Effects of Egg Yolk and Cooling Rate on the Survival of Refrigerated Red Deer (Cervus elaphus hispanicus) Epididymal Spermatozoa     

MR Fernández-Santos  MC Esteso  AJ Soler  V Montoro  JJ Garde 《Reproduction in domestic animals》2006,41(2):114-118

Egg yolk is a common component to sperm refrigeration for most of the deer species, the role of which is to protect sperm membranes against cold shock. In addition, there have been many studies of conservation of ejaculated semen from stags, but few have been reported for epididymal spermatozoa. This work was designed to investigate the combined effects of cooling rates (slow: 0.23 degrees C/min vs rapid: 4.2 degrees C/min) from room temperature to 5 degrees C, and egg-yolk concentration (0, 5 or 20%) in the extender on the survival of Iberian red deer epididymal spermatozoa refrigerated at 5 degrees C. Heterospermic sperm samples were diluted to a final sperm concentration approximately 400x10(6) sperm/ml with a Tris-citrate-fructose (TCF)-egg-yolk diluent. Sperm quality was in vitro judged by microscopic assessments of individual sperm motility [sperm motility index (SMI)], and of plasma membrane (hypo-osmotic swelling test) and acrosome (NAR) integrities. Our results first showed that the presence of egg yolk in the extender significantly improves (p=0.01) the viability and sperm motility after sperm dilution. In addition, acrosome and plasma membrane integrities post-refrigeration did not differ significantly between cooling procedures; however, the SMI differed significantly between cooling procedures (slow: 46.6% vs rapid: 50.0%; p=0.01). Our results also showed that sperm quality was significantly (p<0.01) affected by the combined effects of egg-yolk concentration and cooling procedure, being rapid cooling with 20% of egg yolk the most suitable combination for epididymal sperm refrigeration. In conclusion, egg-yolk improved red deer epididymal spermatozoa characteristics after dilution. Rapid cooling protocol using TCF with 20% egg-yolk significantly improved sperm motility of red deer epididymal spermatozoa after cooling.  相似文献   

Effect of Oviductal Proteins on Structural and Functional Characteristics of Cryopreserved Sperm in Murrah Buffaloes*     

S Imam  MR Ansari  A Kumar  C Singh  VK Bharti  A Kumaresan 《Reproduction in domestic animals》2010,45(2):302-306

The present study was undertaken to elucidate the effect of non‐luteal oviductal proteins on sperm characteristics in Murrah buffaloes. Oviducts from healthy buffaloes were collected immediately after slaughter and the oestrous cycle phase was determined as either luteal or non‐luteal based on ovarian morphology. Non‐luteal oviducts (n = 80) were flushed from the isthmic end of the oviduct with PBS, fluid was centrifuged at 10 000 g at 4°C for 20 min and then dialysed and clarified. The supernatant obtained was lyophilized to concentrate the protein and stored at ?20°C till use. Sixteen good quality ejaculates from four Murrah buffalo bulls were collected using an artificial vagina. After fresh semen analysis, each ejaculate was split into two parts and extended in Tris–citrate–egg yolk glycerol dilutor. Part I of the split ejaculate was treated with non‐luteal oviductal proteins at the dose rate of 1 mg/ml of diluted semen, while part II remained as control. The extended semen was equilibrated for 4 h at 5°C, filled in 0.5 ml French straws, exposed to LN₂ vapour, plunged into LN₂ and then stored at ?196°C. The equilibrated and frozen–thawed semen was evaluated for sperm motility, viability, acrosomal integrity, cervical mucus penetration test and hypo‐osmotic sperm swelling test (HOST). In frozen–thawed semen, the percentage of sperm motility, viability and acrosomal integrity was significantly (p < 0.05) higher in the treatment group compared to the control group. The incorporation of non‐luteal oviductal proteins in the extender increased the ability of sperm to penetrate cervical mucus both after equilibration and the freeze‐thaw process. Similarly, the proportion of sperm with intact plasma membrane, as revealed by HOST values, was also significantly (p < 0.05) higher in the treatment group (32.6%) than the control group (27%) in frozen–thawed semen. It was inferred that incorporation of non‐luteal whole oviductal fluid proteins improved the sperm quality in frozen–thawed semen in Murrah buffaloes.  相似文献   

Use of Vaginal Electrical Resistance to Diagnose Oestrus,Dioestrus and Early Pregnancy in Synchronized Tropically Adapted Beef Heifers     

CD Hockey  ST Norman  JM Morton  D Boothby  NJ Phillips  MR McGowan 《Reproduction in domestic animals》2010,45(4):629-636

The primary objective of this study was to determine whether a single measurement of intravaginal electrical resistance (VER), using the commercially available Ovatec^® probe, can discriminate between dioestrus and oestrus in Bos indicus females, which had been treated to synchronize oestrus. Santa Gertrudis heifers (n = 226) received one of three oestrous synchronization treatments: double PGF_2α 10 days apart, 8‐day controlled internal drug release (CIDR) treatment or CIDR pre‐synchronization + PGF_2α 10 days after CIDR removal. The heifers were inseminated within 12 h following observed oestrus, or, if not observed, at a fixed time approximately 80 h, following the last synchronization treatment. They were palpated per rectum for signs of pregnancy 9 weeks after artificial insemination (AI). Vaginal electrical resistance measurements were taken at the completion of synchronization treatments (presumed dioestrus), immediately prior to AI (oestrus), and then at 3 and 9 weeks post‐AI. Mean VER differed between presumed dioestrus and oestrus (113.7 vs 87.4, p < 0.001). The area under the receiver operating characteristics (ROC) curve was 0.925, indicating that VER was highly discriminatory between dioestrus and oestrus. Vaginal electrical resistance at time of AI was negatively associated with odds of conception when all inseminations were included in the analyses [odds ratio (OR) = 0.97; 95% CI 0.95–1.00; p = 0.018], but not when fixed time AIs were excluded (OR = 1.00; 95% CI 0.97–1.03; p = 0.982). Mean VER readings differed between pregnant and non‐pregnant animals at both 3 weeks (120.5 vs 96.7, p < 0.001) and 9 weeks (124.0 vs 100.3, p < 0.001) post‐AI. However, 3‐ and 9‐week VER measurements were not highly discriminatory between pregnancy and non‐pregnancy (area under ROC curve = 0.791 and 0.736, respectively). Mean VER at time of AI for animals diagnosed in oestrus differed between each of the oestrous synchronization treatments (84.7, 73.6 and 78.9, groups 1–3 respectively, p < 0.001). These findings suggest that measurement of VER may improve accuracy of oestrus diagnoses when selecting cattle for AI following oestrous synchronization programmes involving tropically adapted cattle.  相似文献   

Does the Amount of Progesterone in Intravaginal Implants Used to Synchronise Oestrus Affect the Reproductive Performance of Brahman Heifers Artificially Inseminated at a Fixed Time     

NJ Phillips  G Fordyce  B Burns  P Williams  D Mayer  GA Bo  MR McGowan 《Reproduction in domestic animals》2010,45(6):e392-e397

The study tested the hypothesis that reduced intravaginal implant progesterone (P₄) concentration to synchronise oestrus would increase pregnancy rates to fixed‐time artificial insemination (FTAI) in Bos indicus heifers. Brahman heifers (n = 294; 2 year) were body condition scored (BCS), weighed and scanned for presence of a corpus luteum (CL). Only cyclic heifers were selected and allocated randomly within BCS and 25 kg bodyweight category to one of three P₄ treatment groups. On day 10, heifers received a P₄ implant (CueMate‐1‐pod, 0.78g P₄; CueMate‐2‐pod, 1.56g P₄; or CIDR‐B, 1.9g P₄), 2 mg oestradiol benzoate (ODB) intramuscularly (IM) and 250 ug cloprostenol IM. At day 2, the implant was removed, 250 ug cloprostenol was injected IM and tail paint applied. The heifers received 1 mg ODB 24 h later and were FTAI 48–54 h after implant removal (day 0). Ten randomly selected heifers per group were blood sampled and scanned at days 10, 2, 0 and 6 to define the P₄ profiles pre‐ and post‐FTAI. Heifers were heat‐detected 18–20 days post‐FTAI and oestrous heifers AI’d by the AM/PM rule. Bulls joined the heifers on day 27 post‐FTAI. Transrectal ultrasonography estimated conception date on day 72. Statistical analysis examined the effects of treatment, technician, semen, ovarian status, BCS and liveweight, on pregnancy rate (PR) to FTAI. There was no significant difference (p = 0.362) in PR between treatment groups (CueMate 1‐pod, 36.4%; CueMate 2‐pod, 39.6%: CIDR‐B, 28.3%), but PR was higher in those heifers with increased BCS between FTAI and pregnancy diagnosis (p = 0.005). Thirty‐three per cent of monitor heifers had plasma P₄ concentrations of <1 ng/ml on day 6 after FTAI; only 20% of these conceived vs 60% of heifers with P₄ ≥ 1 ng/ml. In summary, no significant difference in PR was identified between treatments but good BCS and a rising plane of nutrition were critical to PR of these pure grade Brahman heifers in northern Australia.  相似文献   

Antisperm antibodies in repeat‐breeding cows: Frequency,detection and validation of threshold levels employing sperm immobilization,sperm agglutination and immunoperoxidase assay          下载免费PDF全文

SK Srivastava  S Shinde  SK Singh  S Mehrotra  MR Verma  AK Singh  S Nandi  N Srivastava  SK Singh  TK Goswami  SK Bhure  H Kumar  SK Ghosh 《Reproduction in domestic animals》2017,52(2):195-202

Antisperm antibodies have been found in repeat‐breeding(RB) cows, and those causing agglutination and/or immobilization of sperm are considered to be closely related to unexplained infertility. However, a standard protocol for identifying antisperm antibodies (ASA) in cattle is not validated. Therefore, an investigation was undertaken to evaluate sperm immobilization (SIT), sperm agglutination (SAT) and immunoperoxidase (IPT)assays for detection of ASA in serum and their respective threshold levels for confirmation. Animals (heifers, normally breeding, repeat‐breeding and pregnant animals) that were free from IBR, brucellosis and uterine infections (screened by clinical examination) were included in the study. Sperm agglutinating, sperm immobilizing and antisperm antibodies evaluated by respective assay were significantly higher (p < .05) in RB cows compared to other groups. The SIT assay was able to identify 61% of RB caused by ASA, more than those employing SAT and IPT. Furthermore, a dilution rate of 1:5 and 1:80 (confirms 59.0 and 57.0% RB+ve)were sufficient to diagnose ASA by SAT and IPT, respectively. Results indicate the presence of __12.6% clumped spermatozoa and __ 2.6%(cut‐off value) peroxidase‐positive spermatozoa at 1:5 and 1:80 dilutions diagnosed with SAT and IPT, respectively, may be considered as repeaters arising out of ASA. Furthermore, study also showed the presence of lower incidence of ASA positivity in other groups of animals (heifer<normal breeder<pregnant animals) compared to repeaters. Study results show that although IPT is more specific and accurate but SAT and SIT are comparatively simple and cost‐effective assays suitable for detecting ASA under field conditions and thus can be recommended for screening of repeaters.  相似文献   

C, N, and noble gas isotopes in grain size separates of presolar diamonds from efremovka     

AB Verchovsky  AV Fisenko  LF Semjonova  IP Wright  MR Lee  CT Pillinger 《Science (New York, N.Y.)》1998,281(5380):1165-1168

Nanometer-size presolar diamonds from the Efremovka CV3 chondrite were physically separated into several grain size fractions by ultracentrifugation. The coarsest size fraction is the most enriched in carbon-12; the others have broadly similar carbon isotopic compositions. Measurement of noble gases shows that their concentration decreases with decreasing grain size. This effect is attributed to ion implantation. Such an episode could occur in the envelope of a supernova that produced the diamonds, or in interstellar space; in either case, ions with energies above a certain threshold pass completely through the smaller diamond grains without being captured. Concentrations of nitrogen show only minor variations with grain size, indicating a different mechanism of incorporation into the diamonds.  相似文献   

Alteration of oceanic volcanic glass: textural evidence of microbial activity     

MR Fisk  SJ Giovannoni  IH Thorseth 《Science (New York, N.Y.)》1998,281(5379):978-980

The subsurface biosphere may constitute as much as 50 percent of Earth's biomass. Direct and indirect evidence suggests that an extensive biosphere exists in the rocks below the sea floor. This survey of basalts of the Atlantic, Pacific, and Indian Oceans supports the hypothesis that bacteria have colonized much of the upper oceanic crust, which has a volume estimated at 10(18) cubic meters. Although this is the largest habitat on Earth, its low abundance of bacteria constitutes much less than 1 percent of Earth's biomass.  相似文献   

Map-based cloning of the fertility restoration locus <Emphasis Type="Italic">Rfm1</Emphasis> in cultivated barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis>)     

Carine Rizzolatti  Paul Bury  Edouard Tatara  Pierre A. Pin  Nathalie Rodde  Hélène Bergès  Françoise Budar  Hakim Mireau  Jan J. L. Gielen 《Euphytica》2017,213(12):276

Hybridization technology has proven valuable in enhancing yields in many crops, but was only recently adopted in the small grain cereals. Hybrid varieties in barley (Hordeum vulgare) rely on the cytoplasmic male sterility (CMS) system msm1 derived from Hordeum vulgare ssp. spontaneum. The major restorer gene described for the msm1 system is known as Rfm1 and maps to the top of chromosome 6H. To gain further insight into mechanisms underlying male fertility restoration in barley, we used a map-based cloning approach to identify the nuclear gene involved in the restoration mechanism of this hybridization system. Taking advantage of the available genomic resources in barley in combination with a custom-made non-gridded BAC library developed from a restorer line, we cloned and sequenced the Rfm1 restorer locus. The characterization and annotation of the nucleotide sequence for the Rfm1 restorer allele allowed for the identification of the candidate gene for Rfm1. The Rfm1 locus carries a tandem repeat of a gene encoding a pentatricopeptide repeat (PPR) protein. Surprisingly, Rfm1 belongs to the PLS-DYW subfamily of PPR genes known for their involvement in RNA editing in plants organelles, but that to date have not been identified as restorer genes.  相似文献   

Total ear canal ablation and lateral bulla osteotomy in an alpaca     

MR MARRIOTT  AJ DART  C. MACPHERSON  DR HODGSON 《Australian veterinary journal》1999,77(5):301-302

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Tetravalent uranium in calcite   总被引：1，自引：0，他引：1  

NC Sturchio  MR Antonio  L Soderholm  SR Sutton  JC Brannon 《Science (New York, N.Y.)》1998,281(5379):971-973

X-ray absorption spectroscopy and x-ray fluorescence microprobe studies of 35-million-year-old calcite from a Mississippi Valley-type zinc ore deposit indicate substitution of tetravalent uranium for divalent calcium. Thus, tetravalent uranium has a stable location in calcite deposited under reducing conditions. This result validates uranium-series dating methods (including uranium/lead dating) for ancient calcite and shows that calcite provides a sink for uranium in deep groundwater aquifers and anoxic lacustrine and marine basins.  相似文献