Uncommon acute neurologic presentation of canine distemper in 4 adult dogs.

Four uncommon cases of canine distemper (CD) were diagnosed in vaccinated adult dogs. All dogs had acute onset of neurologic signs, including seizures, abnormal mentation, ataxia, and proprioceptive deficits. Polymerase chain reaction for CD virus was positive on cerebrospinal fluid in 2 cases. Due to rapid deterioration the dogs were euthanized and CD was confirmed by postmortem examination.

Rare présentation neurologique aiguë de la maladie de Carré chez 4 chiens adultes. Quatre cas peu communs de maladie de Carré chez des chiens adultes vaccinés. Tous les cas ont présenté un début aigu ou suraigu des signes neurologiques, comportant principalement des crises épileptiques, altération de l'état mental, ataxie, et déficits proprioceptifs. Dans deux cas, la PCR a été positive à la maladie de Carré dans le liquide céphalorachidien. En raison de la progression rapide des signes, les chiens ont été euthanasiés et la maladie de Carré confirmée par la nécropsie.(Traduit par Ana Roman).

Agarose gel electrophoresis of cerebrospinal fluid proteins of dogs after sample concentration using a membrane microconcentrator technique.

BACKGROUND: Cerebrospinal fluid (CSF) is produced in the cerebral ventricles through ultrafiltration of plasma and active transport mechanisms. Evaluation of proteins in CSF may provide important information about the production of immunoglobulins within the central nervous system as well as possible disturbances in the blood-brain barrier. OBJECTIVE: The objective of this study was to measure the concentration and fractions of protein in CSF samples using a membrane microconcentrator technique followed by electrophoresis, and to compare the protein fractions obtained with those in serum. METHODS: CSF samples from 3 healthy dogs and 3 dogs with canine distemper virus infection were concentrated using a membrane microconcentrator having a 0.5 to 30,000 d nominal molecular weight limit (Ultrafree, Millipore, Billerica, MA, USA). Protein concentration was determined before and after concentration. Agarose gel electrophoresis was done on concentrated CSF samples, serum, and serial dilutions of one of the CSF samples. RESULTS: Electrophoretic bands were clearly identified in densitometer tracings in CSF samples with protein concentrations as low as 1.3 g/dL. The higher CSF protein concentration in dogs with distemper was mainly the result of increased albumin concentration. CONCLUSION: The microconcentrating method used in this study enables characterization of the main protein fractions in CSF by routine electrophoresis and may be useful for interpreting the underlying cause of changes in CSF protein concentrations.

Evaluation of electrophoretic profile and albumin quota in the cerebrospinal fluid of dogs with distemper showing or not neurvous signs

The electrophoretic profile of cerebrospinal fluid proteins and albumin quota was studied in healthy dogs and dogs with distemper in either nervous or non-nervous phases. Cerebrospinal fluid (CSF) samples from 30 dogs were collected by puncture of the cisterna magna. The total protein content, the albumin quota, and the electrophoretic fraction of CSF proteins in agarose gel plates were evaluated. Results were similar in healthy dogs and dogs with distemper and no nervous signs, but were significantly increased in the group of dogs with distemper showing nervous signs. The study of CSF protein profile proved useful and contributed significantly on the detection of central nervous system disorders and damages to the blood-brain barrier during the nervous phase of distemper.

Estudaram-se o perfil eletroforético das proteínas liquóricas e a cota de albumina em cães sem e com cinomose na fase neurológica e não-neurológica. A punção da cisterna magna para a obtenção de amostras de liquor realizou-se em 30 cães. Analisaram-se teores de proteínas totais, cota de albumina e fracionamento eletroforético das proteínas liquóricas em gel de agarose. Os resultados foram semelhantes nos cães normais e nos cães com cinomose sem sinais neurológicos e significativamente elevados no grupo de cães com cinomose apresentando sinais neurológicos. O estudo do quadro protéico do líquido cérebroespinhal foi útil e contribuiu significativamente na detecção de lesões ao sistema nervoso central e de danos à barreira hematoencefálica durante a fase neurológica da cinomose.

Cerebellum progesterone concentration decreased in canine distemper virus infection.

Progesterone has neuroprotective effects including augmentation of myelination in the central and peripheral nervous system. This study was designed to determine if demyelinating lesions in the cerebellum resulting from canine distemper virus (CDV) infection are associated with progesterone levels. Progesterone was measured using radioimmunoassay in samples of the cerebellum, corpus callosum, medulla oblongata, parietal, frontal, temporal, and occipital cortices as well as cerebrospinal fluid (CSF) and plasma collected from ten CDV infected and six non-infected dogs. The cerebellum progesterone level was significantly different between CDV infected (0.66+/-0.09 ng/g) and control dogs (1.14+/-0.09 ng/g) (p<0.001); however, no difference was observed for the other CNS regions, plasma and CSF (p>0.05). The cerebellum progesterone level was also significantly different between acute (0.71+/-0.0 5 ng/g) and chronic cases (0.61+/-0.09 ng/g) (p<0.05). The CDV infected cerebella were also categorized histopathologically according to the severity of demyelinating lesions as mild (n=5), moderate (n=2), or severe (n=3) among which the cerebellum progesterone level was significantly different (p<0.05). Progesterone concentration was 0.71+/-0.05 ng/g in mild, 0.65+/-0.10 ng/g in moderate, and 0.56+/-0.07 ng/g in severe cases. In conclusion, progesterone concentration decreases in the cerebellum in CDV infection and the severity of demyelinating lesions is the greatest in cerebella with the lowest progesterone concentrations. The results suggest that local impairment of progesterone metabolism may be associated with the initiation and progression of cerebellar lesions in CDV infection.

Canine distemper virus uses both the anterograde and the hematogenous pathway for neuroinvasion.

Canine distemper virus (CDV), a member of the Morbillivirus genus that also includes measles virus, frequently causes neurologic complications, but the routes and timing of CDV invasion of the central nervous system (CNS) are poorly understood. To characterize these events, we cloned and sequenced the genome of a neurovirulent CDV (strain A75/17) and produced an infectious cDNA that expresses the green fluorescent protein. This virus fully retained its virulence in ferrets: the course and signs of disease were equivalent to those of the parental isolate. We observed CNS invasion through two distinct pathways: anterogradely via the olfactory nerve and hematogenously through the choroid plexus and cerebral blood vessels. CNS invasion only occurred after massive infection of the lymphatic system and spread to the epithelial cells throughout the body. While at early time points, mostly immune and endothelial cells were infected, the virus later spread to glial cells and neurons. Together, the results suggest similarities in the timing, target cells, and CNS invasion routes of CDV, members of the Morbillivirus genus, and even other neurovirulent paramyxoviruses like Nipah and mumps viruses.

Comparison of tissue and fluid samples for the early detection of canine distemper virus in experimentally infected dogs.

The clinical utility of various specimens was examined for the early diagnosis of canine distemper (CD). Seven healthy dogs at 17 weeks of age were experimentally infected with a field isolate of canine distemper virus. The RT-PCR was carried out to detect CDV NP gene. Dogs showed mild fever and leukopenia, however, typical clinical signs of CD were not seen through the experimental period. CDV amplicons were detected more, earlier and for longer period in the conjunctival swabs than in the other samples employed. These results suggested that conjunctival swab samples, which are easy to obtain and non-invasive, would be the most suitable and practical specimen for the early antemortem diagnosis of CDV infection.

Cerebrospinal fluid from a 7-month-old dog with seizure-like episodes.

A 7-month-old dog was presented to the Veterinary Teaching Hospital of the Universidade Estadual de Londrina, Paraná, Brazil with a 1-week history of seizure-like activity and compulsive walking. Neurological deficits included seizures, nystagmus, absence of a menace reaction, depressed postural reactions, spastic tetraparesis, opisthotonos, and spasticity of the thoracic limbs. Cerebrospinal fluid (CSF) evaluation showed severe lymphocytic pleocytosis (554 cells/microL, with 70% lymphocytes) and a protein concentration of 17 mg/dL. The histopathologic findings in cerebrum, cerebellum, and brainstem obtained at necropsy were compatible with acute encephalomyelitis caused by canine distemper virus (CDV). Using reverse transcription-polymerase chain reaction (RT-PCR), CDV RNA was detected in both CSF and fragments of fresh brain tissue. The results indicated that CDV was the agent responsible for the clinical and laboratory presentation. Severe pleocytosis with lymphocyte predominance is an unusual finding in canine distemper and must be differentiated from granulomatous meningoencephalitis. RT-PCR on CSF is a useful, fast, and specific method to diagnose CDV infection in dogs.

Dominating interleukin-10 mRNA expression induction in cerebrospinal fluid cells of dogs with natural canine distemper virus induced demyelinating and non-demyelinating CNS lesions.

Canine distemper virus (CDV) infection in dogs is commonly associated with demyelinating leukoencephalitis (DL). Although the mechanism of primary demyelination in distemper remains undetermined recent studies showed a direct virus-induced cytolysis in early non-inflammatory and immune-mediated mechanisms in inflammatory lesions. To further investigate the pathogenesis of this morbillivirus-induced demyelination the expression of a variety of cytokine mRNA species (interleukin (IL)-1beta, IL-2, IL-6, IL-10, IL-12, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, and interferon (IFN)-gamma in cerebrospinal fluid cells of 12 dogs with CDV encephalitis was investigated employing reverse transcription-polymerase chain reaction (RT-PCR) and these findings were correlated to the type of CNS lesions. Neuropathology revealed the whole spectrum of distemper DL lesions from acute to chronic alterations, however, most plaques lacked active demyelination. Three control animals were devoid of any cytokine expression, whereas in distemper animals IL-10 transcripts were found in nine dogs with acute and chronic lesions. IL-6, TNF, and TGF mRNA was found in six, four, and three animals, respectively. IL-12 and IFN-gamma, suggestive of a TH1-like dominated immune response, were detected only in one animal with chronic lesions. Summarized, TNF and IL-6, associated with disease exacerbation, and IL-10 and TGF, indicative of remission, were often observed simultaneously in distemper DL and could not be assigned to a specific disease stage. However IL-10 mRNA remained the most frequently detected cytokine indicating a stage of inactivity in most animals investigated.

Molecular analysis of the nucleocapsid protein of recent isolates of canine distemper virus in Japan.

We analyzed the molecular properties of the nucleocapsid protein (NP) of canine distemper viruses (CDV), isolated between 1992 and 1995 in Japan. Four CDV field isolates (Yanaka, Ueno, Hamamatsu, and Adachi strains) obtained were antigenically identical. Sequence analysis of entire region of the NP gene of a field isolate, the Yanaka strain, revealed that the NP gene contained 1683 nucleotides and was 93.2% homologous with a laboratory strain, the Onderstepoort strain. The deduced amino acid sequence contained 523 amino acids and was 95.2 and 99% homologous with those of the Onderstepoort and a virulent strain, A75/17 strain, respectively. Since most of the diversities in amino acid sequence occurred in two domains, at the N'- and the C'- termini, we further sequenced 3'-terminal regions of the remaining three field isolates. Based on the sequences, the new CDV isolates had one cluster that distinguished them from the laboratory strain.

Cytological, immunocytochemical and biochemical cerebrospinal fluid investigations in selected central nervous system disorders of dogs.

Cerebrospinal fluid (CSF) obtained from 20 clinically healthy dogs and from 15 dogs affected by neurological disorders were examined for total and differential cell counts, immunocytochemistry for canine distemper virus antigen, total protein concentration and electrophoretic separation, and glucose and enzyme determination. Dogs affected by canine distemper showed an increase in macrophages, presence of specific inclusion bodies, and an increase in total protein concentration and gamma-globulin fraction. In bacterial meningoencephalomyelitis pleocytosis, increase of total proteins with special regard to high molecular weight proteins, elevated levels of lactate dehydrogenase (LDH), alterations of LDH isoenzyme profile, and decrease in glucose levels were detected; inflammatory disorders were more often characterized by an increase in LDH level, while in non-inflammatory disorders (hydrocephalus and spinal cord neoplasia) no variation in LDH levels was detected. Analysis of CSF in dogs appears relatively easy to perform and may help in establishing the condition of the blood-brain barrier as well as in the diagnosis of neurological disorders.

[Cytology of the cerebrospinal fluid of dogs and cats with symptoms of meningitis/meningoencephalitis. Part 3]. / Zytologie des Liquor cerebrospinalis von Hund und Katze bei Symptomen einer Meningitis/Meningoenzephalomyelitis. Teil 3.

A review of the literature of cerebrospinal fluid (CSF) cytology of different forms of meningitis/meningoencephalomyelitis in dogs and cats is given. Eight dogs and three cats with signs of meningitis/meningoencephalitis are presented. Four dogs and one cat improved to normal for 1-3 years. The results of CSF cytology of cases whose etiology could not be determined are compared with those of thirteen dogs with distemper. In 8 of 13 sediments eosinophilic inclusions in monocytes and macrophages were found.

Electrophoretic determination of albumin and gamma globulin concentrations in the cerebrospinal fluid of dogs with encephalomyelitis attributable to canine distemper virus infection: 13 cases (1980-1987).

Thirteen dogs with encephalomyelitis attributable to canine distemper virus infection were classified into 3 groups on the basis of histopathologic evidence of virus-induced lesions in CNS tissue. Analysis of data indicated a similarity within groups when arranged by age, clinical neurologic signs, and alterations in the CSF electrophoretic protein values. Comparison with results in dogs with experimentally induced canine distemper encephalomyelitis suggested that when grouped by age, CSF electrophoretic values are helpful in predicting the CNS histopathologic changes in dogs with naturally acquired canine distemper encephalomyelitis.

Distemper-like disease and encephalitozoonosis in wild dogs (Lycaon pictus).

Clinical signs of a fatal disease resembling those of canine distemper were observed in two groups of captive wild dog (Lycaon pictus) pups 13 days after vaccination with a commercially available combination vaccine for dogs which contained a live attenuated strain of canine distemper virus. Histopathological examination of tissues revealed the presence of intranuclear inclusion bodies in neurons and lesions resembling canine distemper as well as colonies of an Encephalitozoon sp. in the central nervous system and kidneys. Lesions were observed in both organs which resembled those described in other species infected with Encephalitozoon cuniculi.

Production of immunoglobulin G and increased antiviral antibody in cerebrospinal fluid of dogs with delayed-onset canine distemper viral encephalitis.

Sera and cerebrospinal fluid (CSF) from four dogs with delayed-onset canine distemper viral (CDV) encephalitis (old dog encephalitis) were compared with samples from dogs with acute CDV and from recently vaccinated controls. Dogs with old dog encephalitis (ODE) had elevated CSF IgG concentrations (122 micrograms/ml) compared to controls (13 micrograms/ml) without elevated CSF albumin; their CSF IgG index was significantly greater. CSF proteins banding in the alkaline region of isoelectric focusing gels were immunochemically identified as IgG. Detectable viral neutralizing antibody was present in ODE CSF, and formed a larger proportion of IgG in CSF than in serum. Serum samples containing 2 mg IgG bound to all viral polypeptides of both R252 and Onderstepoort CDV isolates by immunoblot analysis. CSF samples of ODE patients bound viral antigens when diluted to contain as little as 5-40 micrograms IgG, while patient serum could be diluted to 40-200 micrograms IgG content compared to serum IgG of 100 micrograms/ml in vaccinated controls. Serial CSF dilutions consistently bound to H and NP polypeptides at the highest dilutions, similar to the binding of serums from recently vaccinated dogs. Thus, dogs with delayed-onset CDV encephalitis have elevated concentrations of CSF IgG, much of which is virus-specific, with an antigen binding pattern similar to that of sera of recently immunized dogs.

Albumin leakage into cerebrospinal fluid of dogs lethally infected with R252 canine distemper virus.

Cerebrospinal fluid (CSF) form nine lethally infected and three convalescent gnotobiotic dogs infected with the R252 strain of canine distemper virus (CDV) was evaluated prior to and following infection. Lethally infected dogs had a mean seven-fold increase in CSF albumin concentration compared to the preinoculation value, not present in dogs destined to survive. Immunochemical examination of tissue from these dogs revealed prominent perivascular localization of albumin. Examination of CSF cells demonstrated mild leukocytosis in both groups at the time when encephalopathic deaths occurred, with decreased lymphocyte percentages, particularly Thy-1-bearing lymphocytes, in lethally infected dogs. These dogs also had more extensive expression of viral antigens in CSF and peripheral blood leukocytes at the time of death than did surviving dogs, and failed to make antibody to viral antigens. The findings link terminal breakdown of the blood-brain barrier and extensive viral antigen expression in CSF leukocytes with experimental CDV infection resulting in death.

Interferon in cerebrospinal fluid. A marker for viral persistence of canine distemper encephalomyelitis.

Interferon (IFN) was measured in serum and cerebrospinal fluid (CSF) of dogs after experimental (intranasal) infection with different strains of virulent canine distemper virus (CDV). Viral strains employed produced neurological changes in dogs that range from acute inflammatory to subacute, delayed demyelinating encephalomyelitis. With few exceptions, first appearance of serum-IFN correlated with the first elevated body temperature 4 days post-infection (p.i.). By 16 days p.i. IFN had disappeared from the serum of all infected dogs. In contrast, IFN was constantly detectable in CSF in dogs with CDV infection of the central nervous system (CNS). It was first detected 5 days p.i., was continuously detectable during the variable preclinical phase and into the period when signs of acute or delayed encephalomyelitis were evident. Dogs from which CDV would be retrieved from CNS tissue at necropsy always had CSF-IFN (up to 56 days p.i.). In contrast, dogs that recovered from infection, substantiated at necropsy by minimal, resolving CNS lesions and non-detectable virus, had IFN in CSF demonstrable for only a brief post-inoculation period. CSF-IFN appears to be a valid marker for CDV persistence in the canine CNS and may have broader applications.

Cerebrospinal fluid cytology in canine neurologic disease.

Samples of cerebrospinal fluid (CSF) of 93 dogs with neurologic diseases were examined by cytomorphologic technique, and the changes in the CSF were correlated with histopathologic examinations of the central nervous system (CNS). It was concluded that CSF examination is a significant aid in obtaining a neurologic diagnosis and that good correlation exists between the CSF changes and the pathologic changes in the CNS. The CSF examination allows making a diagnosis of encephalitis and differentiation between viral and other causes (although in mycotic infection the cell membrane preparation can be used to identify the cause directly), could allow making differentiation between congenital malformations and congenital degenerative disease, and helps in identifying physical spinal cord damage, differentiating it from muscular, neurogenic, or functional disorders clinically presented as spinal ataxia. The CSF cytologic examination can indicate the presence of hemorrhage in the CNS. There is not enough experience available in the diagnosis of brain tumors by means of CSF examination; however, in dogs with lymphosarcoma in the CNS, CSF cytologic changes can be diagnostic.