Uhlmannsiek L, Shen H, Eylers H, Martinsson G, Sieme H, Wolkers WF, Oldenhof H.Cryopreserved semen is routinely shipped in liquid nitrogen. Dry ice could serve as an alternative coolant, however, frozen storage above liquid nitrogen temperatures (LN2, -196 °C) may negatively affect shelf-life and cryosurvival. In this study, we determined critical temperatures for storage of cryopreserved stallion sperm. We evaluated: (i) effects of cooling samples to different subzero temperatures (-10 °C to -80 °C) prior to storing in LN2, (ii) stability at different storage temperatures (i.e., in LN2, dry ice, -80 °C and -20 °C freezers, 4 °C refrigerator), and (iii)...
Fracaro PL, Corcini CD, Souza Gatti NL, Filho JS, Acosta IB, Filho JS, Hartwig FP, Rosa Curcio BD, Varela Junior AS.Glycerol is a cryoprotectant widely used in the freezing of mammalian semen, but no study has demonstrated its optimum concentration and the appropriate exposure time for equine species. Objective: To demonstrate that the exposure time (15, 30, 45, 60, 75 and 90 min) versus concentration (2, 3, 4 and 5%) of the cryoprotectant glycerol influences the freezing success of equine semen. Methods: The ejaculate of 12 stallions were frozen in different glycerol concentrations following different exposure times. The thawed sperm was evaluated for kinetic parameters using a Computer Assisted Semen Anal...
Talluri TR, Jhamb D, Paul N, Singh J, Dedar RK, Mehta SC, Legha RA, Pal Y.The recovery of spermatozoa from the cauda epididymis may be the only option to obtain genetic material from elite stallions that had undergone castration or sudden death due to colic or severe injury. Objective: To evaluate two different protocols for retrieval of stallion epididymal spermatozoa and to evaluate different cryoprotectants on the freezability of the epididymal spermatozoa. Methods: Six epididymides from three stallions were collected immediately after routine castration under general anesthesia. In the first experiment, each epididymis (of two testes) of the same stallion were p...
Abnaroodheleh F, Mosavari N, Pourbakhsh SA, Tadayon K, Jamshidian M. is the main cause of glanders as a dangerous contagious zoonosis disease that is mostly observed in single-hoofed animals, especially horses. Modern molecular techniques have been recently employed to improve epidemiology for identifying and searching for strains of this bacterium at different times and locations. Due to the unknown number of circulating strains and lack of preventive methods, glanders is still observed in the form of epidemics. The present study aimed to evaluate six field isolates plus two laboratory strains of and using the polymerase chain reaction-restriction fragment ...
Sieme H, Oldenhof H, Wolkers WF.Native sperm is only marginally stable after collection. Cryopreservation of semen facilitates transport and storage for later use in artificial reproduction technologies, but cryopreservation processing may result in cellular damage compromising sperm function. Membranes are thought to be the primary site of cryopreservation injury. Therefore, insights into the effects of cooling, ice formation and protective agents on sperm membranes may help to rationally design cryopreservation protocols. In this review, we describe membrane phase behaviour of sperm at supra- and subzero temperatures. In a...
Zinchenko AV, Govorova YS.Critical to the understanding the mechanism of destruction and protection during cryopreservation of biological objects is the knowledge of the conformational transitions of biopolymers experiencing low temperatures in the presence of cryoprotective agents. This information may be derived from the kinetic and thermodynamic parameters of macromolecular thermal denaturation kinetics under different environmental conditions. Objective: The study deals with the influence of cryoprotective agents (glycerol, 1.2-propanediol (1.2-PD), and dimethyl sulfoxide (DMSO)) on thermodynamic and kinetic parame...
Ku0119dzierski W.Leptin is a tissue-derivative adipokine that regulates appetite, food intake and energy expenditure. It is still not clear how exercise affects plasma leptin concentration in horses. The aim of this study was to evaluate the influence of exercise intensity and duration on plasma leptin levels in working horses. A total of 38 horses were prospectively included in the study and grouped according to the type of exercise they performed: dressage (six stallions, group D), jumping (12 stallions, group J), race (12 Thoroughbred horses, six stallions and six mares, group R) and harness (10 light draft...
Medina-Torres CE, Pollitt CC, Underwood C, Castro-Olivera EM, Collins SN, Allavena RE, Richardson DW, van Eps AW.Failure of lamellar energy metabolism may contribute to the pathophysiology of equine laminitis. Tissue microdialysis has the potential to dynamically monitor lamellar energy balance over time. The objectives of this study were to develop a minimally invasive lamellar microdialysis technique and use it to measure normal lamellar energy metabolite concentrations over 24 h. Microdialysis probes were placed (through the white line) into either the lamellar dermis (LAM) (n = 6) or the sublamellar dermis (SUBLAM) (n = 6) and perfused continuously over a 24 h study period. Probes were pl...
Oldenhof H, Gojowsky M, Wang S, Henke S, Yu C, Rohn K, Wolkers WF, Sieme H.The aim of this study was to determine how different membrane-permeable and -impermeable cryoprotective agents modulate tolerance of stallion sperm to osmotic stress and stabilize membranes during cryopreservation. Special emphasis was on hydroxyl ethylene starch (HES), which exposes cells to minimal osmotic stress due to its large molecular weight. Percentages of motile sperm post-thaw were found to be similar when glycerol, sucrose, and HES were used at their optimal concentrations. Percentages of plasma membrane intact sperm after return to isotonic medium were highest for HES. Fourier tran...
Oldenhof H, Friedel K, Akhoondi M, Gojowsky M, Wolkers WF, Sieme H.Stallion sperm exhibits great male-to-male variability in survival after cryopreservation. In this study, we have investigated if differences in sperm freezability can be attributed to membrane phase and permeability properties. Fourier transform infrared spectroscopy (FTIR) was used to determine supra and subzero membrane phase transitions and characteristic subzero membrane hydraulic permeability parameters. Sperm was obtained from stallions that show differences in sperm viability after cryopreservation. Stallion sperm undergoes a broad and gradual phase transition at suprazero temperatures...
Oldenhof H, Friedel K, Sieme H, Glasmacher B, Wolkers WF.Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of free...
Edner AH, Essu00e9n-Gustavsson B, Nyman GC.Muscle metabolism in horses has been studied mainly by analysis of substances in blood or plasma and muscle biopsy specimens. By using microdialysis, real-time monitoring of the metabolic events in local tissue with a minimum of trauma is possible. There is limited information about muscle metabolism in the early recovery period after anaesthesia in horses and especially in the colic horse. The aims were to evaluate the microdialysis technique as a complement to plasma analysis and to study the concentration changes in lactate, pyruvate, glucose, glycerol, and urea during anaesthesia and in th...
Lacitignola L, Fanizzi FP, Francioso E, Crovace A.Proton magnetic resonance spectroscopy (1H NMR) has been successfully used in the study of many biological fluids. The data presented here report on the metabolic profiles of normal equine synovial fluids compared with osteoarthritic (OA) fluids. Twenty-five OA synovial fluid samples and eight normal ones were collected from the forelimb fetlock joint in 22 horses, aged between five and 24 years. 1H NMR spectroscopy was carried out with a Bruker Avance DRX 500 equiped with a cryo-magnet working at 11 Tesla, and 'Mestre-C 4.9.9.6' software was used to analyze the spectra. The study assessed the...
Kardak A, Leibo SP, Devireddy R.The rate at which equine and macaque ovarian tissue sections are first cooled from +25 degrees C to +4 degrees C has a significant effect on the measured water transport when the tissues are subsequently frozen in 0.85 M solutions of glycerol, dimethylsulfoxide (DMSO), or ethylene glycol (EG). To determine whether the response of ovarian tissues is altered if they are suspended in mixtures of cryoprotective agents (CPAs), rather than in solutions of a single CPA, we have now measured the subzero water transport from ovarian tissues that were suspended in mixtures of DMSO and EG. Sections of fr...
Bass LD, Denniston DJ, Maclellan LJ, McCue PM, Seidel GE, Squires EL.Equine embryos (n=43) were recovered nonsurgically 7-8 days after ovulation and randomly assigned to be cryopreserved in one of two cryoprotectants: 48% (15M) methanol (n=22) or 10% (136 M) glycerol (n=21). Embryos (300-1000 microm) were measured at five intervals after exposure to glycerol (0, 2, 5, 10 and 15 min) or methanol (0, 15, 35, 75 and 10 min) to determine changes (%) in diameter over time (+/-S.D.). Embryos were loaded into 0.25-ml plastic straws, sealed, placed in a programmable cell freezer and cooled from room temperature (22 degrees C) to -6 degrees C. Straws were then seeded, h...
Mendes MA, Chies JM, de Oliveira Dias AC, Filho SA, Palma MS.Most commercial recombinant proteins used as molecular biology tools, as well as many academically made preparations, are generally maintained in the presence of high glycerol concentrations after purification to maintain their biological activity. The present study shows that larger proteins containing high concentrations of glycerol are not amenable to analysis using conventional electrospray ionization mass spectrometry (ESI-MS) interfaces. In this investigation the presence of 25% (v/v) glycerol suppressed the signals of Taq DNA polymerase molecules, while 1% (v/v) glycerol suppressed the ...
Sloet van Oldruitenborgh-Oosterbaan MM, Annee MP, Verdegaal EJ, Lemmens AG, Beynen AC.Feeding a high-fat diet increases fat utilisation and may decrease glycogen utilisation resulting in a lower lactate production during moderate exercise. The effects of fat feeding on exercise- and lipid metabolism-associated blood variables were evaluated in 6 Standardbred horses during submaximal exercise. The horses were fed a high- (11.8% fat in total dietary dry matter) or a low-fat diet (1.5% fat) in a cross-over experiment with feeding periods of 4 weeks. At the end of each feeding period, the horses were subjected to a submaximal standardised stepwise exercise test on a treadmill. Bloo...
Alvarenga MA, Landim-Alvarenga FC, Moreira RM, Cesarino MM.The present experiments aimed to examine the substitution of glycerol (G) by ethylene glycol (E) as a cryoprotective agent for stallion spermatozoa. Two different ethylene glycol concentrations (5% and 10%) and also the association of glycerol (2%) and ethylene glycol (3%) (E/G) were studied (Experiment 1). In Experiment 2, two packing systems (0.5 x 4.0 ml) were evaluated using both cryoprotectors. In both experiments, the sperm membrane integrity after freezing was evaluated using transmission electron microscopy. The mean post-thaw motility was 34.25, 36.5, 29.25 and 34.75% for G5%, E5%, E1...
Schott HC, Du00fcsterdieck KF, Eberhart SW, Woody KA, Refsal KR, Coenen M.Incomplete recovery from endurance exercise after an overnight rest period is reflected by persisting weight loss and an elevated plasma aldosterone concentration, even in successful competitors. To determine whether supplementation with high doses of electrolytes, with or without glycerol, enhances recovery, the following were measured in 6 Arabian horses before and after completion of a 60 km treadmill exercise test simulating an endurance ride and after 12, 24, 48, and 72 h of recovery: bodyweight; plasma osmolality; plasma concentrations of protein, electrolytes, aldosterone and cortisol; ...
Bruyas JF, Sanson JP, Battut I, Fiu00e9ni F, Tainturier D.Early (day 6) equine embryos (n=23) were assigned to four treatment groups to assess the cryoprotectant properties of glycerol and ethylene glycol and the effect of adding sucrose during removal of the cryoprotectant: (i) group GG (n=5) embryos were frozen and thawed using 1.5 mol glycerol l(-1) as the cryoprotectant, which was added at 22 degrees C in four steps (0.375, 0.75, 1.125 and 1.5 mol glycerol l(-1)), and removed after thawing in five steps (1.5, 1.125, 0.75, 0.375 and 0.0 mol glycerol l(-1)); (ii) group GS (n=6) embryos were frozen and thawed using 1.5 mol glycerol l(-1) as for grou...
Auer DE, Ng JC, Seawright AA.Topically applied copper phenylbutazone, phenylbutazone, copper salicylate, salicylate and dimethylsulfoxide glycerol (80:20) were investigated as anti-inflammatory agents in rats and horses. Dimethylsulfoxide and glycerol (80:20) or dimethylsulfoxide, ethanol and glycerol (60:20:20) were used as the drug solvents. Subcutaneously administered carrageenin was used to induce inflammatory oedema, either in the paws of rats or the alar fold of the horse. The severity of the oedema and the anti-inflammatory effect of the drugs were assessed by measuring changes in the paw or alar-fold diameters. Co...
Pu00f6su00f6 AR, Viljanen-Tarifa E, Soveri T, Oksanen HE.Effects of graded intensity exercise on plasma lipids was studied in Standardbred and Finnishbred Trotters. The rate of lipolysis indicated by the elevated plasma concentrations of glycerol increased parallel with the intensity of the trot in the Standardbreds, but not as clearly in the Finnishbred trotters. During the exercise plasma triglyceride concentration increased significantly and the increase correlated with the intensity of the exercise as well as the activity of lipolysis. Together with the increase in plasma triglycerides, there was a parallel increase in the pre-beta fraction of l...
Valeri CR, Valeri DA, Gray A, Contreras TJ, Lindberg JR.When equine RBC were frozen with 20% (w/v) glycerol and stored at -150 C for as long as 5 years, there were no adverse effects on freeze-thaw or freeze-thaw-wash recovery or oxygen transport function. The manner in which the glycerol was added to, and removed from, the equine RBC was shown to be an important consideration in ensuring optimal freeze-thaw-wash recovery values.
Snow DH, Summers RJ, Guy PS.The effects of two beta-adrenoceptor antagonists, propranolol (0.2 mg/kg) and metoprolol (0.2 mg/kg) on some physiological and metabolic changes produced by maximal exercise in the horse were investigated. Both drugs reduced the elevation in heart rate seen immediately following exercise and reduced performance as was seen by the increased time taken to perform each gallop. The rise in plasma glucose, glycerol and lactate, and the fall in blood pH seen following exercise were attenuated by both drugs. However, a greater increase in plasma free fatty acids occurred. Exercise produced a nine- to...
Lucke JN, Hall GM.Blood samples were taken from 15 horses before and after a 50-mile ride to examine the changes occurring in some biochemical constituents. There was a significant (P less than 0.05) decrease in plasma potassium, calcium and magnesium concentrations and a rise in inorganic phosphate but there was no alteration in plasma sodium, chloride or protein levels or change in haematocrit. After the ride there was a highly significant (P less than 0.01) fall in blood glucose corresponding with increased lipolysis and a rise in plasma free fatty acids (P less than 0.001) and glycerol (P less than 0.001). ...
Snow DH, Mackenzie G.The effects of prolonged cantering before and after a 10 week training programme were studied in 6 horses. Determinations were carried on on venous blood for packed cell volume, glucose, glycerol, free fatty acids, beta-hydroxybutyrate, 11-hydroxycorticosteroids, pH and pCO2. Exercise caused a slight increase in glucose, lactate and pH, a moderate rise in PCV, glycerol and free fatty acids, and a marked rise in 11-hydroxycorticosteroids. A decrease in venous pCO2 occurred and a slight but not significant decrease in beta-hydroxybutyrate. Training was found to cause no significant difference in...
Snow DH, Mackenzie G.The effects of intermittent maximal exercise (galloping) before and after a 10 week training programme were studied in 6 horses. Determinations were carried out on venous blood for packed cell volume, total plasma protein, glucose, glycerol, free fatty acids, lactate, 11-hydroxycorticosteroids, blood gases and pH. There were marked changes associated with galloping and some of these could be modified with training. The major findings included (i) an elevated blood glucose, (ii) a large increase in glycerol, which was greatest at 30 min post-exercise and was higher following training, (iii) sma...
Anderson MG.The effects of exercise of different intensities on blood concentrations of glucose, lactate, pyruvate, free fatty acids and glycerol were studied in a group of clinically normal horses. Blood lactate, pyruvate and lactate/pyruvate ratio increased during exercise, particularly during galloping. These changes occurred within the first 12-15 seconds of exercise indicating that anaerobic metabolic pathways are brought into use very quickly in the strenuously exercising horse. Since blood glycerol levels were significantly increased during exercise body lipids were also mobilised. At the same time...
Sieme H, Oldenhof H, Wolkers WF.Native sperm is only marginally stable after collection. Cryopreservation of semen facilitates transport and storage for later use in artificial reproduction technologies, but cryopreservation processing may result in cellular damage compromising sperm function. Membranes are thought to be the primary site of cryopreservation injury. Therefore, insights into the effects of cooling, ice formation and protective agents on sperm membranes may help to rationally design cryopreservation protocols. In this review, we describe membrane phase behaviour of sperm at supra- and subzero temperatures. In a...
Lacitignola L, Fanizzi FP, Francioso E, Crovace A.Proton magnetic resonance spectroscopy (1H NMR) has been successfully used in the study of many biological fluids. The data presented here report on the metabolic profiles of normal equine synovial fluids compared with osteoarthritic (OA) fluids. Twenty-five OA synovial fluid samples and eight normal ones were collected from the forelimb fetlock joint in 22 horses, aged between five and 24 years. 1H NMR spectroscopy was carried out with a Bruker Avance DRX 500 equiped with a cryo-magnet working at 11 Tesla, and 'Mestre-C 4.9.9.6' software was used to analyze the spectra. The study assessed the...
Oldenhof H, Friedel K, Sieme H, Glasmacher B, Wolkers WF.Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of free...
Oldenhof H, Gojowsky M, Wang S, Henke S, Yu C, Rohn K, Wolkers WF, Sieme H.The aim of this study was to determine how different membrane-permeable and -impermeable cryoprotective agents modulate tolerance of stallion sperm to osmotic stress and stabilize membranes during cryopreservation. Special emphasis was on hydroxyl ethylene starch (HES), which exposes cells to minimal osmotic stress due to its large molecular weight. Percentages of motile sperm post-thaw were found to be similar when glycerol, sucrose, and HES were used at their optimal concentrations. Percentages of plasma membrane intact sperm after return to isotonic medium were highest for HES. Fourier tran...
Kardak A, Leibo SP, Devireddy R.The rate at which equine and macaque ovarian tissue sections are first cooled from +25 degrees C to +4 degrees C has a significant effect on the measured water transport when the tissues are subsequently frozen in 0.85 M solutions of glycerol, dimethylsulfoxide (DMSO), or ethylene glycol (EG). To determine whether the response of ovarian tissues is altered if they are suspended in mixtures of cryoprotective agents (CPAs), rather than in solutions of a single CPA, we have now measured the subzero water transport from ovarian tissues that were suspended in mixtures of DMSO and EG. Sections of fr...
Snow DH, Mackenzie G.The effects of prolonged cantering before and after a 10 week training programme were studied in 6 horses. Determinations were carried on on venous blood for packed cell volume, glucose, glycerol, free fatty acids, beta-hydroxybutyrate, 11-hydroxycorticosteroids, pH and pCO2. Exercise caused a slight increase in glucose, lactate and pH, a moderate rise in PCV, glycerol and free fatty acids, and a marked rise in 11-hydroxycorticosteroids. A decrease in venous pCO2 occurred and a slight but not significant decrease in beta-hydroxybutyrate. Training was found to cause no significant difference in...
Anderson MG.The effects of exercise of different intensities on blood concentrations of glucose, lactate, pyruvate, free fatty acids and glycerol were studied in a group of clinically normal horses. Blood lactate, pyruvate and lactate/pyruvate ratio increased during exercise, particularly during galloping. These changes occurred within the first 12-15 seconds of exercise indicating that anaerobic metabolic pathways are brought into use very quickly in the strenuously exercising horse. Since blood glycerol levels were significantly increased during exercise body lipids were also mobilised. At the same time...
Pu00f6su00f6 AR, Viljanen-Tarifa E, Soveri T, Oksanen HE.Effects of graded intensity exercise on plasma lipids was studied in Standardbred and Finnishbred Trotters. The rate of lipolysis indicated by the elevated plasma concentrations of glycerol increased parallel with the intensity of the trot in the Standardbreds, but not as clearly in the Finnishbred trotters. During the exercise plasma triglyceride concentration increased significantly and the increase correlated with the intensity of the exercise as well as the activity of lipolysis. Together with the increase in plasma triglycerides, there was a parallel increase in the pre-beta fraction of l...
Snow DH, Mackenzie G.The effects of intermittent maximal exercise (galloping) before and after a 10 week training programme were studied in 6 horses. Determinations were carried out on venous blood for packed cell volume, total plasma protein, glucose, glycerol, free fatty acids, lactate, 11-hydroxycorticosteroids, blood gases and pH. There were marked changes associated with galloping and some of these could be modified with training. The major findings included (i) an elevated blood glucose, (ii) a large increase in glycerol, which was greatest at 30 min post-exercise and was higher following training, (iii) sma...
Medina-Torres CE, Pollitt CC, Underwood C, Castro-Olivera EM, Collins SN, Allavena RE, Richardson DW, van Eps AW.Failure of lamellar energy metabolism may contribute to the pathophysiology of equine laminitis. Tissue microdialysis has the potential to dynamically monitor lamellar energy balance over time. The objectives of this study were to develop a minimally invasive lamellar microdialysis technique and use it to measure normal lamellar energy metabolite concentrations over 24 h. Microdialysis probes were placed (through the white line) into either the lamellar dermis (LAM) (n = 6) or the sublamellar dermis (SUBLAM) (n = 6) and perfused continuously over a 24 h study period. Probes were pl...
Oldenhof H, Friedel K, Akhoondi M, Gojowsky M, Wolkers WF, Sieme H.Stallion sperm exhibits great male-to-male variability in survival after cryopreservation. In this study, we have investigated if differences in sperm freezability can be attributed to membrane phase and permeability properties. Fourier transform infrared spectroscopy (FTIR) was used to determine supra and subzero membrane phase transitions and characteristic subzero membrane hydraulic permeability parameters. Sperm was obtained from stallions that show differences in sperm viability after cryopreservation. Stallion sperm undergoes a broad and gradual phase transition at suprazero temperatures...
Auer DE, Ng JC, Seawright AA.Topically applied copper phenylbutazone, phenylbutazone, copper salicylate, salicylate and dimethylsulfoxide glycerol (80:20) were investigated as anti-inflammatory agents in rats and horses. Dimethylsulfoxide and glycerol (80:20) or dimethylsulfoxide, ethanol and glycerol (60:20:20) were used as the drug solvents. Subcutaneously administered carrageenin was used to induce inflammatory oedema, either in the paws of rats or the alar fold of the horse. The severity of the oedema and the anti-inflammatory effect of the drugs were assessed by measuring changes in the paw or alar-fold diameters. Co...
Alvarenga MA, Landim-Alvarenga FC, Moreira RM, Cesarino MM.The present experiments aimed to examine the substitution of glycerol (G) by ethylene glycol (E) as a cryoprotective agent for stallion spermatozoa. Two different ethylene glycol concentrations (5% and 10%) and also the association of glycerol (2%) and ethylene glycol (3%) (E/G) were studied (Experiment 1). In Experiment 2, two packing systems (0.5 x 4.0 ml) were evaluated using both cryoprotectors. In both experiments, the sperm membrane integrity after freezing was evaluated using transmission electron microscopy. The mean post-thaw motility was 34.25, 36.5, 29.25 and 34.75% for G5%, E5%, E1...
Schott HC, Du00fcsterdieck KF, Eberhart SW, Woody KA, Refsal KR, Coenen M.Incomplete recovery from endurance exercise after an overnight rest period is reflected by persisting weight loss and an elevated plasma aldosterone concentration, even in successful competitors. To determine whether supplementation with high doses of electrolytes, with or without glycerol, enhances recovery, the following were measured in 6 Arabian horses before and after completion of a 60 km treadmill exercise test simulating an endurance ride and after 12, 24, 48, and 72 h of recovery: bodyweight; plasma osmolality; plasma concentrations of protein, electrolytes, aldosterone and cortisol; ...
Mendes MA, Chies JM, de Oliveira Dias AC, Filho SA, Palma MS.Most commercial recombinant proteins used as molecular biology tools, as well as many academically made preparations, are generally maintained in the presence of high glycerol concentrations after purification to maintain their biological activity. The present study shows that larger proteins containing high concentrations of glycerol are not amenable to analysis using conventional electrospray ionization mass spectrometry (ESI-MS) interfaces. In this investigation the presence of 25% (v/v) glycerol suppressed the signals of Taq DNA polymerase molecules, while 1% (v/v) glycerol suppressed the ...
Ku0119dzierski W.Leptin is a tissue-derivative adipokine that regulates appetite, food intake and energy expenditure. It is still not clear how exercise affects plasma leptin concentration in horses. The aim of this study was to evaluate the influence of exercise intensity and duration on plasma leptin levels in working horses. A total of 38 horses were prospectively included in the study and grouped according to the type of exercise they performed: dressage (six stallions, group D), jumping (12 stallions, group J), race (12 Thoroughbred horses, six stallions and six mares, group R) and harness (10 light draft...
Edner AH, Essu00e9n-Gustavsson B, Nyman GC.Muscle metabolism in horses has been studied mainly by analysis of substances in blood or plasma and muscle biopsy specimens. By using microdialysis, real-time monitoring of the metabolic events in local tissue with a minimum of trauma is possible. There is limited information about muscle metabolism in the early recovery period after anaesthesia in horses and especially in the colic horse. The aims were to evaluate the microdialysis technique as a complement to plasma analysis and to study the concentration changes in lactate, pyruvate, glucose, glycerol, and urea during anaesthesia and in th...
Lucke JN, Hall GM.Blood samples were taken from 15 horses before and after a 50-mile ride to examine the changes occurring in some biochemical constituents. There was a significant (P less than 0.05) decrease in plasma potassium, calcium and magnesium concentrations and a rise in inorganic phosphate but there was no alteration in plasma sodium, chloride or protein levels or change in haematocrit. After the ride there was a highly significant (P less than 0.01) fall in blood glucose corresponding with increased lipolysis and a rise in plasma free fatty acids (P less than 0.001) and glycerol (P less than 0.001). ...
Bass LD, Denniston DJ, Maclellan LJ, McCue PM, Seidel GE, Squires EL.Equine embryos (n=43) were recovered nonsurgically 7-8 days after ovulation and randomly assigned to be cryopreserved in one of two cryoprotectants: 48% (15M) methanol (n=22) or 10% (136 M) glycerol (n=21). Embryos (300-1000 microm) were measured at five intervals after exposure to glycerol (0, 2, 5, 10 and 15 min) or methanol (0, 15, 35, 75 and 10 min) to determine changes (%) in diameter over time (+/-S.D.). Embryos were loaded into 0.25-ml plastic straws, sealed, placed in a programmable cell freezer and cooled from room temperature (22 degrees C) to -6 degrees C. Straws were then seeded, h...
Snow DH, Summers RJ, Guy PS.The effects of two beta-adrenoceptor antagonists, propranolol (0.2 mg/kg) and metoprolol (0.2 mg/kg) on some physiological and metabolic changes produced by maximal exercise in the horse were investigated. Both drugs reduced the elevation in heart rate seen immediately following exercise and reduced performance as was seen by the increased time taken to perform each gallop. The rise in plasma glucose, glycerol and lactate, and the fall in blood pH seen following exercise were attenuated by both drugs. However, a greater increase in plasma free fatty acids occurred. Exercise produced a nine- to...
Bruyas JF, Sanson JP, Battut I, Fiu00e9ni F, Tainturier D.Early (day 6) equine embryos (n=23) were assigned to four treatment groups to assess the cryoprotectant properties of glycerol and ethylene glycol and the effect of adding sucrose during removal of the cryoprotectant: (i) group GG (n=5) embryos were frozen and thawed using 1.5 mol glycerol l(-1) as the cryoprotectant, which was added at 22 degrees C in four steps (0.375, 0.75, 1.125 and 1.5 mol glycerol l(-1)), and removed after thawing in five steps (1.5, 1.125, 0.75, 0.375 and 0.0 mol glycerol l(-1)); (ii) group GS (n=6) embryos were frozen and thawed using 1.5 mol glycerol l(-1) as for grou...
Sloet van Oldruitenborgh-Oosterbaan MM, Annee MP, Verdegaal EJ, Lemmens AG, Beynen AC.Feeding a high-fat diet increases fat utilisation and may decrease glycogen utilisation resulting in a lower lactate production during moderate exercise. The effects of fat feeding on exercise- and lipid metabolism-associated blood variables were evaluated in 6 Standardbred horses during submaximal exercise. The horses were fed a high- (11.8% fat in total dietary dry matter) or a low-fat diet (1.5% fat) in a cross-over experiment with feeding periods of 4 weeks. At the end of each feeding period, the horses were subjected to a submaximal standardised stepwise exercise test on a treadmill. Bloo...
Valeri CR, Valeri DA, Gray A, Contreras TJ, Lindberg JR.When equine RBC were frozen with 20% (w/v) glycerol and stored at -150 C for as long as 5 years, there were no adverse effects on freeze-thaw or freeze-thaw-wash recovery or oxygen transport function. The manner in which the glycerol was added to, and removed from, the equine RBC was shown to be an important consideration in ensuring optimal freeze-thaw-wash recovery values.
Zinchenko AV, Govorova YS.Critical to the understanding the mechanism of destruction and protection during cryopreservation of biological objects is the knowledge of the conformational transitions of biopolymers experiencing low temperatures in the presence of cryoprotective agents. This information may be derived from the kinetic and thermodynamic parameters of macromolecular thermal denaturation kinetics under different environmental conditions. Objective: The study deals with the influence of cryoprotective agents (glycerol, 1.2-propanediol (1.2-PD), and dimethyl sulfoxide (DMSO)) on thermodynamic and kinetic parame...
Fracaro PL, Corcini CD, Souza Gatti NL, Filho JS, Acosta IB, Filho JS, Hartwig FP, Rosa Curcio BD, Varela Junior AS.Glycerol is a cryoprotectant widely used in the freezing of mammalian semen, but no study has demonstrated its optimum concentration and the appropriate exposure time for equine species. Objective: To demonstrate that the exposure time (15, 30, 45, 60, 75 and 90 min) versus concentration (2, 3, 4 and 5%) of the cryoprotectant glycerol influences the freezing success of equine semen. Methods: The ejaculate of 12 stallions were frozen in different glycerol concentrations following different exposure times. The thawed sperm was evaluated for kinetic parameters using a Computer Assisted Semen Anal...
Talluri TR, Jhamb D, Paul N, Singh J, Dedar RK, Mehta SC, Legha RA, Pal Y.The recovery of spermatozoa from the cauda epididymis may be the only option to obtain genetic material from elite stallions that had undergone castration or sudden death due to colic or severe injury. Objective: To evaluate two different protocols for retrieval of stallion epididymal spermatozoa and to evaluate different cryoprotectants on the freezability of the epididymal spermatozoa. Methods: Six epididymides from three stallions were collected immediately after routine castration under general anesthesia. In the first experiment, each epididymis (of two testes) of the same stallion were p...
Abnaroodheleh F, Mosavari N, Pourbakhsh SA, Tadayon K, Jamshidian M. is the main cause of glanders as a dangerous contagious zoonosis disease that is mostly observed in single-hoofed animals, especially horses. Modern molecular techniques have been recently employed to improve epidemiology for identifying and searching for strains of this bacterium at different times and locations. Due to the unknown number of circulating strains and lack of preventive methods, glanders is still observed in the form of epidemics. The present study aimed to evaluate six field isolates plus two laboratory strains of and using the polymerase chain reaction-restriction fragment ...
Uhlmannsiek L, Shen H, Eylers H, Martinsson G, Sieme H, Wolkers WF, Oldenhof H.Cryopreserved semen is routinely shipped in liquid nitrogen. Dry ice could serve as an alternative coolant, however, frozen storage above liquid nitrogen temperatures (LN2, -196 °C) may negatively affect shelf-life and cryosurvival. In this study, we determined critical temperatures for storage of cryopreserved stallion sperm. We evaluated: (i) effects of cooling samples to different subzero temperatures (-10 °C to -80 °C) prior to storing in LN2, (ii) stability at different storage temperatures (i.e., in LN2, dry ice, -80 °C and -20 °C freezers, 4 °C refrigerator), and (iii)...