Activities of enzymes in some types of peripheral leucocytes may reflect the differences in nutrient metabolism between dogs and cats

Plasma metabolites and immunoreactive insulin (IRI) concentrations and enzyme activities of some types of peripheral leucocytes were measured to clarify one aspect of the differences in nutrient metabolism between dogs and cats. There were no significant differences in plasma concentrations of glucose, triglyceride, free fatty acids and IRI between dogs and cats. Higher total cholesterol and lower HDL cholesterol concentrations were observed in feline plasma, and H/T ratio (HDL/total cholesterol concentrations) was significantly lower than that in canine plasma. The cytosolic activities of fructokinase (FK), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were significantly higher and the activities of cytosolic malate dehydrogenase (MDH) and mitochondrial glutamate dehydrogenase (GLDH) were significantly lower in feline leucocytes than those in canine leucocytes. Higher activities of FK, PK and G6PD, which regulate the rate of biosynthesis of fatty acids, may reflect the different characteristics in nutrient metabolism in feline tissues from canine tissues.     

Malate dehydrogenase activities are lower in some types of peripheral leucocytes of dogs and cats with type 1 diabetes mellitus

The activities of the enzymes in the malate-aspartate shuttle were measured in peripheral leucocytes of spontaneous type 1 diabetic dogs and cats treated with insulin injections. In the diabetic dogs and cats, fasting plasma glucose concentrations were three- or fourfold greater than the control levels in spite of insulin injections and the activities of cytosolic malate dehydrogenase (MDH), one of pivotal enzymes in the malate-aspartate shuttle, were remarkably lower than the controls. Depressed expression of cytosolic MDH mRNA was confirmed by RT-PCR analysis in the diabetic animals. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M / L ratio) in leucocytes of the diabetic animals was significantly lower than that of normal control animals. The smaller M / L ratio appeared to reflect depression of energy metabolism in the diabetic animals. Intrinsically lower and further decreased MDH activities may be factors that induce insulin resistance observed in diabetic cats.     

斜带石斑白细胞cDNA文库的构建

用Tripure试剂盒提取斜带石斑白细胞总RNA,反转录合成第一链cDNA,进行长距离PCR,蛋白酶K消化,Sfi I酶切,通过CHROMASPIN-400柱,回收大于500bp的cDNA,与λTripIEx2载体连接,体外包装后构建了斜带石斑白细胞的cDNA文库.库容1.65×1.06,重组频率82%,扩增后滴度7.5×109pfu·mL-1.插入片断长度500～2300bp,最多的是在750～1000bp范围.本文库可作为筛选斜带石斑细胞因子基因的重要资源.     

Use of peripheral blood leukocytes in the study of cell-mediated immunity in bovine anaplasmosis—A review

Peripheral blood leukocytes separated by erythrocyte aggregation and density-gradient centrifugation were used in tests to measure the cell-mediated response in bovine anaplasmosis. The leukocyte migration inhibition test was apparently related to the host protective immune response. Leukocyte stimulation by a specific anaplasma antigen elicits positive but low level stimulation indices in cultures of cells collected from cattle exposed to live anaplasma. There appears to be a nonspecific enhancement of blastogenesis in cultures of leukocytes from cattle injected with inactivated-oil fortified Anaplasma marginale vaccine. Cytotoxicity tests indicated that both leukocytes and antibodies are active in clearance of anaplasma infected erythrocytes from the circulatory system.     

Effects of different artificial photoperiods and temperatures on haematological parameters of rainbow trout (Oncorhynchus mykiss)

This study assessed the effects of two artificial photoperiods (LD 24:0 and LD 12:12) and three temperature regimes (9, 11, and 18°C) for 30 days on haematological parameters of trout (Oncorhynchus mykiss) kept in freshwater. Samples were taken at days 7, 14, and 30 during exposure to treatments. A higher mortality (22%) and lower oxygen concentration (<8.0 mg/l) were associated with the combination of photoperiod LD 24:0 and 18°C. The LD 24:0 photoperiod (independently of temperature) increased the haematocrit and the number of erythrocytes at days 7, 14, and 30 (P < 0.01). A temperature of 18°C (independently of photoperiod regimes) diminished the number of total leucocytes, lymphocytes and thrombocytes (P < 0.01). The LD 24:0 photoperiod (also independently of temperature) lowered the number of lymphocytes only after 14 days of experimentation (P < 0.01). Interaction between artificial photoperiod and temperature was only observed at day 14 for polychromatophils (P < 0.01). These results resemble the effects of stress caused by elevated temperatures and the application of continuous light photoperiods, indicating that survival risks may develop in trout farming when this combination is met.     

The effects of floor heating on body temperature, water consumption, stress response and immune competence around parturition in loose-housed sows

The aim of the present study was to study whether floor heating from 12 h after onset of nest building until 48 h after birth of the first piglet had any effect on measures related to body temperature, water consumption, stress response and immune competence in loose-housed sows (n = 23). In conclusion, the present results indicate that floor heating for a limited period around parturition did not compromise physiological and immunological parameters, water intake and body temperature in loose-housed sows. The water intake peaked the day before parturition and the body temperature peaked on the day of parturition. A cortisol peak at parturition, a transient rise in the number of leucocytes and neutrophils and a transient reduction in the number of lymphocytes, erythrocytes and in the PCV value were observed. Around and after parturition some non-specific immunological variables seemed to be stimulated while others seemed to be compromised.     

Differing antimicrobial potency and specificity of peripheral blood and autologous exudative polymorphonuclear leucocytes

Little is known about the antimicrobial potency and specificity of polymorphonuclear leucocytes which actually appear at the sites of bacterial invasion in tissues. In the present work we have compared inflammatory leucocytes induced by intraperitoneal injection of casein in rabbits with autologous peripheral blood cells in killing Escherichia coli serotype 01 and Staphylococcus aureus 502A. The results indicate that inflammatory leucocytes differ significantly from their virgin blood ancestors. While the blood leucocytes were only able to suppress the growth of the gram-negative bacteria, autologous exudative cells killed more than 95% of the test organisms within 1 h of incubation at 37 degrees C. The enhanced microbicidal activity of the inflammatory cells however, was only specific for the gram-negative bacteria, as evidenced by the failure of leucocytes to kill Staph. aureus to the same extent as the peripheral blood cells. In association with the enhanced gram-negative microbicidal activity the inflammatory cells produced chemiluminescence and released two to three times more O2-anions than the peripheral cells. We interpret these observations to mean that chemotactic factors such as casein activate inflammatory cells to increase their oxidative metabolism. Since microbicidal action of leucocytes is thought to proceed in part through oxygen-dependent reactions, the inflammatory leucocytes would be expected to effectively kill bacteria that are highly susceptible to these lethal oxygen metabolites. It cannot therefore be assumed that assessment of the functional capacity of the virgin peripheral blood PMNs would provide information on the functional characteristics of activated leucocytes which actually migrate to and accumulate at inflammatory sites.     

Immune modulation by probiotic strains: quantification of phagocytosis of Aeromonas salmonicida by leukocytes isolated from gut of rainbow trout (Oncorhynchus mykiss) using a radiolabelling assay

A method is described for the rapid and sensitive assay of phagocytosis that utilizes radioactively labelled bacteria. With this method, we observed that phagocytosis of heat-inactivated Aeromonas salmonicida by leukocytes isolated from gut of rainbow trout fed with different viable probiotics (Lactococcus lactis subsp. lactis, Lactobacillus sakei, and Leuconostoc mesenteroides) was significantly higher (P<0.05) after 2 weeks of probiotic-feeding than the control group. However, only phagocytosis of live A. salmonicida by the leukocytes isolated from gut of rainbow trout fed with L. lactis subsp. lactis was significantly higher (P<0.05) than the control group.     

鸡外周血白细胞吞噬试验的研究

[目的]确定鸡外周血白细胞吞噬葡萄球菌的最佳条件。[方法]使用葡萄球菌10145为吞噬材料,以瑞士染色法检测鸡外周血白细胞的吞噬活性。通过正交试验研究吞噬时间、吞噬温度、葡萄球菌浓度对外周血白细胞吞噬百分率(PP)和吞噬指数(PI)的影响,并对其进行形态观察。[结果]细菌浓度是影响白细胞吞噬指数的主要因素,白细胞吞噬指数最高的条件为:温度41℃,时间40 min,菌液浓度为5×106个/ml。各因素对白细胞吞噬指数的影响程度为:细菌浓度>时间>温度。[结论]该研究为白细胞吞噬功能的检测提供了一种较为简便、可靠的方法。     

Tissue chamber model of acute inflammation in farm animal species

A tissue chamber model of acute inflammation for use in comparative studies in calves, sheep, goats and pigs has been established and validated. Tissue chambers were prepared from silicon rubber tubing, of inner diameter 12.7 mm, length 115 mm and volume 15 ml, with 10 holes, each of 6mm diameter, at each end. In each animal two or four chambers were inserted at subcutaneous sites. Six weeks after implantation an acute inflammatory reaction in a single cage was generated by the intracaveal injection of 0.5 ml of 1% carrageenan solution. Serial samples of exudate (injected chamber), transudate (non-injected chamber) and blood were collected for measurement of exudate and transudate leucocyte count, prostaglandin (PG)E(2) concentration in exudate and serum thromboxane (Tx)B(2) concentration. In addition, skin temperature changes over exudate and transudate chambers were recorded. In all four species, carrageenan induced an acute inflammatory response, indicated by increases to peak values followed by return towards baseline in skin temperature, leucocyte count and PGE(2) concentration. For each of these variables in calves, sheep and goats the increases were significantly greater for exudate than for transudate. The degree of intra-species variation in each variable was acceptable. Marked inter-species differences were recorded: skin temperature rise was greatest in calves and least in sheep and goats; exudate PGE(2) concentration was increased in the order sheep>goat>pig>calf; serum TxB(2) concentration was increased in the order calf>goat>sheep>pig and exudate leucocyte count was increased to a greater extent in the pig than in the three ruminant species. The model has advantages over some previously described tissue chamber models of inflammation and will be suitable for use in comparative studies of inflammatory mechanisms and the pharmacokinetics and pharmacodynamics of anti-inflammatory drugs.