Reduced Sperm Quality Research Articles

Acute mild heat stress alters gene expression in testes and reduces sperm quality in mice

Heat stress is a major concern in animal reproduction, as testicular temperature must be 3–5 °C below body core temperature for production of motile and fertile sperm in mammals. Although recent studies concluded that increased temperature per se was the underlying pathophysiology of testicular impairment, more studies are required to better understand the mechanisms. Therefore, our objective was to investigate the impacts of mild acute heat stress on sperm and testes, and based on mRNA, elucidate involvement of StAR, Trp53 and Trp53-dependent intrinsic and extrinsic apoptotic pathways in pathophysiology of testicular heat stress. Forty-eight C57 BCL6 elite male mice were equally allocated into six groups, anesthetized and the distal third of their body immersed in a water-bath at 40 or 30 °C (heat treatment and control, respectively) for 20 min. Intervals from heat exposure (Day 0) to euthanasia were: 8 and 24 h and 7, 14 and 21 d (plus a control group at 14 d). The epididymides were excised, minced and placed in Tyrode albumin lactate pyruvate hepes (TALPH) at 37 °C for 15 min to recover sperm. Based on computer assisted sperm analysis (CASA), heat treatment reduced total and progressive motility ∼40% (P < 0.05) on Days 14 and 21. Furthermore, percentage morphologically normal sperm was significantly decreased on Day 7, with greater reductions on Days 14 and 21, mostly due to increased midpiece defects. Acrosome integrity (FITC PSA) was decreased ∼35% at 8 h (P < 0.05) and reached a nadir on Day 14. There were decreases (P < 0.05) in seminiferous tubule diameter and testicular weight (relative to body weight) on Day 14. Testicular RNA was extracted, reverse-transcribed and cDNA used for PCR. Expression of genes Hspa1b (Hsp70) and Gpx1 had 7- and 10-fold increases (P < 0.001 for each) at 8 and 24 h, respectively, with Hspa1b remaining upregulated at 24 h, whereas StAR peaked at Day 14 (15-fold, P < 0.0001) and had returned to baseline on Day 21. Both Trp53 and Casp8 were upregulated (P < 0.05) on Day 14, whereas Bcl-2 was decreased (P < 0.05) on Days 7 and 14. In conclusion, acute mild heat stress severely reduced sperm quality and based on mRNA, there was upregulation of chaperone and antioxidant systems and Trp53-dependent intrinsic and extrinsic apoptotic pathways, with deleterious effects on sperm, spermatocytes and spermatids. These findings provided insights into the pathophysiology of heat stress and should contribute to development of evidence-based approaches to mitigate effects of testicular heating.

Parameters associated with sperm quality prior to chemotherapy in lymphoma patients.

Sperm quality in lymphoma patients may be reduced even prior to initiation of chemotherapy. The objective of this study was to examine the relationship between lymphoma prognostic factors and sperm quality prior to chemotherapy. A retrospective cohort study was conducted in a Hadassah Medical Center sperm bank and the Hematology department. The cohort included 101 Hodgkin's and 90 non-Hodgkin's lymphoma patients that underwent sperm cryopreservation before chemotherapy between 1998 and 2015. Known lymphoma prognostic factors were compared between patients with normal and impaired sperm parameters. The Prognostic Score Ratio (PSR), an index representing the number of negative lymphoma prognostic measures that found in a lymphoma patient, was additionally calculated and compared between the groups. Among the prognostic factors of lymphoma, the following factors were found to be associated with impaired sperm parameters-low albumin (p<0.001) and haemoglobin (p<0.001) levels, B symptoms (p=0.021) and PSR (p<0.001). Logistic regression showed significant association of albumin and haemoglobin with reduced sperm quality (OR=2.7 and OR=13.5, p<0.05; respectively). To conclude, low albumin and haemoglobin levels are related to reduced sperm quality. The linkage between these prognostic factors and sperm quality may be related to a general inflammatory status.

Protective role of wogonin against cadmium induced testicular toxicity: Involvement of antioxidant, anti-inflammatory and anti-apoptotic pathways

The present study was conducted to identify possible health - promoting effects of wogonin (Wog) on testicular dysfunction in rats caused by cadmium. Pre-treatment of cadmium chloride (Cd: 5 mg/kg b.wt.) administered rats with wogonin (10 mg/kg b.wt) resulted in significant improvement in Cd-induced decrease in body and organ (testes and epididymides) weights. Wogonin treatment significantly improved Cd-induced reduction in sperm quality and quantity, steroidogenic gene (SFI, StAR, CYP11A1, 3β-HSD, CYP17A1 and 17β-HSD) and protein (SF1, StAR and CYP17A1) expressions and serum testosterone levels. Wogonin treatment provided significant protection to Cd-induced aggression in testicular oxidative (elevated levels of MDA) and anti-oxidative (diminished activities of SOD, CAT and GPx) status. Wog significantly up-regulated mRNA levels of Nrf2, NQO1 and HO-1 and down-regulation of Keap1 in cadmium treated testes. Wogonin administration significantly suppressed Cd-stimulated increase in inflammatory reactions (increase in NF-κB p65 DNA, p-IKKβ, TNF-α levels and decrease in IL-10 levels). Wogonin prevented apoptotic damage by enhanced protein distribution of caspase-9, caspase-3, and Bax due to Cd exposure. Furthermore, Wogonin presented significant protection to histo-morphometric changes resulted after Cd administration. Taken together, the findings of this study provided clear evidence of the therapeutic potential of Cd-induced testicular toxicity at least partly due to its antioxidant, anti-inflammatory and anti-apoptotic properties.

Efeito da adição de diferentes antioxidantes em diluentes de criopreservação sobre a qualidade do sêmen e a produção in vitro de embriões bovinos

A qualidade das amostras de sêmen é fundamental para o sucesso dos programas de inseminação artificial em bovinos. Torna-se imprescindível a manutenção das características dos espermatozoides por meio de adequado processo de criopreservação. No entanto, os processos de criopreservação resultam na formação contínua e excessiva de espécies reativas de oxigênio (EROS), que são prejudiciais aos espermatozoides, pois induz a apoptose com consequente redução da qualidade espermática. Dessa forma, têm sido acrescidos aos crioprotetores substâncias antioxidantes como a glutationa e a melatonina, as quais tem como intuito minimizar os efeitos deletérios causados pelas EROS aos espermatozoides, resultando em melhor qualidade espermática, com consequente aumento das taxas de fertilização in vitro. Diante disso, a presente revisão tem por objetivo analisar os efeitos da adição dos antioxidantes glutationa e melatonina em diferentes meios de criopreservação e a produção in vitro de embriões bovinos.

Impact of timing of the anorexigen sibutramine administration on reproductive end-points of male rats.

Sibutramine is a non-selective serotonin-norepinephrine reuptake inhibitor orally administered for weight loss. In a previous study, we showed pharmacological mechanisms involved in the reduction of sperm quality and fertility of rats exposed for 30days to this anorexigen in the light phase of the light-dark (l/d) cycle. It is already known that rodents are nightlife animals, with higher metabolic activity during the dark phase than in the light phase of the light-dark (l/d) cycle. Thus, the present study aimed to investigate whether the deleterious effects on reproductive parameters after sibutramine administration would be enhanced after a shorter period of exposure during the dark phase of the l/d cycle. For this, adult male Wistar rats were treated with sibutramine (10mg/kg/d) or vehicle for 15days during the dark phase of the l/d cycle. Sibutramine treatment decreased final body and reproductive organ weights, as well as serum testosterone levels. Sperm transit time through the epididymis was accelerated, and sperm concentration and motility were diminished in the sibutramine-exposed rats. The decrease in sperm concentration was also verified in the epididymal histological sections. In conclusion, the deleterious effects of sibutramine on reproductive parameters of male rats were enhanced when the exposure occurred in the dark phase of the l/d cycle, even after a short exposure duration. Our results reinforce the impact of timing on drug therapeutic action.

The physiology of male reproduction: Impact of drugs and their abuse on male fertility.

Male factor accounts for about 30-50% of infertility. A common cause of male infertility is drug abuse; either illicit or prolonged use of prescribed drugs. This study provides a review of the physiology of the hypothalamic-pituitary-gonadal axis and recent literature on drugs that have been linked to male infertility and the associated mechanisms. Relevant peer-reviewed papers were assessed online using PubMed/PubMed Central, Scopus, AJOL, Google Scholar and DOAJ databases using Medical Subjects Headings (MeSH) indexes and relevant key word searches. Although drugs are beneficial when used at therapeutic levels, the abuse leads to impairment of hypothalamic-pituitary-gonadal functions, increased sperm DNA fragmentation and apoptosis, and reduced sperm quality. A good knowledge of the physiology of the hypothalamic-pituitary-gonadal axis and the influence of drugs on male fertility will guide healthcare providers in managing cases of infertility.

Spatial arrangement of testicular cells disrupted by transient scrotal hyperthermia and subsequent impairment of spermatogenesis.

The spermatogenesis is temperature-dependent and heat stress have destructive effects on spermatogenesis and reduces sperm quality. Sixteen adult mice were allocated to two groups: hyperthermia and control groups. Scrotal hyperthermia was induced by water bath with 43°C for 30min. Then, the spermatozoon was isolated through the tail region of epididymis for sperm parameters analysis. The testicular tissues were taken for stereological studies, hormonal assay, TUNEL assay and molecular studies. We found a marked decrease in sperm parameters and serum testosterone level in mice induced by scrotal hyperthermia as well as stereological analysis indicated a significant reduction in testicular cells and changes in the spatial arrangement of testicular cells in the scrotal hyperthermia groups compared to the control groups. Moreover, the TUNEL assay results showed that apoptotic cells were enhanced significantly in the group of scrotal hyperthermia compared to the control groups. Furthermore, scrotal hyperthermia caused a reduction in the expression of retinoic acid 8 (STRA8), c-kit and proliferating cell nuclear antigen (PCNA) genes in the scrotal hyperthermia groups compared to the control. According to results, induction of transient scrotal hyperthermia leads to a fluctuation in the spatial arrangement of testicular cells, which finally influences the normal function of spermatogenesis.

Saccharum officinarum juice alters reproductive functions in male Wistar rats.

Infertility is a problem across almost all cultures and societies. Problems in the male partner, especially as a result of unhealthy dietary habits, are the commonest single group of course. Many populations, therefore, tend more toward the use of natural dietary substitutes opined to proffer less risk to reproductive functions and more health benefits. Saccharum officinarum juice (SOJ) is a widely consumed, energy-rich, nutritious substance that has many minerals and enzymes. Saccharum officinarum plant was reported to have anti-thrombosis, anti-inflammatory and immune-stimulatory activities. This study evaluated the reproductive effects of S. officinarum juice in male Wistar rats. A sugarcane press juicer was used to extract S. officinarum juice. Twenty male Wistar rats (100-120 g) grouped into four (n = 5) received 1.0 mL/kg/day distilled water (control), and 1.0, 3.2 and 10.0 mL/kg/day of fresh S. officinarum juice once daily for 8 weeks via gavage. Sperm analysis, histology of testes and epididymides were evaluated by microscopy. Enzyme-linked immunosorbent assay (ELISA) was used in assessing the serum levels of luteinizing hormone, follicle-stimulating hormone and testosterone. Data were analyzed using the analysis of variance at a significance of p < 0.05. SOJ increased fasting blood glucose levels in 3.2 and 10.0 mL/kg groups. The 10.0 mL/kg juice caused a significant increase in testosterone level and sperm count, and it also increased the percentage of aberrant sperm and decreased sperm viability. Saccharum officinarum juice impaired the histological integrity of the testes and epididymides. Thus, S. officinarum juice adversely altered the reproductive functions of male Wistar rats by reducing sperm quality and disrupting testicular architecture.

Effect of Different Media on Short Term Storage of Leaping Mullet (Liza saliens (Risso, 1810)) Sperm

The present study aimed to evaluate the activation characteristics of the leaping mullet spermatozoa and diluted with four different media (D1, D2, D3, D4) for 250 h at 4±2°C. The semen was collected from nine wild fish in Homa Lagoon, İzmir, Turkey in July, during reproductive season. In each trial (n = 9), semen samples were collected and mixed to reduce sperm quality variation to obtain the required gametes for the experiment. The semen was activated with natural seawater (salinity of 34, pH 7.9 and 4±2°C) to determine: motility time, motility rate and viability of sperm. Dilution with inactivation media 1:3 increased motility time of diluted semen than control group. At the end of the experiment, no significant difference was found for the duration of survival in the D2 and D4 groups. The highest survival rate was found in D2 group.

A novel approach for human sperm cryopreservation with AFPIII

Sperm cryopreservation causes different stresses including thermal shock, osmotic damage, and ice crystal formation, thereby reducing sperm quality. Few studies have evaluated the application of AFPs in cryopreservation. The effects of antifreeze protein III (AFP III) on human sperm cryopreservation is not fully understood therefore, we conducted this study to investigate the effects of AFPIII treatment on human sperm parameters following cryopreservation. First, for 20 semen samples the effects of various concentrations of AFPIII (0, 0.01, 0.1, 1, 5, 10 μg/ml) were evaluated. Sperm parameters, such as motility and viability were assessed in order to identify an optimal dose. Next, liquefied 20 semen samples were divided into three aliquots and diluted in glycerol-egg-yolk-citrate (GEYC) cryopreserved without AFPIII (control), with optimal dose of AFPIII, as well as fresh groups. After thawing, samples were evaluated for plasma membrane integrity (PMI), DNA fragmentation index (DFI), reactive oxygen species (ROS), and total antioxidant capacity (TAC) levels. Spermatozoa treatment with 0.01, 0.1 and 1 μg/ml AFPIII increased the sperm motility and viability compared to the control group, but the highest concentrations were ineffective. In conclusion, the results showed that the addition of AFPIII to GEYC at 1 μg/ml improved motility, PMI, viability and TAC, and decreased ROS and DNA fragmentation of cryopreserved human semen compared to the control group.

Three organophosphate flame retardants (OPFRs) reduce sperm quality in Chinese rare minnows (Gobiocypris rarus)

Organophosphate flame retardants (OPFRs) are widespread in the aquatic environment, but the effects of these chemicals on reproductive toxicity are far from clear. In this study, sperm quality in adult male Chinese rare minnows after exposure to tris-(2-butoxyethyl) phosphate (TBOEP), tris-(1,3-dichloro-2-propyl) phosphate (TDCIPP), and triphenyl phosphate (TPHP) was investigated. No obvious change in sperm concentration and vitality was observed after treatments, whereas significant changes in sperm velocity and morphology were found following all treatments (P < 0.05). Moreover, OPFR exposure significantly increased the apoptosis ratios in testis cells. Analysis of the transcriptomic data revealed that Na+/K+ ATPase (NKA) related genes were significantly downregulated, and the NKA enzyme activities after all treatments were significantly inhibited (P < 0.05). However, no obvious change in hormone levels in the groups exposed to TBOEP and TDCIPP was observed. These findings indicate that the OPFR-induced reduction of sperm quality might be due to the effects of OPFRs on NKA enzyme instead of changes in hormone levels.

Effect of refrigeration at -1°C on spermatozoa quality of domestic cats

ABSTRACT: The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P&lt;0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.

Inbreeding depression in sperm quality in a critically endangered amphibian.

Small, isolated populations often experience increased inbreeding and decreased heterozygosity, which increases the potential risk of inbreeding depression. The relationship between inbreeding and sperm health is well-documented in a variety of taxa, but has yet to be explored in amphibians. The dusky gopher frog, Lithobates sevosus, is a critically endangered species with years of documented inbreeding and low genetic variability as a consequence of isolation and population size reduction. This study investigated the effects of inbreeding on sperm quality in captive L. sevosus using an outbred, sister species (Lithobates pipiens) as a standard for comparison. We found L. sevosus to have severely reduced sperm quality in terms of total motility, forward progressive motility, concentration, and viability. Additionally, we observed a significant, negative relationship between total sperm motility and mean kinship within captive-bred individuals. These data serve to enhance our understanding of the role inbreeding plays in amphibians, and to provide valuable insight into new risk factors declining amphibian populations may face.

Glutathione S-Transferases Play a Crucial Role in Mitochondrial Function, Plasma Membrane Stability and Oxidative Regulation of Mammalian Sperm.

Glutathione S-transferases (GSTs) are essential sperm antioxidant enzymes involved in cell protection against oxidative stress and toxic chemicals, preserving sperm function and fertilising ability. Artificial insemination (AI) in pigs is commonly carried out through the use of liquid-stored semen at 17 °C, which not only reduces sperm metabolic activity but also sperm quality and AI-farrowing rates within the 72 h of storage. While one may reasonably suggest that such enzymes are implicated in the physiology and maintenance of mammalian sperm function during liquid-storage, no previous studies conducted on any species have addressed this hypothesis. Therefore, the objective of the present work was to characterise the presence and function of sperm GSTs in mammalian sperm, using the pig as a model. In this regard, inhibition of such enzymes by ethacrynic acid (EA) during semen storage at 17 °C was performed to evaluate the effects of GSTs in liquid-preserved boar sperm by flow cytometry, immunofluorescence, and immunoblotting analysis. The results of this study have shown, for the first time in mammalian species, that the inhibition of GSTs reduces sperm quality and functionality parameters during their storage at 17 °C. These findings highlight the key role of such enzymes, especially preserving mitochondrial function and maintaining plasma membrane stability. In addition, this study has identified and localised GSTM3 in the tail and equatorial subdomain of the head of boar sperm. Finally, this study has set grounds for future investigations testing supplementation of semen extenders with GSTs, as this may improve fertility outcomes of swine AIs.

Распространение основных микотоксинов в кормовом сырье и их характеристика

Mycotoxins are low molecular weight secondary metabolites produced by filamentous fungi. The most relevant groups of mycotoxins found in animal feed are produced by three genera of fungi: Aspergillus, Penicillium, and Fusarium species. Animal feeds are routinely contaminated by mycotoxins from diverse sources. In most cases, the concentrations are low enough to ensure compliance with Maximum Permitted Levels. Poultry, pigs and also aquatic vertebrates are very sensitive to mycotoxins. Ruminants are, however, generally more resistant to the adverse effects of mycotoxins, since the rumen microbiota is capable of degrading mycotoxins. Mycotoxins cause huge economic damage, leading to increased costs for health and veterinary care, as well as reduce livestock production. The effects of long-term exposure of aflatoxins are associated with the reduction of weight gain, decreased milk or egg production, teratogenicity. Ochratoxin A causes renal toxicity and possesses carcinogenic, teratogenic, immunosuppressive and possibly neurotoxic properties. The main trichothecenes mycotoxins are T-2 toxin, deoxynivalenol and diacetoxyscirpenol. Deoxynivalenol is primarily known for causing feed refusal, weight loss. It also causes lesions in the gastrointestinal tract, vomiting, bloody diarrhea and severe dermatitis accompanied by hemorrhaging. Zearalenone and some of its metabolites, can competitively bind to oestrogen receptors leading to reproductive disorders and oestrogenic dysfunction, impairing fertility and increasing the frequency of stillbirths along with reducing sperm quality. Fumonisin causes carcinogenicity and cardiovascular toxic effects.

Long term conjugated linoleic acid supplementation modestly improved growth performance but induced testicular tissue apoptosis and reduced sperm quality in male rabbit.

Conjugated linoleic acid (CLA) is known for its multiple benefits including improvement of growth, increasing lean mass, and anti-carcinogenic effects. However, when used in long-term supplementations CLA does not improve semen parameters in boar and bull and reduces fertility in Japanese quails. The content of unsaturated fatty acids in dietary lipids plays a significant role in spermatogenesis owning the high proportion of unsaturated fatty acids in plasma membrane of sperms. Whether CLA plays a role in testicular tissue and epididymal fat is still unknown. Therefore, in this study we hypothesize that long-term supplementation of equal proportion of CLA isomer mix (c9,t11-CLA and t10,c12- CLA) in rabbit bucks might alter male reproductive potentials. Twelve V-Line weaned male rabbits were used in 26 weeks trial, rabbits were individually raised and randomly allocated into three dietary groups. Control group (CON) received a basal diet, a group received 0.5% CLA (CLA 0.5%), and a group received 1% CLA (CLA 1%). Rabbits were euthanized at the end of the trial and several parameters were evaluated related to growth, semen quality, and testicular and epididymal tissue histopathology and transcriptome. The long-term supplementation of CLA increased feed intake by 5% and body weight by 2–3%. CLA 1% decreased sperm progressive motility. In testicular tissue L-carnitine and α-tocopherol were decreased by CLA supplementation. In epididymal fat, CLA tended to decrease concentration of polyunsaturated fatty acids, the expression of SCD5 gene was upregulated by CLA 1% and CASP3 gene was upregulated by CLA 0.5%. Transcription of PPARG was downregulated by CLA. Feeding 1% CLA also decreased testicular epithelial thickness. Long-term supplementation of CLA modestly enhanced male rabbit growth, but negatively impacted male reproduction, especially at high dose of CLA.

Papaya (Carica Papaya L.) Seed Extract as Male Contraception via Decreasing The Quality of Rat's (Rattus Norvegicus) Sperm

Background. Population growth needs to be controlled so as not to cause new and serious problems. Pregnancy and birth control is a solution to control this sizable population growth. Contraception is a way to prevent pregnancy, by inhibiting the process of ovulation, fertilization and implantation. Nevertheless, most of contraceptive regiments were aimed to woman. Male contraception nowadays is very rare. Papaya seed is one of the male contraceptive candidates which has the potential to be developed as male contraception. Previous studies have shown the potential of papaya seed extract could reduce the weight of reproductive organs of male white rats, where it was believed that there was a decrease in spermatogenesis activity.&#x0D; Methods. This research was an in-vivo experimental research post-test only with control group. The study used 32 male and healthy rats (Rattus norvegicus) Wistar strain with 8-10 weeks of age. Rats were grouped into 1 control group and 4 treatment groups that were randomly selected. The four treatment groups were group 1: control administered 1% 2ml CMC orally one time per day every morning, group 2: were given N-hexane papaya seed fraction, group 3: were given ethyl acetate papaya seed fraction and group 4: were given ethanol-water papaya seed fraction. The three fractions were given to rats orally at a dose of 100 mg / KgBB / day for 24 days. After that, rat's testes and epidydimis were evacuated to observe reproductive organs and sperm quality.&#x0D; Results. Anatomically, Group 2 and 3 were statistically significant in reducing testicular's and epidydimis' weight compared to the control group, but the n-hexane group was more effective in reducing testicular weight than other treatment groups. Furthermore, groups 2, 3 and 4 were statistically significant in reducing sperm quality (motility, viability, amount of sperm cells and normal sperm cell morphology) compared to the control group, but the n-hexane group was more effective in reducing sperm quality than other treatment groups.&#x0D; Conclusion. Papaya seed n-hexane fraction has potential as a male contraceptive by reducing spermatogenesis activity and decreasing sperm quality in white rats.&#x0D; Keyword : papaya seed, male contraception, rat's sperm

Effects of oral administration of titanium dioxide particles on sperm parameters and in vitro fertilization potential in mice: A comparison between nano- and fine-sized particles.

Titanium dioxide particles (TiO2) as the second most widely used materials in consumer products are composed of nano-sized (<100 nm) particles (NPs) and fine-sized (>100 nm) particles (FPs). Toxicological studies on animals have shown that TiO2 NPs exposure can cross the blood-testis barrier and accumulate in the testis resulting in testicular tissue damage and reduction of sperm count and motility. However, there is no information on the toxic effects of TiO2 FPs on male reproductive fertility. Twenty-four adult male mice were randomly divided into three groups including control, TiO2 NPs, and TiO2 FPs (150 mg kg-1 per day). After intragastric administration for 35 days, testicular tissue alterations (seminiferous tubule diameter and germinal epithelial height), sperm parameters (count, motility, viability, morphology, and DNA quality), in vitro fertilization potential, oxidative stress assays such as malondialdehyde (MDA) content, level of glutathione (GSH) and activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in testicular tissue were investigated. The results showed that both sizes of TiO2 caused pathologic changes in the testis and significantly increased MDA level and decreased GSH levels and activities of SOD and GPx in testicular tissue. Moreover, the administration of both sizes of TiO2 significantly decreased all of the sperm parameters and in vitro fertility (fertilization rate and pre-implantation embryos development) compared to control. Administration of TiO2 FPs similar to TiO2 NPs through inducing damages to testis led to a marked reduction in sperm quality, in vitro fertilization, and embryos development in male mice.

Saccharum officinarum molasses adversely alters reproductive functions in male wistar rats

The widely reported anti-androgenic effects of refined sugar led to the exploration of safer alternatives. Saccharum officinarum molasses (SOM), a byproduct of sugar processing is gaining popularity as a substitute. This study investigated the effects of SOM and compared them to those of refined sugar on male reproductive functions. Blackstrap® Saccharum officinarum molasses were subjected to phytochemical screening and proximate analysis and fractionated to obtain methanol (SOMMF) and aqueous (SOMAqF) fractions. Twelve groups (n = 5) of adult male Wistar rats received distilled water (Control); 0.8, 2.5, 7.9 g/kg SOM; 0.0064 g/kg sugar (Dangote®); 0.0064 g/kg sugar+7.9 g/kg SOM; 1.0, 3.2, 10.0 g/kg SOMMF and 0.6, 2.0, 6.4 g/kg SOMAqF, respectively. Administrations were done daily by oral gavage for eight weeks. Sperm profile and testicular and epididymal histology were assessed using microscopy. Serum testosterone was quantified using ELISA. Testicular malondialdehyde (MDA) was assayed by spectrophotometry. Data were analyzed using ANOVA at p < 0.05 significance. Sperm count and viability reduced with 7.9 g/kg SOM, Sugar, 3.2 and 10.0 g/kg SOMMF, 2.0 and 6.4 g/kg SOMAqF. Abnormal sperms increased with 7.9 g/kg SOM, Sugar, 2.0 and 6.4 g/kg SOMAqF. Testosterone level reduced with 6.4 g/kg SOMAqF. Testicular MDA increased with SOM, 3.2 and 10.0 g/kg SOMMF and 6.4 g/kg SOMAqF. Seminiferous tubules and epididymal ducts of 7.9 g/kg SOM, Sugar and SOMAqF-treated rats showed anomalies. Saccharum officinarum molasses altered testicular and epididymal integrity via lipid peroxidation, thus reducing sperm quality and androgen levels in male Wistar rats.

Optimal concentration of Rho-associated coiled-coil kinase (ROCK) inhibitor improved sperm membrane functionality and fertilizing ability of cryopreserved-thawed feline sperm

Sperm cryopreservation induces irreversible loss of viability and fertilizing ability. This study aimed at examining the effects of Rho-associated, coiled-coil kinase (ROCK) inhibitor on quality of frozen-thawed feline sperm. Ejaculated semen from individual cats (n = 6) was examined for the expression of LIMK1 and LIMK2 mediated ROCK cascade. The effects of ROCK inhibitor during cooling and cryopreservation on sperm quality and fertilizing ability were also examined. Feline sperm were treated with different concentrations of ROCK inhibitor (10, 20 and 40 μM) during cooling at 4 °C and cryopreservation. Sperm cooled and conventionally cryopreserved without ROCK inhibitor (0 μM) served as a control group.The ROCK cascade was confirmed in feline sperm as they expressed mRNA of LIMK1 and LIMK2 genes. Cryopreservation significantly reduced sperm quality in terms of viability (91.63 ± 3.96 vs. 60.11 ± 8.93), progressive motility (91.67 ± 3.54 vs. 46.67 ± 8.66) and acrosome integrity (93.49 ± 3.64 vs. 63.81 ± 5.31) for fresh and frozen-thawed sperm, respectively (p < 0.05). The positive effects of ROCK inhibitor on sperm quality were pronounced at 1 and 3 h post-thaw. ROCK inhibitor at 10 μM significantly improved sperm motility and membrane functionality compared to those observed in a control group (0 μM) (p < 0.05). In vitro fertilization revealed that supplement ROCK inhibitor at 10 μM during cryopreservation significantly improved in vitro fertilizing ability of the frozen-thawed sperm (p < 0.05). However, it did not subsequently increase morula and blastocyst rates (p > 0.05). Increased concentrations of ROCK inhibitor to 20 and 40 μM did not further improve the quality of frozen-thawed sperm. In conclusion, an optimal concentration (10 μM) of the ROCK inhibitor added into cooling medium could improve post-thaw sperm quality.