Acid-base indices in ovine blood and cerebrospinal fluid]

By means of the Astrup apparatus (Blood Microsystem BMS 2) acid-base indices in blood and cerebro-spinal fluid of clinically healthy Merino sheep at the age of one year (15 head) and of two or three years (41 head) were determined. Blood collected anaerobically from v. jugularis and liquor collected by means of the suboccipital punction in heparinized syringes and/or caplaris and liquor collected by means of the suboccipital punction in heparinized syringes and/or capillaries were examined immediately after the collection. Following indices were obtained and re-counted according to nomograms: actual pH, partial CO2 pressure--pCO2 base excess--BE, base buffer--BB, standard bicarbonate--SB, actual bicarbonate--AB, total CO2--tCO2 (Tables I, II). Indices of pH, pCO2, BE and BB indicated statistically significant differences in values when comparing liquor with venous blood (in younger ewes). As the values of the sheep cerebro-spinal fluid indices examined are not to be found in literature, the above stated values can be recommended to be used as basic values for a given age group.     

Collection of cerebrospinal fluid from the horse.

Methods for atlanto-occipital and lumbosacral collection of cerebrospinal fluid from the horse are described in detail, and the techniques and complications discussed.     

Evaluation of S100B in cerebrospinal fluid as a potential biomarker for neurological diseases in calves

S100B in cerebrospinal fluid (CSF-S100B) was measured in calves with 20 neurologic and 21 non-neurologic diseases to clarify its utility as a biomarker for neurologic diseases. The median CSF-S100B value in the neurologic disease group (43.0 ng/ml) was significantly higher than that in the non-neurologic disease group (10.2 ng/ml). As CSF-S100B levels in calves with neurologic diseases widely differed, the utility of CSF-S100B as a diagnostic marker for neurologic diseases in cattle remains inconclusive.     

Analysis of cerebrospinal fluid specific gravity

The specific gravity of 71 CSF samples from clinically normal dogs ranged from 1.004 to 1.006, with a mean of 1.005. A 3-year retrospective study of CSF samples obtained from 124 animals suspected of having neurologic disease revealed only 30 samples (24.2%) having a specific gravity greater than 1.006. Diagnostic or prognostic value from CSF specific gravity determination was not recognized.     

A field study of eight ovine vertebral body abscess cases

A vertebral body abscess extending into the epidural space causing spinal cord compression was diagnosed in the cervical spinal cord in four lambs and the thoracic spinal cord in four lambs. The affected lambs were aged between 4 and 10 weeks and clinical signs had been present for 7-21 days before veterinary examination. Treatment with high doses of penicillin did not result in an improvement of the neurological signs and all lambs were destroyed on humane grounds. Collection and analysis of cerebrospinal fluid (CSF) revealed a significant increase in the protein concentration in lumbar compared to cisternal CSF samples. There was a neutrophilic pleocytosis in two lumbar CSF samples. In lambs aged between 4 and 10 weeks with no history of previous locomotor dysfunction, paresis of more than 1 week duration may be suggestive of a vertebral body abscess which has extended into the epidural space causing spinal cord compression. Stagnation of CSF caudal to the lesion results in an increased protein concentration in the lumbar sample.     

Analysis of equine cisterna magna cerebrospinal fluid for the presence of some monoamine neutrotransmitters and transmitter metabolites

Small volumes (0.05 ml) of cisterna magna cerebrospinal fluid (CSF) from 23 neurologically normal horses were analysed for the monoamine neurotransmitters dopamine, norepinephrine, epinephrine, serotonin and their metabolites using high pressure liquid chromatography and electrochemical detection. Two metabolites, homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) were present in all CSF samples. The deaminated and methylated metabolite of dopamine, HVA, was present at a mean concentration of 42.33±3.14 ng/ml of CSF. The deaminated metabolite of serotonin, 5-HIAA, was present at a mean concentration of 45.52±3.65 ng/ml of CSF. A positive correlation was found between the CSF concentrations of HVA and 5-HIAA (r=0.72, p=0.0003). The mean ratio of HVA to 5-HIAA concentrations was 1.07±0.05. The ratios of HVA to 5-HIAA concentrations were found to be more indicative of the serotonergic metabolite 5-HIAA (r=–0.62, p=0.0016) than the dopaminergic metabolite HVA (r=0.11, p=0.60). There was a stronger relationship between the concentrations of 5-HIAA and the ratios of HVA to 5-HIAA in male CSF (r=–0.85, p=0.0006) than in female CSF (r=–0.54, p=0.01).Concentrations of CSF neurotransmitter metabolites were evaluated in horses grouped by age (0–4, 5–9 and 10–13 years). The youngest group of horses had a significantly (p=0.003) greater CSF concentration of HVA than of 5-HIAA. The 10–13-year-old horses had a significantly (p=0.02) lower mean CSF HVA concentration than the 0–4-year-old horses. No age-related differences in CSF 5-HIAA concentrations were detected.     

The management and welfare of some common ovine obstetrical problems in the United Kingdom

This review approaches sheep welfare during the periparturient period from a veterinary practitioner viewpoint. At the most fundamental husbandry level, the general hygiene standards adopted on many sheep farms in the United Kingdom (UK) during the lambing period are unacceptable and require urgent improvement. Concerns also arise from obstetrical conditions attended by farmers which cause pain that can be identified using clinical criteria such as altered demeanour and behaviour of the ewe, and the presence of inflammation of the reproductive tract. Reduced production is evidenced by poor milk production and hungry lambs, the necessity to rear such lambs artificially and, in neglected cases, death of the ewe and her lambs. The correction of dystocia, periparturient vaginal, uterine and rectal prolapses is greatly facilitated after epidural injection of lignocaine, or a combination of xylazine and lignocaine administered by a veterinary surgeon. However, the perceived high professional fees charged for these services, and for Caesarean operations, result in few sheep receiving veterinary care, with such neglect directly contributing to poor welfare standards and an increased ewe mortality rate during the periparturient period. Whilst ending any welfare concern, the destruction of ewes rather than requesting veterinary services purely for economic reasons is a disturbing trend in sheep production in the UK and raises concerns regarding positive interaction between veterinary practitioner and farmer. In this article, veterinary treatments which provide effective analgesia are described and costed for obstetrical conditions in sheep. Recommended treatment protocols are described for the common obstetrical problems in sheep. In addition to appropriate treatment of obstetrical problems and alleviation of pain, if veterinary involvement could reduce ewe periparturient mortality from estimated figures of 4-2% this would save an estimated 400,000 ewes annually in the UK. Sheep welfare could be considerably improved by practising veterinarians providing practical instruction to small groups of clients with funding provided by government agencies.     

Beta-2-microglobulin levels in the cerebrospinal fluid of normal dogs and dogs with neurological disease

BACKGROUND: Cerebrospinal fluid (CSF) analysis is the basis for establishing a diagnosis of central nervous system (CNS) inflammation. However, the information provided by routine CSF analysis is limited. Determination of CSF beta-2-microglobulin (beta2m) concentration has been used diagnostically in humans to identify inflammatory CNS disease; we hypothesized that it may have similar value in dogs. OBJECTIVES: The objective of this study was to measure (beta2m concentration in the CSF of clinically healthy dogs and compare the values to those observed in dogs with inflammatory CNS disease and intervertebral disc disease (IVDD). METHODS: CSF was collected from 10 clinically healthy laboratory dogs and 11 dogs each with inflammatory CNS disease and IVDD. Routine CSF analysis was performed, and (beta2m concentration was measured by ELISA. CSF (beta2m concentration and CSF:serum (beta2m ratio were compared between groups by ANOVA. Linear relationships between CSF total nucleated cell count (TNCC), RBC count, total protein concentration, and (beta2m concentration were assessed by regression analysis. RESULTS: The mean (+/- SD) CSF (beta2m concentration in clinically healthy dogs was 0.36 (+/- 0.05 microg/mL (cisternal) and 0.40 (+/- 0.07 microg/mL (lumbar). Median CSF (beta2m concentration in dogs with IVDD (0.46 microg/mL) and inflammatory CNS disease (0.85 microg/mL) differed from that of controls (0.36 microg/mL; P=.002). The concentration also differed between the 2 disease groups (P=.01). Five dogs with inflammatory CNS disease had CSF:serum (beta2m ratios >1. A correlation was identified between TNCC and (beta2m concentration (r=0.69, P=.0003). CONCLUSIONS: CSF (beta2m concentration is higher in dogs with IVDD and inflammatory CNS disease, with highest values seen with inflammatory disease. This may be attributed in part to the correlation between CSF (beta2m concentration and TNCC, but also may reflect intrathecal immune activation.     

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.     

Review of neurological diseases of ruminant livestock in Australia. VI: postnatal bovine, and ovine and caprine, neurogenetic disorders

This paper, the last in a series of reviews of neurological diseases of ruminants in Australia, discusses a range of neurogenetic disorders of cattle, sheep and goats, including necrotising encephalomyelopathy, glycogen storage disorders, cerebellar abiotrophy and β-mannosidosis. They are categorised by the species and breeds in which they occur.     

Monoclonal antibodies against ovine IgA purified from lung lavage fluid

Ovine secretory IgA (sIgA) has been purified to relative homogeneity by ammonium sulphate precipitation (to 40 per cent w/v) of lung lavage fluid from 3- or 12-month-old lambs, followed by molecular sieve chromatography on a Sephacryl S300 matrix. Three peaks A, B and C with molecular sizes corresponding to 550,000, 400,000 and 165,000 respectively were eluted from the column. Immunoelectrophoresis, radial immunodiffusion and enzyme-linked immunosorbent assays (ELISAs) with class specific antiserum confirmed that peak B contained only IgA. Polyacrylamide gel electrophoresis of peak B under reducing conditions resolved three subunits corresponding to secretory component, heavy and light chains. Hybridomas generated by fusing spleen cells from IgA-primed Balb/C mice with murine myeloma (Sp2/0) were screened for IgA-specific monoclonal antibodies against a panel of ovine IgG2, IgG1, IgA and IgM. One particular clone, F3-4B4, identified as murine IgG1, was monospecific against ovine IgA with no cross reactivity against bovine immunoglobulins. This hybridoma was successfully tested as a serological probe by ELISA profiling to locate IgA containing fractions in the course of immunoglobulin purification from biological fluids.     

Analysis of cerebrospinal fluid from dogs and cats after 24 and 48 hours of storage

OBJECTIVE: To compare differential cell counts and cell characteristics of CSF samples analyzed immediately or after storage for 24 and 48 hours at 4 C with and without the addition of autologous serum. DESIGN: Prospective study. ANIMALS: 36 dogs and 6 cats. PROCEDURE: CSF samples were collected from the cerebellomedullary cistern and divided into 250-microliter aliquots. Slides of CSF samples were prepared by use of cytocentrifugation immediately and after 24 and 48 hours of storage with addition of autologous serum (final concentrations, 11 and 29%). Differential cell counts and number of unrecognizable cells were compared among preparations. RESULTS: Significant differences in the differential cell counts were not detected among samples analyzed before or after storage. Although the number of unrecognizable cells increased with storage time, this did not result in a significant effect on cell distribution or diagnosis. Cells in CSF samples stored with 11% serum more closely resembled cells in fresh samples than did cells in samples stored with 29% serum. CONCLUSIONS AND CLINICAL RELEVANCE: CSF samples collected at veterinary clinics remote from a diagnostic laboratory or during nonoperational hours may be preserved through the addition of autologous serum. Evaluation of such samples is likely to result in an accurate diagnosis for at least 48 hours after collection.     

Analysis of cerebrospinal fluid from the cerebellomedullary and lumbar cisterns of dogs with focal neurologic disease: 145 cases (1985-1987)

Data were obtained from 158 CSF samples from 145 dogs with focal, noninfectious/noninflammatory neurologic disease. The effect of lesion location and the duration and severity of clinical signs were studied. One hundred and twenty-five samples were obtained from the cerebellomedullary cistern (CMC), and 33 were obtained from the lumbar cistern (LC). Intracranial and cervical disease affected the CSF from the CMC more often than did thoracolumbar disease. However, lumbar CSF was more frequently affected by disease anywhere along the neuraxis. For compressive spinal cord disease, the protein concentration at both cisterns was more often high in acute, clinically severe lesions. Intracranial lesions consistently caused abnormalities in CSF from both the CMC (7 of 7; 100%) and LC (2 of 2; 100%). Abnormalities were identified in 16 of 38 (42%) and 5 of 7 (71%) CMC and LC samples, respectively, in dogs with cervical disease. In dogs with thoracolumbar lesions, only 22 of 80 (27.5%) CMC samples were abnormal, compared with 21 of 24 (87.5%) LC samples. These findings suggest that CSF collected cranial to the lesion may be normal or only mildly altered by focal neurologic disease. Fluid obtained caudal to the lesion presumably is more substantially altered because of the predominant caudal flow of CSF. To maximize the yield of diagnostic information from CSF analysis, the fluid should preferably be obtained caudad to the disease site; however, because of problems associated with lumbar puncture, we suggest that CSF from the CMC also be obtained.     

Enrichment of prion protein in exosomes derived from ovine cerebral spinal fluid

Prion diseases are transmissible neurodegenerative disorders affecting humans and a wide variety of animal species including sheep and cattle. The transmissible agent, the prion, is an abnormally folded form (PrP(Sc)) of the host encoded cellular prion protein (PrP(C)). Distribution of the prion protein in the fluids of species susceptible to these diseases is of importance to human health and the iatrogenic spread of prion disease. Aside from blood which is confirmed to be a source of prion infectivity, it is currently unclear which other body fluids harbor a significant transmission risk. In the current study we examined two ovine fluids; pseudo-afferent lymph and cerebral spinal fluid (CSF), for the presence of exosomes and concurrent enrichment of the normal, cellular form of the prion protein (PrP(C)). Here we demonstrate the existence of exosomes in both pseudo-afferent lymph and CSF isolated from sheep. In the CSF derived exosomes we were able to show an enrichment of PrP(C) over unfractionated CSF. This experimental approach suggests that CSF derived exosomes could be used as a novel means of detecting abnormal forms of the prion protein and provide an in vivo link between these vesicles and prion disease pathogenesis.