Heinz body haemolytic anaemia in a dog secondary to ingestion of a zinc toy: a case report

A 6-year-old Labrador retriever was referred for investigation of severe lethargy and suspected immune-mediated haemolytic anaemia. Clinical examination revealed pale mucous membranes and jaundice. Haematology demonstrated large numbers of Heinz bodies and a marked anaemia, which was strongly regenerative. Serum zinc concentrations were markedly elevated. Analysis of a metal toy vomited by the dog 3 days prior to presentation revealed it to be composed of almost pure zinc. A diagnosis of haemolytic anaemia secondary to acute zinc toxicity was made and supportive therapy instigated. There was a subsequent decrease in numbers of Heinz bodies and a rise in the haematocrit, and the dog made an uneventful recovery. Acute zinc toxicity resulting in haemolytic anaemia is rarely observed, and this case was also unusual in that the main clinicopathological finding was the presence of numerous Heinz bodies without other evidence of oxidative damage to red blood cells.     

Hematology of equine fetuses with comparisons to their dams

The Hematologic values of 19 equine fetuses between 202 and 238 days gestation were compared with those of their dams. The red blood cell (RBC) count, hemoglobin concentration, hematocrit, and mean corpuscular hemoglobin concentration were significantly lower in fetal blood, while the mean corpuscular volume, mean corpuscular hemoglobin, and red cell distribution width were significantly higher. Mares had a significantly higher nucleated blood cell count than fetuses, and all nucleated cells were leukocytes (WBC). Most WBC in mare blood were segmented neutrophils and lymphocytes. In contrast, over one-half of the nucleated cells in fetal blood were nucleated RBC, and the majority of WBC in fetal blood were lymphocytes. Mares also had significantly higher plasma protein and fibrinogen concentrations than their fetuses. Mild macrocytosis and mild polychromasia were observed in most fetal blood samples, but not in blood samples from mares. All fetal blood contained reticulocytes, and most samples contained Heinz bodies and Howell-Jolly bodies. The results of this study will contribute to the development of hematologic reference values that may be useful in equine fetal research and, possibly, in the diagnosis of equine fetal disease.     

Morphological lesions in red blood cells from herring gulls and Atlantic puffins ingesting Prudhoe Bay crude oil

Red blood cells from nestling herring gulls and Atlantic puffins that had ingested 10 ml or more of a Prudhoe Bay crude oil/kg body weight/day for four to five days were examined by light and electron microscopy. In stained smears, red blood cells from oil-dosed birds were characterized by anisocytosis, poikilocytosis, reticulocytosis, and Heinz body formation. In transmission electron micrographs, affected cells had intracytoplasmic and intranuclear Heinz bodies, a variety of abnormal cytoplasmic vesicles, degenerate mitochondria, absence of circumferential microtubules, abnormal shape, and crenulation of the plasma membrane. The latter two cell surface anomalies were evident in scanning electron micrographs. Identical lesions were present in red cells from gulls injected with phenylhydrazine. Reticulocytosis was the only change evident in blood from gulls made anemic by hemorrhage. These observations support the hypothesis that the toxicity of ingested Prudhoe Bay oil to red cells was exerted by oxidant chemical compounds.     

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。     

Fluorescein diacetate staining of Anaplasma marginale as a possible measure of viability

Fluorescein diacetate viability-staining technique was applied to Anaplasma marginale. Marginal bodies took up the stain and became fluorescent. When infected red blood cells were incubated at 55 C, the number of fluorescent Anaplasma decreased at a rate similar to the known loss of viability.     

Myospherulosis in the subcutis of a pony.

Myospherulosis was diagnosed in a mature pony. Several parent bodies containing many spherules were observed microscopically in biopsy material from an area of cellulitis. The spherules are altered red blood cells that form as the result of prolonged contact with necrotic fat or petrolatum-based ointments. These structures must be differentiated from fungal elements. The recommended treatment is surgical excision.     

Ultrasonographic and computed tomographic features of rice bodies in an Arabian horse with atlantal bursitis

A 19‐year‐old castrated Arabian male horse presented for evaluation of a firm mass at the dorsal cervical region. Ultrasonography and computed tomography revealed multiple well defined fusiform structures within the atlantal bursa. Multiple glossy smooth, white to yellowish, flattened fusiform structures were removed surgically. These structures were composed of dense fibrin with some leukocytes and red blood cells. The imaging and histopathological features of these structures were similar to chronic ‘rice bodies’ reported in humans with bursitis or tenosynovitis. This is the first veterinary report describing the imaging features of ‘rice bodies’ in a horse with atlantal bursitis.     

Erythropoiesis regulation during the development of ascites syndrome in broiler chickens: a possible role of corticosterone

The ascites syndrome in broiler chickens is attributed to metabolic burdening, which results from intensive genetic selection for rapid growth coupled with exposure to extreme environmental conditions, such as low ambient temperature. These conditions impose on the broilers difficulties in fulfilling tissue demands for oxygen, and the birds exhibit a decrease in blood oxygen saturation and high hematocrit values. It is unknown whether the increase in hematocrit results from alteration in erythropoiesis or from fluid exudation out of the blood system to the abdominal cavity. The present study was conducted to examine the association between abnormal stress response and erythropoiesis process in ascitic broilers. Ascitic chickens revealed a uniquely continuous stress response: expressing an increase (P < or = 0.05) in plasma corticosterone concentration 2 to 3 wk before death. At 5 wk of age, ascitic broilers exhibited an increase (P < 0.05) in hematocrit, blood cell count, and packed cells and blood volumes, with no significant change in plasma volume. These results confirm an accelerated erythropoiesis process in ascitic birds. Increased blood cell production in ascitic birds was matched by an increase (P < 0.05) in the proportion of immature red blood cells (23%) in comparison with broilers that remained healthy (7%), and by decreased (P < 0.05) hemoglobin content relative to red blood cells. We conclude that continually increased corticosterone concentrations, as an inducer of erythropoiesis proliferation and differentiation arrest, in ascitic chickens, resulted in increased production of red blood cells (partially immature) with decreased hemoglobin content; this decrease in hemoglobin might have contributed to enhanced development of hypoxemia and to aggravation of the syndrome.     

Immunohistology of the splenic compartments of the one humped camel (Camelus dromedarius)

The cellular composition of the different splenic compartments is well characterized in several species, but the spleen of the camel has not been studied due to the lack of specific antibodies detecting its leukocyte subsets. Therefore, 5microm frozen sections from 15 camel spleens (0.5-15 years) were studied for acid and alkaline phosphatases and for cross-reaction with antibodies specific for bovine (n=181), swine (n=14) and human (n=6) leukocyte determinants. Fifteen antibodies cross-reacted with camel spleen cells. These included 13 anti-bovine, two anti-human, but no anti-swine antibodies. The lymph follicles mainly consisted of B cells. The germinal centers showed a strong alkaline phosphatase activity. The periarterial lymphatic sheath harbored T lymphocytes. The marginal zone contained gammadelta T cells, CD45R0+, MHC class II DR+, CD44+, IL-A 24+ cells and few macrophages. The red pulp contained B, T, MHC class II DR+, IL-A24+ and gammadelta T cells and few macrophages. The periarterial macrophage sheaths contained many more macrophages than the marginal zone, so they may play a central role in the phagocytosis of the blood born particles. The alkaline phosphatase probably labeled activated B cells, but in contrast to other species no positive cells were found in the marginal zone. In general, lymphocyte compartmentalization in the camel spleen is similar to that in other species except for lower numbers of macrophages and the absence of alkaline phosphatase positive cells in the marginal zone. No age related differences were observed in the splenic compartments.     

Presence of bluetongue virus in the marginal zone of the spleen in acute infected sheep

Bluetongue virus, a member of the genus Orbivirus of the family Reoviridae, is the causative agent of bluetongue, which is a non-contagious Culicoides mediated blood-borne disease. The present study characterizes the pathogenicity of a Taiwan prototype BTV2/KM/2003 in Corriedale sheep inoculated subcutaneously into the ear pinna. Histologically, multifocal petechiated hemorrhage, with mild to moderate inflammation and edema, were present in the contralateral ear pinna, tongue, and facial skin, without remarkable lesions in lymphoid organs. By days post-infection (DPI) 7, viral VP7 antigen, detected by immunohistochemistry, presented in the spleen, chiefly located in the outer rim of <3 cell thickness of marginal zone macrophages bordering the marginal zone and red pulp, and T lymphocytes of the red pulp. By DPI 11, viral signals shifted from the marginal zone to macrophages and small lymphocytes within follicles of the spleen. In situ hybridization with VP7 gene probe detected strong signals in the spleen, chiefly spanning the whole width of 5-10 cell thickness of the marginal zone, including the marginal zone macrophages and marginal zone B cells, as well as macrophages of sheathed capillaries in the red pulp. This study demonstrates molecular as well as morphologic evidence of the presence of bluetongue virus in the marginal zone of the spleen, most likely associated with viremia in acute infection, as previously demonstrated by the authors.     

Nucleic acid distribution pattern in avian erythrocytes and mammalian lymphocytes: Comparative studies by fluorescence microscopy and digital imaging analytical techniques

Nucleated erythrocytes of healthy domestic chicken and ducks, and lymphocytes of healthy Sprague Dawley rats were evaluated for nucleic acid distribution pattern, employing light and fluorescence microscopy procedures, as well as digital imaging analytical methods. The results demonstrate a unique organization of nuclear DNA of mature chicken and duck erythrocytes, as well as immature duck erythrocytes, as delineated spherical nuclear bodies that mostly corresponded with euchromatin zones of the cells in routine Wright-stain blood smears. The nuclear DNA of the rat lymphocytes, on the other hand, was observed as a more diffuse green fluorescing nuclear areas, with punctate variably-sized diffuse areas of RNA red fluorescence. RNA red color fluorescence was also evident in the narrow cytoplasm of the lymphocytes, especially in large lymphocytes, in comparison with the cytoplasm of the mature avian erythrocytes that completely lacked any nucleic acid fluorescence. Nuclear RNA fluorescence was lacking in the mature chicken erythrocytes, compared with those of the mature and immature duck erythrocytes as well as lymphocytes of both avian and rats blood. The significance of these findings lies in the establishment of normal benchmarks for the nuclear and cytoplasmic nucleic acid pattern in eukaryotic cells. These normal benchmarks become valuable in rapid diagnostic situations associated with pathologies, such as the presence of viral nuclear and cytoplasmic inclusion bodies that can alter the nucleic acid pattern of the host cells, and in conditions of cellular abnormal protein aggregations. Variability of cellular nucleic acid pattern can also aid in prognostic assessments of neoplastic conditions.     

Nucleic acid distribution pattern in avian erythrocytes and mammalian lymphocytes: comparative studies by fluorescence microscopy and digital imaging analytical techniques

Nucleated erythrocytes of healthy domestic chicken and ducks, and lymphocytes of healthy Sprague Dawley rats were evaluated for nucleic acid distribution pattern, employing light and fluorescence microscopy procedures, as well as digital imaging analytical methods. The results demonstrate a unique organization of nuclear DNA of mature chicken and duck erythrocytes, as well as immature duck erythrocytes, as delineated spherical nuclear bodies that mostly corresponded with euchromatin zones of the cells in routine Wright-stain blood smears. The nuclear DNA of the rat lymphocytes, on the other hand, was observed as a more diffuse green fluorescing nuclear areas, with punctate variably-sized diffuse areas of RNA red fluorescence. RNA red color fluorescence was also evident in the narrow cytoplasm of the lymphocytes, especially in large lymphocytes, in comparison with the cytoplasm of the mature avian erythrocytes that completely lacked any nucleic acid fluorescence. Nuclear RNA fluorescence was lacking in the mature chicken erythrocytes, compared with those of the mature and immature duck erythrocytes as well as lymphocytes of both avian and rats blood. The significance of these findings lies in the establishment of normal benchmarks for the nuclear and cytoplasmic nucleic acid pattern in eukaryotic cells. These normal benchmarks become valuable in rapid diagnostic situations associated with pathologies, such as the presence of viral nuclear and cytoplasmic inclusion bodies that can alter the nucleic acid pattern of the host cells, and in conditions of cellular abnormal protein aggregations. Variability of cellular nucleic acid pattern can also aid in prognostic assessments of neoplastic conditions.     

Determination of red blood cell potassium content in horses with diarrhoea: A practical approach for therapy

The concentration of potassium in the red blood cells in 15 horses with severe diarrhoea was determined. The red blood cell potassium content (RBCK⁺) was low compared with levels obtained in normal horses. The decrease in the RBCK⁺ was used to evaluate the total body potassium deficit. This calculated amount was administered orally, following each RBCK⁺ determination. In those horses which recovered, RBCK⁺ content returned to normal levels. During treatment, refilling of the red blood cells with potassium occurred only gradually as long as the diarrhoea persisted. In each case, oral loading with the calculated dose resulted in an improvement of the general condition and no side effects were observed. The importance of using RBCK⁺ levels as a diagnostic and therapeutic parameter is discussed.     

Observations on erythrocytic acetylcholinesterase activity in experimentally induced anaplasmosis in calves

Erythrocytic acetylcholinesterase (AChE) activity was assessed in splenectomized bovine calves inoculated with Anaplasma marginale. Other parameters, i.e., haematological values (haemoglobin, packed-cell volume, and total erythrocytic count), percentage parasitaemia and the ensuing host reactions during Anaplasma infection were also studied in relation to AChE activity. There was significant inhibition of red blood cell-AChE activity commencing soon after inoculation of infected blood; this preceded a substantial decrease in haematological values along with high parasitaemia. The changes in AChE activity may possibly be associated with increased cell membrane permeability, thus triggering the entry of A. marginale organisms into red blood cells.     

Malignant stromal cell tumor of the spleen in a WBN/Kob rat

Primary splenic stromal tumors have rarely been reported in rodents. We report the case of a 90-week-old male WBN/Kob rat with a nodular demarcated mass in the spleen, which was kept as a non-treated animal in a long-term animal study. Histopathology revealed round to short spindle-shaped tumor cells arranged in a solid growth pattern. Invasive growth, anisokaryosis, and high mitotic activity (46 per 10 high-power fields [2.37 mm²]) were observed to be multifocal, but most tumor cells showed mild nuclear pleomorphism. The pattern of silver impregnation corresponded to that of the marginal zone of the red pulp. Immunohistochemistry revealed that the tumor cells were double positive for fascin and desmin and focally positive for Iba-1 and OX-6 expression. These characteristics were similar to those observed in fibroblastic reticular cells and dendritic cells in the marginal zone of the red pulp. These findings suggest that the malignant stromal cell tumor of the spleen in this case had characteristics of both fibroblastic reticular cells and dendritic cells.     

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.     

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.     

猪脾脏结构和功能及其免疫关键基因研究进展

脾脏作为猪最大的次级淋巴器官,含有多种免疫活性细胞和免疫因子,是先天性免疫和适应性免疫重要的应答场所,具有广泛的免疫调控功能;同时可清除衰老、损伤红细胞及过滤病原体,并在造血和储藏血细胞方面具有重要作用。这些功能与脾脏的结构密切相关,其中红髓主要作为血液滤过器,执行造血、储血和清除异物的功能;白髓是免疫的主要区域,含有多种免疫细胞,可执行特异性免疫应答调控功能;而边缘区是连接两个区室的重要桥梁,使所有细胞和抗原都能通过其进入脾脏不同部位。不同的发育和免疫关键基因是实现脾脏功能的根本,发育基因的表达保证了其结构的完整,为脾脏执行各种功能提供空间。免疫关键基因的表达确保了免疫反应进行,其中不同模式识别受体基因的表达保证了先天性免疫的吞噬和识别应答,而各种免疫细胞分泌的多种细胞因子和免疫活性物质是获得性免疫反应的基础,实现机体清除和监控抗原的生理过程。随着脾脏研究的深入,对其各种免疫细胞和免疫功能有了新的认识。作者首先阐述了脾脏组织不同区域的生物学功能;然后结合机体免疫应答机制,论述脾脏先天性和特异性免疫功能以及所需免疫细胞和免疫因子的作用与关联;最后简单介绍了脾脏胚胎期和出生后期的组织发育和免疫相关基因,总结了猪脾脏先天性免疫中重要的模式识别受体及其基因家族,为研究猪脾脏先天性免疫功能提供参考。     

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.     

An enzyme-linked antiglobulin test for the detection of erythrocyte-bound antibodies in canine autoimmune haemolytic anaemia

A method has been developed which allows identification and quantitation of red cell-bound immunoglobulins and complement in canine blood. The technique utilizes ELISA methodology and the assay identifies cases of autoimmune haemolytic anaemia which are negative by the Direct Coombs test. Further, the amount of antibody present on the red cells shows a close correlation to the haemoglobin level; suggesting that the degree of sensitization of the RBCs influences their rate of destruction. Favourable response to treatment correlates with a decrease in the levels of bound antibody and complement.