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1.
细粒棘球绦虫——原头蚴在两种细胞培养液中体外培养的初步观察 总被引：1，自引：0，他引：1
通过原头蚴在两种细胞培养液中（RPMI-1640和MEM）培养,观察其存活、生长及发育情况,从而为研究寄生虫发育提供最基本的数据资料。方法：细粒棘球绦虫幼虫——原头蚴培养的细胞培养基（RPMI-1640和MEM）分为4组：Ⅰ组为纯RPMI-1640培养液;Ⅱ组为含10％的小牛血清的1640培养液;Ⅲ组为纯MEM培养液;Ⅳ组为含10％的小牛血清的MEM培养液;并进行对比观察。结果：培养15d的原头蚴的成活率分别为：Ⅰ组69．87％、Ⅱ组80．35％、Ⅲ组50．25％、Ⅳ组60．32％;成囊率分别为：Ⅰ组80．58％、Ⅱ组90．25％、Ⅲ组35．65％、Ⅳ组45．89％;头节外翻率分别为：Ⅰ组95．50％、Ⅱ组98．65％、Ⅲ组60．52％、Ⅳ组70．98％。结论：大多数虫体在早期向囊发育,一部分虫体头节外翻,并伴有规律的伸缩运动,但随时间的延长虫体运动减缓,又向囊蚴发育。通过对细粒棘球绦虫的原头蚴的体外培养,初步表明含有10％小牛血清的细胞培养基RPMI-1640较适合原头蚴的生长发育,在普通培养箱中培养,最适温度在37—40℃,培养液pH=7．2,初步建立了原头蚴体外培养的方法。 相似文献
2.
牛附红细胞体体外培养试验 总被引：31，自引：1，他引：30
用RPMI－1640、M－199、D－MEM3种培养液作为基础培养基，再分别按体积分数加入40％的犊牛血清，采用普通恒温培养箱（37℃）进行牛附红细胞体体外培养。结果表明，牛附红细胞体在RPMI－1640培养基中，每12h更换1次培养液，并补充适量正常红细胞，获得了高达99％的感染率；连续传代培养可达44代以上。 相似文献
3.
以无血清RPMI-1640培养液和DMEM培养液分别灌洗小鼠腹腔，10min后分别吸出灌洗液于100mL／L胎牛血清和DMEM培养液中培养．巨噬细胞吞噬试验检测其活性、台盼蓝测定体外培养细胞的存活率和成层率。结果表明．DMEM培养基体外培养的巨噬细胞存活率和成层率高、吞噬能力强，与RP—MI-1640相比，DMEM可作为一种简单而实用的体外分离培养巨噬细胞的培养基。 相似文献
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5.
将隔离饲养至35日龄麻鸭90只分为3组，每只鸭分别感染包氏毛毕吸尾虫蚴70条，待粪检呈阳性后Ⅰ组和Ⅱ组分别按每只鸭投喂吡喹酮25mg和丙硫苯咪唑30mg,隔日投药1次，Ⅲ组为感染不投药对照组，结果：Ⅰ组鸭转阴率为96．67％；Ⅱ组鸭转阴率为73．33％，组间差异显著（P＜0．05）；Ⅰ组和Ⅱ组鸭从肝门静脉和肠系膜静脉分别检出崩解虫体10条和237条。 相似文献
6.
“健兔灵”对仔兔驱虫增重效果的试验观察 总被引：3，自引：0，他引：3
将８０只６０～７０日龄的仔兔分成Ⅰ、Ⅱ、Ⅲ组,分别饲喂“健兔灵”中药添加剂、投服百球清和复方新诺明,并设对照组进行３０ｄ的驱虫、增重的比较试验。结果试验Ⅰ、Ⅱ、Ⅲ组的球虫转阴率分别为１０％、６０％、０％,球虫卵减少率分别为９７．５％、９８．５％、５３．６％;对照组转阴率和减少率分别为０和－８．５％。经χ２检验,试验Ⅰ、Ⅱ组之间差异不显著（Ｐ＞０．０５）,试验Ⅰ、Ⅱ组与试验Ⅲ组之间差异均极显著（Ｐ＜０．０１）。增重效果表明,试验Ⅰ、Ⅱ、Ⅲ组之间差异均不显著（Ｐ＞０．０５）,试验Ⅰ、Ⅱ、Ⅲ组与对照组之间差异均显著（Ｐ＜０．０５）。 相似文献
7.
雏鸡接种新城疫Ⅱ系苗后,细胞免疫功能表现为首免后有增强趋势,二免后升到极显著水平。采用微量全血培养~3H-胸腺嘧啶核苷(~3H-TdR)掺入法,以植物血凝素(PHA)为刺激原,确定鸡淋巴细胞转化试验的较佳测定条件为:用含5％犊牛血清的RPMI-1640培养液2ml,血量为25μl,刺激管PHA为0.4ml,置41.5℃培养36小时,并在培养结束前18小时加入~3H-TdR。 相似文献
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9.
《畜牧兽医学报》2017,(8)
旨在探索细粒棘球绦虫原头蚴蛋白表达特性,揭示在此发育阶段虫体主要的功能蛋白以及信号通路,为揭示虫体的发育调控、筛选疫苗以及药物靶标奠定基础。本试验采用LC-MS/MS技术对虫体原头蚴表达的蛋白质进行研究,然后对获得数据进行GO、KOG以及KEGG等分析;最后采用qRT-PCR和原位杂交技术对鉴定得到的Wnt信号通路的关键基因β-catenin进行了差异表达和组织定位研究。结果显示,本研究共鉴定得到7 172个肽段,1 197个蛋白质,其中包括多个潜在的抗原蛋白质,如四跨膜蛋白超家族、6-磷酸葡萄糖酸脱氢酶、表皮抗原蛋白和烯醇酶等。信号通路分析结果显示其中632个蛋白参与276个调控通路,诸如与虫体发育紧密相关的Wnt、Notch、Hedgehog、NF-κB、cAMP以及胆酸盐信号通路。对β-catenin的验证结果显示,此基因分布于细粒棘球绦虫原头蚴顶突的小刺以及散在的细胞中,同时其相对转录量在体外培养0~10d的过程中呈递减趋势。综上,本研究解析了细粒棘球绦虫原头蚴的蛋白质表达元件,揭示了原头蚴主要的调控信号通路,为虫体的生长发育机制以及包虫病的有效防控提供了理论依据。 相似文献
10.
为筛选适宜MDCK细胞生长的最佳血清，分析6个批次新生牛血清在两种培养方式即贴壁静置培养条件下MDCK细胞的生长状态、平均集落形成率及微载体悬浮培养条件下MDCK细胞的生长状态、生长速率、生长动力学情况。结果表明6组新生牛血清贴壁静置培养的MDCK细胞，平均集落形成率最大的为第Ⅴ组，最小的为第Ⅰ组，由大到小依次为第Ⅴ、Ⅵ、Ⅲ、Ⅱ、Ⅳ、Ⅰ组；微载体悬浮培养条件下的MDCK细胞，最大增殖密度从大到小依次为第Ⅳ、Ⅲ、Ⅵ、Ⅴ、Ⅱ、Ⅰ组，平均倍增时间由大到小依次为第Ⅵ、Ⅴ、Ⅲ、Ⅱ、Ⅳ、Ⅰ组。因此，通过对不同批次血清贴壁静置培养的MDCK细胞生长状态、平均集落形成率及微载体悬浮培养的MDCK细胞生长状态、生长动力学评价，筛选出适宜细胞生长的新生牛血清批次为第Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ组，而第Ⅰ组新生牛血清对细胞生长的促进效果不明显，细胞生长状态不佳。 相似文献
11.
不同培养液及血清浓度对绵羊孤雌生殖胚胎发育的影响 总被引：1，自引：0，他引：1
体外成熟的绵羊卵母细胞，经5 μmol/L A23187 5 min和2 mmol/L 6-DMAP 4 h激活后，分别在SOFaa、M-199两种培养液中进行培养，试验比较了SOFaa、M-199液分别添加BSA和不同浓度（10%、20%）胎牛血清对绵羊孤雌生殖胚体外发育的影响。M-199+FCS组的卵裂率显著低于SOFaa+BSA和SOFaa+FCS组（P<0.05），SOFaa+BSA组的囊胚率显著低于M-199+BSA、SOFaa+FCS、M-199+FCS三组（P<0.05）；SOFaa+10% FCS与SOFaa+20% FCS组的卵裂率及囊胚率均无显著差异，M-199+10% FCS和M-199+20% FCS组的卵裂率及囊胚率间均无显著差异。结果表明：SOFaa比M-199更适合绵羊孤雌生殖胚体外发育，此外添加10%的FCS即可达到较好的培养效果。 相似文献
12.
屠宰绵羊卵巢卵母细胞的体外培养 总被引：2，自引：0，他引：2
为了使屠宰绵羊卵巢卵母细胞能够用于体外受精,本文着重探索了使卵巢卵母细胞体外培养成熟的方法和条件。实验结果表明,以TCM-199加10％FCS作为基本培养基,培养24～25小时,可以使绵羊卵巢卵母细胞培养成熟,其成熟率可达55.5％(435/784)。如果在培养液内添加hCG(0.02mg/ml)或LH(0.01mg/ml),并且尽可能保持卵丘细胞的完整,则可以使成熟率提高到76.9％(140/182)～82.9％(112/135)。 相似文献
13.
Mbao V Berkvens D Dolan T Speybroeck N Brandt J Dorny P Van den Bossche P Marcotty T 《The Onderstepoort journal of veterinary research》2006,73(3):207-213
Theileria parva sporozoite stabilates are used for immunizing cattle against East Coast fever and in in vitro sporozoite neutralization assays. In this study, we attempted to identify a cheaper freezing medium and quantified the infectivity loss of sporozoites due to refreezing of stabilates, using an in vitro technique. Pools of stabilates prepared using Minimum Essential Medium (MEM), Roswell Park Memorial Institute (RPMI 1640), foetal calf serum (FCS) and phosphate-buffered saline (PBS) were compared. All were supplemented with bovine serum albumin except the FCS. RPMI 1640 was as effective as MEM in maintaining sporozoite infectivity while the infectivity in PBS and FCS reached only 59% and 67%, respectively. In a second experiment, a stabiiate based on MEM was subjected to several freeze-thaw cycles including various holding times on ice between thawing and refreezing. Refrozen stabilate gave an average sporozoite infectivity loss of 35% per cycle. The results indicate that RPMI can be used as a cheaper freezing medium for T. parva stabilates and that refrozen stabilate doses need to be adjusted for the 35% loss of infectivity. 相似文献
14.
An E-rosetting reaction is described which gave 92.1%±2.4 (mean±S.D.) E-rosettes with bovine fetal thymocytes and 48.2%±8.4 with with bovine peripheral blood leukocyte (PBL) preparations. Both culture conditions and culture medium were critical factors in obtaining maximal and reproducible E-rosette numbers. Optimum rosette formation occurred when bovine PBL and neuraminidase treated sheep erythrocytes (nSRBC) were reacted in L-15 culture medium supplemented with 10% fetal calf serum (FCS). Other media including 100% FCS, MEM with 10% FCS, and RPMI-1640 with 10% FCS were less satisfactory. Cultural conditions found to be optimal for enumeration of bovine E-rosettes are similar to those reported as optimal for detection of human T cells. The specificity of rosette formation by bovine thymus derived (T) lymphocytes was shown by demonstration of (1) rosettes and surface membrane immunoglobulins (mIg) on different cells in PBL, (2) rosette formation by the majority of fetal thymocytes, and (3) no inhibition of rosette formation by anti-immunoglobulin serum. Using the E-rosette and mIg assays for presumptive bovine T and B lymphocytes, respectively, it was possible to differentiate from 57.5 to 90% (75.2%±9.3) of cells in bovine PBL preparations, and from 90.2 to 97.5% (94.2%±2.1) of cells in bovine fetal thymocyte preparations into T and B cells. 相似文献
15.
Effect of serum-free co-culture and synchrony of recipients on development of cultured sheep embryos to fetuses 总被引：1，自引：0，他引：1
The percentage of sheep embryos that continued to develop after collection and immediate transfer on d 2 after estrus was similar when phosphate-buffered saline with 10% fetal calf serum (PBSFCS, 45%), physiological saline (50%), or tissue culture medium 199 supplemented with 10% fetal calf serum (M199FCS, 47%) was used to flush embryos from oviducts. Co-culture of sheep embryos for 3 d with oviductal cells tended (P = .1) to reduce the percentage of embryos that developed to fetuses after transfer compared with those embryos transferred immediately. Tissue culture medium 199 supplemented with .3% BSA (M199BSA) was an adequate substitute for M199FCS for culture of sheep oviductal cells if tissue culture wells were pretreated with fibronectin. Estradiol in concentrations from 10 to 1,000 pg/ml and progesterone at concentrations of 1 or 10 ng/ml in M199BSA failed to stimulate embryo development during 3 d of co-culture beyond that seen in co-culture with M199FCS or M199BSA without added steroid. Transfer of sheep embyros co-cultured for 3 d in M199BSA or M199FCS to recipients synchronized with donors resulted in about 19% of the embryos developing to fetuses, whereas transfer to recipients that were in estrus 24 h after donors resulted in 33% of embryos developing to fetuses. The significant (P less than .05) improvement for delayed recipients may reflect the relatively lesser developmental rate of co-cultured embryos compared with that of embryos in vivo. Embryo development into fetuses was similar after co-culture in M199FCS or M199BSA co-cultures; therefore, serum is not required for the co-culture of sheep embryos. 相似文献
16.
Co-culture of ovine ova with oviductal cells in medium 199 总被引：7，自引：0，他引：7
Three experiments were conducted to test the suitability of medium 199 supplemented with 10% fetal calf serum (M199FCS) as a medium for co-culture of one-cell sheep ova with sheep oviductal cells. In Exp. 1, ova were co-cultured for 5 d in 5 ml of M199FCS or in Ham's F10 medium supplemented with 10% fetal calf serum (F10FCS). Co-culture did not increase the number of cleavages at the end of 5 d of culture, but M199FCS supported more cleavages than did F10FCS (P = .016). In Exp. 2, ova were cultured for 1 to 3 d in M199FCS alone or on oviductal, uterine or kidney cell monolayers from ewes 2 d postestrus and transferred to recipients from which they were recovered at 8 d postestrus. Co-culture with oviductal cells improved (P less than .001) the cleavage index of recovered embryos compared with culture in medium alone or co-culture with other cell types. In Exp. 3, monolayers of oviductal cells from ewes 2 d postestrus and from luteal-phase ewes were cultured as in Exp. 2. No difference was observed between the two sources of oviductal cells for their ability to support in vitro development of one-cell sheep eggs for 1 or 2 d. These studies suggest that M199FCS may be a good medium to use in an oviductal cell co-culture system for one-cell sheep ova. Results further suggest that specific secretions of oviductal cells may be important for early embryo development in vivo. 相似文献
17.
Yotsushima K Sakaguchi M Shimizu M Okimura T Izaike Y 《The Journal of reproduction and development》2004,50(4):471-476
The objective of this study was to investigate the influence of fatty acid-free bovine serum albumin (BSA) or fetal calf serum (FCS) on the re-expansion of biopsied blastocysts and post-warm viability of subsequently vitrified embryos. Firstly, blastocysts produced in vitro were biopsied at Day 7 and cultured to allow repair in TCM199 with 0.3% BSA or 5% FCS for 24 h. The re-expansion rates and mean total numbers of cells of the re-expanded embryos after the repair culture with BSA were almost the same as that with FCS. Secondly, after biopsied embryos were similarly cultured for repair with BSA or FCS, re-expanded embryos were selected for vitrification. After warming and exposure to 0.5 M sucrose with 20% FCS in mPBS, the embryos were cultured in TCM199 with 5% FCS for 24 h. The re-expansion rate and mean total number of cells in re-expanded blastocysts in the BSA treatment group (97.4 +/- 2.9% and 106 +/- 42) was significantly higher than that in the FCS treatment group (51.6 +/- 9.1% and 61 +/- 38), respectively (P<0.05 and P<0.01). In conclusion, both FCS and BSA supplementation can be useful for repairing cultures of bovine biopsied blastocysts; but, compared with BSA supplementation, FCS supplementation during repair culture reduces the post-warm viability of biopsied and subsequently vitrified embryos. 相似文献
18.
试验研究了清和促性腺激素对山羊卵丘扩展和卵母细胞核成熟的,卵母细胞与卵丘扩展的关系以及卵丘扩展与卵母细胞核成熟的关系。结果表明：（1）培养24h，添加PMSG组卵母细胞核成熟率显著高于不加激素组（P＜0．05），而培养到27h，2者成熟率之间差异变得不显著，说明添加PMSG卵母细胞核的最终成熟率没有明显影响，但加速了卵母细胞核的成熟进程；（2）M199＋BSA培养27h，卵母细胞核成熟率显著高于培养24h，而M199＋FCS培养27h与培养24h的成熟差异不显著，说明添加BSA时，卵母细胞核体外成熟速度比添加FCS的慢；（3）卵丘扩展良好与扩展不好的卵母细胞核成熟率以及第1极体形态无统计学差异，说明山羊卵母细胞的核成熟可能不依赖于卵丘扩展；（4）山羊的卵丘扩展产不依赖于卵母细胞；（5）将带壁颗粒细胞与不带壁颗粒细胞的COC分开培养发现，2者卵母细胞核成熟度无明显差异，带有壁颗粒细胞的COC卵丘扩展情况明显优于一般COC。 相似文献
19.
The study was carried out from July 2007 to June 2008 in Wolaita Sodo Abattoir to assess the status of hydatidosis in cattle.
Routine meat inspection, hydatid cyst count and characterization were conducted. Out of 400 cattle slaughtered in Wolaita
Sodo Abattoir 64 (16%) animals were found harboring hydatid cysts. Thorough meat inspection in the abattoir revealed that
74 visceral organ were found harboring one or more hydatid cysts. The infection of the lung, liver, spleen and kidney were
found to be 45.94% 45.94%, 6.75% and 1.35% respectively. From the total of 283 hydatid cysts counted 153(54.06%), 17(6.00%),
5(1.76%), 108(38.16%) were found to be small, medium, large and calcified cysts respectively and 170(60.28%), 5(1.76%) and
108(38.16%) were sterile, fertile and calcified cysts respectively. The rate of cyst calcification was higher in the liver
than in the lung while fertility rate was higher among the cysts of the lung. Hydatid cyst viability rate of 40% was observed. 相似文献