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.     

Cytochemical staining characteristics of chicken heterophils and eosinophils

Cytochemistry was used to differentiate chicken heterophils and eosinophils in blood smears and cytospin preparations of isolated leukocytes. Cytochemical staining also was performed to evaluate any alteration of cell staining characteristics after heterophils had been isolated using discontinuous Ficoll-Hypaque gradients. Cytochemical staining reactions of heterophils and eosinophils were the same before and after discontinuous gradient cell separation. Characteristic positive reactions did not occur when chicken heterophils were stained with alkaline phosphatase, peroxidase, Sudan black B, acid phosphatase, alpha-naphthyl acetate esterase, naphthol AS-D chloroacetate esterase, periodic acid-Schiff, or Luna's eosinophil granule techniques. Chicken eosinophils stained positively when peroxidase, Sudan black B, acid phosphatase, and Luna methods were performed. In highly cellular cytospin preparations, peroxidase reactions readily distinguished the few contaminating eosinophils (< 1%) from the heterophils.     

鸡实验性成髓细胞性白血病的细胞化学及超微结构

用鸡成髓细胞性白血病病毒（ＡＭＶ）ＢＡＪ－Ａ株鸡传代血浆毒感染１日龄伊莎鸡雏２０只，在接毒后１９￣２５ｄ于濒死期扑杀、采样，进行细胞化学和电镜检查。结果表明，肝、肾、法氏囊、胸腺及血液的成髓细胞来源于骨髓；其成髓细胞胞浆、胞膜及细胞间隙存在带有囊膜的含核心的病毒粒子。     

Morphologic and immunologic characterization of a canine isthmus mural folliculitis resembling pseudopelade of humans

The clinical, histopathologic and immunopathologic features of a novel form of isthmus mural folliculitis in dogs, which resembles pseudopelade in humans, were characterized. Clinically, dogs exhibited variably distributed foci of alopecia that persisted without treatment or did not respond to immunosuppressive therapy. Histopathologically, mixed mononuclear cell infiltrates, largely lymphocytes, infiltrated the follicular isthmus. Occasionally, inflammation extended above and below the follicular isthmus but did not involve the hair bulb or the epidermis. Severe follicular atrophy and variable atrophy of sebaceous glands occurred in all dogs. Folliculotropic lymphocytes exhibited most commonly CD3 and CD8 (cytotoxic T cells). Autoantibodies specific for the lower hair follicle were detected in the serum of affected patients. Western immunoblotting demonstrated binding of these antibodies to multiple follicular keratinocyte proteins, including hair keratins and trichohyalin. Lack of hair regrowth (in contrast to canine alopecia areata), as well as location of inflammation and extreme atrophy of adnexal units are similar to findings seen in human pseudopelade.     

Bald thigh syndrome of Greyhound dogs: gross and microscopic findings

Bald thigh syndrome (BTS) is a disease limited to Greyhound dogs. It is characterized clinically and grossly by bilateral hair loss on the lateral and caudal thighs. The cause of BTS is unknown but may be associated with hypothyroidism or hyperadrenocorticism. Samples of skin, thyroid glands, and adrenal glands from 43 Greyhound dogs with BTS were examined microscopically. Microscopic changes were characterized by dilatation of follicular infundibula, presence of catagen follicles and epidermal hyperplasia. Changes in the skin from these Greyhound dogs suggest an endocrinopathy as the cause; however, we were unable to confirm which one.     

Grey eosinophils in a Miniature Schnauzer with a poorly differentiated mast cell tumor

A 10‐year‐old female spayed Miniature Schnauzer was presented for investigation of an intra‐nasal mass. The mass was diagnosed by histopathologic examination as an undifferentiated round cell neoplasm with an infiltrate of segmented leukocytes, interpreted as neutrophilic inflammation. The mass was treated with palliative radiation and systemic chemotherapy due to the presence of regional lymph node metastasis. During subsequent monitoring over several months, the peripheral leukocyte concentration was repeatedly within reference intervals to slightly increased with low numbers of toxic neutrophils. Four months after the initial diagnosis, there was a significant leukocytosis of 66 100 cells/μL, and 39 700 cells/μL of the leukocytes had variably mature, lobulated, and hypolobulated nuclei, and grey cytoplasm with clear vacuoles, resembling grey eosinophils. To further characterize these cells, peripheral blood smears from the patient and a canine control with eosinophilia were stained for alkaline phosphatase (AP), peroxidase, and esterase activities, and with Luxol fast blue (LFB). Histopathologic sections of the nasal mass were stained with LFB and immunohistochemically for tryptase. On blood smears, the cytoplasm of the suspected grey eosinophils stained for AP and granules stained with LFB confirmed that there was an eosinophilic lineage. Peroxidase staining was weak, and esterase staining was absent. On histopathologic sections from the nasal mass, the segmented leukocytes contained LFB‐staining granules, indicating an eosinophilic infiltrate was present. Neoplastic cells expressed tryptase, which confirms a mast cell lineage. Our findings suggest that grey eosinophils might be under‐recognized and interpreted incorrectly as toxic neutrophils. This report expands the canine breeds in which these eosinophils have been identified.     

Acute monoblastic leukemia in a FeLV-positive cat

A 1.6-year-old male domestic short hair cat was brought to the Veterinary Medical Teaching Hospital, Kasetsart University, with signs of severe anemia, depression, and general lymph node enlargement. Complete blood count revealed leukocytosis and massive undifferentiated blasts. Testing for antibodies specific to feline leukemia virus (FeLV) was positive, and FeLV nucleic acid was confirmed by nested polymerase chain reaction. Base on cytochemistry and ultrastructure, the cat was diagnosed with acute monoblastic leukemia.     

Hematology and clinical chemistry of adult yellow-headed temple turtles (Hieremys annandalii) in Thailand

Background: Yellow‐headed temple turtles (YHT), Hieremys annandalii, native to Thailand, are protected from exploitation under the Wild Animal Reservation and Protection Act, also listed under Appendix II of the Convention on International Trade of Endangered Species and the International Union for the Conservation of Nature red list. Objectives: The objectives of this study were to describe quantitative, morphologic, and cytochemical features of blood cells and plasma biochemical analytes of clinically healthy YHT. Methods: Blood samples were collected from 40 adult YHT from October 2007 to February 2008. Hematologic and biochemical analyses, cytochemical staining, and ultrastructural evaluation were performed using standard methods. Results: Hematologic results (mean ± SD) included: RBC count, 0.275 ± .094 × 10⁶ cells/μL; WBC count, 11.7 ± 6.6 × 10³ cells/μL; heterophils, 29.4 ± 6.9%; eosinophils, 23.7 ± 5.3%; basophils, 21.2 ± 1.9%; lymphocytes, 14.8 ± 5.9%; and azurophils, 10.7 ± 5.3%. Erythrocytes stained dark red with peroxidase‐staining. Periodic acid‐Schiff stain could not differentiate between thrombocytes and lymphocytes. Thrombocytes contained cytoplasmic vacuoles, similar to mammalian platelets and those of birds and snakes. Heterophils and eosinophils were similar in structure and cytochemical staining characteristics to those of other turtles and reptiles. Structure of basophils was similar to avian basophils. Lymphocytes and azurophils had similar cytochemical staining compared with mammalian lymphocytes and monocytes. Mean MCHC, WBC counts, absolute azurophil counts, and plasma alanine aminotransferase activity were higher in male turtles than in females. Conclusion: Blood characteristics of YHT are species‐specific, and this study can be served as a reference for future clinical studies and medical care of YHT.     

Leukocyte and thrombocyte reference values for channel catfish (Ictalurus punctatus Raf), with an assessment of morphologic, cytochemical, and ultrastructural features

BACKGROUND: Hematology tests are useful to evaluate physiologic disturbances in fish and can provide important information for the diagnosis and prognosis of disease. OBJECTIVES: The primary purpose of this study was to define reference intervals for thrombocytes and leukocytes in healthy channel catfish (Ictalurus punctatus). In addition, the morphologic, cytochemical, and ultrastructural features of blood cells were assessed. METHODS: Blood samples (0.5 mL) were collected into EDTA from 40 clinically healthy catfish on a commercial fish farm in Jaboticabal, Brazil. Thrombocyte, total WBC, and differential WBC counts were determined and reference intervals were calculated as the 25-95th percentiles of data. Thrombocyte and leukocyte morphology was assessed in blood smears stained with May Grünwald-Giemsa-Wright and ultrastructurally by transmission electron microscopy. Cytochemical staining patterns were described using periodic acid-Schiff (PAS), peroxidase, nonspecific esterase, alkaline phosphatase, and toluidine blue. RESULTS: Reference intervals were as follows: thrombocytes 58,802-99,569/microL; total WBCs 27,460-41,523/microL; lymphocytes 5380-11,581/microL; monocytes 2949-7459/microL; neutrophils 12,529-22,748/microL, and basophils 736-2003/microL. Neutrophils were positive for peroxidase and PAS; monocytes were positive for nonspecific esterase; and basophils were positive with toluidine blue. CONCLUSION: The morphologic and staining features of neutrophils and monocytes of channel catfish are similar to those of mammals, and the presence of basophils in this species was verified. These reference intervals and morphologic findings provide a foundation for future investigations on the functions and alterations of blood cells in channel catfish.     

Hematology, leukocyte cytochemical analysis, plasma biochemistry, and plasma electrophoresis of wild-caught and captive-bred Gila monsters (Heloderma suspectum)

Background: The Gila monster (Heloderma suspectum), one of several venomous lizard species in the world, is found within the United States and Mexico and is recognized as an iconic symbol of the American Southwest. Thus, Gila monsters are of growing interest in the captive reptile trade and within zoological and educational institutions. Objectives: The aims of this study were to determine results for CBCs, describe cytochemical reactions in WBCs, and obtain plasma biochemical and protein electrophoresis results from wild‐caught and captive‐bred H. suspectum. Methods: Ventral tail vein blood samples were obtained from 16 captive (14 wild‐caught and 2 captive‐bred) Gila monsters. CBCs, RBC morphometric analysis, plasma biochemical analysis, and protein electrophoresis were performed. Leukocytes were stained for peroxidase, Sudan black B (SBB), chloroacetate esterase, napthyl butyrate esterase, and leukocyte alkaline phosphatase, and blood smears were examined for the presence of hemoparasites. Results: The median (range) PCV was 37% (22–50%) and WBC count was 4.6 × 10³/μL (3.3–6.4 × 10³/μL) with approximately 50% heterophils and fewer lymphocytes, basophils, azurophils, and monocytes in decreasing order. Cytochemical reactions were unique among reptiles with strong staining for peroxidase and SBB in monocytes/azurophils. Biochemical results were similar to those of earlier reports with slight increases in uric acid and urea concentrations. Plasma electrophoretic results indicated that albumin was approximately equal to the combined globulin fractions. Conclusions: Results of blood analysis in healthy wild‐caught and captive‐bred H. suspectum may be useful for monitoring health status in this species.     

雏鸵鸟睾丸的结构及睾丸内神经肽Y的表达

采用光镜和电镜技术,观察雏鸵鸟睾丸的显微和超微结构;采用免疫组织化学方法对NPY在睾丸内的表达进行研究,探明NPY在睾丸内的表达规律.光镜下可见,雏鸵鸟睾丸表面有一层由坚韧的结缔组织构成的白膜,它向睾丸实质内形成的结缔组织不发达,不形成睾丸纵隔和睾丸小隔;雏鸵鸟的曲精小管直径为85～90 μm,生精上皮由支持细胞和1～2层生精细胞构成,此时精原细胞已开始分裂增殖,可见明显的分裂相,初级精母细胞数量少,体积大.电镜下可见,精原细胞胞质内有少量线粒体、游离核糖体和内质网;初级精母细胞富含线粒体、游离核糖体、内质网和溶酶体;支持细胞与精原细胞之间界限清晰,胞质内含较多游离核糖体,其他细胞器不发达;间质细胞的胞核形状不规则,内含较多脂滴.免疫组化显示,NPY在曲精小管周围、睾丸小血管周围、精母细胞和睾丸间质内均有表达,在睾丸小血管周围和精母细胞内表达量较高,而在曲精小管周围和睾丸间质内表达量较低.     

Morphologic and cytochemical characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

A morphologic classification based on the cytochemical characteristics of blood cells of 35 juvenile loggerhead sea turtles (Caretta caretta) is described. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff, and toluidine blue. The morphologic characteristics of erythrocytes were similar to those reported in green turtles. Six types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes, monocytes and thrombocytes. Except for the basophils, the rest of the white blood cells from loggerhead turtles had different cytochemical characteristics compared to blood cells from other sea turtle species. The leukocyte differential count was different from that reported for other sea turtle species. Heterophils were the most numerous leukocytes from these loggerhead turtles, followed by lymphocytes, eosinophils, monocytes and basophils. This paper provides a morphologic classification of blood cells of loggerhead sea turtles that is useful for veterinary surgeons involved in sea turtle conservation.     

德国牧羊犬精子超微结构及超低温冷冻对精子超微结构的影响

为了研究超低温冷冻前后德国牧羊犬精子超微结构的变化,采用扫描电镜和透射电镜对冷冻前后德国牧羊犬精子超微结构变化进行研究,结果显示在扫描电镜下,鲜精整体呈“蝌蚪状”,头部呈扁卵圆形.在透射电镜下精子纵切面头部呈“楔形”,头部细胞膜由外向内分别由质膜、顶体外膜、项体内膜和核膜组成.精子尾部中段横切面由外向内可见线粒体鞘膜、9束外周致密纤维,9对轴丝和2根中央微管.精子头长约为5.40 μm,头宽3.26 μm,颈长1.25 μm,颈宽0.55 μm,尾部中段长11.34 μm,中段直径0.87 μm,尾部长55.70 μm,线粒体螺旋数为44旋.鲜精和冷冻前精子头部、颈部和尾部形态变化差异均不显著(P＞0.05).超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数极显著高于鲜精和冻前精子数(P＜0.01).冻后顶体发生明显形态变化的精子占54.21％,高于发生颈部形态变化(10.61％)和尾部形态变化(28.83％)的精子数.结果表明鲜精和冻前精子形态变化不显著,而超低温冷冻后精子头部、颈部和尾部发生形态变化的精子数显著高于鲜精和冻前精子数.     

Morphologic changes of the anterior corneal epithelium caused by third eyelid removal in dogs

Morphologic changes of corneal epithelium after third eyelid removal were observed in six normal Beagle dogs. The observation was conducted on the 17th, 35th and 72nd week after the removal. Morphologic changes were observed on corneal epithelium layers at 17 and 35 weeks post excision of the third eyelid but they were limited to minor changes including abnormal intercellular adhesion and further exfoliation of superficial cells at 72 weeks. The reduced BUT and evidence of vital positive staining correspond with these morphologic changes. Therefore, these changes are thought to be important findings from a morphologic view point in identifying pathologic symptoms of the cornea using clinical evaluation methods such as BUT and vital staining method. Grossly, no clinical indication of ocular disease was indicated by slit-lamp biomicroscopic observation; however, the BUT shortage and positive finding of vital staining indicated that the corneal epithelium layers after third eyelid removal lacked the essential barrier function. Thus, it would seem appropriate to consider that such a condition of the ocular surface would worsen with excessive exposure of the ocular surface, palpebration disorders and other exogenous factors.