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The effects of magnetic separation on cryopreserved bovine spermatozoa motility, viability and cryo-capacitation status

Faezah SS, Zuraina FM, Farah JH, Khairul O, Hilwani NI, Iswadi MI, et al.

Zygote, 2014 Aug;22(3):378-86.
PMID: 23237064 DOI: 10.1017/S0967199412000597

Cryopreservation is a technique used to preserve cells for long-time storage. It is widely used in agriculture to store male gametes in liquid nitrogen. The aim of this study was to determine the optimum thawing temperature and time for samples subjected to annexin V magnetic-activated cell sorting (AnMACS) as the sperm preparation technique. Pooled semen samples from three ejaculates were divided into two groups. The treatment group was subjected both to AnMACS and to being cryopreserved, whilst the control group was cryopreserved directly without MACS. Post-thaw analysis was carried out for samples thawed at either 20°C for 13 s, 37°C for 30 s, 40°C for 7 s, 60°C for 6 s or 80°C for 5 s. Sperm kinematics, viability and capacitation status were determined for samples subjected to all thawing temperatures described. Results showed that thawing at 37°C for 13 s for MACS-processed samples was a superior option compared with other thawing procedures; there was a significant difference in P < 0.05 values for curvilinear velocity (VCL μm/s) and sperm straightness (STR %) when samples were thawed at 40°C for 7 s, with fewer capacitated spermatozoa (P < 0.05) when samples were thawed at 37°C for 30 s, 40°C for 7 s or 60°C for 6 s. Hence, we can speculate that the use of AnMACS as the sperm preparation technique can somehow enhance sperm cryosurvival rate after cryopreservation, however the fertilization potential of these cells has yet to be determined.

Matched MeSH terms: Sperm Motility*
Causes and consequences of sperm mitochondrial dysfunction

Durairajanayagam D, Singh D, Agarwal A, Henkel R

Andrologia, 2021 Feb;53(1):e13666.
PMID: 32510691 DOI: 10.1111/and.13666

Mitochondria have multiple functions, including synthesis of adenine triphosphate, production of reactive oxygen species, calcium signalling, thermogenesis and apoptosis. Mitochondria have a significant contribution in regulating the various physiological aspects of reproductive function, from spermatogenesis up to fertilisation. Mitochondrial functionality and intact mitochondrial membrane potential are a pre-requisite for sperm motility, hyperactivation, capacitation, acrosin activity, acrosome reaction and DNA integrity. Optimal mitochondrial activity is therefore crucial for human sperm function and semen quality. However, the precise role of mitochondria in spermatozoa remains to be fully explored. Defects in sperm mitochondrial function severely impair the maintenance of energy production required for sperm motility and may be an underlying cause of asthenozoospermia. Sperm mtDNA is susceptible to oxidative damage and mutations that could compromise sperm function leading to infertility. Males with abnormal semen parameters have increased mtDNA copy number and reduced mtDNA integrity. This review discusses the role of mitochondria in sperm function, along with the causes and impact of its dysfunction on male fertility. Greater understanding of sperm mitochondrial function and its correlation with sperm quality could provide further insights into their contribution in the assessment of the infertile male.

Matched MeSH terms: Sperm Motility*
Fulltext Rapid sperm capture: high-throughput flagellar waveform analysis

Gallagher MT, Cupples G, Ooi EH, Kirkman-Brown JC, Smith DJ

Hum Reprod, 2019 07 08;34(7):1173-1185.
PMID: 31170729 DOI: 10.1093/humrep/dez056

STUDY QUESTION: Can flagellar analyses be scaled up to provide automated tracking of motile sperm, and does knowledge of the flagellar waveform provide new insight not provided by routine head tracking?
SUMMARY ANSWER: High-throughput flagellar waveform tracking and analysis enable measurement of experimentally intractable quantities such as energy dissipation, disturbance of the surrounding medium and viscous stresses, which are not possible by tracking the sperm head alone.
WHAT IS KNOWN ALREADY: The clinical gold standard for sperm motility analysis comprises a manual analysis by a trained professional, with existing automated sperm diagnostics [computer-aided sperm analysis (CASA)] relying on tracking the sperm head and extrapolating measures. It is not currently possible with either of these approaches to track the sperm flagellar waveform for large numbers of cells in order to unlock the potential wealth of information enclosed within.
STUDY DESIGN, SIZE, DURATION: The software tool in this manuscript has been developed to enable high-throughput, repeatable, accurate and verifiable analysis of the sperm flagellar beat.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Using the software tool [Flagellar Analysis and Sperm Tracking (FAST)] described in this manuscript, we have analysed 176 experimental microscopy videos and have tracked the head and flagellum of 205 progressive cells in diluted semen (DSM), 119 progressive cells in a high-viscosity medium (HVM) and 42 stuck cells in a low-viscosity medium. Unscreened donors were recruited at Birmingham Women's and Children's NHS Foundation Trust after giving informed consent.
MAIN RESULTS AND THE ROLE OF CHANCE: We describe fully automated tracking and analysis of flagellar movement for large cell numbers. The analysis is demonstrated on freely motile cells in low- and high-viscosity fluids and validated on published data of tethered cells undergoing pharmacological hyperactivation. Direct analysis of the flagellar beat reveals that the CASA measure 'beat cross frequency' does not measure beat frequency; attempting to fit a straight line between the two measures gives ${\mathrm{R}}^2$ values of 0.042 and 0.00054 for cells in DSM and HVM, respectively. A new measurement, track centroid speed, is validated as an accurate differentiator of progressive motility. Coupled with fluid mechanics codes, waveform data enable extraction of experimentally intractable quantities such as energy dissipation, disturbance of the surrounding medium and viscous stresses. We provide a powerful and accessible research tool, enabling connection of the mechanical activity of the sperm to its motility and effect on its environment.
LARGE SCALE DATA: The FAST software package and all documentation can be downloaded from www.flagellarCapture.com.
LIMITATIONS, REASONS FOR CAUTION: The FAST software package has only been tested for use with negative phase contrast microscopy. Other imaging modalities, with bright cells on a dark background, have not been tested but may work. FAST is not designed to analyse raw semen; it is specifically for precise analysis of flagellar kinematics, as that is the promising area for computer use. Flagellar capture will always require that cells are at a dilution where their paths do not frequently cross.
WIDER IMPLICATIONS OF THE FINDINGS: Combining tracked flagella with mathematical modelling has the potential to reveal new mechanistic insight. By providing the capability as a free-to-use software package, we hope that this ability to accurately quantify the flagellar waveform in large populations of motile cells will enable an abundant array of diagnostic, toxicological and therapeutic possibilities, as well as creating new opportunities for assessing and treating male subfertility.
STUDY FUNDING/COMPETING INTEREST(S): M.T.G., G.C., J.C.K-B. and D.J.S. gratefully acknowledge funding from the Engineering and Physical Sciences Research Council, Healthcare Technologies Challenge Award (Rapid Sperm Capture EP/N021096/1). J.C.K-B. is funded by a National Institute of Health Research (NIHR) and Health Education England, Senior Clinical Lectureship Grant: The role of the human sperm in healthy live birth (NIHRDH-HCS SCL-2014-05-001). This article presents independent research funded in part by the NIHR and Health Education England. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. The data for experimental set (2) were funded through a Wellcome Trust-University of Birmingham Value in People Fellowship Bridging Award (E.H.O.).The authors declare no competing interests.

Matched MeSH terms: Sperm Motility*
Fulltext Microtubular Dysfunction and Male Infertility

Gunes S, Sengupta P, Henkel R, Alguraigari A, Sinigaglia MM, Kayal M, et al.

World J Mens Health, 2020 Jan;38(1):9-23.
PMID: 30350487 DOI: 10.5534/wjmh.180066

Microtubules are the prime component of the cytoskeleton along with microfilaments. Being vital for organelle transport and cellular divisions during spermatogenesis and sperm motility process, microtubules ascertain functional capacity of sperm. Also, microtubule based structures such as axoneme and manchette are crucial for sperm head and tail formation. This review (a) presents a concise, yet detailed structural overview of the microtubules, (b) analyses the role of microtubule structures in various male reproductive functions, and (c) presents the association of microtubular dysfunctions with male infertility. Considering the immense importance of microtubule structures in the formation and maintenance of physiological functions of sperm cells, this review serves as a scientific trigger in stimulating further male infertility research in this direction.

Matched MeSH terms: Sperm Motility
Aniline blue test for sperm protamination-a valuable add-on to the bull breeding soundness evaluation

Alfadel F, Yimer N, Hiew MWH

Trop Anim Health Prod, 2023 Feb 10;55(2):76.
PMID: 36764981 DOI: 10.1007/s11250-023-03490-x

Bull breeding soundness evaluation (BBSE) is the most common procedure used to predict bull potential fertility. However, the use of traditional methods for semen evaluation can affect its reliability. The inclusion of additional advanced test in BBSE may increase its accuracy. This study aimed to investigate the correlation between the degree of sperm protamination and BBSE main parameters of scrotal circumference (SC), progressive motility (PM), morphologically normal sperm (NS), and different categories of morphological defects. In addition, to determine the correlation between the three methods used for protamine assessment, five Brangus bulls were subjected to the BBSE. Semen samples were collected via electro-ejaculation and evaluated using traditional methods. Three different methods were used to determine the degree of sperm protamination: aniline blue (AB) staining, chromomycin A3 staining with fluorescent microscope (CMA3-FLM), and CMA3 with flow cytometry (CMA3-FCM). Sperm protamine deficiency assessed using the three methods exhibited significant differences among bulls according to their classification by BBSE, and showed significant negative correlation with semen quality parameters of NS and PM. A significant positive correlation was found between AB positivity and morphological abnormalities. The three methods used for protamine assessment also revealed significant positive correlations. Among the three tests, AB staining was the cheapest and easiest test that offers an objective assessment method for sperm protamination. Hence, it can be concluded that the assessment of protamination using AB staining test might serve as an additional valuable parameter or a replacement whenever detail sperm motility and morphology analyses in conducting BBSE to predict bull fertility are not possible.

Matched MeSH terms: Sperm Motility
Effect of docosahexanoic acid on quality of frozen-thawed bull semen in BioXcell extender

Kaka A, Haron W, Yusoff R, Yimer N, Khumran AM, Sarsaifi K, et al.

Reprod Fertil Dev, 2017 Mar;29(3):490-495.
PMID: 28442061 DOI: 10.1071/RD15089

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen-thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15ngmL(-1) DHA was added. The supplemented semen samples were incubated at 37°C for 15min for DHA uptake by spermatozoa. Later, samples were cooled for 2h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3ngmL(-1) significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3ngmL(-1) concentration of DHA resulted in superior quality of frozen-thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

Matched MeSH terms: Sperm Motility
Antispermatogenic and antifertility effect of Pegaga (Centella asiatica L) on the testis of male Sprague-Dawley rats

Yunianto I, Das S, Mat Noor M

Clin Ter, 2010;161(3):235-9.
PMID: 20589353

Antifertility agents with safety and effectiveness in terms of minimum side effects have always been a subject of debate. Many studies have been conducted on plants to observe the antifertility effect, but majority of them were toxic. Pegaga or Centella asiatica L. is one of the popular herb traditionally consumed raw amongst people in Malaysia. The main objective of the present study was to investigate the effects of Centella asiatica L. extract on rat testis.

Matched MeSH terms: Sperm Motility/drug effects*
Fulltext Predictors of intrauterine insemination success

Zainul MR, Ong FB, Omar MH, Ng SP, Nurshaireen A, Rahimah MD, et al.

Med J Malaysia, 2006 Dec;61(5):599-607.
PMID: 17623962 MyJurnal

Intrauterine insemination (IUI) remains a therapeutic option within means of the majority of infertile couples in Malaysia. Therefore additional information on predictors of IUI success in the local context would provide a more concrete basis for counseling patients on expectations and treatment options. A retrospective analysis of 297 couples who underwent 445 IUI cycles from Jan 2005-Mar 2006 was undertaken. Four fifths were Malay with a mean paternal and maternal age of 35.53 +/- 5.82 (range 24-59) and 33.02 +/- 4.69 (range 21-46) years respectively. Causes of infertility were idiopathic (50%), endometriosis (17%) and anovulation/polycystic ovarian syndrome (15%). Almost 10% were oligoastenoteratozoospermic with another 23% oligozoospermic or astenozoospermic. Combined male and female factors occurred in 26%. A pregnancy rate (PR) of 9.4% per cycle; 14.1% per couple with a cumulative PR of 36.7% per 4 cycles was achieved. Those who became pregnant were significantly younger (31.29 +/- 4.43 vs. 33.21 +/- 4.68 years, p = 0.011) and had more follicles (13.95 +/- 9.72 vs. 11.43 +/- 6.67, p = 0.029) at the time of insemination. PR depreciated with maternal age and semen quality. Maternal and paternal age was inversely correlated to the number of follicles recruited (r = -0.30, p < 0.0005) and progressive sperm motility (r = -0.125, p = 0.013) respectively.

Matched MeSH terms: Sperm Motility*
Aberrant Upregulation of Compensatory Redox Molecular Machines May Contribute to Sperm Dysfunction in Infertile Men with Unilateral Varicocele: A Proteomic Insight

Swain N, Samanta L, Agarwal A, Kumar S, Dixit A, Gopalan B, et al.

Antioxid Redox Signal, 2020 03 10;32(8):504-521.
PMID: 31691576 DOI: 10.1089/ars.2019.7828

Aims:
To understand the molecular pathways involved in oxidative stress (OS)-mediated sperm dysfunction against a hypoxic and hyperthermic microenvironment backdrop of varicocele through a proteomic approach.
Results:
Protein selection (261) based on their role in redox homeostasis and/or oxidative/hyperthermic/hypoxic stress response from the sperm proteome data set of unilateral varicocele (UV) in comparison with fertile control displayed 85 to be differentially expressed. Upregulation of cellular oxidant detoxification and glutathione and reduced nicotinamide adenine dinucleotide (NADH) metabolism accompanied with downregulation of protein folding, energy metabolism, and heat stress responses were observed in the UV group. Ingenuity pathway analysis (IPA) predicted suppression of oxidative phosphorylation (OXPHOS) (validated by Western blotting [WB]) along with augmentation in OS and mitochondrial dysfunction in UV. The top affected networks indicated by IPA involved heat shock proteins (HSPs: HSPA2 and HSP90B1). Their expression profile was corroborated by immunocytochemistry and WB. Hypoxia-inducible factor 1A as an upstream regulator of HSPs was predicted by MetaCore. Occurrence of reductive stress in UV spermatozoa was corroborated by thiol redox status.
Innovation:
This is the first evidence of a novel pathway showing aberrant redox homeostasis against chronic hypoxic insult in varicocele leading to sperm dysfunction.
Conclusions:
Upregulation of antioxidant system and dysfunctional OXPHOS would have shifted the redox balance of biological redox couples (GSH/GSSG, NAD+/NADH, and NADP+/NADPH) to a more reducing state leading to reductive stress. Chronic reductive stress-induced OS may be involved in sperm dysfunction in infertile men with UV, where the role of HSPs cannot be ignored. Intervention with antioxidant therapy warrants proper prior investigation.

Matched MeSH terms: Sperm Motility/physiology
Comparative characteristics of spermatozoa harvested and cryopreserved in culture and cryoprotectant media with or without donor serum proteins

Ata'Allah GA, Adenan NAM, Razali N, Palaniappan K, Saad R, Idris SK, et al.

Reprod Biol, 2017 Jun;17(2):172-179.
PMID: 28511996 DOI: 10.1016/j.repbio.2017.04.004

The objectives of this study is to evaluate the efficacy of protein-free media in the preparation, holding and crypreservation of spermatazoa for use in ART. Normozoospermic semen samples (N=71) were used to compare the effects of media on the survival and quality of spermatozoa when washed and cultured with different media with and without added proteins at 4°C, 15°C, 22°C and 37°C for 0, 4-7 and 24h. Survival and quality of spermatozoa were assessed after freeze-thaw with synthetic cryoprotectant with and without proteins. Ethics/IRB approval was obtained (Ref. 1073.52). Spermatozoa parameters were similar in all media after washing and culture for 24h. Post-thaw survival and quality of spermatozoa was not significantly different 24h after thawing of samples frozen in all cryoprotectant medium. In conclusion synthetic protein-free culture and cryoprotectant media are equal in efficacy to protein-containing media in culture and cryopreservation of spermatozoa . Use of these synthetic media are anticipated to significantly reduce the risk, potentially associated with conventional protein-containing media, of transmission of disease and possibly harmful undeclared proteins to the patient, baby and the healtcare worker. Synthetic media also ensure consistency of quality between batches of media.

Matched MeSH terms: Sperm Motility*
Successful sperm cryopreservation of the brown-marbled grouper, Epinephelus fuscoguttatus using propylene glycol as cryoprotectant

Yusoff M, Hassan BN, Ikhwanuddin M, Sheriff SM, Hashim F, Mustafa S, et al.

Cryobiology, 2018 04;81:168-173.
PMID: 29355519 DOI: 10.1016/j.cryobiol.2018.01.005

This study developed the cryopreservation of brown-marbled grouper spermatozoa for practical application. We examined 32 cryodiluents, developed from four types of cryoprotectants [propylene glycol (PG), dimethyl-sulphoxide (Me2SO), dimethyl-acetamide (DMA) and ethylene glycol (EG)] at four concentrations of 5, 10, 15 and 20% in combination with two extenders [Fetal bovine serum (FBS) and artificial seminal plasma (ASP). Cooling rates were examined by adjusting the height of straws (2.5-12.5 cm) from the liquid nitrogen (LN) vapor and cooled for 5 min before immersion into LN. DNA laddering was used to detect DNA damage in cryopreserved sperm. In fertilization trials, 0.5 g of eggs was mixed with cryopreserved sperm stored for 30 days in LN. The best motility of post-thaw sperm was achieved using 15% PG + 85% FBS (76.7 ± 8.8%); 10% PG + 90% FBS was also effective as cryodiluent. Generally, FBS gave better post-thaw motility compared to ASP. The optimum cooling rate was at 17.6 °C min-1 obtained by freezing at the height of 7.5 cm surface of LN. The results obtained showed that cryopreserved sperm of brown-marbled grouper suffered slight DNA fragmentation, which resulted in significantly lower motility. However, the fertilization (90.9 ± 0.5%), hatching (64.5 ± 4.1%) and deformity rates (3.8 ± 0.2%) obtained from cryopreserved sperm showed no significant difference with fresh sperm. These findings show that the developed protocol for cryopreservation of brown-marbled grouper sperm was viable and will be useful for successful breeding and seed production of brown-marbled grouper.

Matched MeSH terms: Sperm Motility/drug effects
Fulltext Tert-butylhydroquinone attenuates doxorubicin-induced dysregulation of testicular cytoprotective and steroidogenic genes, and improves spermatogenesis in rats

Ujah GA, Nna VU, Suleiman JB, Eleazu C, Nwokocha C, Rebene JA, et al.

Sci Rep, 2021 Mar 09;11(1):5522.
PMID: 33750916 DOI: 10.1038/s41598-021-85026-7

Doxorubicin (DOX) is a broad-spectrum chemotherapeutic drug used in the treatment of cancers. It acts by generating reactive oxygen species in target cells. The actions are, however, not limited to cancerous cells as it attacks healthy cells, killing them. This study investigated the benefits of the antioxidant, tert-butylhydroquinone (tBHQ), on testicular toxicity following DOX therapy. Twenty-four adult male albino rats were assigned randomly into four groups (n = 6), namely: normal control (NC), tBHQ, DOX and tBHQ + DOX groups. tBHQ (50 mg/kg body weight in 1% DMSO) was administered orally for 14 consecutive days, while a single DOX dose (7 mg/kg body weight) was administered intraperitoneally on Day 8. DOX decreased sperm count, motility and viability, and decreased the levels of steroidogenesis-related proteins, and reproductive hormones. Furthermore, DOX decreased the expression of antioxidant cytoprotective genes, and decreased the protein level of proliferating cell nuclear antigen in the testis. Conversely, DOX increased the expression of pro-inflammatory and pro-apoptotic genes in the testis. These negative effects were ameliorated following the intervention with tBHQ. Our results suggest that tBHQ protects the testis and preserves both steroidogenesis and spermatogenesis in DOX-treated rats through the suppression of oxidative stress, inflammation and apoptosis.

Matched MeSH terms: Sperm Motility/drug effects*
Sperm parameters quality and reproductive effects of methanolic extract of Alchornea cordifolia leaves on senescent male rats

Ngaha Njila MI, Massoma Lembè D, Koloko BL, Yong Meng G, Ebrahimi M, Awad EA, et al.

Andrologia, 2019 Oct;51(9):e13359.
PMID: 31353623 DOI: 10.1111/and.13359

The effect of the methanolic extract of Alchornea cordifolia leaves on the fertility of senescent male rats was assessed in this study. 40 rats received daily distilled water, testosterone, 200 and 400 mg/kg of extract of Alchornea cordifolia. The reproductive organs weight, the gonadotropins, testosterone and cholesterol level, the sperm parameters, histology of the testes and epididymis were assessed. The weight of testes and prostate (400 mg/kg) significantly increased (p sperm count and morphology significantly increased at both doses of 200 mg/kg (p sperm motion (PROG, VAP, VSL, VCL) (p

Matched MeSH terms: Sperm Motility/drug effects; Sperm Motility/physiology
The use of the semen analysis in predicting fertility outcome

Arumugam K, Omar SZ

Aust N Z J Obstet Gynaecol, 1992 May;32(2):154-7.
PMID: 1520202

The study investigates the use of the various parameters of the semen analysis in predicting the fertility outcome in 82 infertile couples. The sperm density, % progressive motility, % normal morphology were divided into 'normal' and 'abnormal' based on the criteria proposed by WHO. The subsequent cumulative pregnancy rates were then calculated according to this criteria. A life-table method of analysis was used. All female related fertility factors were excluded. With the exception of a sperm density of less than 20 x 10(6) per ml the other parameters showed no significant correlation with the cumulative pregnancy rates at 12 months or 24 months respectively. We concluded that the semen analysis does not predict the probable outcome of the subsequent rates even when female fertility related factors were excluded apart from a sperm density less than 20 x 10(6) per ml.

Matched MeSH terms: Sperm Motility
Impact of Coenzyme Q10 and Selenium on Seminal Fluid Parameters and Antioxidant Status in Men with Idiopathic Infertility

Alahmar AT, Sengupta P

Biol Trace Elem Res, 2021 Apr;199(4):1246-1252.
PMID: 32572802 DOI: 10.1007/s12011-020-02251-3

Oxidative stress (OS) is a key contributing factor in 30-80% of male infertility cases. To date, several antioxidant treatments have been put forth to manage OS-induced male infertility. This study intended to elucidate the impact of coenzyme Q10 (CoQ10) and selenium on seminal fluid parameters and antioxidant status in infertile men with idiopathic oligoasthenoteratospermia (OAT). In this prospective study, 70 patients with idiopathic OAT were randomly allocated to receive CoQ10 (200 mg/day) or selenium (200 μg/day) for 3 months. Semen quality parameters (following WHO guidelines, 5th edition), total antioxidant capacity (TAC), catalase (CAT), and superoxide dismutase (SOD) activities were compared before and after the treatment. The results of the study showed an increase in sperm concentration with CoQ10 treatment (p sperm motility (p sperm motility (p Sperm concentration, motility, and morphology also correlated significantly with TAC, SOD, and CAT (r = 0.37-0.76). In conclusion, treatment with CoQ10 (200 mg) or selenium (200 μg) could improve sperm concentration, motility, and antioxidant status in infertile men with idiopathic OAT with CoQ10 providing the higher improvement.

Matched MeSH terms: Sperm Motility
Improvement of sperm quality of the depik fish, rasbora tawarensis, after cryopreservation using antioxidant

Muchlisin ZA, Afriani D, Eriani K, Hasri I, Nur FM, Maulida S, et al.

Cryo Letters, 2022;44(1):13-19.
PMID: 36625871

BACKGROUND: The cryopreservation of the sperm of the depik fish, Rasbora tawarensis, has previously been developed. However, the quality of the sperm post cryopreservation was not satisfactory and might be improved through the application of antioxidants.
OBJECTIVE: To determine the most suitable antioxidant for the cryopreservation of the depik fish spermatozoa.
MATERIALS AND METHODS: A completely randomized design with a non-factorial experiment was used and the tested antioxidants were glutathione, beta-carotene, ascorbic acid, and butylated hydroxytoluene (BHT) at 6 % concentrations. All treatments had three replications. The sperms were collected from 10 male fishes and diluted with Ringer solution in a ratio of 1: 20 (v/v, sperm: Ringer solution). Then 5% DMSO and 5 % egg yolk were added to the diluted sperms. Furthermore, 6 % of the tested antioxidants were added to the diluents, and then, cryopreservation was carried out in liquid nitrogen for 14 days.
RESULTS: The ANOVA test showed that the application of antioxidants significantly affected the sperm motility, fertility, and hatching rates of the eggs (P < 0.05). Furthermore, the antioxidants also protected the sperm cells during cryopreservation, with glutathione being the best antioxidant.
CONCLUSION: The application of antioxidants during the cryopreservation of depik fish sperm had a significant effect on motility, fertility and hatchability of eggs post-cryo. Furthermore, glutathione was the most suitable antioxidant. doi.org/10.54680/fr23110110312.

Matched MeSH terms: Sperm Motility
Improvement of sperm quality of the depik fish, rasbora tawarensis, after cryopreservation using antioxidant

Muchlisin ZA, Afriani D, Eriani K, Hasri I, Nur FM, Maulida S, et al.

Cryo Letters, 2023;44(1):13-19.
PMID: 36629837

BACKGROUND: The cryopreservation of the sperm of the depik fish, Rasbora tawarensis, has previously been developed. However, the quality of the sperm post cryopreservation was not satisfactory and might be improved through the application of antioxidants.
OBJECTIVE: To determine the most suitable antioxidant for the cryopreservation of the depik fish spermatozoa.
MATERIALS AND METHODS: A completely randomized design with a non-factorial experiment was used and the tested antioxidants were glutathione, beta-carotene, ascorbic acid, and butylated hydroxytoluene (BHT) at 6 % concentrations. All treatments had three replications. The sperms were collected from 10 male fishes and diluted with Ringer solution in a ratio of 1: 20 (v/v, sperm: Ringer solution). Then 5% DMSO and 5 % egg yolk were added to the diluted sperms. Furthermore, 6 % of the tested antioxidants were added to the diluents, and then, cryopreservation was carried out in liquid nitrogen for 14 days.
RESULTS: The ANOVA test showed that the application of antioxidants significantly affected the sperm motility, fertility, and hatching rates of the eggs (P < 0.05). Furthermore, the antioxidants also protected the sperm cells during cryopreservation, with glutathione being the best antioxidant.
CONCLUSION: The application of antioxidants during the cryopreservation of depik fish sperm had a significant effect on motility, fertility and hatchability of eggs post-cryo. Furthermore, glutathione was the most suitable antioxidant. doi.org/10.54680/fr23110110312.

Matched MeSH terms: Sperm Motility
Deleterious effect of gestagens from wastewater effluent on fish reproduction in aquatic environment: A review

Jenila JS, Issac PK, Lam SS, Oviya JC, Jones S, Munusamy-Ramanujam G, et al.

Environ Res, 2023 Nov 01;236(Pt 2):116810.
PMID: 37532209 DOI: 10.1016/j.envres.2023.116810

Gestagens are common pollutants accumulated in the aquatic ecosystem. Gestagens are comprised of natural gestagens (i.e. progesterone) and synthetic gestagens (i.e. progestins). The major contributors of gestagens in the environment are paper plant mill effluent, wastewater treatment plants, discharge from pharmaceutical manufacturing, and livestock farming. Gestagens present in the aquatic environment interact with progesterone receptors and other steroid hormone receptors, negatively influencing fish reproduction, development, and behavior. In fish, the gonadotropin induces 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) production, an important steroid hormone involved in gametogenesis. DHP interacts with the membrane progestin receptor (mPR), which regulates sperm motility and oocyte maturation. Gestagens also interfere with the hypothalamic-pituitary-gonadal (HPG) axis, which results in altered hormone levels in fish. Moreover, recent studies showed that even at low concentrations exposure to gestagens can have detrimental effects on fish reproduction, including reduced egg production, masculinization, feminization in males, and altered sex ratio, raising concerns about their impact on the fish population. This review highlights the hormonal regulation of sperm motility, oocyte maturation, the concentration of environmental gestagens in the aquatic environment, and their detrimental effects on fish reproduction. However, the long-term and combined impacts of multiple gestagens, including their interactions with other pollutants on fish populations and ecosystems are not well understood. The lack of standardized regulations and monitoring protocols for gestagens pollution in wastewater effluent hampers effective control and management. Nonetheless, advancements in analytical techniques and biomonitoring methods provide potential solutions by enabling better detection and quantification of gestagens in aquatic ecosystems.

Matched MeSH terms: Sperm Motility
Motility, viability and fertility of goldfish Carassius auratus (Pisces: Cyprinidae) post short-term cryopreservation

Nurlaili N, Eriani K, Salma I, Maulida S, Rahayu SR, Handayani LS, et al.

Cryo Letters, 2023;44(3):169-177.
PMID: 37883170

BACKGRUND: Goldfish Carassius auratus is a popular ornamental fish extensively cultured worldwide. Sperm cryopreservation is a common fish breeding method that ensures sperm availability around the year. Studies on cryopreservation of goldfish sperm, especially on the suitability of cryoprotectant types and pre-freezing time, are scarcely available.
OBJECTIVE: To determine the most suitable type of cryoprotectant and pre-freezing for the successful cryopreservation of goldfish sperm.
MATERIALS AND METHODS: A completely randomized design with two factors was utilized in this study. The first factor is the type of cryoprotectants, which included methanol, ethanol, ethylene glycol, glycerol, and DMSO. The second is pre-freezing times of 10, 20, 30, and 40 min at each of the pre-freezing temperatures of 4 degree C, -10 degree C, and -79 degree C, meaning that the total times for the ramping down of temperature were 30, 60, 90 and 120 min, respectively. The Ringer solution and 10% egg yolk were used as extender and extracellular cryoprotectant. The sperm was stored at -179 degree C for 7 days.
RESULTS: The ANOVA test showed that cryoprotectants and pre-freezing significantly affected the motility, viability, and fertility of goldfish sperm after freezing in liquid nitrogen for 7 days (P<0.05). Furthermore, 10% DMSO combined with 15% egg yolk with an pre-freezing time of 20 min can maintain sperm motility, viability, and fertility higher than other treatments, by 79%, 80%, and 33%, respectively. The agarose gel electrophoresis showed no DNA fragmentation in all samples, including fresh sperm.
CONCLUSION: We conclude that 10% DMSO combined with 15% egg yolk and 20 min pre-freezing is the best treatment for goldfish sperm cryopreservation. DOI: 10.54680/fr23310110412.

Matched MeSH terms: Sperm Motility
Effect of butylated hydroxytoluene on cryopreservation of Boer goat semen in Tris egg yolk extender

Memon AA, Wahid H, Rosnina Y, Goh YM, Ebrahimi M, Nadia FM, et al.

Anim. Reprod. Sci., 2011 Nov;129(1-2):44-9.
PMID: 22024366 DOI: 10.1016/j.anireprosci.2011.10.004

The aim of this study was to determine the effect of butylated hydroxytoluene (BHT), a lipid-soluble anti-oxidant added in different concentrations to the Tris egg yolk extenders on semen cytological parameters pre freezing and post thawing (motility, morphology, viability, acrosome integrity and membrane integrity) of Boer goat spermatozoa. A total of 40 ejaculates from four Boer goat bucks were collected using an artificial vagina. Ten replicates of the ejaculates were diluted with a Tris egg yolk based extender which contained various concentrations (0.5mM, 1.0mM, 2.0mM and 3.0mM) of butylated hydroxytoluene while one sample was processed without supplementation of antioxidant and served as control. The diluted semen was cooled at 4°C and loaded into the straw and then stored in liquid nitrogen. It was evident that supplementation of BHT produces positive effect in terms of motility, membrane integrity and acrosome integrity in comparison with the control group in cooled and frozen Boer goat semen. Results showed significant differences in motility, membrane integrity, acrosome integrity and viability of cooled and frozen Boer goat spermatozoa at different concentrations. Motility, membrane integrity, acrosome integrity and viability was significantly higher in all treated groups than the control group (P<0.05) while there was no significant differences (P>0.05) in morphology trait between all group in cooled semen. However, improvement (P<0.05) was observed only in terms of the membrane integrity and acrosome integrity compared to the control and other treated groups in frozen semen. In conclusion, BHT can be used in cryopreservation of Boer goat semen in order to reduce the oxidative stress on spermatozoa.

Matched MeSH terms: Sperm Motility/physiology

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