In Vitro Effect of the Reproductive Hormones on the Oxidative Burst Activity of Polymorphonuclear Leucocytes from Cows: A Flow Cytometric Study

In this study, the effect of reproductive hormones and substances with hormonal activity on the oxidative burst activity of blood polymorphonuclear leucocytes (PMN) high yielding dairy cows was evaluated. Different concentrations of: progesterone, oestradiol 17β, FSH, LH, GnRH, cortisol and PGF2α were incubated in vitro for 4 h with PMN of seven high milk yielding cows, during the period of anoestrous postpartum. Controls were run in parallel in which each hormone was replaced by its solvent. After incubation with hormones the competence of PMN to generate H₂O₂ was monitored by flow cytometry. A down‐regulation on the oxidative burst activity of PMA‐stimulated PMN was observed when cells were incubated with progesterone. Significant (p ≤ 0.001) differences between control and progesterone incubated cells were observed from 6.56 μg/ml. The same predisposition was observed when PMNs were incubated with cortisol. Besides for all concentrations employed, a decrease in the burst activity was observed, only beyond 0.19 mg/ml, statistical differences between the results obtained by the control and the cortisol incubated cells were obtained. Concerning oestradiol 17β, an increase on H₂O₂‐production was observed when PMN were incubated with 15 pg/ml and 45 pg/ml of this steroid (p ≤ 0.05), followed by a depression of the cell’s activity when unphysiological concentrations were employed. Significant (p ≤ 0.05) differences between the obtained with the control and oestradiol 17β incubated cells were observed only in the highest concentration of oestradiol. No statistical differences were observed in the metabolic burst activity of PMN incubated with FSH, GnRH and LH when compared with the results obtained by the control.     

Altered Phagocytic Activity of Macrophages in Oyster Toadfish from a Highly Polluted Subestuary

Abstract

The phagocytic function of macrophages in oyster toadfish Opsanus tau was examined in fish taken from several stations in the highly polluted Elizabeth River, Virginia, during October 1989. Sampling sites selected for this field investigation represented a concentration gradient of 55–96,000 μg/kg for total sediment-bound polycyclic aromatic hydrocarbon. Phagocytic function of macrophages was assessed with a novel technique in which absorbance due to phagocytosis of Congo red-stained yeast cells was determined spectrophotometrically. Results obtained with this new technique were comparable to those obtained with more conventional methods for the assessment of phagocytic function. Thus, we believe that this new technique can be used as a sensitive biological indicator of the effects of environmental degradation on fish immune system function.     

Effect of Oestrous Cycle on the Oxidative Burst Activity of Blood Polymorphonuclear Leucocytes in Cows

Blood polymorphonuclear leucocyte (PMN) oxidative burst activity, plasma cortisol levels, and the total and differential white blood cells counts (WBC) of six cycled dairy cows were evaluated for a period of 24 days, three times a week; on Mondays, Wednesdays and Fridays. The PMN oxidative burst was indirectly evaluated by flow cytometry, measuring the intracellular oxidation of 2′,7′‐dichlorofluorescein diacetate to 2′,7′ dichlorofluorescein (DCF) by H₂O₂‐production. Results are pre‐sented as the mean fluorescence intensity (MFI) of DCF. Cow’s oestrous cycle was evaluated by following the plasma progesterone levels using a radioimmunoassay method. Levels of cortisol in the plasma were measured using a fluorimetric method. The oxidative burst activity of PMN, represented a maximum value (MFI = 117.6 ± 7.4) during the oestrous period. A fall was then observed, in which a steady state was observed during the lutheinic phase of the oestrous cycle, reaching the minimum value [MFI = 73.2 ± 11.2 (p ≤ 0.01)] on the days +8, +9 and +10. No significant variations were observed in the levels of cortisol, or in total and differential WBC, during the whole period. Nevertheless, as far as cortisol levels were concerned, a trend analogous to that of the oxidative burst activity was observed. Our results demonstrated that the oestrous cycle might influence directly, or indirectly, the immune system of cows, by altering the oxidative burst of PMN.     

流式细胞仪测定奶牛全血多形核白细胞的吞噬功能和呼吸爆发作用

试验建立了用流式细胞仪同时测定奶牛全血中多形核白细胞（PMN）吞噬功能和呼吸爆发作用的微量检测方法。奶牛全血用二氢诺丹明处理后，与直径为1．75μm的蓝色荧光乳胶微球培养，PMN内的二氢诺丹明被呼吸爆发过程中产生的还原性物质转化成发绿色荧光的诺丹明（ROD）。用流式细胞仪计数PMN，仪器可在数秒钟内计数几十万个PMN，识别PMN是否吞噬蓝色荧光微球和／或含POD，并可检测每个PMN所吞噬的微球数量。当血液中含细胞松弛素B为1、2．5、5mg/L时，PMN的吞噬功能和呼吸爆发作用均下降。     

Simultaneous Flow Cytometric Analysis of Phagocytosis and Oxidative Burst Activity in Equine Leukocytes

This paper describes a method for simultaneously measuring phagocytosis and oxidative burst activity in equine peripheral blood leukocytes by flow cytometry. Opsonized propidium iodide-labelled Staphylococcus aureus (PI-Sa) was used to measure the uptake of bacteria by equine phacocytes and the oxidative burst activity by oxidation of dihydrorhodamine 123. The requirements to achieve optimal activity of phagocytosis and oxidative burst are described. The advantage of the simultaneous technique is that it provides both independent and comparative values for phagocytosis and the oxidative burst, for the detection of impaired mechanisms of microbial destruction. Furthermore, the technique allows evaluation of opsonization activity in this context.     

The Concentrations of Inflammatory Cytokines and Acute‐Phase Proteins in the Peripheral Blood and Uterine Washings in Cows with Pyometra

The development of pyometra in cows depends largely on the state of local immunity of the uterus. The objective of the study was to evaluate the concentration of the following proinflammatory cytokines: tumour necrosis factor (TNF‐α) and interleukin‐6 (IL‐6); anti‐inflammatory cytokine: interleukin‐10 (IL‐10); and acute‐phase proteins (APPs): haptoglobin (Hp) and serum amyloid A (SAA), in serum and uterine washings in cows with pyometra and healthy animals. The study was performed on 20 cows divided into two groups based on the results of cytological and ultrasonographic tests: a pyometra and a healthy group (10 cows per group). Experimental material consisted of blood serum and uterine washings. The levels of the following cytokines, TNF‐α, IL‐6, IL‐10 and APPs – Hp and SAA, in the study material were determined by ELISA. The results showed that the values of TNF‐α, IL‐6, IL‐10 as well as SAA and Hp were significantly higher in serum of cows with pyometra compared to controls (p < 0.001). The uterine washings had significantly higher levels of IL‐6, IL‐10, and Hp in pyometra cows compared to the control (p < 0.001). Our results indicate that it is possible to monitor the course of pyometra in cows based on the evaluation of the concentration of cytokines and Hp in the serum and uterine washings. Simultaneous evaluation of selected indicators of antagonistic interaction can be helpful in determining the current status of local immunity of the uterus. On this basis, it could be possible to properly select an adjunctive therapy in the form of immunomodulating preparations.     

健康奶牛子宫颈乳酸菌筛选方法的建立及其抑菌活性分析

为筛选用于防制奶牛子宫炎的乳酸菌菌株,本试验选取产后30~50 d的健康奶牛,采集子宫颈口处分泌物,利用MRS选择性培养基进行乳酸菌初步筛选。纯化后的菌株进行革兰氏染色镜检,将具有乳酸球菌或乳酸杆菌典型形态的疑似菌株进行针对性生化鉴定。提取分离菌株的基因组DNA,扩增其16S rDNA全序列并测序,将所得序列与NCBI数据库中的相应序列进行同源性比对分析。采用琼脂扩散法检测分离菌株对致病性金黄色葡萄球菌和大肠杆菌的抑菌活性,并检测分离菌株对山羊上皮细胞的黏附活性。结果显示,经染色镜检、生化鉴定和16S rDNA测序分析,得到13株乳酸菌,其中5株具有较强抑菌活性,6株具有较强黏附上皮细胞的能力,7、12和23号3株兼有良好的抑菌活性和黏附上皮细胞能力。综上,本试验建立了健康奶牛子宫颈中乳酸菌的筛选方法,得到3株具有良好益生特性的乳酸菌,为制备微生态制剂提供了基础材料,对防制奶牛子宫炎具有重要意义。     

猪繁殖与呼吸综合征病毒实验感染SPF猪外周血单核细胞和肺泡巨噬细胞早期凋亡细胞的观察

采用Ａｎｎｅｘｉｎ Ｖ－ＦＩＴＣ／ＰＩ双染色法,用流式细胞仪检测了猪繁殖与呼吸综合征病毒（ＰＲＲＳＶ）实验感染ＳＰＦ猪不同时期外周血单核细胞和肺泡巨噬细胞感染Ａｎｎｅｘｉｎ Ｖ－ＦＩＴＣ＾＋／ＰＩ＾－细胞群（早期凋亡细胞群）。结果显示,ＰＲＲＳＶ感染猪外周血单核细胞和肺泡巨噬细胞Ａｎｎｅｘｉｎ Ｖ－ＦＩＴＣ＾＋／ＰＩ＾－细胞群的表达率均明显高于正常对照猪,感染后２４ｈ表达率达最高值。     

鸡骨髓内单核细胞,巨噬细胞和多核巨细胞发生的形态特点

对雏鸡骨髓内单核细胞，巨噬细胞和多核巨细胞发生的形态结构进行了光镜和电镜观察，从幼稚阶段的单核细胞开始，细胞表面出现突起，从原始阶段开始胞质内出现颗粒。随着细胞向成熟阶段发展，突起和颗粒的数量增多，巨噬细胞的形态、结构与幼稚单核细胞相近。     

孕牛血清早孕因子的生物活性检测及妊娠率分析

采用玫瑰花环抑制试验(RIT)对17头人工授精孕牛血清中早孕因子生物活性进行检测,并进行了妊娠结果回访。结果显示,孕牛在排卵后2 d～7 d内,13头孕牛血清经RIT检测出早孕因子活性,4头孕牛血清中经RIT未检测出早孕因子活性。回访发现8头孕牛娩出胎儿,2头孕牛流产,4头未妊娠,人工受精率为76.47%,妊娠率为58.82%,妊娠至分娩的成功率为47.06%,妊娠早期流产率为11.76%,测出早孕因子活性随访未妊娠的植入期胚泡丢失率为23.08%。用RIT对孕牛血清中早孕因子生物活性的研究,对建立早期妊娠诊断方法和检测早期胚胎发育情况及确定早期胚胎死亡的方法具有重要参考价值。     

猪转移因子对犬中性粒细胞数量和活性的影响

按标准方法提取制备了猪的转移因子（transfer factor, TF）,用吞噬杀伤试验MTT法检测了供试杂种牧羊犬肌肉注射猪TF后外周血中性粒细胞吞噬杀伤活性的变化。试验摸索出MTT法测定犬外周血中性粒细胞吞噬杀伤大肠杆菌的最佳条件为：中性粒细胞浓度1.3×106个/ml、大肠杆菌浓度6×105个/ml 时,大肠杆菌和中性粒细胞混合培养2 h,加入MTT后继续培养4 h。体外试验结果表明,猪TF浓度在0.052～1.56 mg/ml范围内,能够明显促进中性粒细胞吞噬杀菌作用,当猪TF浓度为1.56 mg/ml时,对中性粒细胞杀菌活性的影响最大。体内试验结果表明,注射猪TF后第2 d,外周血中性粒细胞数量最高,中性粒细胞吞噬杀菌能力最强。     

猪繁殖与呼吸综合征病毒感染对猪肺泡巨噬细胞外源性抗原加工递呈相关分子和共刺激分子的影响

采用定量竞争PCR技术对猪繁殖与呼吸综合征病毒（PRRSV）感染后不同时相猪肺泡巨噬细胞（PAM）中外源性抗原加工递呈相关分子SLA-DR、Ii链和SLA-DM分子及共刺激分子CD40、CD80、CD86的mRNA转录动态进行了定量检测。结果表明,PRRSV感染后PAM的SLA-DR mRNA转录水平在3、7和14 d下调,低于对照组;感染后3 d,Ii链、SLA-DM和CD40的mRNA转录水平极显著下调（P〈0.01）;CD80和CD86mRNA转录水平也在感染后3 d明显下调（P〈0.05）。用流式细胞术检测PAM表面的SLA-DR抗原的结果表明,在感染后3 d短暂上升,7 d之后一直低于对照组。由此说明,PRRSV感染早期对猪肺泡巨噬细胞的外源性抗原加工和递呈功能产生显著影响,导致其外源性抗原递呈功能下降,进而影响机体的体液免疫应答。     

In vitro Studies of Group E Streptococci in Swine Leukocytes II. Phagocytic and Bactericidal Properties of Macrophages from Immune and Nonimmune Swine

Macrophages collected from susceptible swine and grown in nonimmune serum phagocytized and digested group E Streptococci (GES). However, macrophages collected from immune swine and grown in nonimmune serum were capable of much higher rates of phagocytosis and intracellular digestion of the organism. The addition of immune serum to cultures of immune macrophages increased the rate of phagocytosis but not the rate of intracellular digestion of GES by the macrophages. These findings are discussed in relation to mechanisms of cellular immunity.     

奶牛隐性乳房炎病原菌的分离与耐药性分析

奶牛乳房炎是奶牛的常见性疾病,一般可分为临床型乳房炎和隐性乳房炎两种类型。隐性乳房炎由于无明显临床症状,不易觉察,可在牛群中普遍流行,给奶牛业造成严重损失。因此,对奶牛隐性乳房炎的防制,具有重要意义。因此,对隐性乳房炎致病菌的分析是防制奶牛隐性乳房炎的关键环节。1材料与方法1.1隐性乳房炎的检测从奶牛场选取无乳房炎临床症状的奶牛采集奶样,用SMT法(上海乳房炎检测法)检测奶牛隐性乳房炎。1.2样品收集对检测有隐性乳房炎的奶牛,常规挤奶消毒后,再用70%酒精棉球消毒乳头和挤奶人员手指。每乳头废弃头三把奶后,收集5ml奶于经…     

奶牛隐性乳房炎的调查分析

在2004年3月-5月间，我们对内蒙古西部地区两个奶牛养殖场和一个奶牛养殖专业村共200头泌乳期奶牛进行隐性乳房炎调查诊断，现将调查情况报告如下。     

蹄病奶牛血液与尿液部分指标的检测及病因分析

针对广西某奶牛场蹄病发病率高的现状,选择34例蹄病奶牛采集其尿液和血液,应用尿常规快速自动分析仪对部分尿液指标和血浆钙、磷水平进行检测与分析。结果表明:尿糖、尿胆红素呈阴性,尿血微量,尿比重、尿pH、尿酮异常者占23.5%(8例),尿蛋白呈阳性者占35.2%(12例);血浆钙平均值为8.3 m g/d l,低于正常参考值(9~12 m g/d l)的下限;血浆磷平均值为6.8 m g/d l,高于正常参考值(4~6 m g/d l)的上限。经过综合分析,认为本病例的发生与亚急性瘤胃酸中毒有关。     

奶牛不孕症问题分析

每一个奶牛场都会遇到繁殖问题,即使经营好的牧场,奶牛的繁殖率也常常达不到正常水平。奶牛不孕症是影响奶牛场繁殖效率的重要因素,而对不孕问题的原因以及寻求相应的解决办法是奶牛饲养管理人员所关心的问题。为此我们全文翻译了美国宾夕法尼亚州大学L.J.Hutchinson博士的这篇     

Pyometra with inguinal herniation of the left uterine horn and omentum in a Beagle dog

Objective: To describe a unique case of pyometra with inguinal herniation of the left uterine horn and omentum. Case summary: A 7‐year‐old, 19 kg, intact female Beagle dog presented for surgical treatment of presumptive pyometra and biopsy of a caudal abdominal mass in the left inguinal mammary gland region. Ventral midline celiotomy was performed, and a distended, fluid‐filled uterus with passage of the distal aspect of the left uterine horn through the left vaginal process into the inguinal canal was identified. The patient recovered uneventfully following ovariohysterectomy and left inguinal herniorrhaphy. New or unique information provided: This is the first documented report of inguinal herniation of a uterine horn associated with a pyometra.     

德国弗莱维赫(Fleckvieh)牛和本地牛杂交F1代母牛生长发育性状测定及分析

[目的]为了研究德国弗莱维赫(Fleckvieh)牛与本地母牛杂交F_1代育成母牛生长发育性状和生长发育规律,为培育北大荒乳肉兼用牛品种,开展选育提高奠定基础。[方法]采用各批次测定体重校正到统一月龄,测定了其初生、4月龄(断奶)、12月龄及18月龄4个阶段的体重、体尺,并对各阶段表型值比较分析。[结果]德国弗莱维赫(Fleckvieh)牛与本地母牛杂交F_1代母牛平均日增重、胸围和体斜长性状,在初生至4月龄断奶阶段生长发育最快;从4月龄断奶至18月龄,与纯种德国弗莱维赫种公牛与澳系(澳大利亚)西门塔尔母牛杂交后代相比,在体重和体尺上有一定差异。[结论]需要不断地改良,以提高后代的生长发育性状,进一步提高生产性能。