Background
Between 30% to 80% of male subfertility cases are considered to be due to the damaging effects of oxidative stress on sperm and 1 man in 20 will be affected by subfertility. Antioxidants are widely available and inexpensive when compared to other fertility treatments and many men are already using these to improve their fertility. It is thought that oral supplementation with antioxidants may improve sperm quality by reducing oxidative stress. Pentoxifylline, a drug that acts like an antioxidant, was also included in this review.  

Objectives
This Cochrane review aimed to evaluate the effectiveness and safety of oral supplementation with antioxidants for subfertile male partners in couples seeking fertility assistance.

Search methods
We searched the Cochrane Menstrual Disorders and Subfertility Group Specialised Register, CENTRAL, MEDLINE, EMBASE, CINAHL, PsycINFO and AMED databases (from inception until January 2014); trial registers; sources of unpublished literature and reference lists. An updated search was run in August 2014 when potentially eligible studies were placed in 'Studies awaiting assessment'.

Selection criteria
We included randomised controlled trials (RCTs) comparing any type or dose of antioxidant supplement (single or combined) taken by the subfertile male partner of a couple seeking fertility assistance with a placebo, no treatment or another antioxidant.

Data collection and analysis
Two review authors independently selected eligible studies, extracted the data and assessed the risk of bias of the included studies. The primary review outcome was live birth; secondary outcomes included clinical pregnancy rates, adverse events, sperm DNA fragmentation, sperm motility and concentration. Data were combined, where appropriate, to calculate pooled odds ratios (ORs) or mean differences (MD) and 95% confidence intervals (CIs). Statistical heterogeneity was assessed using the I2 statistic. We assessed the overall quality of the evidence for the main outcomes using GRADE methods.  

Main results
This updated review included 48 RCTs that compared single and combined antioxidants with placebo, no treatment or another antioxidant in a population of 4179 subfertile men. The duration of the trials ranged from 3 to 26 weeks with follow up ranging from 3 weeks to 2 years. The men were aged from 20 to 52 years. Most of the men enrolled in these trials had low total sperm motility and sperm concentration. One study enrolled men after varicocelectomy, one enrolled men with a varicocoele, and one recruited men with chronic prostatitis. Three trials enrolled men who, as a couple, were undergoing in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) and one trial enrolled men who were part of a couple undergoing intrauterine insemination (IUI). Funding sources were stated by 15 trials. Four of these trials stated that funding was from a commercial source and the remaining 11 obtained funding through non-commercial avenues or university grants. Thirty-three trials did not report any funding sources.

A limitation of this review was that in a sense we had included two different groups of trials, those that reported on the use of antioxidants and the effect on live birth and clinical pregnancy, and a second group that reported on sperm parameters as their primary outcome and had no intention of reporting the primary outcomes of this review. We included 25 trials reporting on sperm parameters and only three of these reported on live birth or clinical pregnancy. Other limitations included poor reporting of study methods, imprecision, the small number of trials providing usable data, the small sample size of many of the included studies and the lack of adverse events reporting. The evidence was graded as 'very low' to 'low'. The data were current to 31 January 2014.

Live birth: antioxidants may have increased live birth rates (OR 4.21, 95% CI 2.08 to 8.51, P< 0.0001, 4 RCTs, 277 men, I2 = 0%, low quality evidence). This suggests that if the chance of a live birth following placebo or no treatment is assumed to be 5%, the chance following the use of antioxidants is estimated to be between 10% and 31%. However, this result was based on only 44 live births from a total of 277 couples in four small studies.

Clinical pregnancy rate: antioxidants may have increased clinical pregnancy rates (OR 3.43, 95% CI 1.92 to 6.11, P < 0.0001, 7 RCTs, 522 men, I2 = 0%, low quality evidence). This suggests that if the chance of clinical pregnancy following placebo or no treatment is assumed to be 6%, the chance following the use of antioxidants is estimated at between 11% and 28%. However, there were only seven small studies in this analysis and the quality of the evidence was rated as low.

Miscarriage: only three trials reported on this outcome and the event rate was very low. There was insufficient evidence to show whether there was a difference in miscarriage rates between the antioxidant and placebo or no treatment groups (OR 1.74, 95% CI 0.40 to 7.60, P = 0.46, 3 RCTs, 247 men, I2 = 0%, very low quality evidence). The findings suggest that in a population of subfertile men with an expected miscarriage rate of 2%, use of an antioxidant would result in the risk of a miscarriage lying between 1% and 13%.

Gastrointestinal upsets: there was insufficient evidence to show whether there was a difference in gastrointestinal upsets when antioxidants were compared to placebo or no treatment as the event rate was very low (OR 1.60, 95% CI 0.47 to 5.50, P = 0.46, 6 RCTs, 429 men, I2 = 0%).

We were unable to draw any conclusions from the antioxidant versus antioxidant comparison as not enough trials compared the same interventions.

Authors' conclusions
There is low quality evidence from only four small randomised controlled trials suggesting that antioxidant supplementation in subfertile males may improve live birth rates for couples attending fertility clinics. Low quality evidence suggests that clinical pregnancy rates may increase. There is no evidence of increased risk of miscarriage but this is uncertain as the evidence is of very low quality. Data were lacking on other adverse effects. Further large well-designed randomised placebo-controlled trials are needed to clarify these results.

Antioxidants for male subfertility

STUDY QUESTION: Is there an association between high levels of sperm DNA damage and miscarriage? SUMMARY ANSWER: Miscarriage rates are positively correlated with sperm DNA damage levels. WHAT IS KNOWN ALREADY: Most ejaculates contain a subpopulation of sperm with DNA damage, also referred to as DNA fragmentation, in the form of double or single-strand breaks which have been induced in the DNA prior to or following ejaculation. This DNA damage may be particularly elevated in some subfertile men, hence several studies have examined the link between sperm DNA damage levels and conception and miscarriage rates. STUDY DESIGN, SIZE, DURATION: A systematic review and meta-analysis of studies which examined the effect of sperm DNA damage on miscarriage rates was performed. Searches were conducted on MEDLINE, EMBASE and the Cochrane Library without any language restrictions from database inception to January 2012. PARTICIPANTS/MATERIALS, SETTING, METHODS: We used the terms 'DNA damage' or 'DNA fragmentation' combined with 'miscarriage', 'abortion' or 'pregnancy' to generate a set of relevant citations. Data extraction was performed by two reviewers. Study quality was assessed using the Newcastle-Ottawa Scale. Meta-analysis of relative risks of miscarriage was performed with a random effects model. Subgroup analyses were performed by the type of DNA damage test, whether the sperm examined were prepared or from raw semen and for pregnancies resulting from IVF or ICSI treatment. MAIN RESULTS AND THE ROLE OF CHANCE: We identified 16 cohort studies (2969 couples), 14 of which were prospective. Eight studies used acridine orange-based assays, six the TUNEL assay and two the COMET assay. Meta-analysis showed a significant increase in miscarriage in patients with high DNA damage compared with those with low DNA damage [risk ratio (RR) = 2.16 (1.54, 3.03), P < 0.00001)]. A subgroup analysis showed that the miscarriage association is strongest for the TUNEL assay (RR = 3.94 (2.45, 6.32), P < 0.00001). LIMITATIONS, REASONS FOR CAUTION: There is some variation in study characteristics, including the use of different assays and different thresholds for DNA damage and the definition of pregnancy loss. WIDER IMPLICATIONS OF THE FINDINGS: The use of methods which select sperm without DNA damage for use in assisted conception treatment may reduce the risk of miscarriage. This finding indicates that assays detecting DNA damage could be considered in those suffering from recurrent pregnancy loss. Further research is necessary to study the mechanisms of DNA damage and the potential therapeutic effects of antioxidant therapy. STUDY FUNDING/COMPETING INTEREST(S): None.

The effect of sperm DNA fragmentation on miscarriage rates: a systematic review and meta-analysis

Do sperm DNA integrity tests predict pregnancy with in vitro fertilization

Computer-assisted sperm analysis (CASA): capabilities and potential developments.

Animal production is a significant source of greenhouse gas (GHG) emissions worldwide. Depending on the accounting approaches and scope of emissions covered, estimates by various sources (IPCC, FAO, EPA or others) place livestock contribution to global anthropogenic GHG emissions at between 7 and 18 percent. The current analysis was conducted to evaluate the potential of nutritional, manure and animal husbandry practices for mitigating methane (CH4) and nitrous oxide (N2O) – i.e. non-carbon dioxide (non-CO2) – GHG emissions from livestock production. These practices were categorized into enteric CH4, manure management and animal husbandry mitigation practices. Emphasis was placed on enteric CH4 mitigation practices for ruminant animals (only in vivo studies were considered) and manure mitigation practices for both ruminant and monogastric species. Over 900 references were reviewed; and simulation and life cycle assessment analyses were generally excluded. In evaluating mitigation practices, the use of proper units is critical. Expressing enteric CH4 energy production on gross energy intake basis, for example, does not accurately reflect the potential impact of diet quality and composition. Therefore, it is noted that GHG emissions should be expressed on a digestible energy intake basis or per unit of animal product (i.e. GHG emission intensity), because this reflects most accurately the effect of a given mitigation practice on feed intake and the efficiency of animal production. EntErIC CH4 MItIgAtIon prACtICEs Increasing forage digestibility and digestible forage intake will generally reduce GHG emissions from rumen fermentation (and stored manure), when scaled per unit of animal product, and are highly-recommended mitigation practices. For example, enteric CH4 emissions may be reduced when corn silage replaces grass silage in the diet. Legume silages may also have an advantage over grass silage due to their lower fibre content and the additional benefit of replacing inorganic nitrogen fertilizer. Effective silage preservation will improve forage quality on the farm and reduce GHG emission intensity. Introduction of legumes into grass pastures in warm climate regions may offer a mitigation opportunity, although more research is needed to address the associated agronomic challenges and comparative N2O emissions with equivalent production levels from nitrogen fertilizer. Dietary lipids are effective in reducing enteric CH4 emissions, but the applicability of this practice will depend on its cost and its effects on feed intake, production and milk composition. High-oil by-product feeds, such as distiller’s grains, may offer an economically feasible alternative to oil supplementation as a mitigation practice, although their higher fibre content may have an opposite effect on enteric CH4, depending on basal diet composition. Inclusion of concentrate feeds in the diet of ruminants will likely decrease enteric CH4 emissions per unit of animal product, particularly when above 40 percent of dry matter intake. The effect may depend on type of ‘concentrate’ inclusion rate, production response, impact on fibre digestibility, level of nutrition, composition of the basal diet and feed processing.

Mitigation of greenhouse gas emissions in livestock production - A review of technical options for non-CO2 emissions

BACKGROUND: Sperm DNA integrity has been shown to be necessary for achieving and sustaining embryo development. The objective was to evaluate the sperm chromatin structure assay (SCSA) as a diagnostic tool in clinical practice for intrauterine insemination (IUI), in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatments. METHODS: A total of 385 semen samples from 234 couples were frozen for SCSA, and smears were prepared for morphology: 48 IUI, 139 IVF and 47 ICSI. The main SCSA variables were DNA fragmentation index (DFI), standard deviation of DFI (SD-DFI) and high DNA stainability (HDS), and the reproductive outcomes were biochemical pregnancy (BP), clinical pregnancy (CP) and implantation ratio (IR). RESULTS: The results showed no significant difference in the fertility variables BP, CP and IR when 27%, and six couples in ICSI treatment achieved CP full-term. DFI >27% had a high prognostic power for predicting no CP for IVF patients, with a specificity of 97%. Couples diagnosed with male infertility had a significantly higher level of DFI compared to couples with idiopathic fertility. Sperm head morphology showed low but significant correlations with the SCSA variables. CONCLUSION: SCSA is a useful tool in andrological diagnosis and contributes with a prognosis for the fertility outcome of conventional IVF. Although full-term pregnancy can be achieved with assisted reproductive techniques with a DFI >27%, the probability of a successful pregnancy may be reduced.

/pdf/the-sperm-chromatin-structure-assay-as-a-diagnostic-tool-in-pgh4c68uzf.pdf

The sperm chromatin structure assay as a diagnostic tool in the human fertility clinic

DNA integrity in sexed bull sperm assessed by neutral Comet assay and sperm chromatin structure assay

Increasing storage time of extended boar semen reduces sperm DNA integrity

Harsh tropical environments impose serious challenges on poorly adapted species. In beef cattle, tropical adaptation in the form of temperature and disease resistance, coupled with acclimatization to seasonal and limited forage, comes at a cost to production efficiency. Prominent among these costs is delayed onset of puberty, a challenging phenotype to manipulate through traditional breeding mechanisms. Recently, system biology approaches, including gene networks, have been applied to the genetic dissection of complex phenotypes. We aimed at developing and studying gene networks underlying cattle puberty. Our starting material comprises the association results of ~50,000 SNP on 22 traits, including age at puberty, and 2 cattle breed populations: Brahman (n = 843) and Tropical Composite (n = 866). We defined age at puberty as the age at first corpus luteum (AGECL). By capturing the genes harboring mutations minimally associated (P < 0.05) to AGECL or to a set of traits related with AGECL, we derived a gene network for each breed separately and a third network for the combined data set. At the intersection of the 3 networks, we identified candidate genes and pathways that were common to both breeds. Resulting from these analyses, we identified an enrichment of genes involved in axon guidance, cell adhesion, ErbB signaling, and glutamate activity, pathways that are known to affect pulsatile release of GnRH, which is necessary for the onset of puberty. Furthermore, we employed network connectivity and centrality parameters along with a regulatory impact factor metric to identify the key transcription factors (TF) responsible for the molecular regulation of puberty. As a novel finding, we report 5 TF (HIVEP3, TOX, EYA1, NCOA2, and ZFHX4) located in the network intersecting both breeds and interacting with other TF, forming a regulatory network that harmonizes with the recent literature of puberty. Finally, we support our network predictions with evidence derived from gene expression in hypothalamic tissue of adult cows.

Gry Boe-Hansen

Papers

The sperm chromatin structure assay as a diagnostic tool in the human fertility clinic

DNA integrity in sexed bull sperm assessed by neutral Comet assay and sperm chromatin structure assay

Increasing storage time of extended boar semen reduces sperm DNA integrity

A single nucleotide polymorphism-derived regulatory gene network underlying puberty in 2 tropical breeds of beef cattle.

Sperm chromatin structure integrity in liquid stored boar semen and its relationships with field fertility.