Flow Cytometric Chromosomal Sex Sorting of Stallion Spermatozoa Induces Oxidative Stress on Mitochondria and Genomic DNA          下载免费PDF全文

CM Balao da Silva  C Ortega‐Ferrusola  JM Morrell  H Rodriguez Martínez  FJ Peña 《Reproduction in domestic animals》2016,51(1):18-25

To date, the only repeatable method to select spermatozoa for chromosomal sex is the Beltsville sorting technology using flow cytometry. Improvement of this technology in the equine species requires increasing awareness of the modifications that the sorting procedure induces on sperm intactness. Oxidative stress is regarded as the major damaging phenomenon, and increasing evidence regards handling of spermatozoa – including sex sorting – as basic ground for oxidative damage. The aim of this study was to disclose whether the flow cytometric sorting procedure increases the production of reactive oxygen species (ROS), and to identify if ROS production relates to DNA damage in sorted spermatozoa using specific flow cytometry‐based assays. After sorting, oxidative stress increased from 26% to 33% in pre‐ and post‐incubation controls, to 46% after sex sorting (p < 0.05). Proportions of DNA fragmentation index post‐sorting were approximately 10% higher (31.3%); an effect apparently conduced via oxidative DNA damage as revealed by the oxyDNA assay. The probable origin of this increased oxidative stress owes the removal of enough seminal plasma due to the unphysiological sperm extension, alongside a deleterious effect of high pressure on mitochondria during the sorting procedure.  相似文献   

Epigallocatechin‐3‐Gallate (EGCG) Reduces Rotenone Effect on Stallion Sperm–Zona Pellucida Heterologous Binding          下载免费PDF全文

M Plaza Dávila  D Bucci  G Galeati  FJ Peña  G Mari  E Giaretta  C Tamanini  M Spinaci 《Reproduction in domestic animals》2015,50(6):1011-1016

Stallion spermatozoa are highly dependent on oxidative phosphorylation for ATP production to achieve normal sperm function and to fuel the motility. The aim of this study was to evaluate the response of equine sperm under capacitating conditions to the inhibition of mitochondrial complex I by rotenone and to test whether epigallocatechin‐3‐gallate (EGCG), a natural polyphenol component of green tea, could counteract this effect. After 2‐h incubation of stallion spermatozoa in modified Tyrode's medium, rotenone (100 nm , 500 nm and 5 μm ) and EGCG (10, 20 and 60 μm ), alone or in combination, did not induce any significant difference on the percentage of viable cells, live sperm with active mitochondria and spermatozoa with intact acrosome. The inhibition of complex I of mitochondrial respiratory chain of stallion sperm with rotenone exerted a negative effect on heterologous ZP binding ability. EGCG at the concentrations of 10 and 20 μm (but not of 60 μm ) induced a significant increase in the number of sperm bound to the ZP compared with that for control. Moreover, when stallion sperm were treated with rotenone 100 nm , the presence of EGCG at all the concentrations tested (10, 20 and 60 μm ) significantly increased the number of sperm bound to the ZP up to control levels, suggesting that this green tea polyphenol is able to reduce the toxicity of rotenone.  相似文献   

Sarcoptic mange in alpacas     

Vine NJ  Keevill G  Foster AP 《The Veterinary record》2010,167(24):946-947

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The effect of fresh seaweed and a formulated diet supplemented with seaweed on the growth and gonad quality of the collector sea urchin,Tripneustes gratilla,under farm conditions     

Abigail J. Onomu  Niall G. Vine  Mark D. Cyrus  Brett M. Macey  John J. Bolton 《Aquaculture Research》2020,51(10):4087-4102

This gonad enhancement study investigates the effect of different fresh and formulated feeds and feeding regimes on the growth and gonad quality of wild‐collected adult sea urchin, Tripneustes gratilla, under farm conditions for over 18 weeks. In the first 12 weeks (phase 1), urchins were fed fresh Ulva rigida (U); a 50:50 mixture of fresh U. rigida and Gracilaria gracilis (UG); fresh G. gracilis (G) and a formulated diet 20U (containing 20% U. rigida), and in the final 6 weeks (phase 2) of the study, diet was changed to a formulated feed (20U diet). By the end of phase 1, urchins fed the 20U diet produced gonads (50.72 ± 5.4 g) that were significantly heavier (p < .001) than the gonads of urchins fed the fresh seaweed diets (U, UG & G). By the end of phase 2, gonad weight of urchins in treatment groups UG‐20U and G‐20U were similar to those fed the 20U‐20U diet. Gonad colour of urchins in the G‐20U treatment became significantly lighter (ANOVA, p = .029) and poorer quality, compared with urchins in the U‐20U group. This gonad enhancement study, conducted on wild‐collected adult T. gratilla, has shown that a formulated feed (20U diet) can enhance gonad growth and produce commercially acceptable gonads. This farm‐based study supports previous findings from aquarium‐based studies by our group and indicates that short‐term sea urchin gonad enhancement can be carried out under farm conditions in South Africa.  相似文献   

Multi-atom resonant photoemission: A method for determining near-neighbor atomic identities and bonding     

A Kay  E Arenholz  S Mun  Garcia de Abajo FJ  CS Fadley  R Denecke  Z Hussain  Van Hove MA 《Science (New York, N.Y.)》1998,281(5377):679-683

Measurements and theoretical calculations are reported for an interatomic multi-atom resonant photoemission (MARPE) effect that permits direct determination of near-neighbor atomic identities (atomic numbers). MARPE occurs when the photon energy is tuned to a core-level absorption edge of an atom neighboring the emitting atom, with the emitting level having a lower binding energy than the resonant level. Large peak-intensity enhancements of 33 to 105 percent and energy-integrated effects of 11 to 29 percent were seen in three metal oxides. MARPE should also be sensitive to bond distance, bonding type, and magnetic order, and be observable via the secondary processes of x-ray fluorescence and Auger decay.  相似文献   

The Impact of Reproductive Technologies on Stallion Mitochondrial Function          下载免费PDF全文

FJ Peña  M Plaza Davila  BA Ball  EL Squires  P Martin Muñoz  C Ortega Ferrusola  C Balao da Silva 《Reproduction in domestic animals》2015,50(4):529-537

The traditional assessment of stallion sperm comprises evaluation of sperm motility and membrane integrity and identification of abnormal morphology of the spermatozoa. More recently, the progressive introduction of flow cytometry is increasing the number of tests available. However, compared with other sperm structures and functions, the evaluation of mitochondria has received less attention in stallion andrology. Recent research indicates that sperm mitochondria are key structures in sperm function suffering major changes during biotechnological procedures such as cryopreservation. In this paper, mitochondrial structure and function will be reviewed in the stallion, when possible specific stallion studies will be discussed, and general findings on mammalian mitochondrial function will be argued when relevant. Especial emphasis will be put on their role as source of reactive oxygen species and in their role regulating sperm lifespan, a possible target to investigate with the aim to improve the quality of frozen–thawed stallion sperm. Later on, the impact of current sperm technologies, principally cryopreservation, on mitochondrial function will be discussed pointing out novel areas of research interest with high potential to improve current sperm technologies.  相似文献   

Membrane Lipids of the Stallion Spermatozoon in Relation to Sperm Quality and Susceptibility to Lipid Peroxidation     

B Macías García  L González Fernández  C Ortega Ferrusola  C Salazar‐Sandoval  A Morillo Rodríguez  H Rodríguez Martinez  JA Tapia  D Morcuende  FJ Peña 《Reproduction in domestic animals》2011,46(1):141-148

Lipids were extracted from ejaculated spermatozoa from seven individual stallions to distinguish neutral lipids (NL) and polar lipids (PL) and determine their variation among stallions and their relationship with sperm quality and sperm susceptibility to lipid peroxidation. The isolated fatty acids were correlated with sperm quality (membrane integrity, mitochondrial membrane potential (ΔΨm) and expression of active caspases) and the sensitivity of the sperm plasma membrane to LPO. The miristic (C14: 0), palmitic (C16: 0), stearic (C18: 0) and oleic (C18: 1n9) acids were predominant among the NLs. Within the phospholipid fraction, the docosapentanoic acid (C22: 5n6) was dominant, albeit varying among stallions. Surprisingly, the percentage of polyunsaturated fatty acids was positively correlated with sperm quality and a low propensity for LPO, probably because these particular fatty acids provide a higher fluidity of the plasma membrane. The stallion showing the poorest sperm membrane integrity plus a high level of LPO in his ejaculate had a lower percentage (p < 0.05) of this fatty acid in his sperm plasma membranes.  相似文献   

Maternal Exposure to High Temperatures Disrupts OCT4 mRNA Expression of Rabbit Pre‐Implantation Embryos and Endometrial Tissue     

F Marco‐Jiménez  C Naturil‐Alfonso  DS Peñaranda  E Jiménez‐Trigos  FJ García‐Diego  JS Vicente 《Reproduction in domestic animals》2013,48(3):429-434

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Urethane anesthesia in adult female rats: preliminary observations     

FJ  Golder  SA Robertson  A Valverde  DC Bolser 《Veterinary anaesthesia and analgesia》2003,30(2):115-115

Urethane is widely used as a rodent anesthetic in the laboratory setting, and is characterized as producing long‐lasting anesthesia. The purpose of this study was to evaluate the quality of anesthesia provided by a single dose of urethane based on the response to a noxious stimulus. If the quality of anesthesia was insufficient to prevent gross purposeful movement (GPM), isoflurane was also administered until no response to noxious stimulation occurred. Five adult Harlan Sprague Dawley rats (6 months of age, 250–300 g) were given urethane (1.4 g kg^?1 IP) and evaluated for 120 minutes post‐injection. If the rats became laterally recumbent by 20 minutes post‐injection, a large hemostat was positioned around the tail and the response to tail clamping was assessed. If no GPM occurred, an additional 20 minutes was allowed to elapse. If the rats were not laterally recumbent or GPM was present, they were placed in a chamber and isoflurane in oxygen was administered. Inspired isoflurane concentrations (ISO) were measured using a S/5 anesthetic gas analyzer (Datex‐Ohmeda Division, Helsinki, Finland) calibrated before each experiment with a standardized calibration gas mixture (DOT‐34 NRC 300/375 m 1014, Datex‐Ohmeda Division, Helsinki, Finland). A period of 20 minutes was allowed for equilibration to inspired ISO. The tail‐clamp stimulus was then re‐applied and the animal's response recorded. If GPM was absent, ISO was lowered by 10–20% and an additional 20 minute interval elapsed. In contrast, if GPM was present, ISO was increased by 10–20%. This procedure was repeated until the ISO required to prevent GPM was determined in duplicate. The position within the estrus cycle influenced pain thresholds in the rats. As such, a vaginal smear was prepared from each rat and the position in the estrus cycle was determined based on vaginal cytology. Rats were euthanatized at the end of the study period. All values were mean ± SD. Four rats became recumbent after urethane injection (time to recumbency: 45 ± 17 seconds). Of these, two rats (one estrus, one metestrus) did not require isoflurane supplementation for the duration of the study. The three remaining rats (two metestrus, one estrus) required isoflurane supplementation. The mean ISO required to prevent GPM was 0.26 ± 0.16%. Position within the estrus cycle did not appear to affect the animal's response to urethane. These results indicate that urethane anesthesia is not long lasting in all rats and provides variable quality of anesthesia. This is of particular concern in the laboratory setting where muscle relaxants are often administered to rats shortly after urethane injection.  相似文献   

Unidentified microsporidian in western pygmy perch Edelia vittata (Nannopercidae) from Australia     

FJ Stephens 《Australian veterinary journal》2009,87(1-2):61-62

Specimens of western pygmy perch, Edelia vittata Castelnau 1873, were submitted for examination because of the presence of black masses in the body wall in the abdominal area. The owner reported that approximately 10% of mature fish in one dam were affected. The masses had no apparent effect on the health of affected fish, but they were unsuitable for sale for aesthetic reasons. After necropsy and histological examination, the masses were found to be microsporidial xenomas within the abdominal cavity. Transmission electron microscopy results suggest that this microsporidian is a previously unreported species, because the melanised xenoma wall and internal structure of the spores differ from previously described species. The epidemiology of the parasite remains largely unknown and highlights the need for further surveys of the disease status and parasites of Australian native fish, especially those of interest to aquaculturists.  相似文献