268 IN VITRO EVALUATION OF FRESH SPERM QUALITY IN TOMCATS: A COMPARISON OF COLLECTION TECHNIQUES

2010 ◽  
Vol 22 (1) ◽  
pp. 291
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
P. Anastasi ◽  
M. Hoogewijs ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Sperm Head ◽  
Lower Percentage ◽  
Rank Test ◽  
Computer Assisted ◽  
Urethral Catheterization ◽  
Group 2 ◽  
Group 1

Semen can be collected from tomcats by urethral catheterization (CT) after medetomidine administration, offering a novel approach to obtain sperm for in vitro fertilization. This study was designed to determine motion characteristics of CT sperm samples by means of computer-assisted sperm analysis (CASA) and to compare sperm quality parameters and in vitro fertilizing capacity of CT spermatozoa with those of spermatozoa retrieved after epididymal slicing (EP). Semen was collected in 17 adult cats by urethral catheterization as reported by Zambelli et al. (2008 Theriogenology 69, 485-490), after which the cat was orchidectomized. Motility (subjective and objective by means of CASA), morphology [eosin/nigrosin (E/N)], plasma membrane integrity (E/N and SYBR14-PI staining), and acrosomal status (FITC-PSA staining) of fresh CT and EP samples were evaluated and compared between both methods with a paired t-test or the Wilcoxon Rank test, dependent on normality of the data. In vitro maturation (24 h), fertilization (18 h), and culture (6h) of grade I to III oocytes were carried out as described by Pope et al. (2009 Theriogenology 71, 864-871). Twenty-four hours after in vitro insemination, fertilization rates were assessed for group 1 (CT; n = 148) and group 2 (EP; n = 159) presumptive zygotes. The distribution of presumptive zygotes between CT and EP over the different developmental stages was compared using Pearson chi-square test. Results showed that total and progressive motility as well as the percentage of normal spermatozoa were higher for EP sperm compared with CT sperm (P < 0.01). Epididymal sperm had a lower percentage of spermatozoa with an intact acrosome (P < 0.01), whereas CT sperm contained more spermatozoa with tail abnormalities (P < 0.01). Other sperm parameters did not differ (P > 0.05) between collection techniques. In group 1, 84% of in vitro-matured oocytes (metaphase II) were penetrated (40.2% cleaved, 24.4% with 2 pronuclei, 12.2% with >2 pronuclei, 7.3% with expanded sperm head), whereas in group 2, penetration rate was 88.5% (42.7% cleaved, 21.8% with 2 pronuclei, 16.7% with >2 pronuclei, 7.3% with expanded sperm head). No difference (P > 0.05) in in vitro fertilizing capacity between spermatozoa collected by means of the 2 methods was found. In conclusion, semen collection by means of CT yields fertilization results similar to epididymal slicing, despite the fact that several sperm variables were different. Because CT is repeatable and easy to perform and does not require a trained male/queen in heat, it may be preferable for routine IVF experiments with fresh spermatozoa. The first author is a research fellow of the Fund for Scientific Research-Flanders (Belgium).

162 EFFECT OF THE SPERM SELECTION WITH ISOLATE® OR PERCOLL® ON SPERM QUALITY AND IN VITRO BOVINE EMBRYO DEVELOPMENT FOR FROZEN - THAWED SEXED AND NONSEXED SEMEN

2012 ◽  
Vol 24 (1) ◽  
pp. 193
Author(s):  
G. A. Bó ◽  
P. Rodriguez Villamil ◽  
G. Moreira ◽  
M. E. Garcia Gomez ◽  
M. Fernandez Taranco ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Sperm Selection ◽  
Production Rates ◽  
Group 4 ◽  
Sexed Semen ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

The aim of this study was to compare the effects of 2 different commercially available spermatozoa separation techniques, Isolate® (Irving-Scientific, Santa Ana, CA, USA) and Percoll® (Nutricell, São Paulo, Brazil), on sperm quality and in vitro embryo production using sexed and nonsexed semen. Oocytes (n = 5046) were obtained from slaughterhouse ovaries and fertilized with frozen-thawed sexed or nonsexed semen from the same 4 Holstein bulls. The experiment was done in 10 replicates, with all treatment groups included. Sperm quality (motility, concentration, morphology and membrane integrity) was evaluated and compared before and after sperm selection by the 2 methods. Oocytes were maturated in TCM-199 supplemented with 0.4% of BSA for 24 h in a controlled atmosphere and then selected and randomly allocated into 4 different groups. Group 1: oocytes fertilized with sexed semen selected by Percoll®; Group 2: oocytes fertilized with sexed semen selected by Isolate®; Group 3: oocytes fertilized with nonsexed semen selected by Percoll®; Group 4: oocytes fertilized with nonsexed semen selected by Isolate®. Fertilization was performed in Fert-TALP medium for 18 h under the same conditions as maturation. Presumptive zygotes were cultured for 7 days in SOF medium in a 39°C humidified incubator with 5% CO2, 5% O2 and 90% N2. Cleavage and blastocyst rates were evaluated on Day 2 and 7, respectively, after fertilization. Proportional data were transformed by square root and then analysed by ANOVA, with type of semen and sperm selection method as the main effects. Regardless of the sperm selection technique, sperm motility and percentage of normal sperm increased (P < 0.005) from the initial post-thaw parameters. For nonsexed semen, Percoll® gradient increased the recovery rate (i.e. final concentration/initial concentration × 100; 57.3 ± 2.7) compared with Isolate® (46.0 ± 1.8; P < 0.05). Furthermore, sperm selected by Isolate® presented significant improvements compared with Percoll® gradient on membrane integrity of sexed (41.0 ± 0.6 vs 38.8 ± 0.8) and nonsexed semen (60.8 ± 1.6 vs 58.8 ± 0.5; P < 0.05). Finally, blastocyst production rates were higher (P < 0.05) for sexed (Group 2: 14.0 ± 1.0) or nonsexed semen (Group 4: 22.0 ± 1.1) selected by Isolate® than for sexed (Group 1: 10.5 ± 1.5) or nonsexed semen (Group 3: 17.0 ± 2.1) selected with Percoll®. In conclusion, selection of both sexed and nonsexed semen for IVF with Isolate® resulted in higher quality sperm and higher embryo production rates than Percoll®.

113 EFFECT OF SPERM COATING ON THE QUALITY OF BOVINE FROZEN - THAWED SPERMATOZOA

2006 ◽  
Vol 18 (2) ◽  
pp. 164
Author(s):  
M. Thys ◽  
A. Van Soom ◽  
J. Dewulf ◽  
T. Rijsselaere ◽  
A. de Kruif
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Univariate Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Freezing And Thawing ◽  
Bovine Sperm ◽  
Sperm Characteristics ◽  
Group 2 ◽  
Group 1

The substantial decrease of sperm quality after cryopreservation remains an important issue in the artificial insemination industry. Sperm coating with Triladyl® (Minitübe, Tiefenbach, Germany) during ejaculation can preserve sperm characteristics and oocyte penetrating capacity of fresh bovine spermatozoa stored in egg yolk diluent for up to 6 days (De Pauw et al. 2003 Theriogenology 59, 1109–1122). Since collecting semen in a tube containing egg yolk-Tris extender (sperm coating) limits the period of contact between spermatozoa and seminal plasma, the present experiment was conducted to assess if this slightly adjusted method of sperm collection could also have a significant effect on bovine sperm quality after cryopreservation. Semen of five young Holstein Friesian bulls was collected by means of an artificial vagina connected to an empty tube (Group 1; five ejaculates per bull) or a tube containing 4 mL of an egg yolk-Tris extender (Groups 2 and 3; each five ejaculates per bull). The semen samples of Group 1 were conventionally diluted in straws (60 × 106 sperm/mL), frozen, and stored in liquid nitrogen. The samples of Group 3 were centrifuged, and after removing diluent and seminal plasma, the sperm pellet was conventionally diluted and processed. The samples of Group 2 were processed without removal of the supernatant. After thawing each ejaculate was analyzed for average path velocity (VAP), beat cross frequency (BCF), and progressive motility (PROG) using CASA (Minitübe, Tiefenbach, Germany). Furthermore, the membrane integrity of each sample was evaluated using fluorescent SYBR®–14/PI staining (BD Biosciences, Erembodegem, Belgium). All parameters were compared among the three groups of sperm using univariate analysis of variance (SPSS 12.0; SPSS, Inc., Chicago, IL, USA). No significant differences could be observed among the three groups for all of the evaluated sperm characteristics (Table 1). A significant effect of the bull could be determined for all analyzed parameters (P ≤ 0.02), except for the percentage of moribund cells. Nevertheless, the group-bull interaction was never statistically significant. Coating bovine sperm with an egg yolk-Tris extender during ejaculation cannot prevent the substantial deterioration of the spermatozoa that occurs during freezing and thawing since this method of sperm collection does not significantly influence the motility parameters or the membrane integrity after thawing. Table 1. VAP, BCF, PROG, and percentage of membrane-intact, dead, and moribund spermatozoa for the three groups of sperm This research was supported by IWT (no. IWT/020727).

180 ASSESSMENT OF BULL SEMEN QUALITY LOADED IN NEW SensiTemp STRAWS USING SEMEN AND IN VITRO PRODUCTION TECHNOLOGIES

2016 ◽  
Vol 28 (2) ◽  
pp. 221
Author(s):  
D. Le Bourhis ◽  
S. Camugli ◽  
P. Salvetti ◽  
L. Schibler ◽  
E. Schmitt
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Cleavage Rate ◽  
Fluid Medium ◽  
And Control

SensiTemp, a new in vitro maturation (IMV) bull straw concept, presents the advantage of colour changing while the straw is thawed. The colour of frozen straws is blue and straws start to become white when the temperature reaches 33°C, with a complete change of colour at 37°C. The objective of this study is to assess sperm quality after thawing of semen frozen in SensiTemp from 2 bulls, by analysing, in experiment 1, sperm motility and membrane integrity using computer-assisted semen analysis (CASA) and flow cytometry (FC), and, in experiment 2, the in vitro embryo production (IVP) using IVP technologies [IVM, IVF, and in vitro culture (IVC)]. The ejaculates of 2 bulls, selected during preliminary experiments on high in vitro fertility, were harvested at CIA L’Aigle, France, and split ejaculates were frozen in experimental (SensiTemp) and conventional (control) straws. In experiment 1 after thawing semen from the 2 types of straws (5 pooled straws each; 2 replicates), motility was assessed using the IVOS CASA system (Hamilton Thorne Inc., Beverly, MA, USA) and membrane integrity was evaluated through FC with Cytosoft software (Millipore-Guava Technologies Inc., Hayward, CA, USA). In experiment 2, IVF was used to evaluate the non-toxicity of SensiTemp and control straws. Cumulus-oocyte complexes (COC; n = 1178; 4 replicates) collected from slaughterhouse ovaries were matured in IVM medium (TCM-199 with bicarbonate, Sigma-Aldrich, Saint Quentin Fallavier, France; 10 µg mL–1 FSH-LH, Reprobiol, Liège, Belgium; and 10% FCS, Thermo Fisher, Illkirch, France) for 22 h. After fertilization, presumptive zygotes of each group (SensiTemp and control for each bull) were cultured in synthetic oviduct fluid medium (SOF, Minitube, Tiefenbach, Germany) with 1% estrous cow serum (ECS) and 0.6% BSA (Sigma-Aldrich, France) up to 8 days. All cultures were conducted at 38.5C in 5% CO2, and 5% O2. The cleavage and blastocysts rates were evaluated on Days 3 and 7, respectively, for each group. Embryo quality was recorded on Day 7 according to the IETS evaluation. Data from each bull were analysed separately using the chi-squared test (P < 0.05). In experiment 1, neither sperm motility from bull 1 (61.2 and 60.5%) and bull 2 (66.2 and 66.5%) nor membrane integrity from bull 1 (58.6 and 52.2%) and bull 2 (61.0 and 61.9%) were different between SensiTemp and control, respectively. Results from experiment 2 showed no difference (P > 0.05) in cleavage rate between SensiTemp and control for the 2 bulls: 92.1 and 91.7% for bull 1 and 94.2 and 94.6% for bull 2 respectively. The blastocysts rate on Day 7 did not differ (P > 0.05) among groups (47.5, 47.1 and 51.3, 50.4% for SensiTemp and control bull 1 and bull 2, respectively) nor the quality of embryos retrieved in the different groups: 25.4, 23.3, and 30.8, 29.6% in grade 1 embryo for SensiTemp and control bull 1 and bull 2, respectively. Those results demonstrate, in vitro, that the new SensiTemp straws were non-toxic and did not affect the semen quality after thawing nor did the SensiTemp straws affect the ability of sperm cells to fertilize oocytes and produce 8-day-old embryos.

12 COMPUTER-ASSISTED SPERM ANALYSIS OF FRESH EPIDIDYMAL CAT SPERMATOZOA AND THE IMPACT OF COOLED STORAGE (4°C) ON SPERM QUALITY

2008 ◽  
Vol 20 (1) ◽  
pp. 86
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
V. De Causmaecker ◽  
J. Dewulf ◽  
...  
Keyword(s):  
Reference Values ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
The Impact ◽  
Motility Parameters

Feline epididymal sperm is commonly used for in vitro fertilization. It also yields the opportunity to conserve genetic material from valuable males that suddenly die. Epididymal sperm quality parameters vary considerably among laboratories, implicating the need for objective evaluation methods. The aim of the present study was to describe reference values of computer-assisted sperm analysis (CASA) parameters of fresh epididymal cat sperm and to assess the effect of prolonged cooled storage (4�C) on various sample characteristics. Epididymides obtained from tomcats after routine orchiectomy (2–4 pairs/replicate) were sliced to release spermatozoa. The sperm suspension was placed on a 2-layer gradient and, after centrifugation, the sperm pellet was recovered. In Experiment 1 (20 replicates), sperm motility parameters were assessed immediately after retrieval (T0) using the Hamilton Thorne analyzer Ceros 12.1 (HTR; Hamilton Thorne Biosciences, Beverly, MA, USA). In Experiment 2, fresh (T0) sperm samples (4 replicates) were evaluated for motility parameters (HTR), acrosomal status (FITC-Pisum sativum agglutinin staining), morphology (eosin/nigrosin (E/N) staining), and membrane integrity (E/N and SYBR�-14-propidium iodide staining; Molecular Probes, Inc., Eugene, OR, USA). After addition (1:2) of a Tris-glucose-citrate diluent containing 20% egg yolk, samples were cooled and reassessed on Days 1 (T1), 3 (T3), 5 (T5), 7 (T7), and 10 (T10). Results were analyzed in a mixed linear model, with replicate as random factor and time as fixed effect (S-PLUS 7.0; Insightful Corp., Seattle, WA, USA). Results of Experiment 1 were as follows (mean � SD): motility (MOT): 80.8% � 23.5; progressive motility (PMOT): 69.9% � 23.2; velocity average pathway (VAP): 98.7 µm s–1 � 24.2; velocity straight line (VSL): 89.3 µm s–1 � 25.4; velocity curved line (VCL): 134.8 µm s–1 � 31.9; amplitude lateral head (ALH): 4.3 µm � 2.0; beat cross frequency (BCF): 34.6 Hz � 7.0; and straightness (STR): 89.6% � 6.6. In Experiment 2, MOT, PMOT, VAP, VSL, VCL, BCF, and the percentage of normal spermatozoa showed a decrease over time (P < 0.05) compared to fresh samples, starting from T1, T3, T5, T7, T5, T3, and T1, respectively. In contrast, STR, ALH, membrane integrity, and the percentage of acrosome-intact spermatozoa were not affected (P > 0.05) by cooled storage. To summarize, we have presented a set of reference values for CASA-parameters of fresh, epididymal cat spermatozoa. Cooled storage impaired most motility parameters and lowered the percentage of normal spermatozoa, but did not influence membrane integrity or acrosomal status. The effect of cooled storage on DNA fragmentation of sperm and its subsequent influence on in vitro embryo development require further investigation.

88 SELECTION OF A CRYOSURVIVAL SPERM POPULATION DOES NOT IMPROVE THE IN VITRO PENETRATING ABILITY OF FROZEN - THAWED BOAR SPERMATOZOA

2008 ◽  
Vol 20 (1) ◽  
pp. 124
Author(s):  
M. L. Perals ◽  
M. A. Gil ◽  
E. M. Garcia ◽  
J. Sanchez-Osorio ◽  
J. M. Vázquez ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Molecular Probes ◽  
Computer Assisted ◽  
Immature Oocytes ◽  
Sperm Analysis ◽  
Boar Spermatozoa ◽  
Casa System ◽  
Selection Of

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.

91 TREATMENT OF GOAT SPERM WITH CATALASE TO IMPROVE POST-THAW QUALITY

2011 ◽  
Vol 23 (1) ◽  
pp. 151
Author(s):  
R. O. C. Silva ◽  
M. Nichi ◽  
E. G. A. Perez ◽  
P. A. A. Góes ◽  
A. Dalmazzo ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Structural Damage ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Thiobarbituric Acid ◽  
Lower Percentage ◽  
Sperm Chromatin ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Sperm Analysis

Semen cryopreservation is extremely important to the use of reproductive biotechnologies in goats. However, studies indicate that cryopreservation may lead to increased oxidative stress which may cause structural damage to biomolecules, DNA, lipids, carbohydrates, and proteins, as well as other cellular components. Previous studies performed by our group indicate fresh goat sperm is highly susceptible to the attack of hydrogen peroxide. Therefore, the treatment with hydrogen peroxide scavengers would be an alternative to improve post-thaw sperm quality in goats. The aim of the present study was to evaluate the efficiency of catalase, an important hydrogen peroxide scavenger, to improve post-thaw quality in cryopreserved goat semen samples. Semen samples from 5 adult goats were collected and cryopreserved (Botu-Bov®, Biotech Ltda.). After thawing, samples were incubated (2 h, 37°C) with 0, 60, 120, and 240 UI mL–1 of catalase. Samples were then analysed for motility using the computer assisted sperm analysis (CASA); the 3–3′ diaminobenzidine stain, as an index of mitochondrial activity; the eosin nigrosin stain, as an index of membrane integrity; the simple stain (Fast green/Bengal rose), as an index of acrosome integrity; sperm chromatin structure assay, as an index of DNA fragmentation; and the measurement of thiobarbituric acid reactive substances (TBARS), as an index of lipid peroxidation. Statistical analysis was performed using the SAS System for Windows (SAS Institute Inc., Cary, NC, USA; least significant differences test and Spearman correlation; P < 0.05). Results showed that catalase treatment after thawing played a role on improving mitochondrial activity. Samples treated with 240 UI mL–1 showed lower percentage of sperm showing low mitochondrial activity when compared with samples treated with 0 and 120 UI mL–1 of catalase (6.5 ± 2.3, 17.2 ± 3.5, and 10.0 ± 1.3%, respectively). However, no effect of catalase was observed on any of the other variables studied. Results indicate that catalase, despite its beneficial effect on mitochondrial activity, does not influence positively on sperm quality after thawing. A hypothesis to explain such results would be that because of seminal plasma dilution with the extender, the antioxidants were also diluted. Therefore, the antioxidant protection would be impaired and the most deleterious reactive oxygen species, as observed in fresh semen, would also be different depending on the semen extender used because sperm are extremely dependent on the extracellular environment due to the reduced cytoplasm and the high content of polyunsaturated fatty acids in the membrane. A study performed by our group confirms such a hypothesis. Possibly, the treatment with catalase would be more effective if performed before cryopreservation. Also, it is possible that the use of different antioxidants would provide better results. Thanks to Nutricell for the media used and CAPES for financial support.

Factors affecting storage of Slovak native rabbit semen in the gene bank

Zygote ◽  
2017 ◽  
Vol 25 (5) ◽  
pp. 592-600
Author(s):  
Barbora Kulíková ◽  
Marta Oravcová ◽  
Andrej Baláži ◽  
Peter Supuka ◽  
Peter Chrenek
Keyword(s):  
Plasma Membrane ◽  
Liquid Nitrogen ◽  
Semen Quality ◽  
Sperm Quality ◽  
Objective Assessment ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Quality Traits ◽  
Plasma Membrane Integrity

SummaryIn this study, fresh and frozen–thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen–thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen–thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen–thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen–thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.

scholarly journals Effect of Splinting in Accuracy of Two Implant Impression Techniques

2014 ◽  
Vol 40 (6) ◽  
pp. 633-639 ◽  
Author(s):  
Erica Dorigatti de Avila ◽  
Fernanda de Matos Moraes ◽  
Sabrina Maria Castanharo ◽  
Marcelo Antonialli Del'Acqua ◽  
Francisco de Assis Mollo
Keyword(s):  
Confidence Interval ◽  
Analysis Of Variance ◽  
Impression Material ◽  
Titanium Screw ◽  
Standard Preparation ◽  
Implant Abutment ◽  
Interface Gaps ◽  
Group 2 ◽  
Group 1

Because there is no consensus in the literature about the need for a splint between copings, the aim of this study was to evaluate, in vitro, the accuracy of 2 impression techniques for implant-supported prostheses. A master cast was fabricated with four parallel implant abutment analogs and a passive framework. Two groups with 5 casts each were formed: Group 1 (squared impression copings with no splint: S) and Group 2 (splinted squared impression copings, using metal drill burs and Pattern resin: SS). The impression material used was polyvinyl siloxane with open trays for standard preparation of the casts. For each cast, the framework was positioned, and a titanium screw was tightened with 10 N·cm torque in analog A, after which measurements of the abutment-framework interface gaps were performed at analogs C and D. This process was repeated for analog D. These measurements were analyzed using software. A one-way analysis of variance (ANOVA) with a confidence interval of 95% was used to analyze the data. Significant differences were detected between S and SS in relation to the master cast (P ≤ 0.05). The median values of the abutment-framework interface gaps were as follows: master cast: 39.64 μm; squared impression copings with no splint: 205.86 μm; splinted squared impression copings: 99.19 μm. Under the limitations of this study, the technique presented for Group 2 produces better results compared with the technique used for Group 1.

The Relationship between Serum Ferritin Levels and 5-Year All-Cause Mortality in Hemodialysis Patients

2021 ◽  
pp. 1-7
Author(s):  
Emre Erdem ◽  
Ahmet Karatas ◽  
Tevfik Ecder
Keyword(s):  
Serum Ferritin ◽  
Hemodialysis Patients ◽  
Rank Test ◽  
Significant Difference ◽  
All Cause Mortality ◽  
Group 3 ◽  
Group 2 ◽  
The Relationship ◽  
Group 1

<b><i>Introduction:</i></b> The effect of high serum ferritin levels on long-term mortality in hemodialysis patients is unknown. The relationship between serum ferritin levels and 5-year all-cause mortality in hemodialysis patients was investigated in this study. <b><i>Methods:</i></b> A total of 173 prevalent hemodialysis patients were included in this study. The patients were followed for up to 5 years and divided into 3 groups according to time-averaged serum ferritin levels (group 1: serum ferritin &#x3c;800 ng/mL, group 2: serum ferritin 800–1,500 ng/mL, and group 3: serum ferritin &#x3e;1,500 ng/mL). Along with the serum ferritin levels, other clinical and laboratory variables that may affect mortality were also included in the Cox proportional-hazards regression analysis. <b><i>Results:</i></b> Eighty-one (47%) patients died during the 5-year follow-up period. The median follow-up time was 38 (17.5–60) months. The 5-year survival rates of groups 1, 2, and 3 were 44, 64, and 27%, respectively. In group 3, the survival was lower than in groups 1 and 2 (log-rank test, <i>p</i> = 0.002). In group 1, the mortality was significantly lower than in group 3 (HR [95% CI]: 0.16 [0.05–0.49]; <i>p</i> = 0.001). In group 2, the mortality was also lower than in group 3 (HR [95% CI]: 0.32 [0.12–0.88]; <i>p</i> = 0.026). No significant difference in mortality between groups 1 and 2 was found (HR [95% CI]: 0.49 [0.23–1.04]; <i>p</i> = 0.063). <b><i>Conclusion:</i></b> Time-averaged serum ferritin levels &#x3e;1,500 ng/mL in hemodialysis patients are associated with an increased 5-year all-cause mortality risk.

scholarly journals A Flexible Multidose GnRH Antagonist versus a Microdose Flare-Up GnRH Agonist Combined with a Flexible Multidose GnRH Antagonist Protocol in Poor Responders to IVF

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Gayem İnayet Turgay Çelik ◽  
Havva Kömür Sütçü ◽  
Yaşam Kemal Akpak ◽  
Münire Erman Akar
Keyword(s):  
Gnrh Agonist ◽  
Gnrh Antagonist ◽  
Poor Responders ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Gnrh Antagonist Protocol ◽  
Group 2 ◽  
Antagonist Protocol ◽  
Group 1

Objective. To compare the effectiveness of a flexible multidose gonadotropin-releasing hormone (GnRH) antagonist against the effectiveness of a microdose flare-up GnRH agonist combined with a flexible multidose GnRH antagonist protocol in poor responders to in vitro fertilization (IVF).Study Design. A retrospective study in Akdeniz University, Faculty of Medicine, Department of Obstetrics and Gynecology, IVF Center, for 131 poor responders in the intracytoplasmic sperm injection-embryo transfer (ICSI-ET) program between January 2006 and November 2012. The groups were compared to the patients’ characteristics, controlled ovarian stimulation (COH) results, and laboratory results.Results. Combination protocol was applied to 46 patients (group 1), and a single protocol was applied to 85 patients (group 2). In group 1, the duration of the treatment was longer and the dose of FSH was higher. The cycle cancellation rate was significantly higher in group 2 (26.1% versus 38.8%). A significant difference was not observed with respect to the number and quality of oocytes and embryos or to the number of embryos transferred. There were no statistically significant differences in the hCG positivity (9.5% versus 9.4%) or the clinical pregnancy rates (7.1% versus 10.6%).Conclusion. The combination protocol does not provide additional efficacy.

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