Antemortem diagnosis of canine neural angiostrongylosis using ELISA

A 5-month-old female Kelpie developed paraparesis, hind limb ataxia and spinal hyperaesthesia 4 days after ovariohysterectomy. Neurological examination demonstrated upper motor neuron signs in the pelvic limbs with lower motor neuron signs in the tail. Cerebrospinal fluid analysis demonstrated an increased protein concentration and marked eosinophilic pleocytosis. The dog was known to have eaten rats, snails and slugs. A tentative diagnosis of neural angiostrongylosis was made and later confirmed using an ELISA based on soluble antigens obtained from larval 4 Angiostrongylus cantonensis. Antibody titres from the patient's serum and CSF were 800 and 6400, respectively. The dog was treated successfully with prednisolone. ELISA testing of serum may provide a non-invasive means for diagnosing neural angiostrongylosis in dogs.     

Equine neural angiostrongylosis

Verminous encephalomyelitis due to Angiostrongylus cantonensis larvae was diagnosed in 2 foals at necropsy. The principal clinical feature was tetraparesis, although history and neurological examination revealed progressive and multifocal neurological disease. At presentation, a tentative diagnosis of parasitic larval migration involving the central nervous system (CNS), presumably due to Strongylus vulgaris, was proposed. Dissection of the spinal cord in one case resulted in recovery of intact larvae of both sexes of A. cantonensis. In both foals, histopathology of the brain and spinal cord revealed nematode sections which were consistent with A. cantonensis larvae.     

A case of canine angiostrongylosis.]

A six-month old bitch presenting a sub-lingual mucocele and hematoma associated with coagulation disorders died four days after the surgical treatment of the mucocele. The necropsy revealed a canine angiostrongylosis, a disease rarely seen in Switzerland. This article summarizes the biology of Angiostrongylus vasorum and describes the lesions and symptoms caused by this cardio-pulmonary helminthosis, as well as its diagnosis and treatment. The connections between angiostrongylosis and coagulation disorders are also discussed.     

Bone marrow contamination of canine cerebrospinal fluid

Bone marrow cells were identified in cytocentrifuge preparations of cerebrospinal fluid (CSF) obtained by lumbar punctures from two dogs. CSF analyses were characterized by normal or increased total cell counts, normal or increased total protein concentration and the presence of erythrocytes. Hematopoietic cells present included both erythroid and myeloid precursors and, in one case, an erythroblastic island was seen. Peripheral blood smear examination confirmed that the hematopoietic cells observed were not the result of blood contamination. Neither case was associated with a difficult tap procedure, nor with a specific disease process. Contamination may have resulted from actual marrow penetration or from extramedullary hematopoiesis in the dura mater. Accurate interpretation of CSF cytology requires the consideration of possible bone marrow contamination when hematopoietic cells are observed.     

Analysis of neurotransmitter metabolite concentrations in canine cerebrospinal fluid

The concentrations of dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindole-3-acetic acid (5-HIAA) in CSF obtained from the cisterna magna of 21 nonneurologically compromised dogs were determined by high-pressure liquid chromatography and electrochemical detection. A rapid method of sample preparation, which involved single filtration through a deproteinizing membrane, was used. Canine CSF obtained in this manner contained 5.78 +/- 0.78 ng of DOPAC/ml, 72.19 +/- 4.09 ng of HVA/ml, and 29.95 +/- 1.67 ng of 5-HIAA/ml. Linear regression analysis between HVA and 5-HIAA yielded a correlation coefficient of 0.4804. The neurotransmitter index, HVA/5-HIAA, was found to be more indicative of the dopaminergic metabolite HVA than the acid metabolite of serotonin, 5-HIAA (correlation coefficient with HVA = 0.5529 vs a correlation coefficient with 5-HIAA = -0.4462). A poor relationship (correlation coefficient = -0.1715) was found to exist between the 2 dopaminergic metabolites DOPAC and HVA in the CSF.     

Proteome analysis of cerebrospinal fluid in healthy beagles and canine encephalitis

We performed proteomics analysis of the cerebrospinal fluid (CSF) of healthy dogs and dogs with meningoencephalitis of unknown etiology (MUE). By comparing two-dimensional electrophoreses (2DE), an upregulated spot was found in MUE dogs. This protein was identified as a neuron-specific enolase (NSE) by analysis with MALDI-TOF mass spectrometry. In comparing dot blots using an antibody against NSE, the NSE levels in the CSF of MUE dogs was significantly higher than that of the controls. NSE is a diagnostic marker of neuroendocrine tumors, brain injury and spinal cord trauma in humans. It seems that the NSE concentration in the CSF is increased by cellular destruction in canine encephalitis. Though elevation of NSE may not be specific in canine encephalitis because the NSE level was increased in other CNS diseases, further study including measurement with serum is necessary.     

Haematological findings in captive dolphins and whales

Objective To examine haematological features in five species of healthy, captive marine mammals.
Animals Twenty bottlenose dolphins ( Tursips truncatus ), seven Pacific white-sided dolphins ( Lagenorhynchus obliquidens ), five Risso dolphins ( Grampus griseus ) and five false killer whales ( Pseudorca crassidens ).
Results and conclusion The red blood cell count was 4.21 × 10¹²/L in bottlenose dolphins, 5.32 × 10¹²/L in Pacific white-sided dolphins, 4.35 × 10¹²/L in Risso dolphins and 4.43 × 10¹²/L in false killer whales. The haemoglobin concentration was 1.51 g/L and packed cell volume 44.7% in bottlenose dolphins; the corresponding values were 1.71 g/L and 48.9% in Pacific white-sided dolphins, 1.72 g/L and 49.4% in Risso dolphins, and 1.52 g/L and 47.8% in false killer whales. The white blood cell count was 7.097 × 10⁹/L in bottlenose dolphins, 5.928 × 10⁹/L in Pacific white-sided dolphins, 5.001 × 10⁹/L in Risso dolphins and 7.921 × 10⁹/L in false killer whales. There were no significant differences in these values among bottlenose dolphins and Pacific white-sided dolphins. The proportion of eosinophils in the differential leukocyte count ranged from 10.3% to 11.5% in bottlenose dolphins, Pacific white-sided dolphins and false killer whales, but was only 0.4% in Risso dolphins. The eosinophilic granules were larger in Risso dolphins and false killer whales than in bottlenose and Pacific white-sided dolphins.     

Reassessment of cytologic values in canine cerebrospinal fluid by use of cytocentrifugation

Using cytocentrifugation, nearly one fourth of canine cerebrospinal fluid (CSF) samples with cell counts in the normal range had abnormalities in cell type or morphologic features. Cerebrospinal fluid samples from 145 dogs with neurologic disorders were evaluated by use of this method. These results indicate that low hemacytometer counts in canine CSF should not be interpreted as normal. By increasing the detection of abnormalities in CSF, cytocentrifugation might improve diagnosis and treatment of canine neurologic disease.     

High resolution protein electrophoresis of 100 paired canine cerebrospinal fluid and serum

This study was performed to investigate the diagnostic relevance of cerebrospinal fluid (CSF) high resolution electrophoresis. The laboratory technique was applied to 100 paired samples of canine CSF and serum, with paired samples tested during the same analytical run, as recommended in human medicine. Ninety four of the dogs had a neurological disease and 6 healthy dogs served as a control group. A strong linear correlation between CSF total protein concentration and the albumin quota (AQ) was found in the control group and in the inflammatory (infectious or noninfectious), neoplastic, and miscellaneous groups: AQ = 0.015 CSF total protein--0.102, r = 0.990. This correlation suggests that an increased CSF total protein concentration can be an indicator of blood brain barrier dysfunction. The highest median AQ value was found in the aseptic suppurative meningitis group, but no statistical differences were found between this and the other groups. The AQ, calculated with this technique, did not provide any additional information. Moreover, although unexpected, the electrophoretic profiles were not characteristic of any particular disease. In conclusion, this study did not confirm high resolution electrophoresis of paired CSF and serum samples to be a valuable ancillary diagnostic tool for canine neurological diseases.     

Evaluation of the ADVIA 120 for analysis of canine cerebrospinal fluid

BACKGROUND: Conventional techniques for canine cerebrospinal fluid (CSF) analysis require large sample volumes and are labor intensive and subject to operator variability. Objective: The purpose of this study was to evaluate the ADVIA120 CSF assay for analysis of canine CSF samples. METHODS: CSF samples collected from 36 healthy control dogs and 17 dogs with neurologic disease were processed in parallel using the automated assay and established manual methods using a hemocytometer and cytocentrifugation. Results for WBC (total nucleated cell) count, RBC count, and differential nucleated cell percentages were compared using Spearman rank correlation coefficients and Bland-Altman bias plots. RESULTS: Correlation coefficients for WBC and RBC counts were 0.57 and 0.83 for controls, and 0.92 and 0.94 for ill dogs, respectively. Coefficients for the percentages of neutrophils, lymphocytes, and monocytes were 0.53, 0.26, and 0.12 for controls and 0.77, 0.92, and 0.70 for dogs with neurologic disease. When data were combined (n=53), correlation coefficients were 0.86 and 0.91 for WBC and RBC counts, and 0.63, 0.43, and 0.30 for neutrophil, lymphocyte, and monocyte percentages. A 9.5% positive bias and 7.0% negative bias were obtained for the ADVIA 120 CSF assay for lymphocytes and macrophages in dogs with neurologic disease with Bland-Altman analysis. A 12.2% positive bias was found for lymphocyte percentage in dogs with neurologic disease. CONCLUSIONS: Manual and automated CSF assays had moderate to excellent correlation for WBC and RBC concentrations, but results were more variable for differential cell percentages. The ADVIA assay may be more useful for assessment of canine CSF with adjustment of cell differentiation algorithms.     

Prevalence and significance of extracellular myelin‐like material in canine cerebrospinal fluid

Background: Myelin‐like material in canine cerebrospinal fluid (CSF) specimens has been attributed to demyelinating or myelomalacic conditions. In our experience, myelin‐like material is observed frequently, especially in lumbar samples, and in a variety of disease conditions. Objectives: The objective of this study was to determine if there are associations between the presence of myelin‐like material and CSF collection site, body weight, underlying disease, and patient outcome. Methods: Wright–Giemsa‐stained cytocentrifuged specimens of CSF from the cerebellomedullary cistern (n=51) and lumbar cistern (n=47) of 98 dogs with neurologic disease were evaluated retrospectively for the presence and amount of extracellular myelin‐like material. Results were compared based on collection site, body weight, type of neurologic disease, and outcome. Results: Myelin‐like material was observed in 20/98 (20%) samples and was more frequently observed in lumbar (17/47, 36%) than cerebellomedullary samples (3/51, 6%) (P=.0028). Samples from dogs <10 kg were more likely to contain myelin (14/36, 39%) compared with dogs ≥10 kg (5/60, 8%) (P=.0052). Larger amounts of myelin‐like material were observed in CSF from dogs with intervertebral disk disease compared with other diseases (P=.045). No association was found between myelin‐like material and outcome. Conclusion: The association of extracellular myelin‐like material in canine CSF samples with sampling site and body weight suggests it is more often an artifact of collection technique and anatomy rather than the result of neurologic disease. Myelin‐like material in CSF is not associated with a poorer prognosis.     

Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.     

Cerebral and conjunctival haemorrhages associated with von Willebrand factor deficiency and canine angiostrongylosis

A case of angiostrongylosis is described in a 14-month-old golden retriever bitch. Conjunctival haemorrhage and neurological signs, referable to a space-occupying cerebral lesion, were associated with defective primary haemostasis caused by low levels of von Willebrand factor. Full clinical recovery followed treatment with desmopressin, fresh whole blood transfusion, fenbendazole and supportive care. The magnetic resonance image of the suspected organising haematoma is described. Similarities to the human condition, acquired von Willebrand syndrome, and a possible role for aberrant larval migration in haematoma formation are suggested.     

Haematological and biochemical findings in cats in Australia with lymphosarcoma

OBJECTIVE: To describe, for the first time, haematological and serum biochemical findings in cases of lymphosarcoma in Australian cats. DESIGN: A prospective multi-institutional study. PROCEDURE: Of 118 affected cats presented to the authors over a 18-month period, 97 were evaluated haematologically and 87 biochemically. Haematological analysis usually included determination of packed cell volume, haemoglobin concentration, red blood cell and leukocyte counts, differential leukcocyte count, reticulocyte count and examination of buffycoat smears for neoplastic cells. Serum biochemical analysis was done primarily with a discrete analyser and included a panel of commonly used analytes. RESULTS: Nonregenerative anaemia was present in 54% (52/97) of cats. Neutrophilia, present in 65% (59/91) of cats, was commonly associated with lymphocytopaenia, eosinopaenia and monocytosis. Of the 13 cats with a secondary leukaemic manifestation, only five had distinct lymphocytosis. Serum biochemical abnormalities either were nonspecific, such as hypoglycaemia in 37% (32/87) of cats, or related to specific tissue involvement, such as hypoalbuminaemia in 76% (31/41) of cats with alimentary involvement and azotaemia in 60% (15/25) of cats with renal involvement. CONCLUSION: It was shown for the first time that haematological and serum biochemical findings are of limited value in diagnosing lymphosarcoma in Australian cats, except if they are leukaemic. Although clinical pathological changes were common, they were nonspecific or related to specific tissue involvement. Their value in assessing response to therapy needs to be examined further. Patient characteristics such as age, breed and sex also had limited effect on laboratory findings and those observed were of little consequence. Additionally, histological and immunophenotypical variations in tumour type had little effect on laboratory findings.     

Eosinophilic cerebrospinal fluid pleocytosis associated with neural Angiostrongylus vasorum infection in a dog

A 1‐year‐old, female intact Pug dog was presented to the Small Animal Teaching Hospital of the University of Liverpool with a 4‐week history of progressive multifocal intracranial signs. Magnetic resonance imaging (MRI) detected multiple hemorrhagic lesions in the brain. The Baermann and zinc sulfate flotation tests with centrifugation, performed on fecal samples, were positive for lungworm larvae and an antigenic test confirmed Angiostrongylus vasorum infection. Anthelmintic treatment was started with a consequent marked clinical improvement. Seventy days later, the dog was clinically normal, and no larvae were detected on the Baermann test. Repeat MRI of the brain revealed marked improvement of the hemorrhagic lesions. Cerebrospinal fluid analysis (CSF) showed marked eosinophilic pleocytosis, and anthelmintic treatment was restarted. A follow‐up CSF analysis 4 months after the first presentation revealed resolution of the eosinophilic pleocytosis. This is the first case report of marked eosinophilic pleocytosis associated with neural A vasorum infection in a dog. The CSF eosinophilic pleocytosis persisted for several weeks after treatment, even in the absence of concurrent clinical signs and with a negative A vasorum Baermann test.     

Characteristics of cerebrospinal fluid associated with canine granulomatous meningoencephalomyelitis: a retrospective study

Cerebrospinal fluid of 22 dogs with histologically confirmed granulomatous meningoencephalomyelitis was analyzed, retrospectively. Seventeen dogs had cisternal CSF analysis, 4 dogs had lumbar CSF analysis, and 1 dog had both. For cisternal CSF, the mean +/- SEM total WBC count was 800.8 +/- 300.9 cells/microliter. The WBC differential count was predominantly lymphoplasmacytic cells, but 13 of the 18 cisternal CSF had polymorphonuclear (PMN) cells, and the mean +/- SEM PMN cell percentage was 18.6 +/- 5.3%. The mean +/- SEM total protein content of cisternal CSF was 255.8 +/- 98 mg/dl. Of 5 cisternal CSF pressures measured, 4 were within the normal range. The mean +/- SEM total WBC count and total protein content of lumbar CSF were 533.4 +/- 256.5 cells/mu/microliter and 163.2 +/- 25 mg of protein/dl, respectively. As with cisternal CSF, the WBC differential count of lumbar CSF was predominantly lymphoplasmacytic cells. Of 5 lumbar CSF, 4 contained PMN cells, but the percentage was less than the PMN cell percentage of cisternal CSF. Although variable, the general pattern of CSF abnormality associated with granulomatous meningoencephalomyelitis was different from the CSF abnormalities commonly seen with viral, bacterial, or mycotic encephalitides.