革胡子鲶血液指标及血细胞发生的观察

测定了革胡子鲶(Clarias leather)的血液指标,红细胞密度为(29.04±2.12)×106个/mm3,白细胞密度为(1.21±0.85)×104个/mm3,血红蛋白含量为(7.64±0.69)g/L,红细胞渗透脆性为(0.46±0.03)%,红细胞比积为(36.69±0.99)%,红细胞沉降率为(3.40±0.56)mm/h。对革胡子鲶外周血进行涂片观察,在涂片上可区分出红细胞、血栓细胞、淋巴细胞、嗜中性粒细胞和单核细胞,但没发现嗜酸性粒细胞和嗜碱性粒细胞。其中,血栓细胞、大淋巴细胞、小淋巴细胞、嗜中性粒细胞和单核细胞在白细胞中所占的比例分别为(3.43±0.74)%、(4.36±0.65)%、(60.09±1.35)%、(16.62±1.29)%、(14.60±0.79)%。另外,在涂片上还发现了未成熟的红细胞。对肝脏、脾脏、头肾和肾脏4种造血组织中血细胞的发生过程进行了观察和测定。观察发现,革胡子鲶的头肾、肾脏和脾脏可以产生各种类型的血细胞,是主要的造血器官。     

外源性IL-6与TNF-α对乳腺炎奶牛乳汁中性粒细胞凋亡的影响

为了明确外源性白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)对奶牛乳腺炎乳汁中性粒细胞凋亡的影响,试验分离乳腺炎奶牛乳汁的中性粒细胞并进行体外培养,以不同浓度的外源性IL-6(0.05 ng/m L、0.1 ng/m L、0.2 ng/m L、0.5 ng/m L)与TNF-α(2 ng/m L、5 ng/m L、10 ng/m L、20 ng/m L)以及IL-6和TNF-α联合液(1∶100,1∶50,100∶1,50∶1,1∶1)对其进行作用,在不同时间点(6,12,24,48,72小时)收集细胞,应用流式细胞仪检测细胞凋亡的情况。结果表明:外源性IL-6对乳腺炎奶牛乳汁中性粒细胞凋亡产生抑制作用;TNF-α在6~48 h范围内对乳腺炎奶牛乳汁中性粒细胞凋亡产生促进作用;IL-6与TNF-α按不同比例混合后共同作用于体外培养的奶牛乳汁中性粒细胞后,IL-6占比多时以抑制细胞凋亡为主,TNF-α占比多时以促进细胞凋亡为主。说明外源性IL-6和TNF-α能影响乳腺炎奶牛乳汁中性粒细胞的凋亡,能为奶牛乳腺炎的预防和治疗提供试验依据。     

金黄色葡萄球菌体外刺激奶牛外周血中性粒细胞对炎性因子分泌的影响

以体外培养的奶牛中性粒细胞为研究对象,用瑞氏姬姆萨染色和台盼蓝拒染法对分离培养的奶牛外周血中性粒细胞进行鉴定和活性检测,ELISA检测金黄色葡萄球菌感染体外培养的奶牛外周血中性粒细胞IL-1β、TNF-α、IL-10及IL-8的分泌情况,Western blot检测金黄色葡萄球菌感染体外培养的奶牛外周血中性粒细胞中NF-κB的激活效应。结果显示,体外分离培养的奶牛外周血中性粒细胞纯度达99%,随着培养时间增加奶牛外周血中性粒细胞活性逐渐降低。ELISA检测结果显示,在3 h～24 h刺激时间范围内,随着金黄色葡萄球菌刺激时间的增加,与空白对照组相比体外培养的奶牛外周血中性粒细胞中IL-10、TNF-α、IL-1β和IL-8的分泌量均显著升高(P<0.001)。Western blot检测结果显示,在15 min～120 min刺激时间范围内,随着SA113感染时间的增加,与空白对照组相比体外培养的奶牛外周血中性粒细胞中IκBα磷酸化水平显著升高(P<0.001)。研究结果表明,成功分离培养了奶牛外周血中性粒细胞;金黄色葡萄球菌(SA113株)刺激体外分离培养的奶牛外周血中性...     

小鼠睾丸间质细胞的分离纯化与体外培养的研究

为了寻求一种快速、高效的体外分离、纯化小鼠睾丸间质细胞的方法,试验采用两种酶(胶原酶Ⅳ和0.25%胰蛋白酶)消化法和6个梯度(70%、65%、60%、55%、50%、45%)的Percoll液对小鼠睾丸间质细胞进行分离、纯化,对分离的细胞进行细胞学观察、台盼蓝染色、3β-羟基类固醇脱氢酶(3β-HSD)染色,采用物理方法、胶原酶Ⅳ+0.25%胰蛋白酶和柠檬酸钠+KCl对睾丸间质细胞进行消化传代培养。结果表明:胶原酶Ⅳ和0.25%胰蛋白酶限时消化能够获得较高活率(90%)的小鼠睾丸间质细胞;经Percoll细胞分离液分离的细胞纯度高达86%,且在34℃、5%CO2条件下培养的细胞生长良好。     

长春地区猫的血象、骨髓象及细胞化学正常值

对16只雄性家猫进行了血液学检测,结果表明,RBC为7.3×10~(12)/L,WBC13.89×10~9/L,Hb105g/L,PCV42.5%,MCV55.187fL;骨髓中,中性杆状核粒细胞为21.82%,晚幼红细胞20.1%,粒红比1.82,骨髓有核细胞数9.1×10~9/L;末梢血及骨髓中的粒细胞PAS均为阳性,铁粒幼红细胞为Ⅰ型或Ⅱ型,末梢血中NAP阴性,嗜酸粒细胞POX为阴性.     

脂联素对酮病奶牛中性粒细胞自噬相关基因mRNA表达的影响

通过模拟向高BHBA的中性粒细胞培养液中添加不同质量浓度的脂联素(ADPN),运用荧光定量PCR技术,分别检测中性粒细胞自噬相关基因mRNA的表达水平。结果显示:添加高BHBA(4.8mol/L)的中性粒细胞培养6h后,与对照组相比,自噬相关基因LC3B、Beclin、ULK1的mRNA表达水平显著升高(P<0.01),SQSTM1(p62)、mTOR的mRNA表达水平显著下降(P<0.01);添加不同质量浓度的ADPN(25,50,100μg/L)后,相对于高BHBA组,中性粒细胞自噬相关基因LC3B、Beclin、ULK1的mRNA表达水平显著下调,且在ADPN 50μg/L时,具有极显著差异性(P<0.01);此外SQSTM1、mTOR的mRNA表达水平显著升高。结果表明:BHBA可增加中性粒细胞自噬相关基因的表达,ADPN可抑制高BHBA引起的中性粒细胞细胞程序性死亡,进一步说明ADPN对调控酮病奶牛中性粒细胞免疫抑制具有重要的价值意义。     

牛精原干细胞的分离和纯化及体外培养的一般特性

采用两步酶消化法制备5月龄的牛生殖细胞悬液,用Percoll不连续密度梯度法分离精原细胞,接种于含10%胎牛血清的DMEM/F12培养基中,37℃,5%CO2饱和湿度培养,观察培养细胞的生长和形态变化。结果5月龄牛的曲细精管主要包含细胞为精原细胞、Sertoli细胞,每克睾丸实质收获生精上皮细胞总数平均为3.18×106个细胞,精原细胞纯化后纯度达69.27%,精原细胞主要分布于27%~35%的Percoll梯度中。牛精原干细胞体外培养6~7 d后开始分裂,20 d后精原干细胞形成小集落。结果表明用两步酶消化、Percoll不连续密度梯度法分离的精原细胞能满足体外培养的需要,可以存活并发生增殖。     

沙尘暴应激对荷斯坦牛红、白细胞及红细胞黏附免疫功能的影响

在春夏之交沙尘暴多发季节,为初步了解沙尘暴对奶牛的影响,在沙尘暴期间和前后随机抽取6头荷斯坦牛,用经典红、白细胞计数、白细胞分类计数及红细胞黏附免疫功能检测方法,检测红、白细胞数及红细胞黏附免疫功能的变化。结果表明。沙尘暴期间红、白细胞总数有所增高;中性粒细胞极显著增多;淋巴细胞却显著下降;C3bR和ICR的受体花环率略有减少。说明沙尘暴所导致的环境低氧使奶牛增加红细胞数以应付缺氧的环境;中性粒细胞为吞噬沙尘颗粒清除异物极显著提高,而具有特异性免疫作用的淋巴细胞却显著下降;同时红细胞黏附免疫功能也有所下降。可见沙尘暴使炎症细胞聚集,特异性免疫细胞显著下降,很可能导致炎症发生。     

小鼠精原干细胞的分离纯化及初步鉴定

用两步酶消化法对昆明白小鼠睾丸精原细胞及精原干细胞进行了体外分离、纯化和初步鉴定。经两步酶消化所获细胞悬液内活细胞、死细胞及细胞团的百分比分别为97.03%、2.97%和2.95%;平均每个睾丸可获得4.53×105个细胞。Percoll不连续密度梯度离心初步纯化后得到4个细胞带,精原细胞主要分布在40%~50%Percoll梯度间,得到的精原细胞密度为3.20×106个/mL。因此,采用两步酶消化,结合Percoll不连续密度梯度离心法分离的昆明白小鼠精原细胞能满足体外培养的需要,还可以作为精原干细胞移植的研究材料。     

用三色间接流式细胞术检测猪外周血白细胞亚群

用三色间接流式细胞术检测猪外周血粒细胞、B细胞和单核细胞亚群。来源于2头猪的12个猪外周血液样品裂解红细胞后,用3种SWC（swine workshop cluster）单抗和不同标记的二抗染色,对全体细胞设门,测定了SWC1、SWC3、SWC8在外周血白细胞上的表达,分析了嗜中性粒细胞（SWCI^＋SWC3^＋SWC8^＋）、嗜酸性粒细胞（SWCI^-SWC3^＋SWC8^＋）、B细胞（SWC1^-SWC3^-SWC8^＋）、单核细胞（SWC1^＋SWC3^＋SWC8^-）在外周血白细胞中的比例。结果显示,细胞分群情况良好,两组测定结果的均值及方差无显著差异,其中嗜中性粒细胞占（32．6±0．8）％,嗜酸性粒细胞占（4．7±0．3）％,B淋巴细胞占（3．7±0．2）％,单核细胞占（2．2±0．2）％。     

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.     

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.     

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.     

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

Tropical Animal Health and Production -     

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.     

Chemotaxis of macrophage in inflammation

Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.     

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