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1.
Catalase activity in equine semen   总被引:1,自引:0,他引:1  
OBJECTIVE: To characterize the activity of catalase in equine semen. ANIMALS: 15 stallions of known and unknown reproductive history. PROCEDURE: Seminal plasma was collected from raw equine semen by centrifugation, and samples of seminal plasma were frozen prior to assay for catalase activity. Tissue samples (n = 3 stallions) from the bulbourethral gland, prostate gland, vesicular gland, and testis were homogenized, and cauda epididymal fluid was collected for determination of catalase activity. Catalase activity was determined as an enzyme kinetic assay by the disappearance of H2O2 as measured by ultraviolet spectrophotometry. RESULTS: Catalase activity in equine seminal plasma was 989.3 +/- 1678 U/ml (mean +/- SEM), and the specific activity of catalase in equine seminal plasma was 98.7 +/- 29.2 U/mg of protein. Specific activity of catalase in tissue homogenates was significantly higher in the prostate gland (954 +/- 270 U/mg of protein) than in the ampulla (59 +/- 5 U/mg of protein), bulbourethral gland (54 +/- 11 U/mg of protein), vesicular gland (39 +/- 3 U/mg of protein), cauda epididymal fluid (11 +/- 3 U/mg protein), or testis (54 +/- 6 U/mg of protein). CONCLUSIONS AND CLINICAL RELEVANCE: Equine seminal plasma contains a high activity of catalase that is derived primarily from prostatic secretions. Procedures such as semen cryopreservation that remove most seminal plasma from semen may reduce the ability to scavenge H2O2 and thereby increase the susceptibility of spermatozoa to oxidative stress.  相似文献   

2.
Preservation of epididymal spermatozoa is an advantageous method to preserve genetic material of endangered species or valuable breeding animals after sudden death and injuries. Despite lower pregnancy rates, fertilization with epididymal sperm has been proven successful. Variable sperm quality after cryopreservation among individual stallions and the usually terminal chance to preserve epididymal sperm are opportunities for the development of a freezing procedure suitable for the majority of stallions. To evaluate the effect of the preservation procedure, we analyzed the sperm motion characteristics after every step of processing. In addition, we investigated the influence of seminal plasma on motility of frozen-thawed semen. We compared three segments of the cauda epididymidis and harvested spermatozoa by retrograde flushing (most caudal part) and mincing (more cranial segments) to augment the number of spermatozoa. During processing, there were differences in sperm motion characteristics depending on the segment of the cauda epididymidis. Distinct increases in motility due to different extenders and the effect of seminal plasma suggest that motion characteristics of raw and frozen-thawed spermatozoa are strongly influenced by microenvironment and must be interpreted with caution.  相似文献   

3.
The aim of the present study was to determine the effects of dietary supplements of vitamins on vitamin status, libido, and semen characteristics in young boars under normal and intensive semen collection. Sixty Landrace, Yorkshire, and Duroc boars were allocated randomly from 6 to 10 mo of age to one of the following diets: 1) basal diet (industry level) for minerals and vitamins (Control, n = 15); 2) basal diet supplemented with vitamin C (ASC, n = 15); 3) basal diet supplemented with fat-soluble vitamins (FSV, n = 15); and 4) basal diet supplemented with water-soluble vitamins (WSV, n = 15). After puberty (approximately 12 mo of age), semen was collected at a regular frequency (three times every 2 wk) for 5 wk. Thereafter, all boars were intensively collected (daily during 2 wk). A recovery period (semen collection three times every 2 wk) followed and lasted for 10 wk. Sperm quality (percentage of motile cells and percentage of morphologically normal cells) and quantity (sperm concentration, semen volume, and total sperm number) were recorded as well as direct and hormone related measurements of boar libido. Blood and seminal plasma samples were taken to monitor vitamin status. High concentrations of B6 (P < 0.05) and folic acid (P < 0.05) were observed in the blood plasma of WSV boars, whereas greater concentrations of vitamin E (P < 0.01) were obtained in FSV boars. In the seminal plasma, folic acid concentrations tended to be greater in WSV boars (P < 0.08). During the intensive collection period, there was a tendency (P < 0.06) for semen production to be greater in WSV boars, the effect being less pronounced (P < 0.10) in FSV boars. During the recovery period, the percentage of motile sperm cells was greater in WSV boars (P < 0.03) and, to a lesser extent, in FSV boars (P < 0.10) compared with Control boars. Sperm morphology and libido were not affected by treatments. These results indicate that the transfer of vitamins from blood to seminal plasma is limited and the dietary supplements of water-soluble and fat-soluble vitamins may increase semen production during intensive semen collection.  相似文献   

4.
Semen samples were collected at weekly intervals for six weeks from eight sexually mature beagles previously shown to produce normal ejaculates. Seminal plasma and sperm fractions were separated by centrifugation and the sodium, potassium, alanine and aspartate aminotransferases, acid and alkaline phosphatase concentrations in the two fractions determined. Regression analysis of the mean weekly values obtained from physical and biochemical examination of the ejaculates showed that sodium ion concentration was highest in seminal plasma. The highest levels of aminotransferases were found in sperm fractions. Those enzymes may be indices of abnormal or damaged spermatozoa. Acid and alkaline phosphatase activity was 100 times greater in seminal plasma than in sperm fractions. Phosphatase concentrations are likely to be dependent on prostate activity. Measurement of acid phosphatase in canine semen therefore may be a useful index of prostate function. The motility of the semen samples was independent of the potassium concentration in seminal plasma. However, there was some evidence of a correlation between sperm motility and the enzyme and sodium content of seminal plasma.  相似文献   

5.
6.
Frequency of semen collection was used to determine methods of stabilising epididymal sperm reserves in young boars in order to measure daily sperm output which then allows estimation of daily sperm production. By collecting from boars 3 times a week (Monday, Wednesday and Friday) most boars reached epididymal stabilisation within 4 weeks and the daily sperm output could be calculated from the following 6 collections. A more rapid method of measuring daily sperm output was achieved by collecting from boars 4 or 5 times during one day and daily for the following 4 days. Daily sperm output measured by this method was not found to be significantly different from the method requiring collections over a 6-week period. Frequency of collection had a significant effect on semen characteristics. Collecting 3 times a week resulted in significantly greater volumes, sperm concentration, total number of sperm in the ejaculate and daily sperm output compared with collecting at intervals of 48 h. A collecting frequency of 48 h resulted in significantly greater volumes, sperm concentration and total number of sperm in the ejaculate but not daily sperm output when compared with collecting every 24 h.  相似文献   

7.
Hyaluronidase release was used as an index of acrosomal membrane damage during cold shock of epididymal boar sperm and ejaculated sperm from intact and vesiculectomized boars. Sperm were also incubated with seminal plasma from intact and vasectomized boars to examine the contributions of male accessory gland secretions. Acrosomal membranes of epididymal sperm were more resistant to cold shock than those of ejaculated sperm. Only 36% of the hyaluronidase released by ejaculated sperm was released by the epididymal sperm in spite of similar hyaluronidase content of the sperm. Preincubation of epididymal sperm in seminal plasma from both intact and vasectomized boars increased resistance to cold shock by 60 to 80%. Initial dilution of epididymal sperm with seminal plasma, rather than Ringer-fructose buffer, was associated with low progressive motility and with retention of cytoplasmic droplets. In contrast, acrosomal membranes of ejaculated sperm from intact and vesiculectomized boars exhibited similar sensitivity to cold shock, releasing hyaluronidase capable of forming .20 and .19 mumol N-acetylglucosamine from hyaluronic acid/10(8) sperm in 8 min. Moreover, seminal plasma from vasectomized boars had no effect on acrosomal sensitivity to cold shock of ejaculated sperm from vesiculectomized boars.  相似文献   

8.
Excretion of porcine parvovirus through the genital tract of boars   总被引:1,自引:0,他引:1  
The putative binding of porcine parvovirus (PPV) to semen components in vitro was examined along with the shedding pattern of PPV in oronasally infected boars. Porcine parvovirus DNA was determined to be bound to spermatozoa that had been incubated in vitro with PPV and washed to remove loosely adherent virus. To determine whether PPV was shed in the semen, four 8-month-old boars, seronegative for PPV, were inoculated oronasally with a virulent strain of PPV. Prior to virus inoculation, a catheter was surgically implanted in the vas deferens for the purpose of collecting cauda epididymal semen free of extrinsic contamination. Epididymal semen specimens were collected prior to inoculation and daily thereafter for 21 days. A fifth boar was inoculated oronasally with PPV, but semen was collected by electroejaculation twice weekly for an equal period of time. Reproductive glands and semen specimens from all boars were examined by nucleic acid hybridization for the presence of viral DNA. All boars seroconverted to PPV, as evidenced by serum antibody titers ranging from 512 to 8,192 hemagglutinating inhibition units/50 microliters. Porcine parvovirus DNA was detected in epididymal semen of 3 of 4 catheterized boars on postinoculation days 5 through 9, but not in semen obtained by electroejaculation. Viral DNA was consistently detected in tissue samples collected on postinoculation days 8 and 21 from the scrotal lymph nodes (4 of 5 boars) and epididymides (3 of 5 boars).  相似文献   

9.
高温对荷斯坦种公牛精液品质及精清生化指标的影响研究   总被引:5,自引:0,他引:5  
以荷斯坦种公牛为试验牛研究了高温对其精液品质和精清生化指标的影响。结果表明:(1)高温可造成种公牛精液品质显著下降。使原精活力、精子密度、活精子百分数和顶体完整率分别比春季下降3·13%(P<0·05)、34·8%(P<0·01)、15·0%(P<0·01)和17·8%(P<0·01),精子畸形率比春季上升25·5%(P<0·01)。(2)夏季荷斯坦种公牛精清睾酮含量仅为4·31pg/mL,极显著低于非高温季节(P<0·01)。(3)高温环境使得荷斯坦种公牛精清钾、钙、镁处于最低水平(P<0·01),精清碱性磷酸酶(ALP)、酸性磷酸酶(ACP)处于最高水平(P<0·01),使精清钠由春季至夏季呈显著下降趋势。  相似文献   

10.
Three normal, sexually mature young boars of Swedish Yorkshire breed were treated with daily intramuscular injections of diethylstilboestrol and/or oestradiolbenzoate for a total period of 13–15 weeks. Whole ejaculates were collected on a dummy sow with an artificial vagina twice a week. The ejaculate was examined as regards semen volume, sperm concentration, sperm motility and sperm morphology. The seminal plasma was analysed for Na, K, Cl, Mg, inorganic phosphate, total protein and fructose. Testicles, epididymides and the accessory sex glands were obtained at slaughter. Material from epididymal segments A, B, Db, Fa and Fb was collected and examined for sperm morphology. Spermatocrit, osmotic pressure and plasma concentrations of Na, K, Gl, total protein and GPC were assessed in material from epididymal segment Fa. The sex glands and the accessory sex glands were examined maeroscopically and microscopically.The semen volume, the sperm concentration and consequently the total sperm count per ejaculate showed a gradual increase during the course of the investigation. This is to be expected as the boars used were comparatively young at the beginning of the experiment. These values were all within normal limits for adolescent boars.Taking all the characteristics examined into consideration no conclusive evidence was found that an exogenic administration of oestrogens to intact boars has any influence on the function of the epididymis and the accessory sex glands.  相似文献   

11.
This study attempted to explain the mechanisms regulating boar fertility by examining seasonal changes in semen characteristics, the composition of seminal plasma and responsiveness of sperm acrosomes to Ca(2+) and the Ca(2+) ionophore A23187 (Ca(2+)/A23187). Sperm-rich and sperm-poor fractions were separately collected from 3 mature fertile Large White boars once a month over a one-year period. During the period of study, ambient temperature and relative humidity were recorded for within the stall in which the boars were kept and the semen characteristics, composition of the seminal plasma of sperm-rich fractions, and occurrence of the acrosome reaction in response to Ca(2+) (3 mM)/A23187 (0.3 microM) were examined. The highest mean maximum and minimum ambient temperatures were recorded in August-September, whereas the lowest mean maximum and minimum ambient temperatures were recorded in December and January, respectively. There was a moderate peak in relative humidity from July to October. The lowest percentages of motile spermatozoa and of spermatozoa with intact acrosomes and highest percentage of spermatozoa with abnormal morphology and strongest agglutination were seen in August-September. The total protein and albumin concentrations were lowest in August-September. Testosterone levels increased gradually as day length decreased after the summer solstice (June) and peaked in October-November. The percentage of acrosome reactions in response to Ca(2+)/A23187 was highest with the quickest response in August-September, as shown by the shortest time required for 50% of relative acrosome reactions. The farrowing rates were lowest in these same 2 months. These results suggest that seasonal infertility in Large White boars may be due, at least in part, to a combination of low motility, abnormal morphology including acrosomal abnormality, and early occurrence of the acrosome reaction in response to stimulus, possibly resulting from a decrease in acrosomal stabilizing proteins in the seminal plasma during summer. These changes may be modulated by heat/humidity stress and/or photoperiod-regulated testosterone.  相似文献   

12.
本试验旨在研究谷胱甘肽过氧化物酶6(glutathione peroxidase 6,GPx6)在大白公猪附睾细胞中的表达和定位,并探究其与公猪繁殖性能的相关性。选取15月龄的公猪和母猪各3头,取睾丸、附睾、前列腺、尿道球腺、精囊腺、卵巢和输卵管,提取蛋白,通过蛋白免疫印迹和免疫组化(IHC)检测组织和细胞中GPx6蛋白的表达和定位;根据评定大白公猪受精能力的数学模型,按照公猪配种胎次≥20胎、3次配种公猪为同一头的标准,筛选并采集符合要求的20头公猪精液,同时,统计相对应的1 279头母猪的生产成绩,计算得到公猪繁殖性能指标(包括窝产总仔数、窝产活仔数、分娩率和繁殖力)。提取精子蛋白和精清蛋白,通过BCA和ELISA检测精子和精清中GPx6蛋白的含量。使用SPSS软件的独立样本t检验及单因素方差分析(one-way ANOVA),进行差异显著性分析,用双变量Pearson进行相关性分析,P<0.05表示差异或相关显著。结果表明,GPx6蛋白在附睾中高表达,IHC结果显示,GPx6蛋白在附睾的顶细胞、基底细胞、晕细胞、主细胞和精子中表达,在肌样细胞中不表达;精清蛋白中GPx6的含量是精子蛋白的7倍,精液中GPx6蛋白的含量与窝产活仔数、窝产总仔数呈负相关关系。结果提示,GPx6在附睾的顶细胞、基底细胞、晕细胞、主细胞和精子中表达,且其在精液中的含量会影响公猪的繁殖性能,这为GPx6对公猪受精能力影响的研究奠定了理论和试验基础。  相似文献   

13.
本试验旨在研究不同精液品质特征长白公猪血清和精浆中元素含量的差异,分析血清和精浆中元素含量对精液品质的影响。基于107头长白公猪的1402次精液品质记录,将公猪群按照精液可利用率划分为3组:低利用率组(利用率<60%,n=21)、中等利用率组(60%≤利用率≤80%,n=27)和高利用率组(利用率>80%,n=59)。采集每头长白公猪的血清和精浆样品,采用电感耦合等离子体质谱法检测血清和精浆中营养元素和毒性元素含量。结果表明:1)低利用率组长白公猪的有效精子数和精子活力显著低于中等利用率和高利用率组(P<0.01),精子畸形率显著高于中等利用率和高利用率组(P<0.01)。2)不同利用率组间长白公猪的血清和精浆元素含量无显著差异(P>0.05);但在血清和精浆元素含量与精液品质参数相关性分析中发现,精浆铅元素含量与精子活力呈显著负相关(P<0.05),与精子畸形率呈显著正相关(P<0.05)。3)对精浆铅含量分组进一步分析发现,精浆铅含量为0μg/L时长白公猪的精子畸形率显著低于精浆铅含量>10.0μg/L时(P<0.05),精子畸形率降低约6.11%。总的来说,长白公猪精浆中毒性元素铅的存在会通过损害精子活力和形态,影响公猪精液品质。  相似文献   

14.
The objective of this study was to investigate the effects of a preparation from herbal extracts (PHE) on libido and semen quality in breeding artificial insemination boars. Ten fertile boars were divided into control and experimental groups according to significant difference of libido. There were no differences in semen quality between groups. Animals were fed a commercial feeding mixture for boars. The feeding mixture for the experimental group was enriched with PHE, which was prepared from Eurycoma longifolia, Tribulus terrestris and Leuzea carthamoides. Duration of the experiment was 10 weeks. Samples of ejaculate were collected weekly. Libido was evaluated according to a scale of 0-5 points. Semen volume, sperm motility, percentage of viable spermatozoa, sperm concentration, morphologically abnormal spermatozoa, daily sperm production and sperm survival were assessed. Amounts of mineral components and free amino acids were analysed in seminal plasma. Significant differences were found in these parameters: libido (4.05 ± 0.22 vs 3.48 ± 0.78; p < 0.001), semen volume (331.75 ± 61.91 vs 263.13 ± 87.17 g; p < 0.001), sperm concentration (386.25 ± 107.95 vs 487.25 ± 165.50 × 10(3) /mm(3); p < 0.01), morphologically abnormal spermatozoa (15.94 ± 11.08 vs 20.88 ± 9.19%; p < 0.001) and Mg concentration (28.36 ± 11.59 vs 20.27 ± 13.93 mm; p < 0.05). The experimental group's libido was increased by 20% in comparison with the beginning of the experiment. Results of this study showed positive effect of PHE on libido and some parameters of boar semen quality.  相似文献   

15.
Alkaline phosphatase activity was recorded in forty ejaculates of the sperm rich fraction of boar semen as 9,790 ± 5,250 Klein-Babson-Read units per 100 ml. of seminal plasma. Acid phosphatase activity in the same ejaculates was 681 ± 304 Babson-Read units per 100 ml. of seminal plasma. No alkaline phosphatase activity was detected in the seminal plasma of vasectomized boars.

The pH of the sperm rich fractions was 7.69 ± 0.33 and the osmotic pressure was 313.56 ± 7.98 milliosmols.

  相似文献   

16.
Membrane vesicles, separated by differential centrifugation from the seminal plasma, were detected in the sperm-rich ejaculate fractions of four boars and three stallions, and in the whole ejaculates of seven rams. The volume and percentage of vesicles, determined by a stereological technique, were higher in the sperm-rich than in the post-sperm-rich fractions of the boar and stallion ejaculates, and no vesicles were detected in the pre sperm-rich fractions. Vesicles were examined by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). For boar, stallion and ram semen, the mean (+/- s.e.m.) vesicle diameters were 130.9 +/- 3.22 (range 18-577), 164.1 +/- 4.42 (range 15-671) and 159.7 +/- 2.92 nm (range 22-986), respectively, although they were not significantly different (p = 0.709). The vesicles had approximately round (TEM) or spherical shape (SEM), were surrounded by single, double or multi-laminar membranes, and were trapped within ample amorphous material, sometimes containing short, flattened membranous elements. The majority of the vesicles had a clear interior but some contained granule-dense material. Ram membrane vesicles, purified from ultracentrifuged plasma by size exclusion chromatography, kept their round shape and the amorphous material was less evident compared with the sections taken before purification. This is the first report to identify seminal plasma membrane vesicles in the different fractions of ejaculated semen in the boar and stallion, and confirms their presence in ram seminal plasma. The origin and function of these vesicles are yet to be elucidated.  相似文献   

17.
This study investigated the epididymis of mature boars (n = 10) by means of B‐mode ultrasound and grey‐scale analysis (GSA) for echogenicity (EG) determination using an ultrasound unit HS 1500V, a linear transducer (frequencies 7.5–9.0 MHz), and standardized unit settings. All boars had their epididymal caput, corpus and cauda scanned six times before and after semen collection, respectively, at weekly intervals. Semen was subjected to spermatological examination including volume, total and forward motility, sperm abnormalities, as well as total sperm count and concentration. The caput and corpus both had a homogeneous fine echotexture. The cauda was homogeneous too but had a marbled echotex‐ture. Echogenicity before and after semen collection was caput > corpus > cauda, respectively (p < 0.001). Echogenicity was higher before than after semen collection for all parts of the epididymis, respectively (p < 0.001). Echogenicity of the caput correlated slightly positively with the total sperm count pre‐collection (r = 0.301; p = 0.020) and with ejaculate volume pre‐ and post‐collection (r = 0.302 vs 0.306; p = 0.017 vs 0.019), and slightly negatively with sperm concentration post‐collection (r = ?0.275; p = 0.034). No relationship was found for EG of the corpus and cauda for any of the ejaculate parameters. In conclusion, B‐mode ultrasound and GSA proved feasible for imaging the epididymis in boars. Single relationships between EG and ejaculate parameters were found and deserve further investigation.  相似文献   

18.
本试验旨在研究骨桥蛋白(osteopontin,OPN)在长白公猪生殖细胞中的表达和定位,并探究其与公猪繁殖性能的相关性。试验采集了长白公猪精液和不同阶段(3日龄、3月龄、6月龄和12月龄)的睾丸组织,通过蛋白印迹的方法检测OPN蛋白在精液和不同月龄睾丸中的表达,通过免疫组化的方法对OPN蛋白在公猪睾丸细胞中进行定位;同时,根据配种胎次≥ 20胎,3次配种公猪为同一头的标准,筛选并采集17头长白种公猪精液,统计相对应的1 388头母猪的生产成绩,计算得到公猪繁殖性能指标(包括窝产总仔数、窝产活仔数、分娩率和繁殖力)。低温离心精液分离得到精子和精浆,丙酮法提取精浆蛋白,Lysis buffer方法提取精子蛋白,最后运用BCA和ELISA的方法检测精子和精浆中OPN蛋白的含量,分析OPN蛋白与公猪繁殖性能的相关性。蛋白印迹结果显示,OPN在精子、精浆和各月龄阶段的长白公猪睾丸中均以两种形式表达(67.4和33.7 ku),且67.4 ku的形式在3月龄公猪睾丸中表达量最高;免疫组化的结果显示,OPN在长白公猪的初级精母细胞、次级精母细胞和精子细胞中表达,在精母细胞、支持细胞和间质细胞中无表达;BCA和ELISA结果显示,精子中的OPN蛋白含量是精浆中的7倍(P<0.05),精液中的OPN蛋白与公猪窝产活仔数显著正相关(P<0.05)。本试验结果表明,OPN在各阶段的长白公猪睾丸中都有表达,且在精子和精浆中也有表达,这可能与公猪的繁殖性能有关,从而为后期OPN蛋白在公猪受精力的研究奠定基础。  相似文献   

19.
Eight adult Landrace boars were housed for 12 mo in one of two social environments. Socially nonrestricted boars were penned adjacent to and allowed minimal physical contact with ovariectomized gilts hormonally induced into estrus every 2 wk. Socially restricted boars were penned behind solid walls to eliminate visual and physical contact with other pigs. All animals were subjected to natural changes in daylength. Semen was collected weekly; gel-free volume, gel weight, sperm concentration and number per ejaculate, sperm motility (percent and type) and semen pH were determined. Total protein, citric acid contents and alkaline phosphatase activity were measured in seminal plasma. Testis length and width and various body temperature measurements were recorded monthly. Except for percent motile sperm and alkaline phosphatase activity, all semen characteristics varied (P less than .05) with month. The pattern of seasonal change in semen volume was modified by social environment (group X month, P less than .05). Sperm numbers were highest in winter and lowest in spring and summer. Ejaculate protein and citric acid contents were highest in fall and winter; decreases in spring were associated with moderate ambient temperatures and increases in daylength (r = -.80, P less than .05). Testicular length for socially nonrestricted boars was maximum in November through January and minimum in April through July, and did not vary as extensively for socially restricted boars. Scrotal temperature was elevated during periods of high ambient temperature, but not to values detrimental to spermatogenic function.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

20.
A study was undertaken to compare the effect on fertility in the fowl of aqueous medium, natural homologous seminal plasma, heterologous turkey seminal plasma and whole turkey semen when whole fowl semen was excessively diluted with these media and inseminated fresh. High dilution with fowl seminal plasma resulted in the best fertility. Dilution with the turkey semen components produced fertility no different from that with aqueous diluent when the dose of spermatozoa was 5 X 5 or 10 X 10(6). The results of this study confirm that 5 X 5 to 10 X 10(6) good quality spermatozoa are sufficient to produce acceptable fertility in weekly inseminations of fresh semen. This enables good quality semen to be highly diluted. However, at high dilution rates there is a need to reconsider the composition of semen diluents, with respect to simulating as yet unknown properties provided by factor(s) in homologous ductus deferens seminal plasma.  相似文献   

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