蛇鮈精子保存液的筛选及其效果研究

旨在研究不同精子保存液对蛇鮈精子活率的影响,并探讨保存蛇鮈精子的最佳保存液。在光镜下观察6种精子保存液保存蛇鮈精子后精子活率的变化,结果表明精子保存液在低温(4℃)保存蛇鮈精子的方法效果较好。6种蛇鮈精子保存液使精子的活率保持为80%的时间有着明显的差异:A液为36 h,B液为4 h,C液为0 h,D液为60h,E液为16 h,F液为6 h,因此在低温(4℃)下,蛇鮈精子的最佳保存液为D液。     

11种灭活型样本保存液对病毒核酸保护效果的比较

收集11种不同厂家的灭活型样本保存液,将甲型流感病毒加入保存液和生理盐水中,在4℃、25℃和37℃条件下保存0 h、2 h、4 h、6 h和24 h后,分别提取各组的病毒核酸,并通过逆转录实时荧光PCR检测病毒载量变化,以探究不同样本保存液对病毒核酸保护效果是否存在差异。结果发现不同样本保存液中的病毒载量随保存时间和温度变化存在显著差异。据此可将11种灭活型样本保存液的保存效果分为四类：(1)优秀产品：在任一温度（4℃、25℃、37℃）条件下,即使保存24h时病毒核酸都未发生明显降解,占比27.2%(3/11);(2)良好产品：低温（4℃）和室温（25℃）情况下核酸未出现降解,但在高温（37℃）条件下6 h后保存效果明显下降,占比27.2%(3/11);(3)合格产品：低温（4℃）条件下对病毒核酸具有较好的保护效果,但在室温（25℃）和高温（37℃）情况下随保存时间延长其保护效果显著下降,占比18.1%(2/11);(4)不合格产品：任一保存条件下对病毒核酸的保护效果都较差,甚至加入病毒后立即导致病毒核酸发生快速降解,占比27.2%(3/11)。以上结果说明不同厂家样本保存液对病毒核酸...     

4℃保存的小鼠屠体卵巢GV卵的发育能力

为了探明雌性动物死亡后GV期卵的可利用性,本研究将ICR系雌性小鼠处死,尸体在4～6℃下分别保存16h、24h和48h,取其卵巢GV期卵,体外成熟后,应用常规法进行体外受精或透明带切割法体外受精,获得2细胞期胚,通过体外培养或胚胎移植观察其发育能力。结果显示,4～6℃下保存16h和24h处理组的体外受精率(分别为31%、33%和21%、24%)与来自新鲜的带有颗粒细胞卵的体外受精率(33%)相比无显著差异。与其相比,保存48h处理组的GV卵已丧失发育能力。此外,体外受精中获得的2细胞胚经体外培养,16h处理组和24h处理组的囊胚率(60%、61%和72%、83%)与新鲜GV期卵处理组的囊胚率(72%)相比差异不显著。获得的2细胞胚经胚胎移植可得到正常幼鼠。上述结果表明,如果雌性小鼠死亡后立即在冷藏温度下(4～6℃)保存,其体内的GV卵在24h以内仍保持发育能力。     

不同温度及时间对液状保存血液质量的影响

目的：研究不同温度对血液液状保存时保存损害机制的影响,并探讨相应的防范措施。方法：取10名健康献血员静脉血,置于CP2D－A保存液中,于0℃和4℃环境条件下,分别于设定的时间（第1周末、第3周和第6周末）内取样检测GSH－Px、LPO、TSH、红细胞膜收缩蛋白、膜脂流动性等指标。结果：固定温度条件下随时间的延长血液过氧化反应增强,保存损害作用增加;同一时期内保存损害作用随温度的降低而减轻,以4℃组血液老化明显。结论：血液过氧化反应随保存期的延长而增加,随保存温度的降低而改善,0℃组保存血液的质量优于4℃组。     

不同温度及时间对保存血液有效期和质量的影响

目的 :研究不同温度对血液液状保存时保存损伤机制的影响 ,并探讨相应的防范措施。方法 :取 10名健康献血员静脉血 ,混合于CP2D A保存液中 ,均分为 2组 ,分别保存于 0℃和 4℃环境条件下 ,分别于设定的时间 (第2 1d和第 4 2d)以shafiq UR rehman法检测膜磷脂 ,同步法检测Na K ATP酶、纯化PDE法检测CaM、分光光度法检测LPO等指标。结果 :固定温度条件下随时间的延长血液过氧化反应增强 ,保存损伤作用增加 ;同一时期内保存损伤作用随温度的降低而减轻 ,以 4℃组血液老化明显。结论 :血液保存损伤作用随保存期的延长而增强 ,随保存温度的降低而改善 ,并指出血液液状保存所存在的问题与展望。     

室温下保存的小鼠屠体卵巢GV卵的发育能力

将ICR系雌性小鼠处死并在10℃、15℃、20℃和25℃下依次保存8、14、24和48 h后,采集其体内的卵巢GV期卵,采用常规方法进行体外成熟和体外受精,获得的2细胞期胚经体外培养或胚胎移植观察其发育能力.其结果,在10℃下保存24 h、15℃下保存14 h、20℃下保存8h和25℃下保存4 h后,其体内附有卵丘细胞的GV卵的体外成熟-体外受精后的2细胞率分别为14%、9%、10%和10%,随着保存温度的提高和保存时间的延长,带有颗粒细胞GV期卵的比率明显降低,同时其GV期卵经体外成熟及体外受精后的2细胞率明显降低.在20℃下保存24 h和25℃下保存14 h时,难以获得形态正常的GV期卵;体外受精获得的2细胞期胚经体外培养,总体上有64%的胚胎发育至扩张囊胚,未见有保存温度和保存时间的显著影响,且利用在15℃保存8 h后的GV卵获得2细胞期胚的移植获得正常新生小鼠.上述结果表明,雌性动物室温条件下死亡后,若能短时间及时采集其体内GV期卵并体外成熟、体外受精,体外培养及胚胎移植技术,就有可能获得新生后代.     

中段尿在常温下保存时间对培养结果的影响研究

目的 探讨临床尿液标本不同放置时间对细菌培养结果的影响,明确尿液标本合理保存时间,加强对标本细菌培养前的保存时间控制.方法 随机选取114份患者尿液标本,比较其在新鲜尿液即刻培养结果和室温(22 ～25℃)的环境中各放置0.5、1、2、3、4和5h等不同时间点细菌培养结果的分离率差异,采用样本率与总体率(新鲜尿液培养的分离率)的差别检验(|u|和P)分析其是否具有统计学意义.结果 新鲜尿液即刻培养与放置0.5h培养结果分离率差异无统计学意义(38.6％);之后随放置时间延长分离率逐渐升高,2h接种标本分离率为43％,差异无统计意义(|u| =0.9489,P ＞0.05);放置3、4和5h后接种标本分离率分别为50.9％、55.3％和63.2％,呈显著增高(|u|值分别为2.627、3.586和5.446,P值均小于0.01).结论 在常温下不同的保存时间对培养结果有着重要影响,尤其在2h后影响更为显著.准确有价值的培养结果,与标本合理的存放时间有极大的关系,加强尿液培养前阶段的保存时间控制非常重要.     

含氢HTK液对大鼠供心保存中心肌细胞凋亡的影响

目的:研究合氢HTK液对供心保存过程中心肌细胞凋亡的影响.方法:SD大鼠随机分4组:对照组(保存液为HTK液)、H1组(保存液为含氢浓度约0.8mM的HTK液),H2组(含氢浓度约0.4mM的HTK液),H3组(含氢浓度约0.2mM的HTK液).采用Langendorff离体大鼠心脏灌注法,心脏分别在各组保存液中冷保存(4℃)6h,Tunel染色测定心肌细胞凋亡率,荧光定量实时PCR检测凋亡相关基因Bcl-2和Bax的mRNA表达,分光光度法检测心肌组织中Caspase-3活性.结果:供心冷保存(4℃)6h后,H1、H2、H3组凋亡指数明显低于对照组.H1、H2、H3组的Bcl-2mRNA表达水平显著高于对照组(P<0.01或P<0.05);对照组的Bax mRNA水平明显高于H1、H2、H3组(P<0.01或P<0.05),对照组caspase-3活性明显高于H1、H2、H3组(P<0.01).结论:舍氢HTK明显抑制供心保存时大鼠心脏心肌细胞凋亡.     

SOD对4℃保存人红细胞损伤的防护作用研究

目的：探讨SOD对延时保存红细胞损伤防护作用的机理。方法：取义务献血人员静脉血,置4℃冰箱保存。保存前后不同时期用标准方法分别检查各项指标,用生物荧光素标记红细胞,测定24h活存率。实验分3组：ACD和／或GMA、抗氧化剂（SOD）组。结果：SOD组保存75d,无氧糖酵解率维持在保存前的86．2％,ATP为56．4％,PK为64．3％,明显高于GMA或ACD组。保存75d整体回输后24h活存率为86．1％,与GMA保存42d(89.1%)结果接近。结论：用抗氧化剂保养液在4℃条件下保存人血红细胞从42d延长到75d,红细胞无氧糖酵解率、ATP、PK和整体回输后24h活存率等均与42d保存方相符,为红细胞的活存提供了理论根据。     

圈养林麝粪便类固醇激素保存时效性研究

用非损伤性采样方法对圈养林麝在三个条件:1)未处理,2)90%甲醇,3)0.9%抗生素,15个时间梯度下(20℃)的粪便中类固醇激素及其代谢物的保存时间及方法进行了比较,结果表明:林麝粪便在室温下(20℃),孕酮及其代谢物48 h有明显的上升趋势;雌二醇、睾酮和皮质醇及其代谢物在18 h均出现显著变化;对比抗生素保存,用甲醇保存是一种较好的保存林麝粪便类固醇激素的方法;然而不同激素用甲醇保存的时间不同,孕酮不宜超过1个月,皮质醇不宜超过半个月,睾酮则不宜超过10天,而雌二醇可以保存2个月之久.     

In vitro storage of unfertilized eggs of the Eurasian perch and its effect on egg viability rates and the occurrence of larval malformations

Ova ageing is the most important factor affecting fish egg quality after ovulation. Long-term storage of fish ova, using cryopreservation and vitrification techniques, has been unsuccessful to date. Instead, short-term in vitro ova storage has been used successfully and optimized in some cultured fish species. In vitro ova storage can drastically improve mass production of larvae and juveniles in the hatcheries by providing the possibility of the synchronous artificial fertilization for different females. To study how long unfertilized eggs of Eurasian perch (Perca fluviatilis L.) can retain their fertilizing ability after stripping, eggs were stored at temperatures of 4°C, 8°C and 12°C for 72 h post-stripping (HPS). The stored eggs of four female perch were separately fertilized at 0 h (i.e. control eggs fertilized before storage) and at 6-hour intervals during the experimental period of 72 h. The embryos reaching the eyed-egg and hatched-larvae stages, eyed-egg mortality and larval malformation rates were recorded as indices of egg quality. The results indicated that the maximum eyed eggs and hatched larvae (86% and 63%, respectively) were observed for eggs fertilized immediately after stripping, whereas the storage of the eggs at 4°C for 48 HPS decreased the eyed-egg and hatched-larvae rates to 46% and 17%, respectively. The use of a higher storage temperature resulted in a more rapid decrease in egg viability: eyed-egg and hatched-larvae rates of 23% and 9%, respectively, were obtained after 48 HPS storage at 8°C and 2% and 1% for eggs stored at 12°C. Eyed-egg mortality and larval malformation rates were not significantly affected by post-stripping ova ageing for at least up to 36 h. Thereafter, both values increased significantly and were measured to be the highest in the most aged ova. The present study demonstrated that stripped Eurasian perch eggs can be stored for at least 12 h at 4°C to 12°C without a significant reduction in their quality.     

Influence of short-term fasting during the luteal phase of the estrous cycle on ovarian follicular development during the ensuing proestrus of ewes

In order to study the effects of storage media and time of storage on the viability of unfertilized eggs of endangered Caspian brown trout (Salmo trutta caspius), the ova of this fish was stored in coelomic fluid and Cortland artificial media at 2–3 °C for 120 h. In this research, Cortland artificial medium was buffered with 20 mM of three different buffers: Hepes (C₈H₁₈N₂O₄S), Tris–HCl (C₄H₁₁NO₃–HCl) and sodium salt Hepes (C₈H₁₇N₂O₄SNa). The pH of these media were adjusted according to natural pH of coelomic fluid. The eggs that stored in these media fertilized at times 0 h (eggs fertilized prior to storage), 48, 72 and 120 h of post-stripping, using fresh and pooled sperm obtained from four to six males. According to the results of present study, time of storage showed a significant (p < 0.05) main effect on eyeing, hatching and eyed eggs mortality rates. Eyeing and hatching rates significantly (p < 0.05) decreased from 97.4 ± 2.1% and 95.1 ± 4.4% at time 0 (eggs fertilized prior to storage) to 77.9 ± 3% and 65.5 ± 5% after 120 h of storage. Within a similar period of time, eyed eggs mortality significantly (p < 0.05) increased from 2.4 ± 2.4% to 17.2 ± 3.9%. No significant (p > 0.05) main effect was found among media buffered with Tris–HCl (82.8 ± 3.2%, 73.4 ± 5.4%, 12.1 ± 4.5%), Hepes (88.2 ± 3.4%, 80.7 ± 5.5%, 9.3 ± 3.4%), sodium salt Hepes (77.8 ± 3.8%, 69.3 ± 5.7%, 12.2 ± 3.9%) and coelomic fluid (84.8 ± 3.8%, 77.7 ± 5.1%, 8.9 ± 2.7%) for eyeing, hatching and eyed eggs mortality rates. There was a negative correlation (r = −0.895, p < 0.001) between eyed eggs mortality and hatching rates. In conclusion, unfertilized eggs of endangered Caspian brown trout can be successfully stored for 48 h without significant loss of fertility. But, storage for 120 h results in the falling of hatching rate. In addition, no significant difference was found between viability rates of ova stored in coelomic fluid and artificial media, 120 h post-storage. It reveals that artificial media could be substituted for coelomic fluid as storage medium at least for 120 h in Caspian brown trout.     

人工扩繁代异色瓢虫卵和成虫最适冷藏条件的探讨

探讨了人工扩繁异色瓢虫时卵和成虫的最适冷藏条件。卵的最适保存温度是 1 0℃ ,冷藏 1 5d内孵化率平均在 60 %以上。成虫的最适保存温度是 1 0℃ ,如将刚羽化的成虫直接在该温度下冷藏 ,冷藏 40d时存活率能保持在 5 0 %左右 ;如将刚羽化成虫先在 1 5℃、0L∶2 4D条件下处理 1 6d,然后置于 1 0℃下冷藏 ,经冷藏 70d后存活率接近 1 0 0 % ,冷藏 90d后存活率仍高于 70 %。表明经一定条件下预处理后再冷藏成虫能保持较高的存活率和较长的存活期 ,可满足人工扩繁时对成虫的冷藏要求。     

光照时间和冷藏对桃小食心虫生长发育和繁殖的影响

研究了光照时间和冷藏对桃小食心虫各虫态生长发育和繁殖的影响.结果表明,随着光照时数的延长,各虫态历期基本呈缩短趋势,光照时间对幼虫滞育率影响最为明显,15 h滞育率最低,无论是光照延长还是缩短,幼虫滞育率都会明显增加.其它重要生命参数如单雌产卵量、幼虫体重、孵化率、脱出率和羽化率等最高点多集中在光照13～17 h,所以,桃小食心虫生长发育和繁殖的最佳光照时间为15 h左右.冷藏时间对蛹历期、成虫寿命和卵历期影响均不明显,但冷藏7 d后羽化率、单雌产卵量及孵化率均呈下降趋势,蛹和卵不易长时间低温存放.     

短时高温对加州新小绥螨发育的影响

为了探究高温逆境条件下加州新小绥螨的生存特性对其种群发育的影响,利用短时高温刺激试验,研究加州新小绥螨卵、幼螨、雌成螨在35、38、42、45 ℃等4个温度下、处理1～8 h后的孵化率、存活率、后期发育历期及繁殖的影响.结果表明: 处理温度越高,时间越长,卵和幼螨的存活率越低,其后续发育历期随处理温度升高和时间增加先缩短后延长.当温度为38 ℃,处理8 h后,卵的后续发育历期最短,为4.1 d.卵在45 ℃下处理2 h以上将不能正常孵化,而幼螨在45 ℃下处理8 h后不能存活;雌成螨产卵期和产卵量基本随温度升高先增加后降低,35 ℃处理8 h后,单雌产卵量最高,为38.9粒;38 ℃处理8 h后,单雌产卵量为36.7粒;45 ℃处理8 h后,单雌产卵量仅为14.5粒.短时高温主要影响加州新小绥螨的孵化率、存活率和后续发育历期,对雌成螨的产卵前期和存活率影响较小.     

短时高温对加州新小绥螨发育的影响

为了探究高温逆境条件下加州新小绥螨的生存特性对其种群发育的影响,利用短时高温刺激试验,研究加州新小绥螨卵、幼螨、雌成螨在35、38、42、45 ℃等4个温度下、处理1～8 h后的孵化率、存活率、后期发育历期及繁殖的影响.结果表明: 处理温度越高,时间越长,卵和幼螨的存活率越低,其后续发育历期随处理温度升高和时间增加先缩短后延长.当温度为38 ℃,处理8 h后,卵的后续发育历期最短,为4.1 d.卵在45 ℃下处理2 h以上将不能正常孵化,而幼螨在45 ℃下处理8 h后不能存活;雌成螨产卵期和产卵量基本随温度升高先增加后降低,35 ℃处理8 h后,单雌产卵量最高,为38.9粒;38 ℃处理8 h后,单雌产卵量为36.7粒;45 ℃处理8 h后,单雌产卵量仅为14.5粒.短时高温主要影响加州新小绥螨的孵化率、存活率和后续发育历期,对雌成螨的产卵前期和存活率影响较小.     

短时高温暴露处理对双尾新小绥螨Neoseiulus bicaudus Wainstein生长发育的影响

【目的】明确新疆本地种捕食螨双尾新小绥螨Neoseiulus bicaudus Wainstein在遇到短时极端高温胁迫后,对其生长发育和种群发展的影响。【方法】利用短时极端高温处理试验,研究双尾新小绥螨卵和成螨分别在38℃、42℃和46℃下,处理2、4和6 h后的孵化率、存活率以及对其未成熟阶段发育历期和生命参数的影响。【结果】卵经过不同时间高温处理后,随着处理温度的升高,处理时间的延长,卵的孵化率逐渐降低,46℃处理2 h的孵化率仅为42.02%,且在46℃处理超过4 h,其卵不能孵化;其各未成熟阶段发育历期有先缩短后延长的趋势,当在38℃处理2 h时,其发育历期最短为4.82 d。雌成螨经过不同时间高温处理后,雌成螨的产卵量、产卵期和寿命随着处理温度的升高、处理时间的延长有逐渐下降和缩短的趋势;42℃,2 h处理下每雌产卵量最低为19.33粒,其产卵期也是最短为10.09 d;38℃,6 h处理下寿命最短为14.68 d。【结论】短时极端高温处理主要影响双尾新小绥螨卵的孵化率、存活率降低和未成熟阶段的发育历期;影响其雌成螨的产卵量和寿命。     

黄粉甲低温贮存对管氏肿腿蜂发育和繁殖的影响

寄主的活动能力和营养状况是影响寄生蜂繁育效果的主要因素。以黄粉甲蛹为寄主,筛选其最佳贮存温度和时间,达到短时间内降低寄主活动能力的同时,保证其有充足营养满足寄生蜂发育所需,对于管氏肿腿蜂的规模化生产有重要意义。在9,3,-3,-9℃条件下,分别贮存黄粉甲TenebriomolitorL.蛹5,15,25,35和45d对管氏肿腿蜂Scleroderma guaniXiaoetWu发育、存活以及增殖的影响。结果表明,寄主活动能力随贮存温度的降低而减弱,有利于管氏肿腿蜂的寄生,寄生率和蜂种存活率不断升高;寄主的不同贮存对寄生蜂卵孵化率影响小于对幼虫存活率影响,而对蛹羽化率则无任何影响。管氏肿腿蜂各阶段历期与寄主贮存温度和时间有着一定关系,-3和-9℃条件下,寄主贮存时间越长,所接寄生蜂产卵前期越短;不同贮存温度和时间的寄主间,寄生蜂卵期、幼虫期和蛹期的持续时间存在一定差异,最短分别为3.39,6.27和15.15d,而对预蛹期无显著差异;幼期的变化趋势同产卵前期。寄主经不同温度和时间贮存后,所繁育的管氏肿腿蜂最高产卵量为33.19粒/雌、最高出蜂量为20.07头/雌、雌雄最高性比可达17.53∶1,分别是CK的2.21,3.17和1.26倍。总体而言,寄主经-9℃贮存15~25d后最有利于繁蜂,其中-9℃贮存15d出蜂量最高。单雌出蜂量较现有报道提高3倍。     

Effect of egg storage duration and egg turning during storage on egg quality and hatching of broiler hatching eggs

In commercial hatcheries, it is common to store eggs before incubation. One practice to improve hatchability consists in egg turning during this storage. This work aims to highlight the effects of turning on the physicochemical aspects of eggs and, consequently, how this turning can influence the hatching of chicks. An experiment was conducted to evaluate the effects of storage duration and egg turning during storage on egg quality, hatchability, and residual analysis. A total of 7 500 hatching eggs were collected from a 55-week-old commercial Cobb500 breeder flock and storage according to the treatments. The experiment was completely randomized in a 3 × 2 factorial design with three storage periods (4, 8, and 12 days) and egg turning (180° turn of eggs once a day) or no turning during storage, totaling six treatments. Regardless of turning, eggs stored for 4 days weighed more than turned eggs stored for 8 and 12 days, which were similar (P < 0.05). Non-turned eggs experienced an increase in relative shell weight with increased storage duration, and non-turned eggs stored for 4 and 8 days differed from non-turned eggs stored for 12 days (P < 0.05). Albumen pH of turned eggs stored for 4 and 8 days was lower than that of non-turned eggs stored for the same durations (P < 0.05). Albumen pH of turned eggs increased as storage duration increased (P < 0.05). Egg turning increased hatching by 2.02% over that of non-turning (P < 0.05). Eggs stored for 12 days, irrespective of turning, had higher late embryonic mortality (P < 0.05) compared to the other treatments. It was concluded that turning eggs during pre-incubation storage was adequate to improve hatchability of fertile eggs. Storing fertile eggs for 12 days is harmful to egg quality and increases embryo mortality even if eggs were turned.     

Fertilization in landlocked sea lamprey: storage of gametes, optimal sperm: egg ratio, and methods of assessing fertilization success

Ovulated, unfertilized eggs of sea lamprey Petromyzon marinus could be stored for 1 day at 15° C without significant loss of fertilizing ability. After 2 days storage most eggs could still be fertilized. Lamprey semen could be stored up to 1 day. Thereafter, a decrease in sperm fertilizing ability occurred, accompanied with a decrease in sperm motility. Unlike teleost fish, sea lamprey eggs could still be fertilized after 1 h contact with water. This extended time of gamete fertility after release into water may help to account for the reproductive success of this species. Maximal fertilization rates were obtained at a sperm: egg ratio of 50 000, a ratio recommended for studies on fertility of individual males. Assessing fertilization success 3 min after fertilization (at cytoplasmic bleb stage) or 5 h after fertilization (at two–cell embryo) was strongly correlated ( r =0·92 and 0·98) with estimation and fertilization success at hatching. These results offer improvement in artificial fertilization techniques under laboratory conditions and provide new information on the biology of fertilization in sea lamprey.