Effect of L-cysteine Supplementation on Cryopreservation of Black Bengal Buck Semen

2021 ◽  
Author(s):  
Krushna Chandra Tudu ◽  
Debajyoti Sarkar ◽  
Ajoy Mandal ◽  
Mohan Mondal ◽  
Subrata Kumar Das ◽  
...  
Keyword(s):  
Treatment Group ◽  
Cell Structure ◽  
Membrane Integrity ◽  
Control Group ◽  
Sod Activity ◽  
Freeze Thaw ◽  
Positive Effects ◽  
Sperm Characters ◽  
Semen Extender

Background: Black Bengal goat is one among the valuable goat breeds of India. Semen preservation and artificial insemination are practical technologies for conservation and improving livestock germplasm. Various biochemical changes that occur during cryopreservation of semen, results in loss of sperm cell structure and their functions. Inclusion of antioxidants to the semen extender has been reported to have positive effects on sperm recovery in various livestock species. The present experiment was carried out to evaluate the effect of antioxidant L-cysteine on post freeze thaw sperm characters, lipid peroxide level and superoxide dismutase (SOD) activity during cryo­preservation of Black Bengal buck semen. Methods: Twenty numbers of semen ejaculates were collected from Black Bengal bucks, antioxidant L cysteine was added to the semen extender @ 0, 1 and 2 mM in control group (CC), treatment group 1 (CT1) and treatment group 2 (CT2), respectively. Semen samples were frozen in liquid nitrogen and measured post freeze-thaw in vitro sperm characters, malondialdehyde (MDA) concentration and SOD activity. Result: Though there was no significant improvement in post thaw sperm motility, functional membrane integrity and viability count, supplementation of L-cysteine significantly improved the percentage of sperm cells with intact acrosome and significantly (P less than 0.05) reduced the level of malondialdehyde. Post thaw SOD activity was found significantly lower in the antioxidant treated groups than the control. It is concluded that supplementation of L cysteine is effective in controlling lipid peroxidation and conserving in vitro sperm characters during freezing of Black Bengal buck semen.

Effect of Butylated Hydroxytoluene and Tocopherol Supplementation on In vitro Sperm Characters during Cryopreservation of Black Bengal Buck Semen

2021 ◽  
Author(s):  
Krushna Chandra Tudu ◽  
Ajoy Mandal ◽  
Mohan Mondal ◽  
Subrata Kumar Das ◽  
Monoj Kumar Ghosh ◽  
...  
Keyword(s):  
Treatment Group ◽  
Butylated Hydroxytoluene ◽  
Control Group ◽  
Sod Activity ◽  
Intact Cells ◽  
Positive Effects ◽  
Sperm Characters ◽  
Group 2 ◽  
Group 1

Background: Black Bengal goat is one of the important goat breeds of India. Cryopreservation of semen and artificial insemination are effective techniques for improving goat breeding programs. Various biochemical and functional changes that occur during freezing-thawing process results in poor post thaw sperm recovery. Supplementation of antioxidants to the extender has been reported to have positive effects on semen cryopreservation in various species. The present study was undertaken to assess the effect of antioxidants butylated hydroxyl toluene (BHT), α tocopherol on post thaw in vitro sperm characters, lipid peroxide level and superoxide dismutase (SOD) activity during cryo­preservation of Black Bengal buck semen. Methods: Semen was collected from bucks by artificial vagina, antioxidant BHT was added to the Tris egg yolk extender @ 0, 1 and 2 mM/ml in control group (BHTC), treatment group 1 (BHTT1) and treatment group 2 (BHTT2), respectively. Similarly, tocopherol was added @ 0, 1 and 2 mg/ml in control group (TFC), in treatment group 1 (TFT1) and treatment group 2 (TFT2), respectively. Each antioxidant was tested with 20 semen ejaculates. Semen samples were frozen in liquid nitrogen and post freeze-thaw in vitro sperm characters, malondialdehyde (MDA) concentration and SOD activity were measured. Result: Post thaw sperm motility, functional membrane integrity and viable count were significantly (P less than 0.05) higher in the BHT and α tocopherol supplemented groups than control groups. Significantly (P less than 0.05) higher acrosome intact cells were recovered in BHTT1, BHTT2 and TFT2 groups than the control and TFT1 groups. Further, BHT supplemented groups had significantly lower level of MDA than the control, while the supplementation of α tocopherol could not control the generation of MDA. Post thaw SOD activity was found significantly lower in the antioxidant treated groups than their respective controls. It is concluded that supplementation of BHT and α tocopherol were found to be more promising in conserving in vitro sperm characters from cryo-damages during freezing of Black Bengal bucks semen.

scholarly journals Hypoxia-preconditioned MSCs Have Superior Effect in Ameliorating Renal Function on Acute Renal Failure Animal Model

2019 ◽  
Vol 7 (3) ◽  
pp. 305-310 ◽  
Author(s):  
Agung Putra ◽  
Dannis Pertiwi ◽  
Meidona Nurul Milla ◽  
Ulfah Dian Indrayani ◽  
Durotul Jannah ◽  
...  
Keyword(s):  
Renal Failure ◽  
Animal Model ◽  
Renal Function ◽  
Acute Renal Failure ◽  
Treatment Group ◽  
Creatinine Level ◽  
Histological Analysis ◽  
Control Group ◽  
Positive Effects

BACKGROUND: Acute renal failure (ARF) is a serious disease characterised by a rapid loss of renal functions due to nephrotoxic drug or ischemic insult. The clinical treatment approach such as dialysis techniques and continuous renal enhancement have grown rapidly during past decades. However, there is yet no significant effect in improving renal function. Hypoxia-preconditioned mesenchymal stem cells (HP-MSCs) have positive effects on the in vitro survival and stemness, in addition to angiogenic potential. AIM: In this study, we aimed to analyse the effect of HP-MSCs administration in improving renal function, characterised by blood urea nitrogen (BUN) and creatinine level. METHODS: A group of 15 male Wistar rats weighing 250 g to 300 g were used in this study (n = 5 for each group). Rats were randomly distributed into 3 groups: Vehicle control (Veh) as a control group, HP-MSCs and normoxia MSCs (N-MSCs) as the treatment group. Renal function was evaluated based on the BUN and creatinine levels using the colourimetric method on day 5 and 13. The histological analysis using HE staining was performed on day 13. RESULTS: The result showed there is a significant decrease in BUN and creatinine level (p < 0.05). The histological analysis of renal tissue also showed a significant decrease between Veh and treatment group (p < 0.05). CONCLUSION: Based on this study, we conclude that HP-MSCs have a superior beneficial effect than N-MSCs in improving renal function in an animal model of gentamicin-induced ARF.

Preservation of Black Bengal buck semen in soybean lecithin based chemically defined extender

2018 ◽  
Author(s):  
Pangdun Konyak ◽  
Ajoy Mandal ◽  
Mohan . ◽  
C. Bhakat ◽  
S. K. Das ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Soybean Lecithin ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Control Group ◽  
Initial Dilution ◽  
Nitrogen Vapor ◽  
Sperm Characters

This experiment was conducted with the aim to study the effect of replacing egg yolk with soybean lecithin (SL) for cryopreservation of Black Bengal buck semen. Sexually matured Black Bengal buck (n = 5) were used and the ejaculates were obtained using an artificial vagina method. The semen samples were pooled and diluted in Tris extender with 5% Glycerol containing either 15% egg yolk (control group) or SL at different concentrations (1% SL, 1.5% SL and 2% SL). The semen samples were filled in straws and cooled gradually to 5 °C. Semen straws were equilibrated for 3 hours at 5°C and were frozen in static liquid nitrogen vapor and stored in liquid nitrogen. Semen samples were evaluated after initial dilution, after completion of equilibration and after freeze thawing for in vitro sperm characters such as sperm motility, functional membrane integrity and malondioldehyde (MDA) concentration. Semen samples preserved in extender containing 1% SL was able to maintain in vitro sperm characters similar to the extender containing egg yolk. However, significant (P less than 0.05) reduction in all semen parameters was observed as the concentration of soybean lecithin increased above 1% level. It is concluded that an extender containing soybean lecithin @ 1% with 5% Glycerol can be used for replacing egg yolk for cryopreservation of Black Bengal buck semen.

Mechanism of the adverse effect of hyaluronidase used for oocyte denudation on early development of bovine embryos

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Shiori Ashibe ◽  
Kanade Irisawa ◽  
Ken Yokawa ◽  
Yoshikazu Nagao
Keyword(s):  
Treatment Group ◽  
Assisted Reproductive Technologies ◽  
Cumulus Cells ◽  
Reproductive Technologies ◽  
Control Group ◽  
Free Calcium ◽  
Parthenogenetic Development ◽  
Early Embryos ◽  
Bovine Oocytes

Summary Hyaluronidase is widely used in animal and human assisted reproductive technologies (ARTs) to remove cumulus cells around oocytes. However, adverse effects of hyaluronidase treatment, such as increased rates of degeneration and parthenogenesis, have been found after treatment of human and mouse oocytes. Currently, the mechanism(s) of the detrimental effects are unclear. The present study was initiated to identify the mechanism of adverse responses to hyaluronidase treatment in bovine oocytes and early embryos. Cumulus cells were removed from cumulus–oocyte complexes (COCs) with or without hyaluronidase and the oocytes were subjected to intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF). Significantly lower rates of blastocyst formation were obtained in the hyaluronidase treatment group after ICSI (22.4%) and IVF (21.2%) compared with the non-hyaluronidase control groups: 36.1% after ICSI and 30.4% after IVF. Next, we examined the effect of hyaluronidase on parthenogenetic development rates and on the cytoplasmic levels of free calcium ions (Ca2+), reactive oxygen species (ROS) and reduced glutathione (GSH). No differences in parthenogenesis rates were found between treated and untreated groups. Ca2+ levels in oocytes from the hyaluronidase treatment group indicated using mean fluorescence intensity were significantly higher (68.8 ± 5.3) compared with in the control group (45.0 ± 2.5). No differences were found in the levels of ROS or GSH between the treated and untreated groups. We conclude that hyaluronidase might trigger an increase in Ca2+ levels in oocytes, resulting in a decreased potential for normal embryonic development.

scholarly journals Can Dog-Assisted Intervention Decrease Anxiety Level and Autonomic Agitation in Patients with Anxiety Disorders?

2021 ◽  
Vol 10 (21) ◽  
pp. 5171
Author(s):  
Dorota Wołyńczyk-Gmaj ◽  
Aleksandra Ziółkowska ◽  
Piotr Rogala ◽  
Dawid Ścigała ◽  
Ludwik Bryła ◽  
...  
Keyword(s):  
Heart Rate ◽  
Treatment Group ◽  
Trait Anxiety ◽  
State Anxiety ◽  
Brief Symptom Inventory ◽  
Anxiety Symptoms ◽  
Control Group ◽  
Resting Heart Rate ◽  
Full Data ◽  
Positive Effects

Few studies have explored the influence of an Animal-Assisted Intervention on patients with mental disorders. We investigated it’s impact on anxiety symptoms. We divided 51 patients with anxiety symptoms into two groups—treatment group, that went for a short 15–20 min’ walk with a dog, his handler and a researcher and control group, that went for a walk only with a researcher. We used State-Trait Anxiety Inventory (STAI), Visual Analogue Scale (VAS) of fear, Beck Depression Inventory (BDI), Ford Insomnia Response to Stress (FIRST), Brief symptom Inventory (BSI) and VAS of satisfaction after trial to assess. We also checked the resting blood pressure and resting heart rate before and after performing psychological tests while sitting. We have obtained full data of 21 people from the research group and 26 people from the control group. After the intervention, the treatment group reported lower anxiety levels as a state (Mean (M) = 34.35; Standard Deviation (SD) = 6.9 vs. M = 40.94; SD = 8.6) and fear (M = 1.05; SD = 1.0 vs. M = 2.04; SD = 2.2) than the control group. After a walk with a dog, trait anxiety (M = 34.35; SD = 6.9 vs. M = 46.3; SD = 9.6), state anxiety (M = 48.9; SD = 7.2 vs. M = 53.9; SD = 7.8), fear (M = 1.05; SD= 1.0 vs. M = 2.57; SD = 2.3) and resting heart rate (M = 71.05; SD = 12.3 vs. M = 73.67; SD = 13.1) decreased significantly, while walking without a dog only reduced state anxiety (M = 47.24; SD = 11.0 vs. M = 40.94; SD = 8.6). Multivariate analysis of variance showed that after the walk, state anxiety was significantly lower in the treatment group than in the control group, F(1.35) = 6.706, p <0.05, η2 = 0.161. Among those who walked with a dog, the intervention also led to significant decreases in fear and resting heart rate, F(1.44) = 11.694, p < 0.01, η2 = 0.210 and F(1.45) = 8.503; p < 0.01; η2 = 0.159, respectively. For anxious patients, a short walk with a dog is more beneficial than a walk without one. We found significant positive effects of a dog’s company on vegetative arousal and mental comfort. This is another study confirming the possible therapeutic effect of the animal on anxiety symptoms. Further research is required, especially in the large groups of patients, as recommendations on the use of Animal Assisted Interventions (AAI) are needed.

scholarly journals The Effect of Adding Different Levels of Curcumin and Its Nanoparticles to Extender on Post-Thaw Quality of Cryopreserved Rabbit Sperm

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1508 ◽  
Author(s):  
Sameh Abdelnour ◽  
Mahmoud Hassan ◽  
Amer Mohammed ◽  
Ahmad Alhimaidi ◽  
Naif Al-Gabri ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Positive Influence ◽  
Control Group ◽  
Curcumin Nanoparticles ◽  
Total Antioxidant ◽  
Semen Extender ◽  
Apoptosis Process ◽  
Structural Levels

The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (−196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.

scholarly journals Antibacterial Activity of Lumbricus Rubellus Earthworm Extract Against Porphyromonas Gingivalis as the Bacterial Cause of Periodontitis

2019 ◽  
Vol 7 (6) ◽  
pp. 1032-1036 ◽  
Author(s):  
I Gusti Agung Ayu Dharmawati ◽  
Tjokorda Gde Bagus Mahadewa ◽  
I Putu Eka Widyadharma
Keyword(s):  
Antibacterial Activity ◽  
Treatment Group ◽  
Lumbricus Rubellus ◽  
Control Group ◽  
Inhibitory Zone ◽  
Control Group Design ◽  
Zone Diameter ◽  
Significant Difference ◽  
The Mean

AIM: The purpose of this study was to determine the antibacterial activity of Lumbricus rubellus earthworms through inhibitory zone diameter to the growth of the bacterium Phorphyromonas gingivalis as the cause of periodontitis. METHODS: This was an experimental study with randomised posttest-only control group design. The study was conducted at the Microbiology Research Center laboratory at the Faculty of Dentistry, Airlangga University, Indonesia. The study was conducted in vitro, the sample size was calculated using the Federer formula as many as four agar plates containing bacteria Phorphyromonas gingivalis, with each plate given five different treatments: control (ethanol), Lumbricus rubellus earthworm extract (ECT) with concentrations of 50%, 25%, 12.5%, and 6.25% respectively. The data in the form of inhibition zone diameter (measured in millimetres) obtained were tested using One-Way ANOVA. RESULTS: The mean diameter of the inhibitory zone extract of Lumbricus rubellus earthworm on the growth of Phorphyromonas gingivalis bacteria in the treatment group had significant differences (p < 0.05). The mean inhibition zones between controls and the ECT treatment group (ECT 50%, ECT 25%, ECT 12.5%) were statistically different (p < 0.05), in contrast with ECT 6.25% (p > 0.05) which did not show significant difference with the control group (p > 0.05). CONCLUSION: Lumbricus rubellus earthworm extract with a concentration of 50% has the largest diameter of the inhibitory zone on the growth of the Phorphyromonas gingivalis bacteria. The 6.25% earthworm extract showed no antibacterial activity against the growth of Phorphyromonas gingivalis bacteria.

The effects of chronic administration of ghrelin on rat sperm quality and membrane integrity

2009 ◽  
Vol 59 (2) ◽  
pp. 159-168 ◽  
Author(s):  
Arash Kheradmand ◽  
Majid Taati ◽  
Homayoon Babaei
Keyword(s):  
Plasma Membrane ◽  
Treatment Group ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Chronic Administration ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Significant Difference

AbstractAlthough ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the effects of chronic administration of ghrelin on the motility, plasma membrane integrity and concentration of rat spermatozoa. 45-d male Wistar rats were scheduled for the study and were divided into control and treatment groups. In the treatment group, 1 nmol of ghrelin was administered as sc injection for 10 consecutive days or vehicle (physiological saline) to the control rats. Sperm collection was achieved by killing of the rats on days 15, 25 and 50 after first injection. Total sperm motility and forward progressive movement did not exhibit significant difference during the experiment, although, there was a tendency for greater motion rate on d 15 and 25 in the treated rats compared to the control group. Plasma membrane integrity (HOS-reacted spermatozoa) was significantly higher in the treated animals, especially on day 15 as well as day 25, because of possible antioxidant properties of ghrelin. This value was statistically higher on day 15 than that of day 25 (P <0.05). Likewise, there was a significant correlation between the FPM (P <0.0001, r = 0.79) and TSM (P <0.01, r = 0.52) with the HOS test percentage in the treatment group. It was not observed statistically difference in the sperm concentration between groups during all of the experimental days. In conclusion, chronic administration of ghrelin (similar to induced by energy deficiency such as fasting) increased the integrity of sperm membrane, however, the sperm motility and concentration did not display any alterations.

scholarly journals Transformasi Genetik Tanaman Kentang (Solanum tuberosum L.) dengan Gen acvB Menggunakan Vektor Agrobacterium tumefaciens

2021 ◽  
Vol 11 (1) ◽  
pp. 63
Author(s):  
DARWIN SILALAHI ◽  
I GEDE PUTU WIRAWAN ◽  
MADE SRITAMIN
Keyword(s):  
Solanum Tuberosum ◽  
Agrobacterium Tumefaciens ◽  
Treatment Group ◽  
Genetic Transformation ◽  
Solanum Tuberosum L ◽  
Control Group ◽  
Shoot Height ◽  
Specific Pcr ◽  
Convenient Technique

Agrobacterium tumefaciens Mediated Genetic Transformation of acvB Gene in Potato (Solanum tuberosum L.). Genetic transformations are now routinely applied to plant mediated by Agrobacterium tumefaciens as the most convenient technique. This study aimed to prove the success of A. tumefaciens mediated genetic transformation in potato. A. tumefaciens LBA (pBI 121) and explant of potato shoot were used in this study. Explants were grown in vitro on Murashige and Skoog media. Transformation was implemented using smear technique by smearing A. tumefaciens to injured explant. Experimental groups consisted of two groups: control group which did not receive transformation treatment and treatment group receiving transformation treatment. Explant growth was observed through the presence of shoots, branches and the shoot height. Explants in the treatment group resulted in a higher number of shoots, branches, and shoot heights compared to control. Phenol compounds appear in explant epidermal tissue, indicating the wounds produced by A. tumefaciens infection, thus the gene predicted to be transformed. Identification by PCR is needed to prove the existence of the acvB gene in potato plants genome, using acvB specific PCR primer as the marker, such as (5?-CCCT CTAG AGAC CCGC GCCA AGGCG-3?) and (5?CGCG TCGA CCTT GTCG GAAAG -3?) with 540-bp in base pair size produced.

scholarly journals Effect of oocyte vitrification before and after in vitro maturation towards Bcl-2, Bax and Bcl-2/Bax ratio expression

2017 ◽  
Vol 24 (2) ◽  
pp. 56
Author(s):  
Zakiyatul Faizah ◽  
Haryanto Aswin ◽  
Hamdani Lunardhi
Keyword(s):  
Treatment Group ◽  
Ethylene Glycol ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Basic Medium ◽  
Control Group ◽  
Oocyte Vitrification ◽  
Expression Ratio ◽  
Before And After

Objectives: to compare the expression of Bcl-2, Bax and Bcl-2/Bax ratio in cumulus cell and oocyte between vitrified oocyte pre and post in vitro maturation.Materials and Methods: Maturation was operated in medium TC 100 µl for 24 hours. Vitrification begins with washing oocyte in PBS basic medium supplemented of 20% serum for 1-2 minutes, followed by equilibration medium PBS + 20% serum + 10% ethylene glycol for 10-14 minutes, then transferred to 20% serum + PBS + 0.5 M sucrose + 15% ethylene glycol + PROH 15% for 25-30 seconds. Thawing is processed by submerging the oocytes in the media: 1). PBS + 20% serum + 0.5 M sucrose, 2). PBS + 20% serum + 0.25 M sucrose, and 3). PBS + 20% serum + 0.1 M sucrose. Imunocytochemistry observed the expression of Bcl-2, bax and Bcl-2/bax ratio.Results: Bcl-2 expression on oocyte in control group differed significantly with treatment group, Bcl-2 expression on cumulus in control group differed significantly with treatment 1 group. Bax expression on oocyte in control group differed significantly with treatment group. Bax expression on cumulus in control group differed significantly with treatment group. Bcl-2/Bax expression ratio on oocyte and cumulus did not differ significantly in all groupConclusion: No difference Bcl-2/Bax expression ratio on oocyte and cumulus between vitrified oocyte pre and post in vitro maturation.

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