92 PROCESSING OF POSTMORTEM BOVINE EPIDIDYMAL SPERM AFTER COOLING THE TESTES FOR 24 HOURS

2008 ◽  
Vol 20 (1) ◽  
pp. 126 ◽  
Author(s):  
J. R. Saenz ◽  
C. A. Guerrero ◽  
D. Paccamonti ◽  
B. Eilts ◽  
K. R. Bondioli ◽  
...  
Keyword(s):  
Room Temperature ◽  
Cold Shock ◽  
Membrane Integrity ◽  
Sperm Parameters ◽  
Epididymal Sperm ◽  
Mean Values ◽  
Circular Pattern ◽  
Plastic Bags ◽  
Cold Room ◽  
Medial Section

Harvesting and cryopreservation of epididymal sperm from postmortem animals is one way to save gametes from genetically valuable males. Before sperm can be properly harvested from the epididymides, usually the testes need to be cooled and transported to a semen laboratory. Previously, it has been recommended to harvest epididymal sperm from cooled testes in a 4°C cold room. However, a walk-in cold room is not always available. The objective of this study was to hold the testes for 22 h at 4°C, and then remove the epididymal sperm at room temperature (22 to 23°C; Treatment A) or in a cooled environment (4°C; Treatment B). Testicles within the scrotum were collected from sexually mature mixed breed bulls (n = 11) at a local abattoir, placed into plastic bags, and transported (2 h) in a Styrofoam ice chest (pre-cooled with frozen gel packs) to the laboratory. Each pair of testes was then removed from the ice chest and placed into a refrigerator at 4°C for 22 h. After 22 h of cooling, each testicle of the pair was removed from the scrotum and randomly assigned to either Treatment A or Treatment B. The sperm were flushed in a retrograde flow out of a small incision made at the medial section of the cauda epididymides. Total motility (TM) and progressive motility (PM) were determined using an inverted Nikon Diaphot microscope. Membrane integrity (MI) was determined using SYBR 14 and propidium iodide staining under a microscope with epifluorescence detection capability. Sperm in a Triladyl® one-step extender were frozen in 0.5-mL plastic straws in the vapor 2 cm above the LN2. A paired t-test was used for statistical analyses. In summary, there was significant decrease (P ≤ 0.05) from pre-freeze (PF) to post-thaw (PT) for all of the sperm parameters within each treatment (Table 1). However, there were no significant differences between PF and PT sperm parameters between treatments for any of the parameters measured. Post-thaw sperm from all samples had a tendency to swim in a circular pattern after warming, which is a known sign of sperm affected by cold shock. It is likely that during their cold storage, testes were cooled too quickly, inducing cold shock. In summary, cooling bull testes for 24 h and processing them at room temperature produced results similar to those for processing the testes in a cool environment. The PT parameters in this study suggest that these sperm could be used for IVF and/or ICSI procedures. Table 1. Pre-freeze and post-thaw mean values (±SEM) for bull epididymal sperm parameters

168 POST-THAW VIABILITY OF EPIDIDYMAL SPERM FROM WHITE-TAIL DEER THAT WAS FROZEN USING GLYCEROL OR DMSO AS THE CRYOPROTECTANT

2009 ◽  
Vol 21 (1) ◽  
pp. 183
Author(s):  
M. A. Stout ◽  
J. S. Saenz ◽  
G. T. Gentry ◽  
S. P. Leibo ◽  
K. R. Bondioli ◽  
...  
Keyword(s):  
Treatment Group ◽  
Sperm Morphology ◽  
Membrane Integrity ◽  
Slow Cool ◽  
Epididymal Sperm ◽  
Cold Environments ◽  
Normal Sperm ◽  
Nitrogen Vapor ◽  
Dmso Treatment ◽  
Medial Section

Cryoprotectants are compounds that protect and maintain the viability of cells when being subjected to cold environments. In addition to glycerol, DMSO has been evaluated for post-thaw epididymal sperm viability in various endangered species, but has not yet been assessed in White-tail deer. Testicles (in the scrotum) were collected from hunter harvested sexually mature White-tail bucks (n = 7) during the peak rutting season. Testicles within the scrotum, once removed from the postmortem buck, were placed in a plastic Ziploc bag and then into a Styrofoam ice chest (at ambient temperature) and transported to the Embryo Biotechnology Laboratory (EBL). Upon arrival at the EBL, sperm were flushed in a retrograde flow out of a small incision at the medial section of the cauda epididymides. Epididymal sperm were flushed from the epididymis using a non glycerol extender. Sperm from both epididymides of each buck were pooled and allowed a slow cool to 4°C. Upon reaching 4°C, the pooled epididymal sperm were equally divided and either 6% glycerol (A) or 6% DMSO (B) was added to each sperm sample. After the addition of the cryoprotectant sperm were loaded into 0.5-mL plastic straws and cryopreserved using liquid nitrogen vapor. Total motility (TM) was determined subjectively using an inverted Nikon Diaphot microscope. Membrane integrity (MI) was determined using SYBR 14 and propidium iodide staining under a microscope equipped with epiflurescence. Morphology was determined using an eosin-nigrosin staining. A paired t-test was used for statistical analyses. There were no differences between the glycerol and DMSO treatments for any of the three parameters measured (Table 1). However, post-thaw total motility and membrane integrity were significantly lower for both treatment groups (A and B) when compared with pre-freeze values. Furthermore, the DMSO treatment group had significantly lower normal morphology when compared with pre-freeze values, but was not different than the glycerol treatment group. Our results indicate that cryopreservation of epididymal sperm from White-tail deer with either 6% glycerol or 6% DMSO yield similar results for total motility, membrane integrity, and normal sperm morphology. Table 1.Pre-freeze and post-thaw total sperm motility, membrane integrity and normal sperm morphology for White-tail deer epididymal sperm Tony Vidrine of the Louisiana Department of Wildlife and Fisheries.

scholarly journals Evaluation of a panel of spermatological methods for assessing reprotoxic compounds in multilayer semen plastic bags

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
M. Schulze ◽  
F. Schröter ◽  
M. Jung ◽  
U. Jakop
Keyword(s):  
Membrane Integrity ◽  
Diglycidyl Ether ◽  
Mitochondrial Activity ◽  
Prolonged Storage ◽  
Prolonged Incubation ◽  
Plastic Bags ◽  
Plastic Materials ◽  
Bisphenol A Diglycidyl Ether ◽  
Semen Preservation ◽  
Boar Spermatozoa

AbstractThe increase of fertility performance in sows is one of the biggest achievements in pig production over the last 30 years. Nevertheless, pig farms using artificial insemination (AI) repeatedly experienced in recent year’s fertility problems with dramatic consequences due to toxic compounds from plastic semen bags. In particular, bisphenol A diglycidyl-ether (BADGE) present in multilayer plastic bags can leach into the semen and could affect the functionality of the spermatozoa. Former studies could not find any alterations in spermatozoa based on the exposure to BADGE. The aim of the study was to evaluate effects of BADGE on boar spermatozoa using an extended panel of spermatological methods. In spring 2019, a large drop in farrowing rates from 92.6 ± 2.3% to 63.7 ± 11.1% in four sow farms in Croatia was detected. In migration studies, BADGE could be identified as a causal toxic compound and leached into the extended semen in concentration of 0.37 ± 0.05 mg/L. Detailed spermatological studies showed that significant predictors for effects on spermatozoa were different levels of motility and kinematic data after a prolonged storage time, thermo-resistance test (prolonged incubation time), mitochondrial activity, membrane integrity and fluidity. No serious effects were observed for sperm morphology and DNA fragmentation. These results provide new insights into the development of a new quality assurance concept for a detailed spermatological examination during testing of plastic materials for boar semen preservation. It could be shown that boar spermatozoa are an excellent biosensor to detect potential toxicity and fertility-relevant compounds.

Laboratory formation of border ice and frazil slush

1977 ◽  
Vol 4 (2) ◽  
pp. 153-160 ◽  
Author(s):  
T. O'D. Hanley ◽  
B. Michel
Keyword(s):  
Room Temperature ◽  
Variable Speed ◽  
Water Ice ◽  
Air Temperatures ◽  
Cold Room ◽  
Rate Of Increase ◽  
Steel Tank ◽  
Cylindrical Steel ◽  
Variable Speed Motor ◽  
Water Speed

Ice formation under controlled conditions was studied in a cold room using a cylindrical steel tank 120 cm in diameter and 76 cm deep. Paddles turned by a variable speed motor moved the water about the axis of the tank. Twenty-one thermistors were used to record temperatures above and below the surface of the water. Ice was allowed to form with the cold room temperature held at −2 °C, −5 °C, −10 °C, and −20 °C and with water speeds (measured 4 cm from the tank wall) from 0 to 73 cm/s.At zero water speed ice formed first as needles randomly oriented over the surface. With flowing water, border ice width increased linearly with time, but independently of water speed. The rate of increase of the border width w fits the equation dw/dt = (4/9)|Ta|0.68 where Ta is the Celsius air temperature.Frazil was never observed at water speeds less than 24 cm/s, but was always formed at this or greater speeds, regardless of cold room temperature. The volume of slush accumulated was greater at greater water speeds and at colder air temperatures. Observed slush volumes are compared with semiempirical values of the ice mass obtained from theoretical equations. The data suggest that growth of ice crystals began when the water was supercooled by approximately 0.02 °C.

Effects of B Vitamins, Liver and Yeast on Growth under Cold Room and Room Temperature Conditions

1948 ◽  
Vol 35 (1) ◽  
pp. 39-48 ◽  
Author(s):  
E. D. Smith ◽  
B. H. Ershoff ◽  
R. J. Winzler ◽  
H. J. Deuel
Keyword(s):  
Room Temperature ◽  
B Vitamins ◽  
Temperature Conditions ◽  
Cold Room

THE SUSCEPTIBILITY OF SPERMATOZOA TO TEMPERATURE SHOCK

1959 ◽  
Vol 19 (3) ◽  
pp. 211-220 ◽  
Author(s):  
R. G. WALES ◽  
I. G. WHITE
Keyword(s):  
High Temperatures ◽  
Seminal Plasma ◽  
Room Temperature ◽  
Cold Shock ◽  
Differential Staining ◽  
Temperature Shock

SUMMARY The susceptibility of bull, ram, rabbit, dog, human and fowl spermatozoa to cold shock and high temperatures has been assessed. Motility and differential staining were used as criteria. Ram and bull spermatozoa were increasingly affected by cold shock at temperatures below 15° C; other spermatozoa were, however, little affected. Epididymal ram spermatozoa, particularly those with an attached kinoplasmic droplet, were more resistant than ejaculated ones; the addition of seminal plasma had little effect. Second ejaculates from bulls were slightly more resistant than first ejaculates. Washing bull or fowl spermatozoa free of seminal plasma did not influence their susceptibility to cold shock. Five min at 50° C severely depressed the motility of all spermatozoa except those of the fowl which were, however, completely immobilized at 55° C. Most spermatozoa took up stain more readily when mixed with it at high temperatures than when brought back to room temperature and then mixed; this is due to an increase in the toxicity of the stain at high temperatures.

scholarly journals Oxidative stability of tilapia feeds containing Saccharomyces cerevisiae and Spirulina platensis

2020 ◽  
Vol 50 (1) ◽  
Author(s):  
Thiago Luís Magnani Grassi ◽  
Juliana Sedlacek-Bassani ◽  
Elisa Helena Giglio Ponsano
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Vitamin E ◽  
Spirulina Platensis ◽  
Nile Tilapia ◽  
Room Temperature ◽  
Control Diet ◽  
Thiobarbituric Acid ◽  
Thiobarbituric Acid Reactive Substances ◽  
Oxidative Rancidity ◽  
Plastic Bags

ABSTRACT: The aim of this study was to evaluate the effect of the inclusion of microbial biomass on the oxidative rancidity of tilapia rations stored for 12 months. Treatments included a control diet and diets supplemented with either 0.01% vitamin E, 0.25 and 0.5% of Saccharomyces cerevisiae and 0.25 and 0.5% of Spirulina platensis. Experimental diets were stored in the dark inside plastic bags at room temperature (25 °C) for 12 months. The oxidative rancidity was measured as thiobarbituric acid reactive substances (TBARS). It was concluded that the inclusions of Spirulina platensis at 0.25% (1.734 ± 0.206) and 0.5% (1.629 ± 0.181) and Saccharomyces cerevisiae at 0.5% (1.459 ± 0.305) minimized the oxidative rancidity in comparation to control diet (2.843 ± 0.109) of Nile tilapia until 12 months of storage.

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.

Effect of sweat loss on body fluids

1964 ◽  
Vol 19 (6) ◽  
pp. 1119-1124 ◽  
Author(s):  
Stanislaw Kozlowski ◽  
Bengt Saltin
Keyword(s):  
Body Weight ◽  
Water Loss ◽  
Evans Blue ◽  
Room Temperature ◽  
Extracellular Fluid ◽  
Sweat Loss ◽  
Average Decrease ◽  
Plastic Bags ◽  
Inulin Space ◽  
Additional Water

Six healthy men were studied under normal conditions and after dehydration caused by sweating produced 1) in a sauna at 80 C, 2) by hard muscular work at 18 C, and 3) by mild exercise at room temperature 38 C. The dehydration period lasted for 2.5@#X2013;3.5 hr. Sweat was collected in impermeable plastic bags around the forearm. Body weight, Evans blue space, and apparent inulin space were determined before and about 90 min after the dehydration period. The average decrease in body weight was 3.1 (4.1%), 3.1, and 3.5 kg for 1, 2, and 3, respectively. The reduction in apparent inulin space was 1.4, 0.2 and 1.3 liters, respectively. The decrease in Evans blue space paralleled the reduction in apparent inulin space. In the three conditions no significant differences were found in calculated total loss of electrolytes. Water liberated from combustion of fat and carbohydrates plus water previously stored with glycogen can account for up to 1.1 liters of the intracellular water loss during the hard exercise (2). The additional water loss from the cells is discussed in light of electrolyte shifts. dehydration; intracellular fluid; extracellular fluid; plasma volume Submitted on January 20, 1964

Effects of cold exposure and dehydration on renal function in black-tailed prairie dogs

1977 ◽  
Vol 42 (2) ◽  
pp. 295-299 ◽  
Author(s):  
J. D. Hamilton ◽  
E. W. Pfeiffer
Keyword(s):  
Renal Function ◽  
Room Temperature ◽  
Sodium Concentration ◽  
Urine Concentration ◽  
Cynomys Ludovicianus ◽  
Prairie Dogs ◽  
Body Temperatures ◽  
Concentration Gradients ◽  
Plasma Urea ◽  
Cold Room

Black-tailed prairie dogs (Cynomys ludovicianus) were deprived of food and water for several weeks during the fall and winter in a cold-room hibernaculum (Ta 5–8 degrees C), and for several days at room temperature during the summer. Body temperatures (Tb) were determined periodically in nine animals by radiotransmitters implanted in the abdomen. Animals deprived of food and water in the summer were killed when maximum urine concentration was achieved. Eight animals in the winter were active when killed after 7–35 days in the hibernaculum with Tb between 18 and 36 degrees C. Five animals that became torpid periodically in the winter were killed after 19–42 days in the hibernaculum when their Tb indicated torpor (Tb less than 13 degrees C). Active animals in the summer and winter possessed pronounced renal corticomedullary urea and sodium concentration gradients. Torpid animals lacked these gradients and had lower urine and plasma osmotic concentrations than active animals. Plasma urea values and terminal osmolal U/P ratios were lowest in torpid prairie dogs.

Acetate-1-C14 metabolism of white fat from hamsters in cold exposure and hibernation

1963 ◽  
Vol 205 (5) ◽  
pp. 905-908 ◽  
Author(s):  
Joan Baumber ◽  
Arliss Denyes
Keyword(s):  
Cold Exposure ◽  
Room Temperature ◽  
Fat Pad ◽  
Golden Hamsters ◽  
Epididymal Fat ◽  
Cold Room ◽  
White Fat

Incorporation of C14 from acetate-1-C14 into lipid and CO2 by epididymal fat from golden hamsters kept at room temperature, acclimated to 5 ± 1 C, in hibernation and arousing from hibernation, was measured in vitro at 37 C. Summer and winter series were compared. The C14O2 production by tissue from control and acclimated animals was similar but the C14O2 production of tissue from hibernating and arousing hamsters was significantly greater than that from acclimated animals. There was a large increase in the lipid-C14 of tissue from cold-acclimated animals and this increase persisted into hibernation but was slightly depressed in tissue from arousing animals. Many acclimated and all hibernating hamsters had involuted testes and a greater incorporation of C14 into lipid than those with noninvoluted testes. A greater percentage of hamsters hibernated in the cold room during the winter and at this time the incorporation of C14 into lipid by the fat pad was greater than in the summer.

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