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1.
Testicles were isolated from thirty five apparently healthy dromedary camels (Camelus dromedarius), aged between 5 to 18 years, in a local slaughterhouse during the rutting season. Epididymal fluid was collected from one epididymis for determination of twelve biochemical and antioxidant parameters using ELISA commercial kits. Spermatozoa were harvested from each region of the other epididymis (head, body and tail) and stored in SHOTOR®, Green buffer® + 20% egg yolk and INRA-96® extenders at 5 and 30 °C. Results revealed that, in the epididymal fluid, concentrations of testosterone, glucose, albumin, total protein, cholesterol, fatty acids, iron, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were 5.19 ± 1.69 ng/mL, 3.10 ± 0.41 mmol/L, 6.26 ± 1.26 g/dL, 0.50 ± 0.07 mg/dL, 1.74 ± 0.09 mmol/L, 6.62 ± 0.81 nmol/ul, 926.20 ± 100.18 ug/dL, 51.17 ± 7.74 mIU/ml, and 143.16 ± 18.67 mIU/ml, respectively. The antioxidants activities of catalase, glutathione peroxidase (GPx) and superoxide dismutase (SOD) in the epididymal fluid were 121.55 ± 6.57 nmol/min/ml, 59.35 ± 10.98 nmol/min/ml and 0.18 ± 0.03 U/ml, respectively. Epididymal sperm motility and concentration were higher (P < 0.05) in the body and tail than the head. The viability indices of total and forward sperm motility, at 5 and 30 °C, obtained from the tail region were superior (P < 0.05) in both SHOTOR® and INRA-96® extenders than Green buffer extender. It may be concluded that INRA-96® extender is the best for storing dromedary epididymal spermatozoa at 5 and 30 °C.  相似文献   

2.
The present study was conducted in the Laboratory of Animal Physiology and Biotechnology, Department of Animal Production, Faculty of Agriculture, Mansoura University, Egypt. The present investigation aimed at studying effects of ages, pubertal stages and seasons of the year on testosterone concentrations in blood plasma and tissue homogenate of the testes. The testes used in the current study were collected from a total of 104 one-humped male camels (Camelus dromedarius). Samples were taken from pre (1–3.5 years) and post (3.5–13 years) pubertal camels. Testes were studied for a two consecutive seasons. The freshly prepared homogenate of the testicular tissue and blood plasma were used for determining the concentrations of testosterone in plasma and testicular extract. The concentrations of testosterone in blood plasma and testicular tissue were significantly increased during the breeding season compared with that of non-breeding season; the concentration of testosterone was higher in testicular tissue than in blood plasma.Testosterone concentrations in plasma and testicular tissue were increased in breeding than in non-breeding season. In addition, the testosterone concentrations were closely related with seasonal changes, stage of puberty and advancing age.  相似文献   

3.
Caffeine supplementation during oocyte IVM has been reported to improve preimplantation embryo development and the quality of in vitro–produced blastocysts in a range of species; but no studies have been done in camels. The present study investigated the effect of caffeine supplementation during dromedary camel oocyte IVM on nuclear maturation rates, IVF events, and subsequent preimplantation development. Cumulus–oocyte complexes obtained at slaughter were matured in vitro in caffeine supplemented medium either for 30 hours (caffeine 30 hours) or in the medium without caffeine supplement for 24 hours and then transferred to freshly prepared IVM medium supplemented with 10 mM caffeine for another 6 hours (caffeine 6 hours). Cumulus–oocyte complexes matured for 30 hours in the medium without caffeine supplement were used as a control. Matured oocytes were fertilized in vitro by epididymal spermatozoa of mature male camels collected from a local slaughterhouse. Eighteen hours after insemination, presumptive zygotes were cultured in modified KSOMaa medium for 7 days. Maturation and fertilization rates were significantly higher in the caffeine 6-hour group compared with the control group (P < 0.05), whereas IVM of oocytes in caffeine-supplemented medium for 30 hours did not affect these parameters (P > 0.05). Interestingly, IVM of oocytes in caffeine supplemented medium for 6 hours significantly (P < 0.05) increased the frequencies of blastocyst development by more than two-fold when compared with control (27.78% vs. 11.76%). In conclusion, culturing dromedary camel oocytes in maturation medium without caffeine for 24 hours and then in the medium supplemented with 10 mM caffeine for 6 hours during 30-hour IVM can significantly improve frequencies of nuclear maturation, fertilization rate, and subsequent preimplantation development.  相似文献   

4.
Haemaphysalis longicornis is an important vector of various pathogens in domestic animals and humans. The tick is a unique species with bisexual and parthenogenetic races. Although mating induces oocyte development, it is possible in the parthenogenetic race to complete oogenesis without copulation. Here we examined the developmental process of oocytes from unfed to the oviposition period in parthenogenetic H. longicornis. We classified the developmental stages of oocytes into five stages: stage I, germinal vesicle occupies more than half of the cytoplasm; stage II, germinal vesicle occupies less than half of the cytoplasm; stage III, germinal vesicle migrates from the center in the oocyte to the vicinity of the pedicel cells; stage IV, the cytoplasm is filled with yolk granules of various sizes; stage V, the cytoplasm is occupied by large yolk granules. Oocytes at the unfed period were undeveloped and classified as stage I. Stage I and II oocytes were observed at the rapid feeding period, indicating that oocyte development began after the initiation of blood feeding. All developmental stages of oocytes were observed at the pre-oviposition period. At 10?days after the beginning of the oviposition period, the ratios of stage I and II oocytes were higher than those of the previous period, suggesting that the ovarian development and activity may be continuing. Based on these findings, we propose classification criteria for the oocyte development in the parthenogenetic H. longicornis. The criteria will be useful for understanding the mechanisms of tick reproduction and transovarial transmission of pathogens.  相似文献   

5.
Most porcine oocytes used in studies on embryo biotechnology and the in vitro production of embryos are currently obtained from the ovaries of slaughtered gilts. The duration and temperature during ovary transportation and handling might, therefore, affect the recovery of culturable COCs, chromatin configuration and developmental competence of oocytes. The effects of ovary storage temperature on chromatin configuration and in vitro maturation of porcine oocytes were examined in this study. Ovaries collected from a slaughterhouse were stored in vitro for 8 h under different temperatures. The results showed that more culturable COCs were isolated from the ovares stored at 39℃ than that from ovaries stored at 31℃ or 20℃ and before storage. Thirty-one centidegree was the best storage temperature in terms of cumulus expansion, nuclear maturation and morphology of the first polar body after in vitro maturation culture. The ability of cumulus expansion was completely lost in COCs derived from ovaries stored at 39℃ for 8 hours. Ovary storage (at both 31℃ and at 20℃ ) increased the proportion of oocytes with the GVc configuration in which chromatin condensed into a single big clump at the nucleolus and the functional significance of this configuration needs further investigations [ Acta Zoologica Sinica 51 (5): 919 923, 2005].  相似文献   

6.
Research into an optimal cryoprotectant, its concentration and equilibration time underlies the successful cryopreservation of dromedary camel spermatozoa. This study assessed the cryo-efficiency of different cryoprotectants, their concentration and equilibration time and any interactions. In experiment 1, semen samples (n = 4 males; 2 ejaculates/male) were frozen using Green Buffer containing one of four cryoprotectants (3% glycerol, ethylene glycol, methyl formamide, dimethyl sulfoxide) and using 4 equilibration times (10 min, 0.5, 1 and 2 h). Glycerol and ethylene glycol provided the best motility recovery rates and different equilibration times were not significant for any cryoprotectant nor were any interactions noted. However different equilibration times were pertinent for improved kinematic parameters BCF and VSL. In experiment 2, glycerol and ethylene glycol were evaluated at 4 concentrations (1.5, 3, 6, 9%) with 0.5 h equilibration (n = 4 males, 3 ejaculates/male). Sperm motility recoveries, kinematics and acrosome status were assessed. Higher values for LIN and STR were found with ethylene glycol. At 0 and 1 h post thaw 3 and 6% of either cryoprotectant resulted in better motility values than 1.5%. Acrosome integrity was compromised at 9% cryoprotectant. There were interactions between cryoprotectant and concentration in total motility at 0 and 1 h. For glycerol, total motility recoveries were best at 3–9%; for ethylene glycol 1.5–6% were best at 0 h and 3–6% at 1 h. In conclusion, 3–6% glycerol or ethylene glycol offered the best cryoprotection for camel sperm while different equilibration times were not critical.  相似文献   

7.
There is a clinical demand for efficient cryopreservation of cloned camel embryos with considerable logistic and economic advantage. Vitrification of in vivo derived embryos has been reported in camels, but there is no study on vitrification of cloned embryos. Moreover, whether characteristic differences between cloned and in vivo derived embryos imply different vitrification requirement is unresolved. Here, we compared survival, re-expansion and pregnancy rates of cloned embryos vitrified using two commercial vitrification kits (Cryotec and Kitazato), developed basically for human embryos, and a vitrification protocol developed for in vivo camel embryos (CVP). Cloned embryos responded dynamically to vitrification-warming steps in commercial kits, with a flat shrinkage in the final vitrification solution and a quick re-expansion to the original volume immediately after transferring to the isotonic warming solution. Contrarily, full shrinkage was not observed in CVP method, and majority of embryos were still collapsed post-warming. The immediate re-expansion was highly associated and predictive of higher survival and total cell number, and also better redox state of embryos vitrified by Cryotec and Kitazato kits compared to CVP method. Importantly, while 30% blastomere loss, verified by differential dye exclusion test, was tolerated in vitrified embryos, >50% blastomeres loss in non-expanded blastocysts implied the minimal essential cell survival rate for blastocoelic cavity re-expansion in vitrified cloned camel blastocysts, irrespective of vitrification method. A protocol-based exposure of embryos to cryoprotectants indicated that cryoprotectant toxicity, per se, may not be involved in lower cryosurvival of embryos in CVP vs. Cryotec and Kitazato. The initial pregnancy rates were numerically higher in Cryotec and Kitazato frozen transfers compared to fresh transfer (56.3, 60 and 33.3%, respectively), and importantly, a higher percentage of established pregnancies in vitrified groups passed the critical 3 months period of early embryonic loss compared to sibling fresh clone pregnancies (50, 40, and 10%, respectively). Results confirmed the suitability of Cryotec and Kitazato kits for vitrification of cloned camel embryos and that vitrification may improve pregnancy outcome by weeding out poor competent embryos.  相似文献   

8.
9.
It was hypothesized that optimal sperm recovery rate (RR) without damage to the sperm would be obtained after centrifugation without a cushion solution. Semen collected three times from six light breed stallions was extended to 25 × 106 sperm/mL and centrifuged at CON (noncentrifuged), 900NC (no-cushion), 900C (cushion), 1800NC, and 1800C × g for 10 minutes. Sperm concentration, motility (TM and PM), and intact plasma membranes (PLM) and acrosomes (ACR) pre- and postcentrifugation (D0) and after 24 hours (D1) of cooling were evaluated. The RR in the CON (100 ± 0.0), 900NC (93.7 ± 2.9), and 1800NC (96.7 ± 2.6) groups was significantly higher than the 900C (68.7 ± 4.6) and 1800C (79.6 ± 3.5) groups. The D0 TM and PM were not different between the CON, 900NC, 900C, and 1800C, but were lower for the 1800NC group. The D1 TM and PM of the 900NC (75.2 ± 3.8 and 71.1 ± 4.1) and 900C (76.2 ± 3.7 and 72.4 ± 4.0) groups were significantly higher than the 1800NC (71.7 ± 4.1 and 67.3 ± 4.4) and 1800C (71.6 ± 4.1 and 67.2 ± 4.4) groups, and the CON (66.2 ± 4.5 and 60.0 ± 4.8) group was significantly lower than the other groups. The D1 PLM of the CON, 900NC, 900C, 1800NC, and 1800C groups were not different. The ACR on D1 was significantly lower for the CON (93.0 ± 2.4) group compared with all other groups. Optimal RR preserving sperm integrity was obtained in the 900NC group.  相似文献   

10.
This study investigated the capacity of healthy oocytes derived from follicles of different size to undergo normal fertilization and early embryonic development in vitro and full-term development in vivo. Ovaries were collected from a local abattoir and dissected and classified as follows: group A, greater than 4-8 mm (large); group B, greater than 2-4 mm (medium); and group C, greater than 1-2 mm (small). Oocytes were isolated by puncturing the follicular wall and pressing of the follicle. Only healthy-looking cumulus-oocyte complexes (COC) were used for in vitro maturation. Oocytes were fertilized in vitro by frozen/thawed semen from one bull. Approximately one-fourth of all oocytes was fixed and stained 15-20 h after fertilization, to determine penetration rates. The remaining eggs were transferred to culture medium and were cultivated for up to 9.5 days. Cleavage was observed 65 h and 7 days after fertilization. Expanded, hatching, and hatched blastocysts were fixed and stained after 9.5 days of culture. A total of 86 blastocysts derived from group A and B oocytes was nonsurgically transferred to synchronized recipients 7-8 days after onset of culture. A total of 6.624 follicles were dissected from 265 ovaries, and 1,485 oocytes were isolated from 1,671 group A follicles, 3,509 oocytes from 3,862 group B follicles, and 965 oocytes from 1,091 group C follicles. The fertilization rate, rate of normal fertilization, rate of polyspermy, and rate of other abnormal fertilization features were as follows: group A, 84.9%, 43.2%, 34.1%, 7.6%; group B, 83.6%, 44.8%, 31.1%, 7.8%; and group C, 61.7%, 13.1%, 33.7%, 19.1%.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
Two experiments were conducted to determine the effects of storage on equine ovaries or isolated oocytes. Ovaries were collected at an abattoir and were maintained at room temperature during collection and transport (3-9h total). After arrival at the laboratory, ovaries were divided into three groups: immediate oocyte collection (control), storage at room temperature overnight (15-18 h) before oocyte collection, or storage at 4 degrees C overnight before oocyte collection. Collected oocytes were cultured in maturation medium for 24h. There was a significant increase in the proportion of oocytes classified as having compact cumuli in the two storage groups when compared with the controls. For oocytes originally having expanded cumuli, the rate of maturation to MII was significantly higher in the control group (72%) than in either storage group, and the maturation rate for oocytes from ovaries stored at room temperature (27%) was significantly higher than that for ovaries stored at 4 degrees C (10%). A similar trend was seen for oocytes originally having compact cumuli (24, 11, and 3% in MI-II for control, room temperature, and cold groups, respectively). In Experiment 2, we evaluated the effect of different packaging systems on the maturation of horse oocytes within a portable incubator. Use of 1 ml of equilibrated maturation medium in a 1 ml glass vial was associated with maturation equivalent to that for standard incubation.  相似文献   

12.
需氧厌氧培养方式对甲型副伤寒沙门菌检出率的影响   总被引:3,自引:0,他引:3  
目的探讨需氧和厌氧血培养方式的选择对甲型副伤寒沙门菌检出率的影响。方法使用mini VI-TAL自动荧光血培养仪或BacT/Alert 3D培养仪对18684例疑似血流感染患者血液(含骨髓)标本进行需氧和(或)厌氧培养(其中仅做需氧培养935例,仅做厌氧培养16例,需氧和厌氧配对培养17733例,每瓶注入约5 ml血液)。结果18684例血液标本,共分离到甲型副伤寒沙门菌3888例(20.81%)。在17733例需氧和厌氧配对培养中检出3613例,其中仅需氧阳性406例占11.24%(406/3613),仅厌氧阳性405例占11.21%(405/3613),需氧和厌氧培养均阳性者2802例占77.55%(2802/3613),需氧和厌氧均生长的阳性报警时间分别为(22.56±13.22)h和(26.69±15.80)h,仅需氧阳性的报警时间为(32.85±23.33)h,仅厌氧阳性的报警时间为(34.46±18.44)h。结论需氧和厌氧血培养方式获得甲型副伤寒沙门菌的阳性率相同,采用需氧和厌氧瓶配对培养可提高阳性检出率,只做厌氧或需氧培养将有11.24%或11.21%阳性病例被漏检。  相似文献   

13.
Song Y  Wang C  Wang C  Lv L  Chen Y  Zuo Z 《Animal reproduction science》2009,110(3-4):306-318
The present study was undertaken to examine the effect of administered recombinant mouse leptin on the recovery of regressed ovary in fasted ducks. Twenty-eight ducks were divided into five groups: fed ad libitum (control; n = 5), fasted control (FC; n = 5), fasted + low dose of leptin (F + L; n = 5), fasted + medium dose of leptin (F + M; n = 5) and fasted + high dose of leptin (F + H; n = 3). All four fasted groups were fasted for 2 days and then ad libitum and the ducks were treated with leptin at doses of 0 (control and FC), 50 (F + L), 250 (F + M) and 1000 (F + H) μg/kg body weight/day on day 3–5. Results showed that a moderate dose of leptin (250 μg/kg body weight/day) injected during the re-feeding period: (i) promoted the recovery of the regressed ovary as evidenced by an increase in ovary weight and recovery of yellow hierarchical follicles; (ii) elevated the plasma 17β-estradiol (E2) level; (iii) increased the mRNA levels of ovary follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR) and estrogen receptor-β (ER-β). Furthermore, the results also showed that a high dose of leptin (1000 μg/kg body weight/day) may have a negative effect on the recovery of the regressed ovary. In conclusion, this study indicates that, in ducks, leptin may be involved in the recovery of the regressed ovary caused by 2 days of fasting. This effect may be related to increased plasma E2 levels and stimulation of the mRNA levels of ovarian FSHR, LHR and especially ER-β.  相似文献   

14.
应用ELISA法检测纯化狂犬病疫苗制备过程中的病毒回收率   总被引:5,自引:0,他引:5  
应用ELISA和病毒毒力滴度测定两种方法对狂犬病疫苗纯化过程中各步样品进行检测,并应用NIH效力测定法对ELISA法测定的结果进行验证。结果显示,ELISA法具有灵敏度高、特异性强、重复性好、简便易行等优点,是一种较理想的病毒回收率测定方法。  相似文献   

15.
Mares (n = 39) were classified according to age as young (less than 1.5 yr, n = 17) or old (more than 1.5 yr, n = 22) and sacrificed. Ovaries were measured and weighed, and the number of follicles and CL were counted. Follicle size and distribution were recorded (external: > 20 mm, < 20 mm; internal: > 5 mm, < 5 mm). External follicles were aspirated while internal follicles were sliced. The number and Type of oocytes recovered using each method were recorded. Oocyte recovery rates (oocytes/ovary) resulted in a mean of 0.92 oocytes by aspiration and 1.36 oocytes by additional slicing. The mean numbers of available follicles (8.11 and 5.02) oocytes recovered (4.94 and 3.02) and oocytes selected per ovary (3.29 and 2.32) were not significantly different in the young or old mares, respectively.  相似文献   

16.
Effect of preservation on wet weight biomass of chironomid larvae   总被引:1,自引:1,他引:0  
Preservation of four species of chironomid larvae in either 10% formalin or 70% ethanol caused changes in the preserved wet weight expressed as a percentage of the original live wet weight. The magnitude of the change varied with species, preservative, preservation time and the nature of the water used for preservative dilution. Failure to take these factors into account can lead to errors in wet weight measurements as great as 47%. The use of io% formalin prepared with filtered habitat water appears to be the most suitable preservative for chironomid larvae.  相似文献   

17.
Lin C  Tsai S 《Theriogenology》2012,77(6):1257-1261
Understanding chilling sensitivity and chilling injury of coral oocytes, in the presence and absence of a cryoprotectant, is important in developing cryopreservation protocols, as well as for short-term storage and transport (e.g., for species conservation). The objective of this study was to investigate the chilling sensitivity of hard coral (Echinopora spp.) oocytes and the effectiveness of methanol (as a cryoprotectant) in protecting these oocytes during short-term, low temperature preservation. Oocytes were exposed to 0.5, 1, or 2 m methanol at 5, 0, or −5 °C for 1, 2, 4, 6, 8, 16, or 32 h, and their quality determined based on adenosine triphosphate (ATP) content. Methanol at 0.5 m was the most effective means to reduce chilling-induced reduction in ATP concentrations. Coral oocytes can be stored at room temperature for 4 h in filtered nature seawater with no detrimental effect on oocyte quality; however, in the present study, oocyte survival was extended for 8 h by addition of methanol in low concentrations (0.5 or 1 m) at low temperatures (5 and 0 °C). These findings should enhance conservation efforts and facilitate low-temperature transport of endangered and threatened coral species.  相似文献   

18.
The effect of 6-dimethylaminopurine (6-DMAP) on germinal vesicle breakdown (GVBD) and maturation in bovine oocytes was investigated in this study. This puromycin analog has been shown to be an inhibitor of phosphorylation. Whereas GVBD occurred in nearly all oocytes (96.8%, 120/124) in control medium, presence of 6-DMAP (2 mM) blocked this process almost completely, irrespective of the presence (98.3% GV, 349/355) or absence (97.1% GV, 165/170) of cumulus cells. When lower concentrations of 6-DMAP were used (100-500 microM), GVBD was observed in 87.9% of oocytes, but their maturation was arrested at late diakinesis-metaphase I stage. The inhibition of GVBD was fully reversible, but most of the metaphase II plates were abnormal (80%). To assess whether the action of 6-DMAP is different from the inhibitors of protein synthesis, metaphase II oocytes were exposed to either cycloheximide or 6-DMAP, respectively. Whereas in cycloheximide-supplemented medium approximately 80% of the oocytes were activated, parthenogenetic activation was much less frequent after incubation in 6-DMAP (14.5%). Fusion studies showed that, even if GVBD occurs in 6-DMAP supplemented medium, the level of the maturation-promoting factor (MPF) is decreased. These experiments may indicate the importance of phosphorylation for GVBD in cattle oocytes.  相似文献   

19.
Summary The effect of clomiphene citrate on the rabbit ovary was studied in mature nulliparous rabbits pretreated with three consecutive doses ranging from 0.01–10.0 mg/kg per day. With increasing doses a trend of decrease in mean ovarian weight (mg/kg body weight) is observed 2 days after termination of treatment. Five days later a significant increase occurs, which then subsides again to control values on day 12 after termination of treatment. During this period, no matings or injections of luteinizing hormone were performed to trigger ovulation; consequently no ovulations are observed. Folliculogenesis appears as normally; number and morphology of follicles are within normal ranges. No endogenous, spontaneous gonadotropin surges are detected in blood serum up to the 7th day after termination of treatment (2 and 10 mg doses). The surface epithelium of the ovary resembles normal germinal epithelium; however, after treatment with high doses a secretory-like activity is observed, accompanied with ultrastructural changes.Supported by the Deutsche Forschungsgemeinschaft, Special Program on Biology and Clinics of Reproduction. (Grant Be 524/7-7).Visiting Scientist from the Department of Obstetrics and Gynecology, Hadassah University Hospital, Jerusalem, Israel.  相似文献   

20.
Summary The development of the telotrophic ovary in the Staphylinid beetle,Creophilus maxillosus was examined. Cells, termed chordoblasts were identified in the germarium of 1-day-old pupae. Each of the chordoblasts undergoes a series of synchronous mitoses. Owing to the precise control of the cleavage plane, which is vertical to the long axis of the ovariole, each of the chordoblasts gives rise to a linear chain of sibling chordocytes. Extra DNA synthesis within each sibling string is usually limited to the most posterior chordocyte only, this being an oocyte progenitor.Divisions of the oocyte progenitor are differential mitoses in which the extra DNA material is transported preferentially towards the posterior pole of the spindle. As extra DNA synthesis and preferential segregation of this material result in gradual increase of this DNA in the nuclei of oocyte progenitors, cytokinesis of these cells becomes highly unequal, the larger of the two cells produced at each differential mitosis being as a rule the posterior cell, i.e. the oocyte progenitor of the next cell generation. As a resul of the series of differential mitoses each chordoblast gives rise to a number of nurse cells and only one definitive oocyte.It is suggested that somatic prefollicular tissue plays a decisive role in oocyte determination in the Coleopteran telotrophic ovary.This word was supported in part under Contract DPKBN/52/76-II. 1. 3. 10. with the Polish Academy of Science  相似文献   

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