猪淋巴细胞转化增殖试验反应的最佳条件探讨

为了确定猪淋巴细胞转化增殖试验反应的最佳条件,研究以东北民猪为试验猪,采用MTT法研究不同浓度犊牛血清(FCS)、淋巴细胞、植物血凝素P(PHA-P)对猪淋巴细胞增殖的影响。结果表明:当培养时间为60 h、犊牛血清浓度为10%、淋巴细胞浓度为2×106个/m L、PHA-P刺激的最佳浓度为12.5μg/m L时,可显著刺激T淋巴细胞的增殖。     

MTT法检测番鸭外周血淋巴细胞转化功能的研究

为了建立番鸭淋巴细胞转化检测的MTT法 ,筛选细胞浓度、刀豆蛋白ConA浓度和培养时间 3个参数对试验条件进行了研究。结果确定了MTT法检测番鸭淋巴细胞转化能力的最佳培养条件 ,细胞浓度 5× 1 0 6个 /mL在 2 0 μg/mLConA的RMPI 1 640完全培养基 40℃培养 60h。试验表明 ,该方法可用于番鸭体外细胞免疫功能的检测     

鱼类外周血淋巴细胞的分离和体外培养

为了研究细胞因子在鱼类免疫系统中的重要作用,试验应用不同浓度的淋巴细胞分离液分别对鲤鱼、草鱼、鲫鱼外周血淋巴细胞进行分离.结果发现:浓度为1.085g/mL的淋巴细胞分离液可将鱼类外周血中淋巴细胞有效地分离出来,并且鲫鱼的淋巴细胞分离效果比鲤鱼好,草鱼次之;同时采用浓度为30,50,100 μg/mL的刀豆蛋白(Con...     

应用CCK8法检测鸡淋巴细胞活性的检测最佳条件研究

为了研究应用CCK-8法检测鸡淋巴细胞增殖的体外培养最佳条件,试验设置了不同种板细胞数、血清浓度、培养时间、孵育时间和检测波长等条件对其进行比较研究。结果表明:1 750 r/min转速的改良鸡外周血淋巴细胞分离方法效果最佳;在体外增殖培养试验中,CCK-8加样4 h后在450 nm波长处检测OD值最理想,培养时间以44 h为最佳;在细胞数量一致的情况下,胎牛血清浓度为5%或者为10%差异不显著(P0.05)。当细胞悬液浓度为1×10~6个/m L时,加入100,50,25,12.5μL不同细胞数,4组间比较均差异显著(P0.05),并且随着细胞悬液浓度的增加OD值增加;最佳铺板细胞的细胞悬液浓度为2.0×10~6~3.0×10~6个/m L,取50~100μL为宜,即最佳细胞铺板数量控制在每孔1.5×10~5~2.0×10~5个/100μL。说明应用CCK-8法检测鸡外周血淋巴细胞活性的最佳条件除种板细胞数不同外,在检测波长、血清浓度、加样后孵化时间等方面与其他动物试验差别不大。     

硫酸化酵母葡聚糖对鸡淋巴细胞的影响

为研究硫酸化酵母葡聚糖(SCG)对鸡细胞免疫的影响,将7种浓度SCG单独或与Con A、LPS加入到鸡脾脏淋巴细胞的培养体系中,培养48 h时用MTT法测定淋巴细胞增殖的变化;将5种浓度的SCG分别与Con A加入到鸡脾淋巴细胞的培养体系中,培养24 h后收集细胞培养上清液,测定IL-2和IFN-γ的含量。结果显示,SCG浓度在在250~2 000μg/m L时,SCG+Con A、SCG各剂量组随着多糖浓度的增加其A570值先增加后降低(P0.05);SCG+LPS各剂量组的A570随着多糖浓度的增加而降低,2 000μg/m L剂量组显著低于其他三组(P0.05);均具有显著的量效关系。SCG浓度在62.25~1 000μg/m L时,多糖组脾脏淋巴细胞体外培养上清液中IL-2与IFN-γ浓度,随着多糖浓度的增加其促进淋巴细胞分泌细胞因子的能力先增加后降低,具有明显量效关系;且均在250μg/m L时效果最好。结果表明,SCG在合适剂量能单独或协同Con A、LPS刺激鸡T、B淋巴细胞增殖,促进淋巴细胞分泌IL-2和IFN-γ,从而增强鸡的细胞免疫。     

中药复方多糖对TLR4抗体阻断的不同MHCB-LβⅡ基因型鸡淋巴细胞中相关因子的影响

为探究中药复方多糖对不同MHC B-LβII基因型鸡的免疫调节作用受体的影响,采用PCR-SSCP方法将黄麻肉鸡按不同MHC B-LβII基因型分组,于28日龄在各组随机取5只试验鸡,采集外周血,分离淋巴细胞,与终浓度为0.5、1、2μg/m L的TLR4抗体共培养1 h后,分别加入终浓度为200、100、50、25、0μg/m L的中药复方多糖共培养24 h,采用ELISA法检测不同MHC B-LβII基因型鸡外周血淋巴细胞培养上清液中IL-6、TNF-α和NF-κB的质量浓度。结果表明:加入TLR4抗体后,中药复方多糖促进淋巴细胞分泌IL-6、TNF-α和NF-κB的能力受到抑制,并且随着TLR4抗体浓度的增加,抑制作用增强。试验表明TLR4是中药复方多糖的免疫调节作用受体之一,中药复方多糖可能通过与淋巴细胞表面TLR4结合,经TLR4/NF-κB信号通路活化转录因子NF-κB,促进IL-6和TNF-α的分泌发挥其免疫调节作用。     

杂交水牛核型分析以及淋巴细胞分裂周期的研究

水牛分河流型和沼泽型两大类,两者的染色体数目不同,杂交后代之间杂交会产生3种染色体核型,不同染色体核型的杂交水牛其繁殖性能存在差异,为筛选具有偶数倍染色体数的水牛,提高群体的繁殖水平,通常要对水牛的染色体进行核型分析。实际培养中发现水牛的淋巴细胞培养有其特殊性,本研究以人类外周血淋巴细胞培养分析核型法为基础,通过改良和优化该方法以期用于杂交水牛。研究结果表明,犊牛和成年母牛的淋巴细胞在培养过程中添加100U/m L和50U/m L的肝素,淋巴细胞培养周期分别为72h和90h,秋水仙素处理浓度均为0.06μg/m L的条件下,做核型分析才可各自获得较多目数的良好分裂相。改良的外周血淋巴细胞培养分析核型法可以用于杂交水牛的核型分析,效果良好。     

酵母多糖对鲤鱼脾脏淋巴细胞分泌功能的影响

为了研究酵母多糖对鲤鱼脾脏淋巴细胞的分泌功能是否有影响,试验用淋巴细胞分离液从鲤鱼脾脏中分离淋巴细胞,向淋巴细胞悬液中加入刀豆蛋白A,然后加入不同浓度的酵母多糖,培养48 h后,对酵母多糖是否影响淋巴细胞分泌白细胞介素-2(IL-2)和γ干扰素(IFN-γ)进行了研究。结果表明:50 mg/L酵母多糖组与对照组相比,IL-2和IFN-γ的分泌差异不显著(P 0. 05);500 mg/L酵母多糖和100 mg/L酵母多糖组与对照组相比,差异极显著(P0. 01)。说明添加合适浓度的酵母多糖,可以促进体外培养的鲤鱼脾脏淋巴细胞分泌IL-2和IFN-γ。     

天门冬多糖对小鼠脾脏淋巴细胞体外增殖的影响

本试验旨在研究天门冬多糖对小鼠脾脏淋巴细胞体外增殖的影响。从正常小鼠分离脾脏淋巴细胞体外培养,在细胞培养体系中分别加入终浓度为50、100、200、400 mg/L天门冬多糖或配合刀豆素(5 mg/L)作用24、487、2 h,以MTT法测定细胞的增殖程度。结果表明,天门冬多糖可显著促进体外培养的小鼠脾脏淋巴细胞增殖,并与ConA有协同作用。     

不同刺激剂PHA和ConA对绒山羊淋巴细胞有丝分裂中期化的影响

试验旨在探索使较高比例的淋巴细胞富集在有丝分裂G2/M期的最佳条件。运用植物血凝素(PHA)和刀豆蛋白A(ConA)对淋巴细胞进行刺激使其增殖,培养一定时间后加秋水仙素对淋巴细胞进行同步化处理,用流式细胞仪检测G2/M期的细胞数量进行比对,观察试剂的最佳作用浓度和加秋水仙素的最佳时间。结果表明,采用PHA和ConA刺激淋巴细胞增殖的最佳作用浓度是60和5 μg/mL,且淋巴细胞经ConA刺激培养45 h再加秋水仙素处理5 h G2期的比例最高为58.38%。结果提示,就绒山羊的淋巴细胞来说,ConA刺激绒山羊淋巴细胞增殖的效果比PHA好,且摸索出细胞G2/M期的时间点至关重要。     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ

An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein     

Avian brood parasitism — a growing research area in behavioral ecology (part Ⅱ)

正We are pleased to publish the second special issue on avian brood parasitism and to be responsible guest editors for the two special issues of Chinese Birds (Vol. 3, No. 4, 2012 and Vol. 4, No. 1, 2013), entitled "Avian Brood Parasitism - A Growing Research Area in Behavioral Ecology". The first issue was published in December 2012. The goal of the two special issues is to publish accumulated knowledge and some of the recent developments in     

Internal Parasites and Haematological Values in Cattle Slaughtered in Buea Subdivision of Cameroon

Tropical Animal Health and Production -