scholarly journals Tropical summer induces DNA fragmentation in boar spermatozoa: implications for evaluating seasonal infertility

2019 ◽  
Vol 31 (3) ◽  
pp. 590 ◽  
Author(s):  
Santiago T. Peña Jr. ◽  
Felicity Stone ◽  
Bruce Gummow ◽  
Anthony J. Parker ◽  
Damien B. B. P. Paris

Summer infertility continues to undermine pig productivity, costing the pig industry millions in annual losses. The boar’s inefficient capacity to sweat, non-pendulous scrotum and the extensive use of European breeds in tropical conditions, can make the boar particularly vulnerable to the effects of heat stress; however, the link between summer heat stress and boar sperm DNA damage has not yet been demonstrated. Semen from five Large White boars was collected and evaluated during the early dry, late dry and peak wet seasons to determine the effect of seasonal heat stress on the quality and DNA integrity of boar spermatozoa. DNA damage in spermatozoa during the peak wet was 16-fold greater than during the early dry and nearly 9-fold greater than during the late dry season. Sperm concentration was 1.6-fold lower in the peak wet than early dry whereas no difference was found across several motility parameters as determined by computer-assisted sperm analysis. These results demonstrate that tropical summer (peak wet season) induces DNA damage and reduces concentration without depressing motility in boar spermatozoa, suggesting that traditional methods of evaluating sperm motility may not detect inherently compromised spermatozoa. Boar management strategies (such as antioxidant supplementation) need to be developed to specifically mitigate this problem.

Reproduction ◽  
2004 ◽  
Vol 128 (2) ◽  
pp. 171-179 ◽  
Author(s):  
Harald Schmidt ◽  
Günter Kamp

Hyperactivity, a form of sperm motility characterized by vigorous flagellar movements, has been proposed as essential for fertilization in mammals. The objective of the present study was to establish a method for inducing hyperactivityin vitroin boar spermatozoa and to define threshold values to differentiate between hyperactive and non-hyperactive spermatozoa by computer-assisted sperm analysis (CASA) as a prerequisite for analyzing the energy metabolism during hyperactivity. In TALP-HEPES medium, non-frozen boar spermatozoa were stimulated to hyperactivity by 50 μmol l−1Ca2+within 15 min at 37 °C if 5 μmol l−1of the Ca2+ionophore A23187 was present. If 25% seminal plasma was present, boar spermatozoa required higher Ca2+concentrations (about 700 μmol l−1) for hyperactivity. Under both conditions, immobilization and head-to-head agglutination were low so that hyperactive spermatozoa could be analyzed for at least 40 min. The transition from normal to hyperactive movement was characterized by an increase in flagellar beat angle from 49° ± 12° to 200° ± 36° (n= 32) and a decrease in flagellar curvature ratio from 0.89 ± 0.04 to 0.47 ± 0.11 (n= 32). For quantification of hyperactive boar sperm, kinematic parameters of hyperactive and non-hyperactive spermatozoa were measured by CASA and statistically evaluated (receiver operating characteristic (ROC) curve analysis). The threshold values of the following four parameters were well suited for differentiating between hyperactive and non-hyperactive boar spermatozoa (ROC curve analysis: >50% specificity at 100% sensitivity). Hyperactive boar spermatozoa showed mean lateral head displacement >3.5 μm, curvilinear velocity >97 μm s−1, linearity <32% and wobble <71%. According to this multiparametric definition, induction of hyperactivity increased significantly (P< 0.0001) the fraction of hyperactive spermatozoa in semen samples from 5.1 ± 4.3% (n= 13) to 48.3 ± 6.6% (n= 7) in the absence and to 44.2 ± 7.6% (n= 10) in the presence of 25% seminal plasma, while the overall percentage of motile spermatozoa did not change significantly.


2013 ◽  
Vol 25 (8) ◽  
pp. 1134 ◽  
Author(s):  
Aimee J. Silla

Anurans exhibit a greater reproductive diversity than any other vertebrate order. However, studies investigating the effects of the external fertilisation environment on fertilisation success are limited to aquatic-breeding species. This study investigated the effects of fertilisation medium osmolality, sperm concentration and short-term oocyte storage on fertilisation success in a terrestrial-breeding anuran, Pseudophryne guentheri. Split-clutch experimental designs were used to determine optimal fertilisation conditions. To determine the effect of short-term sperm storage, sperm viability was assessed using fluorescence microscopy and percentage sperm motility and velocity quantified with a computer-assisted sperm analysis system. Fertilisation success was highest in media ranging in osmolality from 25 mOsm kg–1 to 100 mOsm kg–1, representing a broader range and higher optimal osmolality than previously reported for aquatic breeders. High rates of fertilisation (>75%) were achieved in relatively low sperm concentrations (2.5 × 104 mL–1). Oocytes stored in isotonic solutions (200 mOsm kg–1) retained fertilisation capacity (32%) after 8 h of storage, while sperm suspensions maintained motility (≥26%) for 13 days. Additional studies on terrestrial-breeding anurans will be required to ascertain whether the optimal fertilisation conditions reported reflect adaptations to achieve fertilisation in a terrestrial environment.


2012 ◽  
Vol 24 (1) ◽  
pp. 213 ◽  
Author(s):  
C. A. Burroughs ◽  
K. M. Evans ◽  
R. W. Lenz ◽  
G. E. Seidel

We evaluated sex-sorting parameters and post-thaw motility for sperm stored with or without seminal plasma for 8 h before sorting. One first ejaculate was collected from each of 6 bulls routinely collected via artificial vagina; ejaculates contained at least 70% motile and 75% morphologically normal sperm and sperm concentrations ranged from 0.75 to 2.21 × 109 sperm mL–1. Ejaculates were divided into 2 samples and centrifuged at 1000 × g for 15 min. Seminal plasma from 1 sample was replaced with TALP (pH 7.4) to a sperm concentration of 1.4 × 109 sperm mL–1. The seminal plasma/sperm admixture of the other sample (control) was suspended to initial ejaculate sperm concentration. Both samples were stored for 8 h at 16°C before being subjected to standard sex-sorting procedures. Sperm were analyzed and bulk sorted on a MoFlo SX (XY Inc., Navasota, TX, USA) flow cytometer/cell sorter for percentage of live-oriented cells, percentage of membrane-impaired sperm (cell membranes permeable to red food colouring, which were discarded during sorting) and resolution between X- and Y-bearing sperm populations (peak to valley ratio). Sorted sperm were frozen according to standard procedures and post-thaw motility was determined immediately after thawing using computer-assisted sperm analysis. Treatments were compared using a paired t-test. Control sperm stored with seminal plasma resulted in a higher percentage of live-oriented cells (55%) versus those stored without seminal plasma (51%; P = 0.02). The percentage of membrane-impaired sperm was lower for control sperm (19%) than that of samples without seminal plasma (28%; P < 0.001). Resolution was greater for sperm stored without seminal plasma (34%) than for control sperm (10%; P = 0.04). Post-thaw, both total and progressively motile sperm were higher for samples without seminal plasma (63 and 53%, respectively) compared with those of the control samples (52 and 45%, respectively; P < 0.04). In conclusion, sperm stored for 8 h without seminal plasma had greater resolution between X- and Y-bearing populations and higher post-thaw motility than control sperm. However, these samples had a higher percentage of membrane-impaired sperm that were removed during sorting. Long-term storage of sperm in their seminal plasma before sex-sorting appears to be detrimental to post-sorting, post-thaw sperm motility.


2007 ◽  
Vol 19 (1) ◽  
pp. 280
Author(s):  
M. Sansegundo ◽  
J. C. Gardon ◽  
F. Garcia-Vazquez ◽  
J. Gadea ◽  
C. Matas

The motion ability of mammalian spermatozoa is acquired during their epididymal transit but observed only upon dilution with seminal plasma (SP) at the time of ejaculation (Yanahimachi 1994 in The Physiology of Reproduction, New York: Raven Press). The bicarbonate present in seminal plasma activates multiple sperm functions, some of which are essential for the initiation of motility. Sperm hyperactivity has been observed in vitro in various mammalian species, especially if capacitation of spermatozoa was induced with Ca2+ and bicarbonate media, such as TALP (Harrison et al. 1996 Mol. Reprod. Dev. 45, 378–391). Computer-assisted sperm analysis (CASA) is a tool for the objective assessment of sperm motility. The aim of this study was to determine if there are differences in motility parameters of ejaculated (EJ) and epididymal (EP) boar spermatozoa under different treatments. Ejaculated and epididymal sperm cells from 10 different boars in each group were used. The sperm treatments were: washed in Dulbecco&apos;s PBS supplemented with 0.1&percnt; BSA (PBS-BSA), washed on a Percoll gradient (PG), and unwashed (UW: Control); the sperm samples were incubated in TALP medium at 38.5&deg;C and 5&percnt; CO2 during the analysis. Motion parameters were determined using a computer-assisted sperm analysis (CASA) system. A 7-&micro;L drop of the sample was placed on a warmed (37&deg;C) slide. At least 4 fields per sample were evaluated, with a minimum of 100 spermatozoa counted per sub-sample. The CASA-derived motility characteristics studied were motility (MOT, &percnt;), progressive motility (PM, &percnt;), curvilinear velocity (VCL, &micro;m s&minus;1), straight-line velocity (VSL, &micro;m s&minus;1), average path velocity (VAP, &micro;m s&minus;1), linearity of the curvilinear trajectory (LIN, ratio of VSL/VCL, &percnt;), straightness (STR, ratio of VSL/VAP, &percnt;), amplitude of lateral head displacement (ALH, &micro;m), wobble of the curvilinear trajectory (WOB, ratio of VAP/VCL, &percnt;), and beat cross-frequency (BCF, Hz). Data were analyzed by ANOVA. If we evaluated all of the data together (EJ vs. EP), EP sperm after treatment showed a higher motility (PM: 38.20&percnt;; MOT: 74.23&percnt;) than EJ sperm (PM: 29.27&percnt;; MOT: 63.24&percnt;), and all of the motion parameters related to velocities and ALH were higher in EP (VCL: 86.02; VSL: 41; VAP: 57.94; ALH: 3.21) than in EJ (VCL: 69.70; VSL: 34.67; VAP: 48.16; ALH: 2.54). No differences were found for LIN, STR, WOB, and BCF. The treatments significantly affected the VCL and ALH, with lower values for the PG treatment. When VCL was lower and the VSL and VAP were similar, consequently the LIN and WOB were significantly higher for the PG group. STR also was higher for the PG group. In conclusion, when both groups of sperm were incubated in TALP medium, the EJ sperm showed a decrease in the majority of motion parameters when compared with EP sperm. This work was supported by MEC (AGL2006-03495/GAN) and Fundaci&oacute;n S&eacute;neca (03018/PI/05).


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nikolas Zuchowicz ◽  
Jonathan Daly ◽  
Jessica Bouwmeester ◽  
Claire Lager ◽  
E. Michael Henley ◽  
...  

AbstractThe declining reproductive viability of corals threatens their ability to adapt to changing ocean conditions. It is vital that we monitor this viability quantitatively and comparatively. Computer-assisted sperm analysis (CASA) systems offer in-depth analysis used regularly for domestic and wildlife species, but not yet for coral. This study proposes quality control procedures and CASA settings that are effective for coral sperm analysis. To resolve disparities between CASA measurements and evaluations by eye, two negative effects on motility had to be resolved, slide adhesion (procedural) and sperm dilution (biological). We showed that the addition of bovine serum albumin, or caffeine, or both to fresh sperm reduced adhesion in the CASA cassettes, improved motility and motile sperm concentration (P < 0.0001), yet these additions did not affect measurements of total sperm concentration. Diluting coral sperm reduced sperm motility (P = 0.039), especially from heat-stressed corals. We found CASA concentration counts comparable to haemocytometer and flow cytometer measures (P = 0.54). We also found that motile sperm per egg is a useful predictor of fertilisation success, using cryopreserved sperm. Standard measurements of coral reproductive characteristics inform our understanding of the impacts of climate change on reef populations; this study provides a benchmark to begin this comparative work.


2021 ◽  
Vol 2 ◽  
Author(s):  
Mathieu Bonneau ◽  
Nausicaa Poullet ◽  
David Beramice ◽  
Laurent Dantec ◽  
Laurianne Canario ◽  
...  

Behavior is a good indicator of animal welfare, especially in challenging environments. However, few studies have investigated how pig behavior changes during heat stress. The current study is a proof-of-concept using Convolutional Neural Network (CNN) models to monitor pig behavior in order to investigate the differences in behavioral response to heat stress of two contrasted breeds: Large White (LW), selected for high performance, and Creole (CR), adapted to tropical conditions. A total of 6 slaughter pigs (3 CR and 3 LW; 22 weeks of age) were monitored from 8:30 to 17:30 during 54 days. Two CNN architectures were used to detect the animal (Yolo v2) and to estimate animal's posture (GoogleNet). Pig postures estimated by the neural network showed that pigs spent more time lying on their side when temperature increased. When comparing the two breeds, as temperature increases, CR pigs spent more time lying on their side than LW pigs, suggesting that they use this posture to increase thermoregulation and dissipate heat more efficiently. This study demonstrates that neural network models are an efficient tool to monitor animal behavior in an automated way, which could be particularly relevant to characterize breed adaptation to challenging environments.


2008 ◽  
Vol 20 (1) ◽  
pp. 87
Author(s):  
C. O. Hidalgo ◽  
A. Rodríguez ◽  
C. Díez ◽  
D. Martín ◽  
M. Carbajo ◽  
...  

The Bermeya goats are an endangered autochthonous breed distributed in the north of Spain. To ensure their genetic diversity and long-term survival, morphological and functional parameters of the semen must be known in order to preserve the current genetic stock in a germplasm bank. The aim of this work was to establish basic characteristics and post-thaw survival of Bermeya goat's semen obtained by electro-ejaculation, that is not well described in the literature. The semen was collected by electro-ejaculation from 7 bucks, 1 to 7 years old, twice per week, for 9 weeks (n = 83). Fresh semen was evaluated for volume (V), concentration (C), motility, morphology, functional integrity of the sperm (spz) membranes (hypoosmotic swelling test; HOST), and acrosome integrity rate (NAR). Individual and progressive sperm motility were analyzed by means of a computer-assisted sperm analysis system (CASA: SCA 2002�, Microptic, Barcelona, Spain) immediately after dilution with the extender at 37�C, and after cooling to 4�C; five fields per sample (diluted to 204 � 106 spz mL–1) were evaluated under a phase contrast microscope (100�). The NAR and morphological abnormalities of sperm head, midpiece, tail, and cytoplasmic droplets were determined by counting 100 spz under 1000�. For freezing, ejaculates with at least 80% motile spz were diluted at 32�C with Krebs-Ringer solution containing 20% egg yolk and 14% glycerol to a final concentration of 400 � 106 spz mL–1, cooled to 4�C for 90 min, aspirated into 0.25-mL plastic straws (IMV�, L'Aigle, France), frozen at 7 cm above liquid nitrogen (LN2) phase for 10 min, and then plunged into the LN2. Straws were thawed in a water bath at 39�C for 30 s for post-thaw survival analysis. Data were analyzed by the GLM and FREQ procedures (SAS; SAS Institute, Inc., Cary, NC, USA) and expressed as means � standard error. Fresh semen characteristics were: V = 1.7 � 0.1 mL; C = 2619 � 106 � 153 spz mL–1; total and progressive motility were 89.0 � 2.1% and 66.9 � 2.1%, respectively. Percentages of head abnormalities were 4.8 � 0.5; midpiece: 3.8 � 0.7; tail: 4.7 � 1.0; cytoplasmic droplets: 8.3 � 0.7; intact acrosome: 91.8 � 0.6; and membrane integrity: 49.2 � 2.1. At 4�C, the % of total motile spz was 62.6 � 1.6, and the post-thaw survival rate was 46.3 � 1.5. There were only individual differences (P < 0.001) between bucks on sperm concentration, head abnormalities, and cytoplasmic droplets. In conclusion, our results indicate that semen quality is related to each individual animal and that electro-ejaculation allows collection of semen of satisfactory quality to use as fresh and for cryopreservation. However, the validity of our results for possible future sperm banking of endangered Bermeya goats semen must be confirmed by field trials.


2008 ◽  
Vol 20 (1) ◽  
pp. 124
Author(s):  
M. L. Perals ◽  
M. A. Gil ◽  
E. M. Garcia ◽  
J. Sanchez-Osorio ◽  
J. M. Vázquez ◽  
...  

Boars can be classified as good or bad sperm freezers according to their sperm cryosurvival. Different sperm selection techniques, such as PureSperm� (PS; MidAtlantic Diagnostics, Inc., Mount Laurel, NJ, USA), have been developed to improve functional competence of spermatozoa. The aim of this experimental study was to assess the ability of PS for improving the in vitro penetrating ability of frozen–thawed boar spermatozoa from good and bad sperm freezers. The sperm-rich fractions from two boars, good (Boar A) and bad (Boar B) freezers, were extended in a lactose/eggyolk/ glycerol/Equex Stem (Noba Chemical Sales, Inc., Scituate, ME, USA) mixture (1 � 109 sperm mL–1), dispensed into 0.5-mL straws, and frozen using a programmable cell freezer. After thawing (1.200�C min–1), semen from each boar was split into two aliquots of 500 µL. One aliquot was used as the control. The second was placed into a tube of PS gradient (90%/45%) and centrifuged at 425g for 20 min; the pellet re-suspended in 1 mL of BTS and re-centrifuged at 320g for 10 min (PS sample). Control and PS samples were diluted in supplemented TCM-199 (TCMm; Roca et al. 1998 Reprod. Fertil. Dev. 10, 479–485) at 200 � 106 sperm mL–1. Sperm survival (SV) was assessed afterTCMm dilution according to progressive sperm motility (PSM, %) using a computer-assisted sperm analysis (CASA) system (ISAS�), and plasma and acrosome membrane integrity (PMI; %) by flow cytometry (SYBR�-14/PE-PNA/PI; Molecular Probes, Leiden, The Netherlands). A homologous in vitro penetration (hIVP) assay, using immature oocytes (20 oocytes/2 mL TCMm supplemented with caffeine and calcium lactate), was used to assess sperm penetrating ability (Martinez et al. 1993 Theriogenology 40, 547–557). A total of 960 immature oocytes were inseminated (200 � 103 sperm/oocyte) in 3 batches. After 18 h of co-incubation at 39�C under 5% CO2 in air, the oocytes were washed, mounted on slides, fixed with ethanol:acetic acid (3:1, v/v) for 48 h, stained with 1% lacmoid, and examined under a phase contrast microscope (�400). Oocytes with swollen or unswollen heads of sperm found in the vitellus were considered as penetrated. Sperm penetrability ability (SPA) was assessed according to penetration rate (PR) and the mean number of sperm per oocyte (S/O). Data were analyzed using a PROMIXED model and expressed as mean � SEM. Boar A showed better (P ≤ 0.01) results for both SV and SPA parameters than boar B, independent of sperm treatment. PureSperm improved (P ≤ 0.05) PSM and PMI in both boar A (control v. PS: 48.0 � 5.8 v. 66.5 � 3.6 and 63.1 � 7.7 v. 88.4 � 1.3, respectively) and boar B (12.3 � 1.2 v. 22.2 � 3.7 and 44.3 � 3.5 v. 58.7 � 7.0, respectively). However, no differences (P ≥ 0.05) were observed in PR and S/O in either boar A (71.2 � 3.4 v. 78.3 � 3.1 and 5.0 � 0.4 v. 5.2 � 0.4, respectively) or boar B (34.3 � 3.6 v. 37.3 � 3.9 and 1.5 � 0.1 v. 1.5 � 0.1, respectively). In conclusion, under our laboratory conditions, PureSperm selection improves sperm quality but not in vitro penetrating ability of frozen–thawed spermatozoa of both good and bad sperm freezers. This work was supported by CICYT (AGF2005-00760), Madrid, Spain.


2020 ◽  
Vol 4 (1) ◽  
pp. 293-298
Author(s):  
Tasha R Gruhot ◽  
Lea A Rempel ◽  
Brett R White ◽  
Benny E Mote

Abstract Semen quality has a dramatic impact on reproductive efficiency in the swine industry, influencing both conception rate and litter size. The objective of this study was to assess whether the presence of varicocele hinders semen quality in both thermoneutral and heat stress (HS) conditions. At approximately 6 mo of age, ultrasonography was used to measure left and right pampiniform plexus area in order to detect varicocele in maternal line boars at the University of Nebraska–Lincoln. Between 10 and 12 mo of age, semen was collected from each boar (n = 28) twice weekly. Boars were collected under thermoneutral conditions, were then heat stressed for 7 d to exacerbate any semen quality issues, and semen was collected post-HS for 6 wk. Sperm characteristics were determined by computer-assisted semen analysis. The presence of varicocele had a significant effect on sperm concentration (P = 0.04) and trended toward significance for mean sperm head area (P = 0.06) throughout the duration of the study. An interaction existed between varicocele and collection time point at weeks 2–5 post-HS for distal droplet percentage, suggesting that boars with varicocele were possibly more susceptible to heat-stress-induced semen quality issues than boars without varicocele. Moreover, semen quality was reduced in boars with versus without varicocele under both thermoneutral and HS conditions. Therefore, detection of varicocele by ultrasound could represent a potential marker of fertility in young boars or as a component trait in selection indices for fertility.


2013 ◽  
Vol 57 (3) ◽  
pp. 429-432 ◽  
Author(s):  
Anna Domosławska ◽  
Sławomir Zduńczyk ◽  
Wojciech Niżański ◽  
Tomasz Janowski

Abstract Semen quality parameters of infertile and fertile dogs were compared. Sperm concentration and semen motility parameters were measured by the Hamilton-Thorne Semen Analyser IVOS 12.3. The spermatozoal morphology and the percentage of live spermatozoa were examined microscopically. Forty-six dogs of various breeds were examined. Twenty dogs had a conception failure within last year. These dogs had a history of prior normal fertility. Twenty six fertile dogs served as control. All animals underwent a clinical examination as well as ultrasonography. Sperm concentration was significantly lower in infertile dogs than in fertile dogs. For most determined motility parameters (MOT, PMOT, VAP, VSL, VCL, BCF, RAPID, STATIC) significant differences between infertile and fertile dogs were found. The percentage of spermatozoa with normal morphology also significantly differed between both groups. Ultrasonography of the prostate gland and testes revealed no pathological conditions. The testicular degeneration was assumed to be a possible cause of infertility in these dogs. The present study showed that the most sperm CASA motility parameters were significantly lower in infertile dogs in comparison to the fertile ones, and confirmed the usefulness of the Hamilton-Thorne Semen Analyser for a quick and objective analysis of sperm concentration and motility in dogs.


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