scholarly journals Aseptic Technology for Cryoprotectant-Free Vitrification of Human Spermatozoa by Direct Dropping into Clean Liquid Air: Apoptosis, Necrosis, Motility, and Viability

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Mengying Wang ◽  
Evgenia Isachenko ◽  
Plamen Todorov ◽  
Gohar Rahimi ◽  
Peter Mallmann ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Microbial Contamination ◽  
Good Alternative ◽  
Human Spermatozoa ◽  
Progressive Motility ◽  
Cooling Agent ◽  
Late Apoptosis ◽  
Apoptosis And Necrosis ◽  
Motility Rate

This study aimed to compare the quality of human spermatozoa vitrified by direct plunging into liquid nitrogen vs. liquid air. Spermatozoa were divided into three groups: fresh spermatozoa (Group F) were used as a control. Spermatozoa suspension (20 μl) was vitrified in open granules by direct dropping into liquid nitrogen (Group LN) or clean liquid air (Group LA). After warming at 37°C, the progressive motility rate of Group F was reduced from 65.9 ± 2.5% to 34.0 ± 1.9% (Group LN) and 38.1 ± 2.3% (Group LA), respectively (P1-2,3 < 0.05). The reductions in viability were 65.6 ± 2.2%, 29.0 ± 1.8%, and 36.6 ± 2.6% for Groups F, LN, and LA, respectively (P1-2,3 < 0.05). Comparing spermatozoa vitrified in liquid nitrogen vs. liquid air, no significant differences were detected in motility (34.0 ± 1.9% vs. 38.1 ± 2.3%), viability (29.0 ± 1.8% vs. 36.6 ± 2.6%), early apoptosis (13.8 ± 1.5% vs. 14.3 ± 1.8%), late apoptosis (45.5 ± 1.8% vs. 43.7 ± 2.2%), and necrosis (19.5 ± 2.0% vs. 15.0 ± 1.8%; p>0.01 for all respective differences). There was a statistical tendency for increasing rates of “progressive motility” and “viability” and decreasing rates of “apoptosis” and “necrosis” when comparing spermatozoa vitrified in liquid air vs. liquid nitrogen. It is concluded that cryoprotectant-free vitrification by the direct dropping of human spermatozoa in a clean cooling agent (liquid air) is a good alternative to the use of nonsterile liquid nitrogen and can be used to cool cells while minimising the risk of microbial contamination.

scholarly journals Cholesterol-loaded cyclodextrin plus trehalose improves quality of frozen-thawed ram sperm

2019 ◽  
Vol 64 (No. 03) ◽  
pp. 118-124 ◽  
Author(s):  
ME Inanc ◽  
S Gungor ◽  
C Ozturk ◽  
F Korkmaz ◽  
I Bastan ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Mitochondrial Membrane ◽  
Improve Quality ◽  
Progressive Motility ◽  
Computer Aided ◽  
Acrosome Integrity ◽  
Ram Sperm ◽  
Additive Control ◽  
Artificial Vagina

The objective of this study was to determine effects of supplementing Tris-based semen extenders with either cholesterol-loaded cyclodextrin (CLC) or 7-dehydrocholesterol loaded cyclodextrin (7-DCLC) plus trehalose (T) for cryopreservation of ram semen. Semen was collected with an artificial vagina from five Merino rams (2–3 years of age) during the non-breeding season. Ejaculates were pooled, divided into eight equal portions, diluted with a standard Tris-based extender containing: no additive (control); T (50 mM); or T (50 mM) + 1.5, 2.5 or 3.5 mg of either 7-DCLC or CLC. Semen was chilled from 37°C to 4°C, placed in 0.25 ml French straws, held 5 cm above liquid nitrogen for 12 minutes, then plunged into liquid nitrogen. After thawing, a computer-aided semen analyzer system (CASA) was used to assess motility, whereas plasma membrane and acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were assessed with flow cytometry. Sperm supplemented with 2.5 mg and 3.5 mg CLC + T had the highest (P &lt; 0.05) total and progressive motility (65.2 ± 4.7 and 19.0 ± 1.0% respectively, mean ± SEM), albeit with no significant differences from sperm with 1.5 or 3.5 mg CLC + T. Sperm with 2.5 mg CLC + T had the highest (P &lt; 0.05) PMAI (59.3%; not different from 3.5 mg CLC + T) and highest (P &lt; 0.05) HMMP (64.6%; not different from 1.5 or 3.5 mg CLC + T). The lowest ALH value, 2.8 ± 0.3 µm was in the 2.5 mg 7-DCLC + T group; otherwise, there were no significant differences among groups for any other CASA end point. In conclusion, adding CLC + T to a tris-based extender optimized quality of frozen-thawed ram semen. Therefore, extenders including CLC + T have potential to improve quality of frozen-thawed ram sperm.

scholarly journals Effect of Oral Administration of Selenium and Vitamin E on the Quality of Fresh, Refrigerated and Frozen Semen in French Bulldog Breed Dogs

2017 ◽  
Vol 45 (1) ◽  
pp. 7
Author(s):  
Marcelo George Mungai Chacur ◽  
Mariana Grandis Ripari de Souza ◽  
Camila Dutra de Souza ◽  
Camila Pires Cremasco
Keyword(s):  
Vitamin E ◽  
Citric Acid ◽  
Oral Administration ◽  
Liquid Nitrogen ◽  
Sperm Concentration ◽  
Oral Supplementation ◽  
Frozen Semen ◽  
French Bulldog ◽  
Fresh Semen

Background: New methodologies have been developed seeking to maximize pregnancy rate in female dogs created in commercial kennels, and also in order to maintain the quality of canine semen after dilution, refrigeration or freezing. One of the main factors that generate damage to sperm is oxidative stress, to minimize sperm damage, selenium and antioxidants like vitamin E are administered, by oral administration, seeking to improve the quality of semen. The objective was to study the effect of vitamin E and selenium, by oral administration, in the quality of fresh, refrigerated and frozen semen in adult dogs French Bulldog breed.Materials, Methods & Results: Semen samples were collected from 5 adult dogs, French Bulldog breed, being 2 semen drawing before the daily oral supplementation with vitamin E and selenium (ESE®) and semen drawing at 20, 40 and 60 days after the beginning of oral supplement. The ejaculated samples were diluted in TRIS - fructose citric acid (3.28 g TRIS-hydroxy-methyl-amino-methane, 1.78 g of citric acid monohydrate and 1.25 g of D - fructose, dissolved in 100 mL of distilled water and added of 20% egg yolk and 6% of glycerol. The characteristics evaluated in fresh semen were: volume (mL), color, appearance, concentration (x106 / mL), sperm motility (%), sperm strength (1 to 5) and morphology (%). For refrigerated and frozen semen were analyzed: sperm motility (%), sperm strength (1-5) and morphology (%). Diluted semen samples were centrifuged at: 1500 g/10 min and “pellets” formed by sperm of each ejaculated, detached from the tube wall were diluted homogeneously in the diluent TRIS type up to the final volume of 1.5 mL. After that, packaged in 0.5 mL French straws, kept under refrigeration at 5ºC/4 h, placed in nitrogen vapor at -120ºC/15 min, and dipped in liquid nitrogen at -196ºC and then stored on identified rachis and stored in liquid nitrogen container until the time of thawing in  water bath at 37°C/30 s for semen microscopic analysis. Data from fresh, refrigerated and frozen semen were statistically analyzed by analysis of variance and the average compared by 5% of Tukey test. Fresh semen sperm concentration differed (P < 0.05) between the samples, rising after 40 days after the beginning of oral supplementation with selenium and vitamin E. For the spermatic strength, better score (P < 0.05) was observed at collection 4, in 40 days after the beginning of oral supplementation to dogs. For fresh and refrigerated semen, the total defects, defects of head, acrosome and tail did not differ (P > 0.05) between the samples. Total sperm defects and minor head and tail defects did not differ (P > 0.05) between the samples in post-thawing. Regarding the acrosome defects after thawing, there was a significant reduction (P < 0.05) in samples performed 40 and 60 days after the beginning of oral supplementation with selenium and vitamin E.Discussion: Attention should be paid for what purpose the extenders within the refrigeration or freezing biotech will be used. The managed supplement, by oral administration, containing selenium and vitamin E, influenced beneficially raising the sperm concentration in fresh semen and decreasing the acrosome defects in frozen semen. Oral administration of supplementation with selenium and vitamin E is recommended for improving the quality of fresh and frozen semen in dogs.

Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

2017 ◽  
Vol 29 (3) ◽  
pp. 490 ◽  
Author(s):  
Asmatullah Kaka ◽  
Wahid Haron ◽  
Rosnina Yusoff ◽  
Nurhusien Yimer ◽  
A. M. Khumran ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Optimum Level ◽  
Docosahexanoic Acid ◽  
Normal Morphology ◽  
Bull Semen ◽  
Acrosome Integrity

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

scholarly journals A Comparison of Cell Wall Disruption Techniques for the Isolation of Intracellular Metabolites from Pleurotus and Lepista sp.

2006 ◽  
Vol 61 (5-6) ◽  
pp. 347-350 ◽  
Author(s):  
Rilka M. Taskova ◽  
Holger Zorn ◽  
Ulrich Krings ◽  
Henning Bouws ◽  
Ralf G. Berger
Keyword(s):  
Secondary Metabolites ◽  
Liquid Nitrogen ◽  
Cell Walls ◽  
Intracellular Metabolites ◽  
Bead Milling ◽  
Nucleic Acid Isolation ◽  
Cell Wall Disruption ◽  
Bead Mill ◽  
Pleurotus Sapidus

Different techniques were compared for their effectiveness in the disruption of the rigid cell walls of Basidiomycetes. Grinding under liquid nitrogen, stirred glass bead milling and enzymatic cell lysis were applied to the mycelia of Pleurotus sapidus and Lepista irina grown submerged. Each of the disruption procedures was evaluated by testing the quantity and quality of released intracellular metabolites: DNA, RNA, enzymes, and secondary metabolites. The most suitable method for nucleic acid isolation was grinding under liquid nitrogen, while bead mill homogenization was the superior technique for isolation of active enzymes. A new effective method is proposed for isolation of secondary metabolites with the aid of bead milling of fungal mycelia.

Analysis of Micro Structure and the Resistivity of Cu / Ni Thin Coat as a Low Temperature Sensor Using Electroplating Method Assisted with Magnetic Field outside of the Ion Flow

2021 ◽  
Vol 885 ◽  
pp. 141-147
Author(s):  
Winda Noor Santi ◽  
Moh. Toifur
Keyword(s):  
Magnetic Field ◽  
Low Temperature ◽  
Liquid Nitrogen ◽  
Micro Structure ◽  
Medical Field ◽  
Cold Temperatures ◽  
Ion Flow ◽  
Resistance Temperature Detector ◽  
Temperature Detector

Preservation of materials using liquid nitrogen media has been widely used. One of them is used in the medical field, namely cryonic technology. Cryonics is a method of preservation at cold temperatures using a cryoprotectant in liquid nitrogen. To maintain the quality of the material, a sensor that can detect the temperature of liquid nitrogen is needed. Low temperature sensors with Cu and Ni based Resistance Temperature Detector with layers (RTD) have been made, but these sensors have a layer of Ni deposits that are not yet homogeneous. So quality improvement is needed by adding an external magnetic field. Based on this, the aim of this research is to synthesize a thin layer of Cu / Ni using electroplating method assisted by external magnets parallel to the ion currents

scholarly journals Sperm cryopreservation of lane snapper Lutjanus synagris(Linnaeus, 1758)

2015 ◽  
Vol 75 (3) ◽  
pp. 662-669 ◽  
Author(s):  
EG Sanches ◽  
IR Oliveira ◽  
PCS Serralheiro ◽  
VR Cerqueira
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Equilibration Time ◽  
Dilution Ratio ◽  
Sperm Cryopreservation ◽  
Ph Values ◽  
Cryopreserved Sperm ◽  
Time Density ◽  
Motility Rate

AbstractThis study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%), five cooling rates (110, 90, 60, 45 e 30°C –min), nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes) e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20) on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P<0.05) was achieved by combining extender with pH 8.2 with 10% concentration of dimethylsulfoxide and cooling rate 60°C –min, 1 minute of equilibration time and 1:3 (v/v) dilution ratio. The use of cryopreserved sperm presented fertilization rates >60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.

scholarly journals The use of oxybutynin for treating facial hyperhidrosis

2011 ◽  
Vol 86 (3) ◽  
pp. 451-456 ◽  
Author(s):  
Nelson Wolosker ◽  
José Ribas Milanez de Campos ◽  
Paulo Kauffman ◽  
Marco Antonio Munia ◽  
Samantha Neves ◽  
...  
Keyword(s):  
Quality Of Life ◽  
Good Alternative ◽  
Emotional Disturbances ◽  
Thoracic Ganglion ◽  
Compensatory Hyperhidrosis ◽  
Thoracic Sympathectomy ◽  
Serious Emotional Disturbances ◽  
Important Complication ◽  
Before And After

BACKGROUND: Facial hyperhidrosis is a disease that may lead patients to serious emotional disturbances. Video-assisted thoracic sympathectomy provides excellent resolution of facial hyperhidrosis, but is associated with certain complications. The most frequent and important complication is compensatory hyperhidrosis. Especially in patients who have undergone resection of the second thoracic ganglion, the risk of severe compensatory hyperhidrosis is higher, which may cause dissatisfaction with the procedure. OBJECTIVE: The aim of this study was to evaluate the efficacy of the use of low doses of oxybutynin in treating facial hyperhidrosis as well as the level of patient satisfaction with its use. METHODS: 25 patients with facial hyperhidrosis were treated with oxybutynin. The patients underwent 2 evaluations: before and after treatment. These evaluations were used to assess the patients' clinical improvement and quality of life. RESULTS: We observed that more than 75% of the patients evolved with an improvement in facial hyperhidrosis, and 52% of them presented a great improvement. CONCLUSION: Treatment of facial hyperhidrosis with oxybutynin is a good alternative to sympathectomy, since it presents good results and improves quality of life, in addition to not exposing patients to the risk of experiencing the side effects of sympathectomy

scholarly journals Microbial quality of beef and hygiene practices in small and medium slaughterhouses and butcheries in Uganda

2020 ◽  
Author(s):  
Juliet Kyayesimira ◽  
Wangalwa Rapheal ◽  
Grace Kagoro Rugunda ◽  
Lejju Julius Bunny ◽  
Morgan Andama ◽  
...  
Keyword(s):  
Microbial Contamination ◽  
Microbiological Quality ◽  
Microbial Load ◽  
Microbial Quality ◽  
E Coli ◽  
Hygiene Practices ◽  
Safe Level ◽  
Microbiological Methods ◽  
Beef Carcass

Abstract Background If hygiene practices along the beef processing nodes at small and medium enterprise (SME) slaughter houses and butcheries are not observed, they may pose a health risk due to microbial contamination. In SME slaughterhouses and butcheries, the risk may be higher due to transmission of foodborne pathogens. This study determined the hygienic practices and microbial quality risk among meat handlers (MH) in SME slaughterhouses and butcheries. Methods Assessment of microbiological quality of beef was carried out at slaughter houses and butcher shops in the districts of Western, Central and Eastern regions of Uganda. A cross sectional study was conducted from June 2017 to January 2018 using observation checklists to record unhygienic practices among the various actors. Microbial load at slaughter and butchery was determined from a total of 317 swab samples collected from carcass, tools, protective clothing and hands of meat handlers. The microbiological quality of beef was evaluated using standard microbiological methods. The samples were inoculated into differential and selective media. Results Butcheries had the highest microbial load on beef carcass ranging from 4.76 log 10 cfu/cm 2 to 7.90 log 10 cfu/cm 2 Total Viable Counts (TVC) while Total Coliform Counts (TCC) ranged from 1.42 log 10 cfu/cm 2 to 3.05 log 10 cfu/cm 2 , E. coli ranged from 0.68 log 10 cfu/cm 2 to 1.06 log 10 cfu/cm 2 and Staphylococcus aureus ranged from 3.25 log 10 cfu/cm 2 to 4.84 log 10 cfu/cm 2 . Salmonella was absent in all the samples analysed. Results of overall microbial quality of beef in Uganda indicated that only TCC (1.60±0.26 log 10 cfu/cm 2 ) of the beef carcass samples at slaughter houses was not significantly above the safe level (p = 0.693). Overall microbial load (TVC, TCC, E. coli and S. aureus ) at butcheries were significantly (p < 0.05) above the safe level. Butcheries of Mbale district had the highest percentage (70%) of beef carcass samples above the TCC safe levels whereas butcheries of Mbarara district had the highest percentage (40%) of beef carcass samples above the E. coli safe levels. TVC from hands and clothes at butchery across the three study districts varied significantly (p=0.007) with the highest counts (7.23 log 10 cfu/cm 2 ) recorded from personnel clothes and lowest (5.46 log 10 cfu/cm 2 ) recorded from hands. On the other hand, swab samples picked from chopping board and working table at the butchery did not show significant variation in TVC, TCC, E. coli and S. aureus microbial loads across the three study districts. Conclusion Hygienic handling of carcasses after slaughter is critical in preventing contamination and ensuring meat safety in informal meat trading sectors in Uganda. Handling practices of beef at Ugandan slaughterhouses and butcheries are not hygienic hence not up to standard and they contribute to microbial contamination of beef posing a risk to consumers. The distribution stage is the most critical period, during which the quality of meat can easily be compromised.

scholarly journals A STUDY TO ACCESS THE PHYSIO-CHEMICAL PARAMETERS OF WATER QUALITY OF SELECTED PONDS FROM THREE DISTRICTS OF TAMIL NADU, INDIA

2021 ◽  
Vol 21 (2) ◽  
Author(s):  
Kamalasan Prathap- Mathan ◽  
Joseph Babila- Jasmine ◽  
Muthukumar Thilagavathi
Keyword(s):  
Tamil Nadu ◽  
Microbial Contamination ◽  
Renewable Resource ◽  
Chemical Parameters ◽  
Chemical Examination ◽  
Physiochemical Parameters ◽  
The Earth ◽  
Living Organisms ◽  
Physical And Chemical

Water is the most abundant and non-renewable resource in the earth, which play an important role in all living organisms. A study on physiochemical parameters of Manur, Vallanadu and Sundarapandiapatinam ponds from Tirunelveli, Thootukudi and Ramanathapuram districts of Tamil Nadu, India, has addressed the seasonal changes. During the month of January to April samples from Manur was chemically portable, Vallanadu sample was physically portable, Sundarapandiapatinam samples were physically and chemically not portable. During the month of May to August Vallanadu and Manur samples were portable by its physical and chemical examination, Sundarapandiapatinam samples were non portable by its physiochemical property. September to December the samples from all the three ponds were non portable because of exceeding the permissible limit. All the samples were bacteriologically unsafe in nature because of its microbial contamination. This implies the water bodies are not fit for domestic and drinking purpose, thus proper management has to be done by the society and implement government guidelines to save the natural resources from manmade activities.

scholarly journals Assessment of quality in specific fractions of Large White Yorkshire boar semen

2021 ◽  
Vol 52 (2) ◽  
Author(s):  
K. G. Ambily ◽  
Malati Naik ◽  
Hiron M. Harshan ◽  
C. Jayakumar ◽  
M. P. Unnikrishnan ◽  
...  
Keyword(s):  
Cold Shock ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Membrane Cholesterol ◽  
Large White ◽  
Progressive Motility ◽  
Sperm Membrane ◽  
Boar Semen ◽  
Quality Assessments

Boar semen is voluminous and ejaculated as jets or fractions of pre-sperm, sperm rich (SRF) and post-sperm rich fractions. Recent studies have reported more resilient characteristics of sperm in initial portions of SRF towards cold shock and cryopreservation. The present study was conducted to assess the quality of specific fractions of SRF, namely, first 10mL of SRF (F1) and rest of SRF (F2) in Large white Yorkshire (LWY) boar semen. Ejaculates were collected using gloved-hand technique and were subjected to quality assessments of volume, pH, sperm progressive motility, concentration, plasma membrane integrity, abnormality, acrosome integrity and sperm membrane cholesterol. Upon statistical analysis, significant differences were noticed in volume, pH, sperm concentration and sperm membrane cholesterol between fractions of the ejaculate.

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