Gheorghe Solcan

An exploratory study using a statistical approach as a platform for clinical reasoning in canine epilepsy

Links between deficits identified on neurological examination, age of seizure onset and the presence of structural forebrain disease have often been postulated in dogs presenting with a history of seizures. The purpose of this study was to assess the influence of such factors on the likelihood of structural or functional brain disease, via a thorough history taking process and interictal neurological examination. Four hundred and four dogs with seizures due to intracranial causes were included. Data including age, sex, neuter status, time until diagnosis, age of seizure onset in years, type of seizure, seizure symmetry, seizure severity, interictal neurological deficits, MRI changes and side effects associated with antiepileptic drugs were extracted from medical files. Two hundred and fifty-eight dogs were diagnosed with epilepsy of unknown origin (EUO), 11 with symmetrical structural lesions and 135 with asymmetrical structural lesions. Multinomial analysis demonstrated that dogs that were older at seizure onset were significantly more likely to have an asymmetrical structural lesion than EUO (OR 95% CI: 1.4-1.8). Dogs that had single seizures rather than cluster seizures were less likely to have asymmetrical structural lesions than dogs with EUO (OR 95% CI: 0.2-0.7). Dogs with abnormal neurological examinations were 16.5 times more likely to have asymmetrical structural lesions (OR 95% CI: 8.5-32.1) and 12.5 times more likely to have symmetrical structural lesions (OR 95% CI: 3.0-52.3) than EUO. These findings support the importance of considering interictal neurological deficits and seizure history in clinical reasoning.

The hepatoprotective effect of sea buckthorn (Hippophae rhamnoides) berries on induced aflatoxin B1 poisoning in chickens

The leaves and berries of sea buckthorn (SB; Hippophae rhamnoides; family Elaeagnaceae) are medically claimed as having phytoantioxidant, antiinflammatory, and anticancerous properties in humans. This study evaluated the hepatoprotective activity of oil from SB berries against toxicity induced by aflatoxin B1 (AFB1) in broiler chickens. The toxicity of AFB1 led to lower total serum proteins and specifically reduced albumin (P < 0.001). Serum aspartate aminotransferase increased from 191.14 ± 11.56 to 218.80 ± 13.68 (P < 0.001). When chickens were simultaneously dosed with AFB1 and an extract of SB berries, subsequent histology of the liver showed a significant reduction of necrosis and fatty formation compared with chickens treated with AFB1 alone. Immunohistochemical results indicated that COX2, Bcl-2, and p53 were highly expressed in the liver of AFB1-treated chickens and their expression was significantly reduced by SB oil supplementation. The levels of AFB1 residues in chickens livers were significantly reduced by SB oil from 460.92 ± 6.2 ng/mL in the AFB1 group to 15.59 ± 6.1 ng/mL in the AFB1 and SB oil group. These findings suggest that SB oil has a potent hepatoprotective activity, reducing the concentration of aflatoxins in liver and diminishing their adverse effects.

The 1H NMR Profile of Healthy Dog Cerebrospinal Fluid

The availability of data for reference values in cerebrospinal fluid for healthy humans is limited due to obvious practical and ethical issues. The variability of reported values for metabolites in human cerebrospinal fluid is quite large. Dogs present great similarities with humans, including in cases of central nervous system pathologies. The paper presents the first study on healthy dog cerebrospinal fluid metabolomic profile using 1H NMR spectroscopy. A number of 13 metabolites have been identified and quantified from cerebrospinal fluid collected from a group of 10 mix breed healthy dogs. The biological variability as resulting from the relative standard deviation of the physiological concentrations of the identified metabolites had a mean of 18.20% (range between 9.3% and 44.8%). The reported concentrations for metabolites may be used as normal reference values. The homogeneity of the obtained results and the low biologic variability show that the 1H NMR analysis of the dog’s cerebrospinal fluid is reliable in designing and interpreting clinical and therapeutic trials in dogs with central nervous system pathologies.

Screening diabetic cats for hypersomatotropism: performance of an enzyme-linked immunosorbent assay for insulin-like growth factor 1

Screening diabetic cats for feline hypersomatotropism (HS) is currently dependent on using a radioimmunoassay (RIA) for measurement of growth hormone or insulin-like growth factor 1 (IGF-1), both of which require radioactivity, are costly and have limited availability. Performance of an enzyme-linked immunosorbent assay (ELISA) using anti-human IGF-1 antibodies was assessed. Total IGF-1 was determined in diabetic cat samples across a wide range of IGF-concentrations using a previously validated RIA (serum: 92 cats; plasma: 31 cats). Repeat IGF-1 measurement was then performed using the ELISA-system. Mean IGF-1 recovery after serial dilution proved satisfactory with a correlation coefficient of 0.96 (serum) and 0.97 (plasma). Appropriate precision was established [intra-assay coefficient of variation (CV) 9.5 ± 2% (serum) and 13.6 ± 7% (plasma); inter-assay CV 11.4 ± 4% (serum) and 7.6 ± 6% (plasma)] and significant effect of hyperlipidaemia, haemoglobinaemia, bilirubinaemia and storage was excluded, with the exception of an increase in serum IGF-1 when left at room temperature for more than 24 h. ELISA concentrations correlated significantly with RIA concentrations (serum Pearson r2: 0.75; plasma: 0.83, P <0.001). Receiver operating characteristics analysis showed an area under the curve of 0.99 (serum) and 0.96 (plasma), and indicated high diagnostic accuracy for categorising a diabetic cat correctly as suspicious for HS at a serum IGF-1 cut-off of 997 ng/ml (sensitivity, 100%; specificity, 88.1%). The current study is the first to validate an easy-to-use and economical IGF-1 ELISA for the screening for HS among diabetic cats, which is important given the suspected significant prevalence of HS-induced diabetes mellitus.

EFFECT OF OCHRATOXIN A ON THE INTESTINAL MUCOSA AND MUCOSA-ASSOCIATED LYMPHOID TISSUES IN BROILER CHICKENS

The immunotoxic effect of ochratoxin A (OTA) on the intestinal mucosa-associated lymphoid tissue and its cytotoxic action on the intestinal epithelium were studied in broiler chickens experimentally treated with the toxin. From the 7th day of life, 80 male broiler chickens (Ross 308) were randomly divided into four groups of 20 birds each. The three experimental groups (E1-3) were treated with OTA for 28 days (E1: 50 μg/kg body weight [bw]/day; E2: 20 μg/kg bw/day; E3: 1 μg/kg bw/day) and the fourth group served as control. Histological examination of the intestinal mucosa and immunohistochemical staining for identification of CD4+, CD8+, TCR1 and TCR2 lymphocytes in the duodenum, jejunum and ileocaecal junction were performed, and CD4+/CD8+ and TCR1/TCR2 ratios were calculated. OTA toxicity resulted in decreased body weight gain, poorer feed conversion ratio, lower leukocyte and lymphocyte count, and altered intestinal mucosa architecture. After 14 days of exposure to OTA, immunohistochemistry showed a significant reduction of the lymphocyte population in the intestinal epithelium and the lamina propria. After 28 days of exposure, an increase in the CD4+ and CD8+ values in both the duodenum and jejunum of chickens in Groups E1 and E2 was observed, but the TCR1 and TCR2 lymphocyte counts showed a significant reduction. No significant changes were observed in Group E3. The results indicate that OTA induced a decrease in leukocyte and lymphocyte counts and was cytotoxic to the intestinal epithelium and the mucosa-associated lymphoid tissue, altering the intestinal barrier and increasing susceptibility to various associated diseases.

Clinical reasoning in feline epilepsy: Which combination of clinical information is useful?

We sought to identify the association between clinical risk factors and the diagnosis of idiopathic epilepsy (IE) or structural epilepsy (SE) in cats, using statistical models to identify combinations of discrete parameters from the patient signalment, history and neurological examination findings that could suggest the most likely diagnosis. Data for 138 cats with recurrent seizures were reviewed, of which 110 were valid for inclusion. Seizure aetiology was classified as IE in 57% and SE in 43% of cats. Binomial logistic regression analyses demonstrated that pedigree status, older age at seizure onset (particularly >7years old), abnormal neurological examinations, and ictal vocalisation were associated with a diagnosis of SE compared to IE, and that ictal salivation was more likely to be associated with a diagnosis of IE than SE. These findings support the importance of considering inter-ictal neurological deficits and seizure history in clinical reasoning.

Ultrastructural lesions and immunohistochemical analysis of Bcl-2 protein expression in the kidney of chickens with experimental ochratoxicosis

A study was conducted to evaluate the nephrotoxic effect of ochratoxin A (OTA) in broiler chickens. Forty Ross 308 broilers (6 days old) were divided into two groups: one group received daily, by gavage, ochratoxin A at a daily dose of 50 μg/kg body weight for up to 21 days, while the control group received only diluent (sunflower oil). After 21 days, the chickens were euthanised and the kidneys removed for analysis by histopathology and immunohistochemistry to detect an anti-apoptotic marker (Bcl-2), and by transmission electron microscopy. Macroscopically the kidneys were enlarged, showing degeneration and gout deposits. Histologically, glomerulonephrosis and tubulonephrosis were common lesions in all chicks. In two of the five chicks exposed to OTA for 21 days, focal tubular cell proliferation, multiple adenoma-like structures and Bcl-2-positive epithelial cells were identified in layers of the renal papilla and in convoluted tubules. Transmission electron microscopy of the proximal convoluted tubules identified abnormal forms of mitochondria. The nephrotoxic effect of ochratoxicosis in chickens is probably due to carcinogenic changes induced in the epithelial tissues.

Interictal cardiac autonomic nervous system disturbances in dogs with idiopathic epilepsy

Autonomic nervous system (ANS) activity in the interictal period (InIp) in dogs with presumed idiopathic epilepsy (pIE) was assessed using heart rate variability (HRV) analysis. The HRVs obtained from 28 pIE dogs with interictal epileptic discharges (InIEd; 11 with treatment and 17 without treatment) detected on electroencephalography (EEG) were compared with those obtained from 13 healthy dogs. On electrocardiographic (ECG) study, the P wave dispersion (PWD; P<0.001), P max (P=0.004) and corrected QT interval (QTc; P=0.025) were significantly increased in the pIE group. On the basis of HRV analysis, the pIE dogs had an increased activity of the parasympathetic component of the ANS, including the percentage of R-R interval (pNN50%) that differs more than 50ms (P=0.011) and high frequency band (HF; P=0.041). Administration of phenobarbitone had no influence on the ANS pattern when pIE subgroups were compared (P>0.05). In InIp, dogs elicited specific conductibility delays of the electrical impulses (increased PWD and QTc interval); these delays are considered to be risk factors for developing severe arrhythmias, such as atrial fibrillation and ventricular tachycardia. When compared with human beings, a different ANS pattern characterised by increased parasympathetic activity was observed, which may influence the therapeutic approach of IE in dogs.

Evaluation of central vestibular syndrome in dogs using brainstem auditory evoked responses recorded with surface electrodes

ABSTRACT The present study aimed to analyse the wave morphology, amplitude, latency, and intervals of the brainstem auditory evoked responses (BAERs) in dogs with central vestibular syndrome (CVS) recorded with surface electrodes. Ten dogs with CVS were examined by mono- and binaural stimulation, using the Neuropack electrodiagnostic system, with stimulus intensities of 90 dBSPL. BAERs examinations revealed morphological changes of waves I, II, III, and V and decreased amplitudes of all waves in 7/10 dogs. P values obtained were = 0.014 for wave I amplitude, 0.031 for II, and III and 0.032 for V. Comparing the latencies of waves I, II, III, and V generated by right and left monoaural stimulation in dogs with CVS, we did not observe significant differences (P>0.05). No statistical differences were observed for BAERs latencies of the waves recorded after binaural and monaural stimulation (left or right). As far as we know, this is the first study of BAERs using surface electrodes, obtained from dogs with CVS. Keywords: dog, brainstem auditory evoked responses, electrophysiology, central vestibular syndrome, surface electrodes RESUMO

Brainstem auditory evoked responses in healthy Argentine Mastiff dogs recorded with surface electrodes

RESUMOEvocado auditivo de respostas – BAER ) foi gravado utilizando eletrodos de superficie em sete caes Mastiff argentino saudaveis para estimulacoes monoaural e binaural em intensidades sonoras de 90 dB NPS . As latencias e amplitudes das ondas I, III e V bem como os intervalos InterWave I- III , III -V e IV foram calculados. As diferencas estatisticas foram obtidas para as ondas III e latencias V, amplitudes das ondas III e intervalos I- III e I-V entre mono e estimulacao binaural. Por serem uma das racas geneticamente predispostas a surdez congenita, obtendo valores de referencia BAER, os caes Mastiff argentino tem uma importância clinica especial.