酵母细胞壁在异育银鲫饲料中添加效果的研究

本试验通过添加酵母细胞壁(YCW)研究其在异育银鲫(Carassius auratus gibelio)饲料中的应用效果。结果表明,饲料中添加0.10%酵母细胞壁饲喂15d后,试验组血清SOD活性极显著高于空白对照组(P<0.01);饲喂30d后试验组血清溶菌酶活性显著高于空白对照组(P<0.05);饲喂60d后试验组背肌中RNA/DNA较空白对照组提高12.18%(P>0.05);饲喂60d后试验组前肠皱褶高度极显著高于空白对照组(P<0.01)。表明饲料中添加0.10%酵母细胞壁对异育银鲫的非特异性免疫力具有明显的促进作用,对其背肌中RNA/DNA和肠道发育也具有一定的促进作用。     

Cytochemical properties of chicken blood cells resembling both thrombocytes and lymphocytes

Generally accepted criteria were used to identify typical nucleated thrombocytes and typical small lymphocytes in chicken-blood smears subjected to modified-Wright staining. Other cells, here referred to as "intermediate cells," were difficult to classify because in some aspects they resembled thrombocytes while they also had features typical of small lymphocytes. The "intermediate cells" had small, round or oval nuclei with coarsely condensed chromatin, characteristic of both thrombocytes and small lymphocytes. In addition, "intermediate cells" had moderately abundant cytoplasmic volumes, typical of thrombocytes but blue cytoplasm lacking both granules and vacuoles, which is characteristic of small lymphocytes. It made little difference to the thrombocyte count whether these cells were classified as thrombocytes or small lymphocytes; however, this decision made a substantial difference to the lymphocyte count in some chicken-blood smears. Most "intermediate cells" (351 of 410 cells examined) were nonfluorescent after treatment with formaldehyde gas. Furthermore, most "intermediate cells" failed to acquire characteristic pigments when subjected to either Grimelius staining (179 of 204 cells examined) or periodic acid-Schiff staining (173 of 206 cells examined). Typical small lymphocytes reacted in the same way, failing to fluoresce after gaseous formaldehyde treatment (65 of 65 cells examined) and failing to react during Grimelius staining (41 of 44 cells examined) or periodic acid-Schiff staining (21 of 21 cells examined). In contrast, almost all typical thrombocytes became fluorescent in response to gaseous formaldehyde (709 of 718 cells examined) and gave positive reactions when subjected to Grimelius staining (381 of 382 cells examined) or periodic acid-Schiff staining (322 of 326 cells examined). These findings suggested that "intermediate cells" should be classified as lymphocytes in differential cell counts.     

小鼠精原干细胞的形态结构及其细胞化学和免疫组化特性

应用光镜、电镜观察了 ７～８ ｄ 小鼠精原干细胞的形态结构特点;应用细胞化学、免疫组织化学方法研究了体外培养小鼠精原干细胞的细胞化学和免疫组织化学特性。结果显示,精原干细胞呈圆形,直径（９±１） μｍ ,核大,呈圆形或轻微卵圆形,胞质较少;核内常染色质占绝对优势,异染色质极少;胞质内核糖体、线粒体较丰富,其他细胞器不发达;精原干细胞 Ａ Ｋ Ｐ染色呈阳性或强阳性,胞质内脂滴很少或没有;精原干细胞表达特异的 ｃｋｉｔ 受体;体外培养的小鼠精原干细胞常呈不完全的胞质分裂,细胞间由细胞间桥相连而呈链状或团状的细胞群丛,细胞群常由偶数细胞组成。     

Principles of peripheral blood mononuclear cell apheresis in a preclinical canine model of hematopoietic cell transplantation

BACKGROUND: Preclinical studies of peripheral blood mononuclear cell (PBMC) transplantation conducted in a well-established canine hematopoietic cell transplantation (HCT) model have been successfully translated to human patients over the past 5 decades. OBJECTIVE: We retrospectively investigated the safety and feasibility of PBMC apheresis in the canine model of HCT by analyzing apheresis parameters, cell yields, and the impacts of donor-related and apheresis-related variables on collection yields and donor stability. ANIMALS: One hundred and twenty dogs that underwent PBMC aphereses were evaluated. METHODS: Aphereses were performed with a COBE Spectra blood separator and a central dual-lumen catheter, with or without recombinant canine granulocyte colony-stimulating factor (rcG-CSF) stem cell mobilization. RESULTS: Aphereses from dogs not given rcG-CSF yielded an average volume of 280 +/- 42 mL containing an average of 15,086 +/- 9,834 leukocytes/mL. Aphereses from dogs given rcG-CSF yielded an average volume of 261 +/- 55 mL containing an average of 39,711 +/- 24,488 leukocytes/mL. Higher pre-apheresis white blood cell (WBC) counts correlated with higher apheresis WBC yields (R=0.50, P<.0001). The correlations of collection time, inlet volume, and collection flow rate on WBC yields were statistically significant but only weak to moderate in magnitude (R=0.34, P=.0001; R=0.38, P=.0006; R=0.26, P=.002, respectively) as were the correlations of collection time and inlet volume on collection volumes (R=0.30, P=.002; R=0.42, P<.0001, respectively). All dogs recovered promptly after PBMC aphereses and catheter removal, without complications. CONCLUSIONS AND CLINICAL IMPORTANCE: These data may be useful for translating PBMC apheresis technology to the field of veterinary oncology for the treatment of dogs with hematologic malignancies.     

Differential cytokine expression in T cell subsets from bovine peripheral blood

Although distinct cytokine expression in T cell subsets is well understood in mice and humans, limited information is available on bovine T cell subsets. In the present study, we analyzed the mRNA expression of 10 kinds of cytokines and CD25 expression in CD4⁺, CD8⁺, WC1⁺ and WC1^-γδ T cell subsets in bovine peripheral blood by Concanavalin A (Con A) stimulation. CD25 expression was significantly increased in CD4⁺, CD8⁺ and WC1⁺γδ T cells, but not in WC1^-γδ T cells by Con A stimulation. In CD4⁺ T cells, the mRNAs of Interleukin (IL)-2, IL-6, IL-10, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, TNF-β and transforming growth factor (TGF)-β were expressed in control cultures, and IL-3, IL-4 and granulocyte-macrophage colony stimulating factor (GM-CSF) were newly expressed when the cells were stimulated with Con A. CD8⁺ T cells expressed the mRNAs of IL-6, TNF-α, TNF-β and TGF-β in control cultures, and newly expressed those of IL-2, IFN-γ and GM-CSF, but did not express those of IL-3, IL-4 or IL-10 after Con A stimulation. The cytokine expression profile of WC1⁺γδ T cells was similar to that of CD8⁺ T cells. However, WC1^-γδ T cells did not express any cytokine mRNA except TGF-â mRNA. These results will contribute to elucidate the participation of T cell subsets in immune responses against infectious disease in cattle.     

日粮中血粉替代鱼粉对草鱼生长和肌肉氨基酸组成的影响

试验旨在研究血粉替代部分鱼粉对草鱼生长及肌肉营养成分的影响。以含鱼粉12%日粮为对照组,以血粉分别替代对照组饲料中25%、50%和75%的鱼粉,配制3种试验饲料,投喂平均体质量194.0 g的草鱼64 d。结果表明,各试验组草鱼摄食量下降,增重率显著低于对照组(P<0.05),但25%血粉替代组草鱼饲料系数、肝体比、脏体比、肌肉粗脂肪的含量与对照组无显著性差异(P>0.05),而50%和75%血粉替代组的饲料系数、肌肉粗脂肪明显升高;与对照组相比较,各血粉替代组的粗蛋白含量、粗灰分含量、必需氨基酸总量和鲜味氨基酸总量均无显著性差异(P>0.05)。综合分析表明,血粉代替草鱼日粮中鱼粉的量为25%时,可以获得较好的效果。     

Hematology, cytochemistry and ultrastructure of blood cells in fishing cat (Felis viverrina)

Hematological, cytochemical and ultrastructural features of blood cells in fishing cat (Felis viverrina) were evaluated using complete blood cell counts with routine and cytochemical blood stains, and scanning and transmission electron microscopy. No statistically significant difference was found in different genders of this animal. Unique features of blood cells in this animal were identified in hematological, cytochemical and ultrastructural studies. This study contributes to broaden hematological resources in wildlife animals and provides a guideline for identification of blood cells in the fishing cat.     

Comparative cell morphology in the peripheral blood film from exotic and native animalsa

Haematological investigation is an important part of disease diagnosis. This is particularly so when investigating individual animal disease. It may also be important when investigating diseases in groups of animals, but the opportunity to perform necropsies to directly detect disease processes often diminishes its usefulness. Haematological investigation is essentially similar for all species. The presence of nucleated erythrocytes and thrombocytes in nonmammals requires alteration of haemoglobin measurement and cell counting. In addition, it may cause some confusion in identification of cells in peripheral blood films. Examination of blood films is an important component of haematological investigation and provides useful information on erythroid, leukocytic and platelet/thrombocytic alterations. Interpretation of alterations is essentially similar for all species. However, cell identification can at times be difficult. There are five basic leukocytes in all species: neutrophil (mammals) or heterophil (nonmammals), eosinophil, basophil, lymphocyte and monocyte. In nonmammals it may be difficult at times to distinguish between heterophils and eosinophils. In addition, lymphocytes may be confused with thrombocytes. However, a common-sense approach to the examination of the peripheral blood film will minimise this confusion.     

Comparative clinical study of canine and feline total blood cell count results with seven in‐clinic and two commercial laboratory hematology analyzers

Background: A CBC is an integral part of the assessment of health and disease in companion animals. While in the past newer technologies for CBC analysis were limited to large clinical pathology laboratories, several smaller and affordable automated hematology analyzers have been developed for in‐clinic use. Objectives: The purpose of this study was to compare CBC results generated by 7 in‐clinic laser‐ and impedance‐based hematology instruments and 2 commercial laboratory analyzers. Methods: Over a 3‐month period, fresh EDTA‐anticoagulated blood samples from healthy and diseased dogs (n=260) and cats (n=110) were analyzed on the LaserCyte, ForCyte, MS45, Heska CBC, Scil Vet ABC, VetScan HMT, QBC Vet Autoread, CELL‐DYN 3500, and ADVIA 120 analyzers. Results were compared by regression correlation (linear, Deming, Passing‐Bablok) and Bland–Altman bias plots using the ADVIA as the criterion standard for all analytes except HCT, which was compared with manual PCV. Precision, linearity, and carryover also were evaluated. Results: For most analytes, the in‐clinic analyzers and the CELL‐DYN performed similarly and correlated well with the ADVIA. The biases ranged from ?0.6 to 2.4 × 10⁹/L for WBC count, 0 to 0.9 × 10¹²/L for RBC count, ?1.5 to 0.7 g/dL for hemoglobin concentration, ?4.3 to 8.3 fL for MCV, and ?69.3 to 77.2 × 10⁹/L for platelet count. Compared with PCV, the HCT on most analyzers had a bias from 0.1% to 7.2%. Canine reticulocyte counts on the LaserCyte and ForCyte correlated but had a negative bias compared with those on the ADVIA. Precision, linearity, and carryover results were excellent for most analyzers. Conclusions: Total WBC and RBC counts were acceptable on all in‐clinic hematology instruments studied, with limitations for some RBC parameters and platelet counts. Together with evaluation of a blood film, these in‐clinic instruments can provide useful information on canine and feline patients in veterinary practices.     

Effect of immune-mediated erythrocyte agglutination on analysis of canine blood using a multichannel blood cell counting system

Blood from six dogs with in vitro immune-mediated erythrocyte agglutination resulted in analytical errors in directly measured counting and sizing functions on a multichannel blood analysis system with histogram capability. Errors in the directly measured values, mean cell volume (MCV), and erythrocyte count were attributed to agglutinated erythrocyte particles that persisted during the relatively short reagent contact time of the analysis. Agglutinated particles less than 240 fl were visible on erythrocyte histograms and resulted in a false low erythrocyte count and false high MCV. Agglutinated cell particles greater than 240 fl were not present on the histogram scale. Because these latter particles exceeded the upper threshold, they did not influence determination of MCV, but resulted in a further decrease in the erythrocyte count. As a result, the other dependent erythrocyte indices were in error. These included false low hematocrit and false high mean corpuscular hemoglobin concentration (MCHC), when compared to corrected reference blood values. Similar errors occurred when analyzing blood samples that were agglutinated in vitro by incubating erythrocytes with incompatible plasma. The counting and sizing errors observed with electronic counting techniques were eliminated or greatly reduced by incubating blood in cell counting diluent for 10 minutes followed by analysis on a single channel counter with attached particle size analyzer. Error in erythrocyte measurement on a multichannel system may be anticipated if there is overt erythrocyte agglutination in a blood sample, an abnormally high MCHC is reported by the system, or subpopulations of large volume (agglutinated cells) are observed on a volume distribution histogram.     

L-苹果酸和柠檬酸联合对异育银鲫生长性能、抗氧化能力、血液和血清生化相关指标的影响

试验旨在研究柠檬酸(CA)和L-苹果酸(MA)联合对异育银鲫幼鱼生长性能、血清学指标、血液学指标和抗氧化指标的影响。选取7.5 g重复,每个重复16尾鱼。这5个组分别饲喂基础饲料(Ⅰ组),以及基础饲料中分别添加0.2%CA(Ⅱ组)、0.2%CA+0.1%MA(Ⅲ组)、0.2%CA+0.2%MA(Ⅳ组)、0.2%CA+0.4%MA(Ⅴ组),试验周期为60 d。结果表明:相比Ⅱ组,Ⅲ、Ⅳ组增重率和特定生长率显著升高(P0.05),最高值均出现在Ⅳ组,Ⅳ组白细胞总数(WBC)显著提高(P0.05),Ⅲ、Ⅳ组红细胞总数(RBC)、血红蛋白(HGB)含量均显著升高(P0.05),Ⅲ、Ⅳ、Ⅴ组平均血红蛋白浓度(MCH)显著升高(P0.05);但是相比于Ⅰ组,Ⅱ、Ⅲ、Ⅳ、Ⅴ组血小板(PLT)浓度均显著降低(P0.05);相比于Ⅱ组,Ⅲ、Ⅳ、Ⅴ组ALT、AST显著升高(P0.05),Ⅳ、Ⅴ组乳酸脱氢酶(LDH)活性显著提高(P0.05),Ⅳ组碱性磷酸酶(AKP)活性显著升高(P0.05),Ⅲ、Ⅳ组总蛋白(TP)、白蛋白(ALB)浓度显著升高(P0.05),AKP、ALB、TP最高值均出现在第Ⅳ组;与Ⅱ组相比,随着L-苹果酸浓度增高,Ⅲ、Ⅳ、Ⅴ组异育银鲫超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性均出现先上升后下降的趋势,最高值均出现在第Ⅲ组;结果证明适量添加柠檬酸和L-苹果酸可以显著提高异育银鲫的生长性能和抗氧化能力,但可能诱导肝损伤和引起血小板降低症。     

Hematologic reference intervals for koi (Cyprinus carpio), including blood cell morphology, cytochemistry, and ultrastructure

BACKGROUND: Hematologic data are used routinely in the health care of humans and domestic mammals. Similar data for fish are largely fragmentary or have not been collected. OBJECTIVES: The primary purpose of this study was to determine hematologic reference intervals for koi, an ornamental strain of the common carp (Cyprinus carpio). Secondarily, the morphology, cytochemical reactions, and ultrastructure of koi blood cells were characterized. METHODS: A CBC was performed manually on heparin-anticoagulated blood specimens using Natt and Herrick's diluent and a Neubauer-ruled hemacytometer. Leukocyte differential counts were done on Wright-Leishman- and Diff-Quik-stained blood smears. Cytochemical reactions of koi leukocytes were determined using commercial kits. Transmission electron microscopy was performed to characterize the ultrastructural features of koi blood cells. RESULTS: Hematologic reference intervals were established for healthy koi for PCV (30-34%), hemoglobin concentration (6.3-7.6 g/dL), RBC count (1.7-1.9 X 10(6)/ microL), WBC count (19.8-28.1 X 10(3)/ microL), RBC indices, and differential leukocyte counts. Lymphocytes were the predominant leukocyte (accounting for up to 80% of all leukocytes), whereas eosinophils were rare. Basophils were positive with PAS staining. Naphthol AS-D chloroacetate esterase activity was observed only in eosinophils. alpha-Naphthyl butyrate esterase and beta-glucuronidase activities were positive in monocytes. Some lymphocytes were reactive for alpha-naphthyl butyrate esterase and acid phosphatase activity. Ultrastructurally, leukocytes, erythrocytes, and thrombocytes were identified on the basis of cytoplasmic organelles and granule appearance. CONCLUSION: Hematologic reference intervals and knowledge of the cytochemical reactions and ultrastructural characteristics of koi leukocytes will help standardize hematologic studies in this species.     

Glycophorin A-knockout mice, which lost sialoglycoproteins from the red blood cell membrane, are resistant to lethal infection of Babesia rodhaini

Recent in vitro-based studies using several Babesia spp. have suggested that sialic acids and/or sialoglycoproteins on host red blood cells (RBCs) play an important role in their invasion of RBCs. In the present study, we analyzed the RBC characteristics of glycophorin A (GPA)-knockout mice and studied their in vivo susceptibility to lethal infection of Babesia rodhaini for the first time. In immunoblot and lectin blot analyses, glycoproteins containing O-linked oligosaccharides terminated with alpha2-3-linked sialic acids disappeared from the RBCs of GPA homozygous ((-/-)) mice. Flow cytometric analysis showed a remarkable reduction of Maackia amurensis lectin II binding to the surface of GPA(-/-) RBCs relative to control RBCs, indicating an appreciable loss of alpha2-3-linked sialic acids on the RBC surface of GPA(-/-) mice. Importantly, while B. rodhaini caused lethal infection in wild-type mice, the infected GPA(-/-) mice showed inhibition of parasite growth and eventually survived. These results indicate that RBC sialoglycoproteins lost in GPA(-/-) mice are involved in the in vivo growth of B. rodhaini, probably functioning as essential molecule(s) for the parasite invasion of host RBCs in the blood circulation.     

Age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle

It is well known that the immune system changes with age during development and maturation in Holstein cattle. But age-related changes in leukocytes and T cell subsets in peripheral blood of Japanese Black cattle still remain unclear. The aim of the present study was to investigate comparative changes of leukocytes (granulocytes, monocytes, B cells and T cells) and T cell subsets (CD4⁺, CD8⁺, γδ, CD8⁺γδ and WC1⁺γδ T cells) in Japanese Black cattle aged 0.5, 1, 2, 6, 18 and 36–41 (adult) months on flow cytometry using specific monoclonal antibodies for the cell surface markers. T cell proportion was approximately 40% in 2-month-old cattle and decreased to 20.6% in adults. In contrast, B cell proportion significantly increased from 7.4% to 28.2% with age. In T cell subsets the percentage of CD4⁺ T cells significantly increased from 40.5% to 60%, but that of WC1⁺γδ T cell subset significantly decreased with age. The percentages of CD8⁺ and CD8⁺γδ T cells did not change. The present study details the proportional changes in leukocyte and T cell subsets with age in the peripheral blood of Japanese Black cattle and these findings are similar to those described for Holstein cattle.     

Canine PHA-stimulated adherent cell enhance interferon-gamma production and proliferation of autologous peripheral blood mononuclear cells

Dendritic cells are specialized antigen‐presenting cells with immuno‐modulating functions that are attractive for clinical applications for cancer immunotherapy. This study examined immunostimulatory functions of phytohemagglutinin (PHA)‐stimulated adherent cells (PHA‐Ad cells) from peripheral blood mononuclear cells (PBMCs) in dogs. PHA‐Ad cells enhanced interferon‐γ from autologous PBMC in vitro. PHA‐Ad cells also stimulated antigen‐independent proliferation of peripheral blood lymphocytes. These results suggest that PHA‐Ad cells from PBMC possess a stimulatory function to evoke anti‐tumour immunity and that they demonstrate potential for therapeutic applications in dogs.     

琼胶多糖对异育银鲫生长、体组成和抗氧化能力的影响

为评估琼胶粉替代异育银鲫饲料中淀粉对异育银鲫生长和抗氧化性能的影响,试验以不含琼胶粉的基础饲料为对照组饲料,以1％(D1组)、3％(D2组)、5％(D3组)琼胶粉替代淀粉配制成的饲料为试验组饲料(所有饲料均等氮等脂),饲喂规格一致的异育银鲫幼鱼[初始重(10.32±0.16)g],每组3个重复,每个重复25尾试验鱼,饲...     

牛膝多糖对脂多糖免疫应激仔猪血液生化指标和白细胞分类计数的影响

试验旨在探讨牛膝多糖(ABPS)对脂多糖(LPS)刺激的断奶仔猪血液生化指标和白细胞分类计数的影响。选用48头(28±3)d、体重(8.45±0.14)kg的杜洛克×长白×大白断奶仔猪,采用2×2因子设计,两个因子分别为不同日粮处理(0或500mg/kgABPS)和免疫应激(注射LPS或生理盐水)。试验期28d。在第14天和第21天,每日粮组的一半猪注射100μg/kgBW的LPS,另一半注射生理盐水作对照。注射后3h,采血测定血液生化指标,并进行白细胞分类计数。结果表明:LPS刺激提高了第14天和第21天血清中碱性磷酸酶、谷草转氨酶(GOT)的活性及谷草转氨酶与谷丙转氨酶的比值(P0.05),并降低了第21天血清总蛋白和白蛋白的含量(P0.05);ABPS提高了第14天血清总蛋白和球蛋白的含量(P0.05)。在第14天,LPS刺激降低了白细胞、淋巴细胞、单核细胞和中性粒细胞的含量及单核细胞比例(P0.001)。在第21天,LPS刺激降低了白细胞、淋巴细胞的含量及比例,而提高了中性粒细胞比例(P0.001)。ABPS提高了白细胞(14d:P0.10)和中性粒细胞(14d:P0.05;21d:P0.05)的含量及中性粒细胞比例(14d:P0.05;21d:P0.10),降低了淋巴细胞比例(14d:P0.05;21d:P0.10)。结果提示,LPS可引起仔猪血液生化指标和白细胞分类计数发生显著变化,ABPS在一定程度可缓解上述现象,表现出一定的改善免疫和缓解炎症的作用。