Morphologic changes in chicken cells after in vitro exposure to Mycoplasma gallisepticum

Mycoplasma gallisepticum (MG) was used to expose chicken peripheral blood lymphocytes (PBLs), red blood cells (RBCs), heterophils, and chicken tumor cells (MSB-1 and HD-11 cells). Incubation of PBLs with MG for 3 hr resulted in extensive clumping of lymphocytes. Incubation of the MSB-1 cells with MG also caused clumping of the cells, with many of the cells showing perforations and others showing capping of the surface projections. Incubation of RBCs with MG resulted in an altered cell surface morphology, a decrease in cell size, and perforation. There were no discernible changes on the surface of the heterophils and the HD-11 cells. However, the HD-11 cells appeared to have a decreased ability to attach to the surface of the plastic and to have a decreased ability to respond to chemoattractant fMLP after 24 hr of incubation. These results suggest that, under the conditions used, MG caused certain damage to peripheral blood cells and a significant decrease in chemotactic response in the HD-11 cells.     

Newcastle disease virus-induced damage to embryonic tracheae and red blood cells

Newcastle disease virus (NDV)-induced damage to the embryonic chicken trachea, the tracheal explants, and red blood cells (RBCs) was studied by scanning electron microscopy. In the tracheae, NDV caused deciliation and depletion of the epithelial cells. In the tracheal explants, the same pathologic changes could be seen within 6 hr of virus infection. In RBCs, a perforation and a change in surface morphology were observed.     

Generation of neuronal-like cells from umbilical cord blood-derived mesenchymal stem cells of a RFP-transgenic cloned cat

Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are multipotent adult stem cells, which can differentiation into cells of connective tissue and neural lineages. This study investigated the potential for neuronal differentiation of red fluorescent protein (RFP)-transgenic cat UCB-derived MSCs. The cells were cultured in pre-induction medium for 24 hr and in neuronal-induction medium for 72 hr. Immunofluorescent staining showed that 6.85% of the total cells were beta III-tubulin-positive, 3.37% were neurofilament light (NF-L)-positive and 7.04% were neurofilament medium (NF-M)-positive. A beta III-tubulin band was detected by western blot analysis. Our results demonstrate that RFP-transgenic UCB-derived MSCs can be differentiated into neuronal cells in vitro. Thus, RFP-transgenic MSCs could provide alternative tracing material for stem cell transplantation.     

Immunostimulating and growth-promoting effects of sugar cane extract (SCE) in chickens

Polymorphonuclear cells of the peripheral blood in the chicken significantly increased their phagocytosis when cultured with sugar cane extract (SCE; 250-1,000 microg/ml) for 24 hr. Chickens orally administered SCE (500 mg/kg/day) for 3 or 6 consecutive days at 1 week of age showed significantly higher body weight and gain in body weight/day and a lower food conversion ratio within the growing period of 6 weeks than physiological saline-administered control chickens. Furthermore, oral administration of SCE also resulted in significantly higher immune responses against sheep red blood cells and Brucella abortus. These results suggest that SCE has immunostimulating and growth promoting effects in chickens.     

Malaria in an eastern screech owl (Otus asio)

Owls are frequent carriers of blood parasites but clinical malaria infections are rare. Various stages of Plasmodium subpraecox were seen in 90% of the erythrocytes of an Eastern screech owl (Otus asio) showing symptoms consistent with malaria 1 wk after admission for traumatic injuries. An additional unidentified blood parasite, either a Plasmodium or a Haemoproteus spp. was found in small numbers of red blood cells on blood films examined at admission and at day 7 postadmission. Combined infestation, trauma-induced stress, and iatrogenic corticosteroid administration are possible factors that could have induced disease. Oral treatment with mefloquine at 30 mg/kg, repeated after 12, 24, and 48 hr, proved successful in eliminating both organisms and signs of clinical disease.     

豚鼠醛化红细胞在人流感病毒血凝活性测定中的应用

在流感疫情监测中,血凝试验是最常用的检测方法.针对鸡红细胞不能凝集流感病毒"O"相毒株,新鲜红细胞需现采现配、麻烦、费时,易溶血、不易保存等问题,本试验对豚鼠醛化红细胞制备方法的筛选和制备条件的优化进行研究,并以新鲜豚鼠红细胞为对照,用于流感病毒不同型别血凝试验.结果表明,戊二醛固定法制备的豚鼠醛化红细胞不溶血、结果稳定性好、保存期长、血凝活性与新鲜豚鼠红细胞基本一致,可以替代新鲜豚鼠红细胞用于血凝试验.     

Correlation between ability of Ornithobacterium rhinotracheale to agglutinate red blood cells and susceptibility to fosfomycin

Twenty five freeze-dried isolates of Ornithobacterium rhinotracheale were used for the determination of minimum inhibitory concentrations (MIC) against the antibiotic fosfomycin (Fosbac, produced by Bedson SA, consisting of a 25% mixture of fosfomycin). The same isolates were tested for their ability to haemagglutinate chicken red blood cells. Ten of the 25 isolates were found to be susceptible to fosfomycin (MIC values below 128 ug/ml). All of these isolates were able to agglutinate red blood cells. This is the first report on the ability of O. rhinotracheale to agglutinate red blood cells. The remaining 15 isolates were resistant to fosfomycin (MIC values above 128 ug/ml). Only five of these isolates were found to have the ability to agglutinate red blood cells. There appears to be a correlation between the ability of O. rhinotracheale isolates to agglutinate red blood cells and their susceptibility to fosfomycin. The ability of certain isolates of O. rhinotracheale to agglutinate red blood cells, raises the questions of differences in virulence between the isolates which can agglutinate red blood cells and those which cannot and the use of this ability to agglutinate red blood cells as an alternative method for serotyping O. rhinotracheale.     

Evaluation Of An Additive Solution For Preservation Of Canine Red Blood Cells

The effect of an additive preservative solution on canine red blood cell posttransfusion viability (PTV) and on selected canine red blood cell biochemical parameters was studied. One unit (450 mL) of blood was collected from 6 clinically normal dogs into the anticoagulant citrate phosphate dextrose, centrifuged, and the plasma removed. The red blood cells were then suspended in 100 mL of a saline, adenine, dextrose, and mannitol solution and stored at 4°C. Aliquots were removed for study at 1, 10, 20, 30, 37, and 44 days. The 24-hour PTV of autologous red blood cells was determined using a sodium chromate (⁶¹Cr) label. Red blood cell concentrations of 2,3-diphosphoglycerate (2,3-DPG), adenosine-5'-triphosphate (ATP), and pH were also determined. Canine red blood cell PTV, pH, ATP, and 2,3-DPG concentrations decreased during storage ( P < .05). The PTV decreased from 94% using day 1 red blood cells to 80% and 75% using day 37 and day 44 red blood cells, respectively ( P < .05). Although the mean PTV of the day 44 stored units equaled the Food and Drug Administration (FDA) minimum standard for human red blood cells, the PTV was substandard in 75% of the day 44 units. The FDA standard was exceeded in 83% of the day 37 units. It was concluded that 37-day-old canine red blood cells preserved with a saline, adenine, dextrose, and mannitol solution are of acceptable quality for transfusion.     

鹿茸醇提物对用环磷酰胺处理的小白鼠红细胞免疫功能的影响

结果表明 ,鹿茸醇提物组 (即实验组 )小白鼠红细胞的C3b受体酵母菌花环率为( 1 3 5± 0 2 ) %,对照组为 ( 1 0 1± 0 1 ) %,两组相比较 ,差异显著 (P <0 0 5) ;试验组小白鼠红细胞的免疫复合物酵母菌花环率为 1 1 3± 0 2 %,对照组为 1 6 3± 0 3%,两组相比较 ,差异显著 (P <0 0 5)。这一试验结果提示 ,鹿茸醇提物可增强由环磷酰胺诱导的免疫功能低下的小白鼠红细胞的免疫功能 ,即鹿茸醇提物对于环磷酰胺对机体的副作用具有一定的对抗作用。     

当归对鸡血液指标的影响研究

为探讨当归对鸡血液指标的影响,试验用高、中、低3个剂量的当归煎剂分别给正常和贫血鸡进行灌服,测定试验鸡的红细胞数、血红蛋白含量、红细胞压积和血沉值,并与正常及贫血不用药组进行比较.贫血鸡模型利用环磷酰胺制造.结果发现:当归能显著提高正常鸡的红细胞数、血红蛋白含量、红细胞压积和血沉值;同时当归能够逆转环磷酰胺所致鸡的红细胞数、血红蛋白含量、红细胞压积和血沉值,使其恢复至正常水平.     

Physiologic effects of furosemide in combination with water restriction when administered at 4 and 24 hours prior to high-intensity treadmill training

Although controversial, due to its reported effectiveness in attenuating bleeding associated with exercise-induced pulmonary hemorrhage (EIPH), furosemide is currently a permitted race day medication in most North American racing jurisdictions. The objective of this study was to assess the efficacy of furosemide in reducing the presence and severity of EIPH when administered 24 hr prior to strenuous treadmill exercise. Eight exercised Thoroughbred horses received saline or 250 mg of furosemide either 4 or 24 hr prior to high-speed treadmill exercise in a balanced 3-way cross-over design. Blood samples were collected for determination of furosemide, lactate, hemoglobin, blood gas, and electrolyte concentrations. Heart rate and pulmonary arterial pressure were measured throughout the run and endoscopic examination and bronchoalveolar lavage (BAL) performed. Horses were assigned an EIPH score and the number of red blood cells in BAL fluid determined. Although not significantly different, endoscopic EIPH scores were lower in the 4-hr versus the 24-hr and saline groups. RBC counts were not significantly different between the treatment groups. Pulmonary arterial pressures were significantly increased at higher speeds; however, there were no significant differences between dose groups when controlling for speed. A small sample size and unknown bleeding history warrant a larger-scale study.     

Evaluation of Canine Red Blood Cells Stored in a Saline, Adenine, and Glucose Solution for 35 Days

An additive solution for the storage of red blood cells was evaluated for use in dogs. Blood collected from 6 dogs was processed into packed red blood cells and stored for 35 days in the additive solution Nutricel (Miles, Inc, Pharmaceutical Division, West Haven, CT). Packed red blood cells stored in citrate-phosphate-dextrose-adenine (CPDA-1; Fenwal Laboratories, Baxter Health Care Corp, Deerfield, IL) also were evaluated for comparison. Red blood cell 2,3-diphosphoglycerate (2,3-DPG) concentration, adenosine triphosphate (ATP) concentration, percentage hemolysis, and pH were determined. The red blood cell post-transfusion viability (PTV) after 35 days of storage was assessed with both single-labeled chromium 51 (⁵¹Cr) and double-labeled technetium 99m/chromium 51 (^99mTc/⁵¹Cr) techniques. Mean ATP concentration and percentage hemolysis of the cells stored in Nutricel were 1.1 μmol/g hemoglobin (Hb) and 0.28% respectively and did not differ significantly (P < .05) from the values of 1.0 μmol/g Hb and 0.33% from the CPDA-1-stored red blood cells. The mean pH of red blood cells stored in Nutricel was 6.19, which was significantly lower than the pH of 6.47 for cells stored in CPDA-1. The mean 2,3-DPG concentration of red blood cells stored in Nutricel was significantly higher at 10.1 μmol/g Hb than the 2,3-DPG concentration of 3.4μmol/g Hb for cells stored in CPDA-1. The mean PTV of canine red blood cells stored in Nutricel for 35 days was 85% with ⁵¹Cr and 90% with ^99mTc/⁵¹Cr. This was significantly higher than the mean PTVs of 38% and 36% for the CPDA-1 stored cells as assessed with ⁵¹Cr and ^99mTc/⁵¹Cr techniques, respectively. It was concluded that 35-day-old canine red blood cells stored in Nutricel are of acceptable quality for transfusion purposes.     

高剂量铜对昆明小鼠生长和血细胞数的影响

将50只30日龄健康昆明小鼠随机分为对照组、试验Ⅰ、Ⅱ、Ⅲ和Ⅳ组共5组,每组10只,雌雄各半,分笼饲养。对照组、试验Ⅰ、Ⅱ和Ⅲ组每天分别按硫酸铜0、20、50、100 mg/kg体重灌胃1次,试验Ⅳ组按100 mg/kg体重灌胃的同时饮用500 mg/L VC溶液,其余各组自由饮用蒸馏水。分别在试验开始及第7、14、21和28天称重,尾静脉采血测定血常规指标。试验结果显示,整个试验期间,与对照组相比,试验Ⅰ组的体重、红细胞数量、血红蛋白含量和白细胞数量均无显著差异(P>0.05);试验Ⅱ组在试验第28天时红细胞数量显著降低(P<0.05),白细胞数量显著增加(P<0.05),体重和血红蛋白含量均无显著差异(P>0.05);试验Ⅲ组在试验第28天小鼠体重和红细胞数量均极显著降低(P<0.01),血红蛋白含量显著降低(P<0.05),白细胞数量无显著变化(P>0.05)。与试验Ⅲ组相比,试验Ⅳ组第21和28天的小鼠体重显著增加(P<0.05),但红细胞数量、血红蛋白含量和白细胞含量均无显著差异(P>0.05)。研究结果表明,高剂量铜能显著抑制小鼠的生长,降低血液中红细胞、白细胞数量及血红蛋白含量。VC对高铜引起的体重下降有部分逆转作用,对高铜引起的红细胞数量、血红蛋白含量和白细胞数量均无显著影响。     

The involvement of polymorphonuclear leukocytes in the pathogenesis of bronchopneumonia in calves. I. Activated granulocyte induced lipid peroxidation in red blood cells.

The influence of free radicals generated by polymorphonuclear leukocytes from bronchopneumonic calves on lipid peroxidation in red blood cells was studied. Incubation of granulocytes with red blood cells causes a rise in malondialdehyde production in these cells in linear dependence on the granulocyte number. Incubation of red blood cells with malondialdehyde causes a generation of adduct moiety of this compound with phosphatidylserine and phosphatidylethanolamine. This adduct is also present in erythrocytes which have been incubated with neutrophils of bronchopneumonic calves. Cytochrome c loaded erythrocyte ghosts showed a high reduction rate of this compound during incubation of ghosts with neutrophils of diseased calves. It is suggested that neutrophils of bronchopneumonic calves are in an activated state in the blood-stream, and free radicals generated by these cells are capable of peroxidizing the red blood cell membrane lipids and causing cross-links between phospholipid moieties in erythrocyte membranes.     

Effects of recombinant bovine granulocyte-macrophage colony-stimulating factor on bovine peripheral blood neutrophil functions in vitro and in vivo

Effects of recombinant bovine granulocyte-macrophage colony-stimulating factor (rboGM-CSF) on bactericidal activity of bovine peripheral blood neutrophils in vitro and in vivo were studied. In in vitro experiment, bovine blood neutrophils were cultured for 9 hr in media containing 0.005, 0.05 or 0.5 microg/ml of rboGM-CSF. Neutrophils treated with rboGM-CSF showed significantly higher luminol-dependent chemiluminescence (LDCL) than control cells. In in vivo experiment, neutrophils isolated from cows injected 5.0 microg/kg of rboGM-CSF showed significantly higher Nitrobluetetrazolium (NBT) reduction value than that from control cows 24 hr post injection. Total leukocyte counts of cows injected rboGM-CSF sharply decreased 6 hr post injection and recovered to normal level 2 days post injection. Body temperature of these cows rose 6 hr post injection and back to normal level at 24 hr post injection. It was suggested that rboGM-CSF enhanced bactericidal activity of bovine neutrophils both in vitro and in vivo.     

Exit of Anaplasma marginale from bovine red blood cells.

Incubation of Anaplasma marginale-infected bovine red blood cells in a flow culture system resulted in a decrease of observable marginal bodies. The decrease was twice the degree of hemolysis, indicating that marginal bodies may leave red blood cells without concomitant lysis of the host cell.     

Bronchoalveolar lavage cytology in cynomolgus monkeys and identification of cytologic alterations following sequential saline lavage

Total and differential cell counts were determined on cytolytic specimens obtained by fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) of five normal cynomolgus monkeys. Total nucleated cell counts ranged from 100 to 430 cells/microliters. Macrophages were approximately 91% of total nucleated cells, while lymphocytes were 3%, neutrophils 4%, and eosinophils 2% of the initial BAL from each monkey. Less than 1% of the cells were mast cells and ciliated or nonciliated epithelial cells. The effects of repeated saline BAL on pulmonary cell populations were evaluated. Saline lavage of individual lung lobes resulted in a marked rise in circulating blood neutrophils at 4 hr after BAL; there was a similar rise in neutrophils in lavage fluids 24 hr after the initial lavage. Differential and total cell counts of both blood and lavage fluid returned to normal if subsequent lavages were spaced at 48-hr intervals. Lymphocytes were not present in saline-lavaged lung lobes, and protein levels of lavage fluids did not rise significantly. BAL produced a transient, reversible, intra-alveolar influx of neutrophils which was preceded by mobilization of bone marrow-stored neutrophils. Neutrophilia in the lavage fluid and blood was not detectable if lavage and blood sampling procedures were done at 48-hr intervals (which did not alter Ia antigen expression among BAL cells). These observations indicate that BAL is a valid method for sampling and assessing pulmonary cellular and fluid constituents if the procedures are done at intervals of at least 48 hr.     

Phenotypic and molecular characterization of isolates of Ornithobacterium rhinotracheale from chickens and pigeons in Taiwan

Forty Ornithobacterium rhinotracheale (ORT) strains were isolated from 28 chickens and 12 pigeons for the first time in Taiwan. All isolates reacted positively in the p-nitrophenyl-beta-D-galactopyranoside (PNPG) and oxidase tests, showing an API 20NE identification system biocode 0-0-2-0-0-0-4. All the pigeon isolates and 85.7% (24 of 28) of the chicken isolates belonged to serotype A. Compared to the ORT ATCC 51464 strain, 14.3% (4 of 28) of chicken isolates and 58.3% (7 of 12) of pigeon isolates showed smaller colonies after 72 hr incubation. Most of the chicken isolates (22 of 28), but none of the pigeon isolates, could agglutinate chicken and pigeon red blood cells. There appears to be a correlation that ORT isolates with a larger colony size tend to be more able to agglutinate red blood cells than the ORT isolates with a smaller colony size. A majority of isolates was sensitive to amoxicillin, ampicillin, ceftiofur, penicillin, and oxytetracycline. The 16S ribosomal RNA (rRNA) sequences of 23 Taiwanese ORT isolates showed high identity (98%-100%) to sequences in GenBank. Phylogenetic analysis of these sequences showed that pigeon isolates formed a distinctive cluster, while chicken isolates and all other 16S rRNA sequences obtained from GenBank belonged to another two clusters. The results indicate that pigeon ORT isolates are different from most chicken isolates in regard to a number of phenotypic and molecular traits.     

Lowered Antioxidant Status of Red Blood Cells in Post-Parturient Haemoglobinuria of Buffaloes

The antioxidant status of the red blood cells of buffaloes (n = 20) suffering from post-parturient haemoglobinuria (PPH) was assessed by comparing their tocopherol (vitamin E) and reduced glutathione contents with those of red blood cells from apparently healthy buffaloes (n = 20). The red cell tocopherol content of the diseased buffaloes (1.76±0.11 g/ml) was significantly (p<0.01) lower than that of healthy buffaloes (2.45±0.14 g/ml). This may be the first report comparing the concentration of tocopherol in the red blood cells of buffaloes suffering from PPH and apparently healthy buffaloes.There was a drastic reduction in the reduced glutathione content in the red cells of haemoglobinuric buffaloes (23.74±2.86 mg%) compared to the healthy control buffaloes (73.71±3.87 mg%). The diseased buffaloes also exhibited severe hypophosphataemia. These findings suggest that an impaired or insufficient antioxidant potential of the red blood cells in this disease in buffaloes is associated with the phosphorus deficiency.     

Cytotoxic antibodies to chicken adipocytes and their precursors: lack of tissue specificity

1. Antisera raised in sheep against chicken adipocyte plasma membranes recognised adipocyte, liver and red blood cell membranes in enzyme immunoassays (EIA). 2. These antisera were cytotoxic when incubated with both adipocytes and their precursors grown in culture, and with red blood cells. 3. Adsorption of antisera with liver membranes completely abolished the response to liver and red blood cell membranes in the EIA. The response in adipocytes was reduced to only about 6% of the titre present in the unadsorbed sample. 4. Adsorption of antisera with liver membranes abolished the cytotoxic response to red blood cells and to adipocyte precursors, but some response to adipocytes still remained. 5. These results are in contrast to the rat for which adipocyte specific antisera were obtained which represented about 75% of the unadsorbed titre.