scholarly journals Alpha-tocopherol improves sperm quality by regulate intracellular Ca2+ intensity (influx/efflux) of Simmental bull cattle sperm

Author(s):  
Hermin Ratnani ◽  
Suprayogi TW ◽  
Sardjito T ◽  
Susilowati S ◽  
Azura S

Background: The effects of α-tocopherol on intracellular Ca2+ intensity in semen cryopreservation by regulate intracellular Ca2+ intensity have not been reported yet. Objective: The research was conducted to evaluate the effect of supplementation α-tocopherol into egg yolk skim milk extender on sperm quality and intracellular Ca2+ intensity. Methods: Semen samples were collected and supplemented with respectively 0mM (P0); 0.5mM (P1); 1mM (P2); 1.5mM (P3) and 2mM (P4) α-tocopherol in extender before cryopreservation processes. Post-thawing sperm was evaluated for motility, viability, and abnormality using Phase Contrast Microscope (200x) with eosin-nigrosine staining, and intracellular Ca2+ intensity of the best result dose was evaluated using Confocal Laser Scan Microscope (400x) with Fluo-3 Staining. Results: The results showed there was a significant difference (P≤0.05) in sperm motility and viability between P0; P1 with P2; P3; P4. The Motility and viability between groups P0; P1 and P3; P4 showed no significant difference (P≥0.05), while P2 with P3; P4 showed significant difference (P≤0.05). There was a significant difference (P≤0.05) in sperm abnormality of P0; P1 with P2; P3; P4. The abnormality between P0; P1 and P2; P3 showed no significant difference (P≥ 0.05), while P2; P3 showed a significant difference with P4 (P≤0.05). The best result in sperm quality was supplementation with 1.5mM α-tocopherol. Ca2+ intracellular intensity: 142.76 ± 21.8 au (P0) and 176.06 ± 61.43 au (P3). Conclusions: It was concluded that 1.5mM α-tocopherol is the best dose to improve sperm quality by regulating intracellular Ca2+ intensity on Simmental bull cattle.

2021 ◽  
pp. 2073-2084
Author(s):  
Suherni Susilowati ◽  
Imam Mustofa ◽  
Wurlina Wurlina ◽  
Indah Norma Triana ◽  
Suzanita Utama ◽  
...  

Background and Aim: Kacang buck sperm is cryosensitive due to the seminal plasma of semen itself. Meanwhile, bull seminal plasma contains the insulin-like growth factor-1 (IGF-1) complex, which is cryoprotective. The addition of the crude protein of Simmental bull seminal plasma increased the quality of post-thawed semen of Kacang buck. The study was conducted to determine the effects of Simmental bull seminal plasma with IGF-1 on the fertility of post-thawed Kacang buck semen. Materials and Methods: Buck semen was diluted in the following skim milk-egg yolk extender preparations: Without the addition of Simmental bull seminal plasma IGF-1 complex protein (T0); with the addition of 12-μg Simmental bull seminal plasma IGF-1 complex protein (T1); and with the addition of 24-μg Simmental bull seminal plasma IGF-1 complex protein (T2). The extended semen was packed in 0.25-mL straws and frozen. Post-thawed semen fertility was evaluated based on the following variables: Sperm motility, viability, intact plasma membrane (IPM), malondialdehyde (MDA) levels, capacitation status, and acrosome reaction. The difference in each variable among the groups was evaluated using analysis of variance, followed by Tukey's honestly significant difference test, at a 95% level of significance. Meanwhile, principal component analysis (PCA) was used to identify the principal component of semen fertility among the seven parameters. Results: The T1 group showed the highest sperm motility, viability, IPM, and percentage of incapacitated sperm and the lowest MDA levels, percentage of capacitated sperm, and acrosome reaction. PCA revealed that sperm motility had a moderate to very robust correlation with other variables and is the most crucial parameter, accounting for 80.79% of all variables. Conclusion: The IGF-1 complex in Simmental bull seminal plasma was useful for increasing the fertility of post-thawed Kacang buck semen, and sperm motility was the principal component of semen fertility.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ameena Sultana ◽  
Heena Zainab ◽  
Pramod Jahagirdar ◽  
Deepa Hugar ◽  
Shaimaa

Abstract Background Age estimation is an important factor in forensic science for human identification. Teeth are considered to play a vital role as they resist decomposition at death unlike other tissues. This resistance and the gradual structural changes that take place throughout the life of an individual have made teeth useful indicators for age estimation. Dental cementum shows continuous apposition throughout the life of an individual. Tooth cemental annulation is a microscopic method for the determination of an individual’s age based on the analysis of incremental lines of cementum. Light microscopy as well as specialized microscopic methods have been employed to enhance the assessment of the cemental annulations. Periodontal disease is the most common dental problem affecting millions of people. Assessing the efficiency of the tooth cemental annulations method in periodontally diseased teeth is an important requisite. This study aims at assessing and evaluating the tooth cemental annulations in normal and periodontally diseased teeth using phase contrast microscopic method for age determination. Results A total of 60 teeth were included in the study and out of which 30 teeth were normal (sound teeth without any associated pathologies) and 30 were periodontally involved teeth respectively. Longitudinal ground sections were prepared and observed under phase contrast microscope. Measurements were made using an image analyzer software. The total width of the cementum was divided by the distance between two incremental lines. The eruption age of the tooth was then added to this to obtain the chronologic age for each individual. The results in the present study showed that tooth cemental annulations are applicable to periodontally sound teeth as well as in periodontally diseased teeth. There was no significant difference of estimated age from the actual age in both periodontally sound and periodontally diseased teeth. Normal teeth showed a reliability value of 92% and periodontally compromised teeth showed 96% respectively. There was no substantial influence of periodontal health on the estimated age. Conclusions The study concludes that the use of phase contrast microscopy in conjunction with image enhancement procedures improves the accuracy of age estimation and may serve as a reliable aid in forensic identification.


2007 ◽  
Vol 19 (1) ◽  
pp. 128
Author(s):  
C. Tamargo ◽  
C. Díez ◽  
J. De La Fuente ◽  
M. Carbajo ◽  
J. M. Benito ◽  
...  

The need to conserve farm animal biodiversity is accepted by many countries through the ratification of the convention of biological diversity, and sperm quality is known to be an important criterion in the evaluation of breeding soundness. The aim of this work was to characterize the semen of a local breed of ponies 'Asturcon' (maintained free over the mountains all year around) before its incorporation into a germplasm bank. Semen was obtained from six stallions (6–17 years of age) using an artificial vagina, 3 days/week, during 12 weeks. Immediately after collection, gel-free semen was evaluated for volume, sperm concentration, and motility. Semen motility was again evaluated after equilibration/refrigeration. For evaluation of individual (IM) and progressive motility (PM) rates, semen was diluted (20 � 106 spermatozoa/mL) and analyzed with a CASA System (SCA; Microptic S.L., Barcelona, Spain). Five fields per sample were evaluated (minimum 500 spermatozoa/sample) under a phase contrast microscope (100�). Semen samples were subjected to a hypo-osmotic swelling test (HOS) test to detect the presence of swollen tails in a 100 mM citrate–fructose solution. Percentages of altered acrosomes and morphological abnormalities were determined by counting 100 spermatozoa (1000�). Then, semen was diluted and centrifuged for 10 min at 600g. After the supernatant was discarded, the pellet was re-suspended in freezing medium (skim milk extender containing 2% egg yolk and 2.5% glycerol) to a final concentration of 100 � 106 spermatozoa/mL, and equilibrated/cooled (60 min) to 4�C. Statistical analysis was carried out by means of the GLM and CORR procedures and Duncan test for means (SAS Institute, Inc., Cary, NC, USA). A significant effect between males (P < 0.05) on semen quality, such as volume of the ejaculate, sperm concentration, and morphological abnormalities, were detected among stallions. On the other hand, positive and significant correlations were found between the sperm motility immediately after collection and after equilbration/refrigeration (r = 0.73; P < 0.05); moreover, sperm motilities (both fresh and refrigerated) correlated with the results of the HOS test (r = 0.56; P < 0.001, and r = 0.27, P < 0.05, respectively). These preliminary results confirm that the sperm of the Asturcon ponies breed can be collected and will survive the equilibration/refrigeration procedures. Conservation and development of local breeds is important because they represent a unique source of genes for improving health and performance traits of industrial breeds. However, complementary studies on the ability of the stallion sperm to survive freezing/thawing procedures in rates higher than 30% are needed to ensure that genetic banks are correctly created. This work was performed in collaboration with ACPRA and Dep�sito de Sementales de Santander (Spain), and supported by RZ2004-00031-C02-01.


2021 ◽  
Vol 4 (2) ◽  
pp. 249
Author(s):  
Syuhuud Arumbinang Wajdi ◽  
Budi Utomo ◽  
Rimayanti Rimayanti ◽  
Erma Safitri ◽  
Tri Wahyu Suprayogi ◽  
...  

The purpose of this research was to determine the best dosage of Moringa oleifera Aqueous extract in egg yolk skim milk extender for post thawed Limousin Bull sperm quality on aspect viability, and the level of. The treatment was divided into five groups: egg yolk and skim milk diluter (P0), 2,5% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P1), 5% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P2), 10% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P3), 20% M. oleifera aqueous ekstract in 4 ml egg yolk skim milk (P4). The sperm quality was observed post thawing. The data were analyzed using ANOVA and Duncant Test. The best sperm motility showed on P2 with 43b ± 5,70, the best sperm viability showed on P3 with 58,20b ± 8,72 and than the lowest level of malondialdehyde showed on P4 with 5,434a ± 1,034. In conclusion addition of M. oleifera on dose 10% can increase quality of Limousin Sperm Post Thawed.


2020 ◽  
Vol 9 (3) ◽  
pp. 69
Author(s):  
Satya Alysa Cahya Puspita ◽  
Suherni Susilowati ◽  
Trilas Sardjito ◽  
Abdul Samik ◽  
Indah Norma Triana ◽  
...  

Spermatozoa in fresh semen of Sapudi ram has a limited life span. The storage of semen in cold temperatures (5 °C) is intended to prolong the spermatozoa's life. However, storage in cold temperatures can lead to increased production of reactive oxygen species (ROS). This condition reduces the quality of spermatozoa. The purpose of this study was to determine the effect of alphatocopherol supplementation in skim milk-egg yolk extender on viability, motility, and plasma membrane integrity of Sapudi ram spermatozoa. Fresh semen derived from Sapudi ram was divided into four treatment groups. Control treatment (P0): semen was added in the extender of skim milkegg yolk without alpha-tocopherol. Three other treatments: P1, P2, and P3 semen were added in skim milk-egg yolk extender with the supplementation of 0.25, 0.5, and 1 gram alpha-tocopherol/ 100 mL extender, respectively. The results showed that the viability, motility, and integrity of the spermatozoa plasma membrane decreased gradually according to the storage length. Supplementation of skim milk-egg yolk extender with 0.5 gram of alpha-tocopherol/100 mL (P2) was able to maintain spermatozoa quality longer (p <0.05) than the control group. It can be concluded that alpha-tocopherol with a concentration of 0.5 g/100 mL of skim milk-egg yolk extender effectively maintains the quality of Sapudi ram spermatozoa in storage at 5 ° C.


TAPPI Journal ◽  
2010 ◽  
Vol 9 (10) ◽  
pp. 7-15
Author(s):  
HANNA KOIVULA ◽  
DOUGLAS BOUSFIELD ◽  
MARTTI TOIVAKKA

In the offset printing process, ink film splitting has an important impact on formation of ink filaments. The filament size and its distribution influence the leveling of ink and hence affect ink setting and the print quality. However, ink filaments are difficult to image due to their short lifetime and fine length scale. Due to this difficulty, limited work has been reported on the parameters that influence filament size and methods to characterize it. We imaged ink filament remains and quantified some of their characteristics by changing printing speed, ink amount, and fountain solution type. Printed samples were prepared using a laboratory printability tester with varying ink levels and operating settings. Rhodamine B dye was incorporated into fountain solutions to aid in the detection of the filaments. The prints were then imaged with a confocal laser scanning microscope (CLSM) and images were further analyzed for their surface topography. Modeling of the pressure pulses in the printing nip was included to better understand the mechanism of filament formation and the origin of filament length scale. Printing speed and ink amount changed the size distribution of the observed filament remains. There was no significant difference between fountain solutions with or without isopropyl alcohol on the observed patterns of the filament remains.


2021 ◽  
pp. 096739112110055
Author(s):  
Gunce Ozan ◽  
Meltem Mert Eren ◽  
Cansu Vatansever ◽  
Ugur Erdemir

Surface sealants are reported to ensure surface smoothness and improve the surface quality of composite restorations. These sealants should also reduce the bacterial adhesion on composite surfaces however, there is not much information regarding their performance on bulk-fill composite materials. The aim of this study was to evaluate the effect of surface sealant application on surface roughness and bacterial adhesion of various restorative materials. Disc-shaped samples were prepared from a compomer, a conventional composite and three bulk-fill composites. Specimens of each group were divided into two groups (n = 9): with/without surface sealant (Biscover LV, [BLV]). Surface roughness values were examined by profilometry and two samples of each group were examined for bacterial adhesion on a confocal laser scanning microscope (CLSM). Bacterial counts were calculated by both broth cultivation and microscopic images. Results were analyzed with one-way ANOVA and Bonferroni/Dunn tests. Following the BLV application, there was a decrease in the surface roughness values of all groups however, only Tetric N-Ceram Bulk and Beautifil-Bulk groups showed significantly smoother surfaces (p < 0.001). There were no significant differences among material groups without BLV application. Evaluating bacterial adhesion after BLV application, conventional composite had the lowest values among all followed by the compomer group. Beautifil-Bulk had significantly the highest bacterial adhesion (p < 0.05), followed by Tetric N-Ceram Bulk group. Without BLV application, there was no significant difference among bacterial adhesion values of groups (p > 0.05). CLSM images showed cell viability in groups. Bulk-fill composites showed higher bacterial adhesion than conventional composite and compomer materials. The surface sealant was found to be highly effective in lowering bacterial adhesion, but not so superior in smoothing the surfaces of restorative materials. So, surface sealants could be used on the restorations of patients with high caries risk.


2004 ◽  
Vol 16 (2) ◽  
pp. 226 ◽  
Author(s):  
F. Martinez-Pastor ◽  
F. Olivier ◽  
T. Spies ◽  
L. Anel ◽  
P. Bartels

Biological Resource Banks represent a potentially valuable tool for species conservation. It is, however, necessary to understand the species-specific cryopreservation process and its consequences for spermatozoa to aid in the development of assisted reproduction as a future conservation tool. The aim of this study was to assess the in vitro functionality of white rhinoceros Cerathoterium simum epididymal spermatozoa both before and after cryopreservation. Testes from a harvested white rhino bull were removed and transported at 5°C to the laboratory within 4h. The cauda epididymis was dissected out and flushed with 2mL of Tris-citrate egg yolk extender (fraction A, Biladyl, Minitüb, Germany). A 0.1mL aliquot was removed for analysis and the balance (9mL; 2mL fraction A+7mL sperm sample) mixed with an additional 27.2mL of Tris-citrate egg yolk with glycerol (fraction B, Bidadyl). The extended sample was allowed to cool to 4°C over a 6-h period before an additional 29.2mL of cooled fraction B were added (final sperm concentration=150×106mL−1). Sperm samples were loaded into 0.25-mL straws and frozen over LN2 vapor (4cm for 20min) for later assessment. Sperm straws were thawed by placing the straws in water at 37°C for 30s. Pre-freeze and post-thaw evaluations were carried out in the same manner. Media used included: HEPES for washing (20mM HEPES, 355mM sucrose, 10mM glucose, 2.5mM KOH) and HEPES saline (197mM NaCl, instead of sucrose). An aliquot was diluted with HEPES (washing) and centrifuged for 5min at 600×g; the pellet was resuspended in HEPES saline. Sperm motility (total motility %, TM;; and progressive motility %, PM) was assessed using phase contrast microscopy (×200; 37°C). Sperm plasma membrane status was assessed using the fluorescent dye, propidium iodide (50ngmL−1 in HEPES saline;; 10min, RT). Percentage of cells with plasma membranes intact (unstained;; PMI) was recorded. Mitochondrial status was assessed with the fluorescent dye, JC-1 (7.5μM in HEPES saline;; 30min, 37°C). The % of cells with an orange-stained midpiece was recorded (active mitochondria;; MIT). Resilience to hypoosmotic shock (HOS test) was assessed by diluting a sample in 100mOsm/kg HEPES saline (1:20; 15min, RT). An aliquot was stained with PI to assess plasma membrane status (HOSPMI), and the rest was fixed with formaldehyde, and % coiled tails (positive endosmosis;; HOST) was estimated using phase contrast microscopy (×400). Evaluations of PMI, MIT and HOSPMI were performed using fluorescence microscopy (×400, 450–490nm excitation filter). The results indicated that quality was good pre-freezing (TM: 60%; PMI: 86%; MIT: 100%), except for a PM value of 15%. After thawing, although there was a drop in TM (30%), there was no decrease in PM (20%). Our in vitro functional assessment indicated a loss of quality between the pre-freeze and post-thaw evaluations, but PMI and MIT maintained their pre-thaw levels (60% and 72%, respectively). The HOS test, which indicates plasma membrane integrity, decreased from the pre-freeze level (91%) to a post-thaw value of 70%. HOSTPMI was 72% pre-freeze, and decreased to 54% post-thaw. In conclusion, epididymal spermatozoa from the white rhino may retain its functionality after cryopreservation in a commerically available cryo-extender (Bidadyl). The use of assisted reproduction techniques could someday play a role in the management and conservation of the white rhinoceros and related species.


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