/343 (8.5%) washed semen samples. Ureaplasmas adherent to the surface of spermatozoa were demonstrated by indirect immunofluorescent antibody testing. U. parvum serovar 6 (36.6%) and U. urealyticum (30%) were the most prevalent isolates in washed semen. A comparison of the semen andrology parameters of washed semen ureaplasma positive and negative groups demonstrated a lower proportion of non-motile sperm in the washed semen ureaplasma positive group. Conclusion(s): Ureaplasmas are not always removed from semen by a standard ART washing procedure and can remain adherent to the surface of spermatozoa.