A multicenter prospective evaluation of owner medication adherence for feline cardiovascular disease in the referral setting.

OBJECTIVE: The aim of this paper is to evaluate owners' chronic medication adherence for management of feline cardiovascular disease in the small animal referral setting. ANIMALS, MATERIALS AND METHODS: A questionnaire-based study of owners at five multispecialty, small animal referral centers was conducted. Owners completed a written survey evaluating demographics, degree of medication adherence, and difficulties encountered for medication adherence. Owners were free to decline participation in the study. RESULTS: Fifty-four questionnaires were available for review. The most common diagnosis was hypertrophic cardiomyopathy (n = 31, 57.4%). Clopidogrel was the most cited medication that was difficult to administer consistently (n = 13, 24.0%) although twenty owners (37.0%) reported no difficulty consistently administering medications. "Taste of medication" (n = 14, 25.9%) was the most reported reason for difficulty medicating their cat, and most owners (n = 36, 66.7%) stated twice daily was the highest frequency of heart medications they feel they can consistently administer. Fifty owners (92.6%) met the criteria for medication adherence. CONCLUSIONS: Chronic medication adherence in this study population was high. Clopidogrel was the most difficult medication to consistently administer, and twice a day dosing was the highest frequency of medication administration most owners could achieve. Cardiologists should be aware of these factors when determining optimal treatment protocols for the management of cardiovascular disease in cats.

Evaluation of owner medication adherence for canine cardiovascular disease in the referral setting.

OBJECTIVE: To evaluate owners' medication adherence for management of cardiovascular disease in the small animal specialty setting. ANIMALS, MATERIALS, AND METHODS: A questionnaire-based study of owners at a single, multispecialty, small animal referral center was conducted. Owners completed a written survey evaluating demographics, degree of medication adherence, and difficulties encountered for medication adherence. Owners were free to decline participation in the study. RESULTS: Ninety-six questionnaires were completed during the study period. Fifty-four respondents (56.3%) reported treating their pet for over one year for cardiovascular disease, and myxomatous mitral valve disease was the most common diagnosis (76.0%). Twenty-nine (30.2%) respondents felt they could consistently administer a maximum of three medications daily, and 46 (47.9%) respondents reported twice-daily dosing was the highest frequency of administration that could be consistently achieved. Twenty-two respondents (22.9%) reported that their pet required daily medications for at least one other chronic condition besides cardiac disease. Seventeen respondents (17.7%) reported at least occasionally missing medications, although only one (1.0%) reported it was more than 20% of medications per week. CONCLUSIONS: Medication adherence was high in this study population. Almost one-half of owners surveyed reported being unable to consistently administer more than three medications daily, and approximately one-quarter of patients were treated for at least one additional chronic condition requiring daily medications, in addition to cardiac disease. Clinicians should be aware of these factors when determining optimal treatment protocols for the management of cardiovascular disease in dogs.

Bioremediation and biovalorisation of olive-mill wastes.

Olive-mill wastes are produced by the industry of olive oil production, which is a very important economic activity, particularly for Spain, Italy and Greece, leading to a large environmental problem of current concern in the Mediterranean basin. There is as yet no accepted treatment method for all the wastes generated during olive oil production, mainly due to technical and economical limitations but also the scattered nature of olive mills across the Mediterranean basin. The production of virgin olive oil is expanding worldwide, which will lead to even larger amounts of olive-mill waste, unless new treatment and valorisation technologies are devised. These are encouraged by the trend of current environmental policies, which favour protocols that include valorisation of the waste. This makes biological treatments of particular interest. Thus, research into different biodegradation options for olive-mill wastes and the development of new bioremediation technologies and/or strategies, as well as the valorisation of microbial biotechnology, are all currently needed. This review, whilst presenting a general overview, focus critically on the most significant recent advances in the various types of biological treatments, the bioremediation technology most commonly applied and the valorisation options, which together will form the pillar for future developments within this field.

Acute necrotising pulmonary vasculitis and pulmonary hypertension in a juvenile dog.

A five-month-old female Jack Russell terrier was presented for investigation of acute lethargy, anorexia, coughing, respiratory distress and weakness. Examination findings included cyanosis, a grade 3 of 6 systolic heart murmur and prolonged capillary refill time. Radiography and echocardiography revealed severe pulmonary hypertension, cor pulmonale and right-sided heart failure. Indirect measurement of the systolic pulmonary artery pressure estimated pressures over 100 mmHg. Despite treatment the patient died. Post-mortem examination did not identify a congenital cardiovascular anomaly. Histopathology confirmed acute necrotising pulmonary arteritis and immunohistochemistry failed to identify any immune complex or complement deposition.

Comparison of radioimmunoassay and chemiluminescent assay methods to estimate canine blood cortisol concentrations.

BACKGROUND: Non-radioactive assay methods are widely used in commercial laboratories to measure canine blood cortisol concentrations, despite a paucity of published validity data of these tests compared with the traditional 'gold standard' radioimmunoassay. OBJECTIVES: To compare a commercial chemiluminescence assay with radioimmunoassay for blood cortisol measurement, determine the effect of storage on the radioimmunoassay, and determine the impact of any differences on clinical decisions. METHODS: The study included 54 client owned dogs undergoing adrenal function testing. Fresh plasma or serum samples (n=170) were assayed for cortisol using radioimmunoassay (RIA1). Samples (n=196) were also frozen and stored in batches, and assayed by chemiluminescence and radioimmunoassay (RIA2). RESULTS: Overall, there was a strong correlation (r2=0.967, P<0.001) between RIA2 and chemiluminescence concentrations without significant difference between means. Strong correlations were present for RIA2 and chemiluminescence at concentration subgroups of >400 nmol/L (r2=0.869, P<0.001), <100 nmol/L (r2=0.790, P<0.001), and <40 nmol/L (r2=0.738, P<0.001). Significant differences between means were present for RIA2 and chemiluminescence concentrations in the <100 nmol/L, and <40 nmol/L (P<0.001) groups. Despite a significant difference in RIA1 and RIA2 results overall, there was no significant difference between RIA1 and RIA2 for any of the concentration groups. In seven cases, discrepant RIA2 and chemiluminescence results may have altered clinical decisions. CONCLUSIONS: Although RIA and chemiluminescence cortisol concentrations appear highly correlated, a significant difference may exist for concentrations less than 100 nmol/L in stored canine sera. Results of chemiluminescence cortisol assays should be interpreted with caution unless the specific assay method in the laboratory has been adequately validated in dogs.

Four cases of snake envenomation responsive to death adder antivenom.

Death adder envenomation is rare in humans and there is only one brief report previously in dogs. This paper details three cases of canine common death adder (Acanthophis antarcticus) envenomation and one case of bardick (Echiopsis curta) envenomation which were responsive to death adder antivenom. The available literature on death adder envenomations is also reviewed. The main clinical sign in the four dogs was severe lower motor neuron paralysis. There was no clinical evidence of coagulopathy or myopathy. Use of a snake venom detection kit was essential for selection of appropriate antivenom. Death adder and bardick envenomation in dogs potentially has a good prognosis if sufficient antivenom is administered and intensive supportive care is available.

Primary hypoparathyroidism in dogs: a retrospective study of 17 cases.

OBJECTIVE: To evaluate the clinico-pathological findings, response to treatment and prevalence of complications in dogs with primary hypoparathyroidism. DESIGN: Retrospective study of 17 dogs presenting to the University of Melbourne Veterinary Clinical Centre and Murdoch University Veterinary Hospital over a 15 year period (1990 to 2004). Case records were evaluated for signalment, body weight, diet type, historical and clinical findings, serum total calcium, phosphate, albumin and parathyroid hormone concentrations, urinary fractional excretion ratios of calcium and phosphate, electrocardiogram (ECG) results, treatments administered, outcome and period of follow-up. RESULTS: The most common breeds identified were St Bernard (three dogs), Chihuahua (two dogs), German Shepherd (two dogs) and Jack Russell Terrier (two dogs). Three dogs were cross bred. Seizures, muscle tremors and fasciculations, stiff gait, tetany, muscle cramping, behavioural change and hyperventilation were the most common clinical signs. Vomiting, inappetence, diarrhoea, abdominal pain, hyperthermia, facial pruritus, ataxia, weakness, cataracts, and circling also occurred with less frequency. The mean duration of observed clinical signs preceding diagnosis was 33 days (median 13 days, range 1 to 173 days). All dogs had marked hypocalcaemia with normal or mildly increased serum albumin concentrations. Mean phosphate concentrations were significantly higher in inappetent dogs (P = 0.049). Mean serum calcium concentrations were significantly lower in dogs with cataracts compared to those without (P = 0.046). There were no other significant relationships between serum calcium or phosphate concentrations and the clinical presentation or outcome. No significant correlations were identified between the presence of a particular clinical sign and the duration of clinical signs. ECGs were obtained in four dogs and all exhibited QT interval prolongation due to a ST-segment prolongation. Sixteen of 17 dogs were treated successfully for hypocalcaemia and discharged from hospital. Acute management included parenteral calcium gluconate (10 dogs) and intravenous anticonvulsants (five dogs). Chronic therapy included oral vitamin D analogues and calcium supplementation. Treatment complications occurred in two dogs and included acute renal failure (one dog) and iatrogenic tissue necrosis following subcutaneous calcium administration (one dog). The mean follow-up period was 14.5 months (median 13 months, range 0 to 39 months). Twelve dogs were alive at the last follow up and two dogs were euthanased for unrelated reasons. The type of vitamin D analogue used was not associated with outcome. CONCLUSION: Primary hypoparathyroidism was an uncommon diagnosis in dogs. Saint Bernards, cross bred dogs, German Shepherd dogs and Terrier breeds were most commonly affected. Neurological signs were the most common presenting clinical signs, although alimentary signs may have been more common than previously reported. Dogs with primary hypoparathyroidism appeared to have a good prognosis following initiation of calcium supplementation and vitamin D therapy. Complications of treatment were uncommon and could be minimised with regular monitoring.

The positional specificity of a desaturase in the psychrophilic bacterium Micrococcus cryophilus (ATCC 15174).

The positional specificity of the desaturase activity in the psychrophilic bacterium Micrococcus cryophilus (ATCC 15174) is shown to be delta9. The desaturase is inhibited by sterculic acid. Small amounts of delta8, delta10 and delta11 isomers are present. The implications of these findings for fatty acid metabolism in M. cryophilus are discussed. It is suggested that the temperature-dependent chain length change, known to occur in the phospholipid fatty acids of this bacterium, is not mediated by either a temperature-dependent change in desaturase substrate specificity or the induction of new desaturase enzymes with novel positional specificity. It is concluded that the control by temperature of fatty acid chain length is mediated by either a temperature-dependent change in the products of fatty acid synthetase or a temperature-sensitive palmitate elongase.

The effect of salinity on the phase behaviour of total lipid extracts and binary mixtures of the major phospholipids isolated from a moderately halophilic eubacterium.

The effects of molar NaCl concentrations on the phase behaviour of the total lipid extracts and binary mixtures of the major phospholipids, namely phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), isolated from the moderately halophilic eubacterium, Vibrio costicola, grown in 1 M and 3 M NaCl containing media have been studied using X-ray diffraction and freeze-fracture electron microscopy. The effect of both the PE/PG ratio and alterations in fatty acid composition were examined by using binary mixtures which mimicked the PE/PG ratio found in the native bacterial membranes. We show that the samples exhibited complex phase behaviour, including the formation of non-bilayer phases, which depend upon the salinity of both the bacterial culture medium and the suspending solution. The total lipid from bacteria cultured in 1 M NaCl-containing medium and dispersed in 1 M NaCl exhibited a mixture of L alpha and hexagonal-II phases at the optimum growth temperature of the organism (i.e., 30 degrees C), whereas the same lipid dispersed in 3 M NaCl showed only a hexagonal-II phase down to a temperature of +3 degrees C. The total lipid extracted from 3 M NaCl cultures showed only lamellar phases over the temperature range studied (+50 degrees C to -50 degrees C), but the phase transition temperatures of the various lamellar phases were generally higher when the lipid was dispersed in 3 M compared with 1 M NaCl. The phase behaviour of the binary mixtures was similar but not identical to that of the corresponding total lipid extracts and it is suggested that the minor lipid components (diphosphatidylglycerol, lysophosphatidylethanolamine and lysophosphatidylglycerol) play a part in determining the phase behaviour of the native membranes. These results show that the PE/PG ratio and fatty acid composition of the individual phospholipids, which are normally regulated by Vibrio costicola in vivo in response to culture medium salinity, are both important in maintaining a stable bilayer structure within the membrane.

Polyunsaturated fatty acids in the psychrophilic bacterium Shewanella gelidimarina ACAM 456T: molecular species analysis of major phospholipids and biosynthesis of eicosapentaenoic acid.

The production of eicosapentaenoic acid [20:5omega3; EPA] from Shewanella gelidimarina (ACAM 456T) was investigated with respect to growth temperature and growth on sole carbon sources. The percentage and quantitative yield of EPA remained relatively constant at all growth temperatures within or below the optimal growth temperature region. At higher growth temperatures, these values decreased greatly. Growth on differing sole carbon sources also influenced the percentage and amount of EPA produced, with the fatty acid composition influenced by provision of potential acyl chain primers as sole carbon sources. The highest amounts of EPA occurred from growth on propionic acid and L-leucine respectively, while the highest percentage of EPA occurred from growth on L-proline. Monounsaturated fatty acid components and EPA were concentrated in phosphatidylglycerol (PG), while the proportion of branched-chain fatty acids was elevated in phosphatidylethanolamine (PE); the two major phospholipid classes. Specific associations of EPA with other acyl chains were identified within cellular phospholipid classes. The association of EPA with 17:1 and 18:0 acyl chains in phospholipid species was specific to PG, whereas the association of EPA with i13:0/13:0 and 14:0/i14:0 was specific to PE. Such acyl chain 'tailoring' is indicative of the important role of EPA in bacterial membrane adaptive responses. EPA was also a large component (22%) of a non-esterified fatty acid (NEFA) fraction within the total lipid extract of the bacterium. This may point toward a particular role of NEFA in polyunsaturated fatty acid (PUFA) metabolism. The formation of EPA was investigated by labelling with L-[U-14C]serine and sodium [1-14C]acetate. The accumulation of radiolabel within unsaturated intermediates (di-, tri- and tetraunsaturated fractions) was low, indicating a rapid formation and derivatisation of these components. Similar results were found for the unsaturated fatty acid fractions of both PE and PG using sodium [1-14C]acetate radiolabel. The regulation of triunsaturated fatty acid components may be a potential control site in PUFA biosynthesis.

The role of substance P and calcitonin gene-related peptide in neurogenic plasma extravasation and vasodilatation in the rat.

Immunohistochemistry combined with retrograde tracing has been used to show that of the afferent neurons supplying the dorsomedial surface of the hind paw, approximately 30% contain substance P and 50% calcitonin gene-related peptide immunoreactivity. Stimulation of the saphenous nerve causes plasma extravasation and antidromic vasodilatation in this area of skin. The roles of calcitonin gene-related peptide and substance P released from peripheral afferent endings in mediating these effects were examined using immunoneutralization. In pilot experiments, the binding of radiolabelled peptide to the immunoglobulin fraction of calcitonin gene-related peptide and substance P antisera was characterized in quasi-physiological conditions. Systemic administration of either substance P or calcitonin gene-related peptide antibodies caused a significant decrease (P less than 0.05) in plasma extravasation measured by the Evans Blue method in response to topical application of mustard oil (0.5%) to the skin, or of capsaicin (5 microM) to the saphenous nerve. Topical application of mustard oil also produced a 52.9 +/- 5.1% increase in skin red cell flux. This increase was significantly decreased by both substance P and calcitonin gene-related peptide antibodies. The results suggest that both peptides are involved in mediating neurogenic inflammatory responses.

Immunization with calcitonin gene-related peptide reduces the inflammatory response to adjuvant arthritis in the rat.

The influence of circulating antibodies to calcitonin gene-related peptide on the inflammatory response was examined in rats with adjuvant-induced arthritis. Rats were immunized with alpha calcitonin gene-related peptide conjugated to thyroglobulin, and circulating antibodies were identified by their capacity to bind radiolabelled rat alpha or human calcitonin gene-related peptide. In unimmunized rats and rats immunized with thyroglobulin alone, the secondary lesions (characterized as paw swelling, nodules on ears and tail, and inflamed nose) produced after adjuvant-induced arthritis were similar. However, at 21 days, when these lesions were maximal, the animals immunized with calcitonin gene-related peptide showed decreased numbers of lesions. An additional marker of disease activity, namely alpha 1 glycoprotein levels in plasma, was also measured. Again, plasma alpha 1 glycoprotein levels were similar in rats that were unimmunized or received thyroglobulin alone, but at 21 days were significantly reduced in animals immunized with calcitonin gene-related peptide. In contrast, the initial foot swelling seen in the first few days after injection of adjuvant was not significantly different in the various groups. The results suggest that antibodies to calcitonin gene-related peptide are able to reduce the severity of the adjuvant arthritis syndrome, and that this peptide contributes to the inflammatory response seen in the later stages of the disease model.

Ultrastructural study of selenite-induced nuclear cataracts.

The formation of the selenite-induced cataract was investigated by examining the ultrastructure of the cataract with transmission electron microscopy (TEM). A lacy, or honeycomb, appearance of the nuclear cataract seen by light microscopy was resolved by TEM to be due to the aggregation or precipitation of cytoplasmic material. Despite severe intracellular changes the fibers retained their close apposition to one another. These results are consistent with the hypothesis that a major lesion in selenite-induced nuclear cataracts is the formation of insoluble cytoplasmic aggregates.

Relationship of radiologic position to the diagnostic yield of fiberoptic bronchoscopy in bronchial carcinoma.

The radiologic position of tumors in 100 patients with bronchial carcinoma was compared with the diagnostic yield from fiberoptic bronchoscopy. A new method for dividing the chest x-ray film into three areas (hilar, perihilar, and peripheral) was based upon the expected range of vision of the fiberoptic bronchoscope. At bronchoscopy without fluoroscopy, only eight (36 percent) of 22 radiologically peripheral tumors were diagnosed, compared with 31 (94 percent) out of 33 hilar tumors (p less than 0.001) and 34 (76 percent) out of 45 perihilar tumors (p less than 0.01). The results show that in hilar and perihilar tumors, fiberoptic bronchoscopy gives a high diagnostic yield, whereas in peripheral tumors the yield is poor. This method allows the clinician to judge from the chest x-ray film whether a tumor is likely to be visible bronchoscopically. Thus, in radiologically peripheral tumors, as defined by our method, fluoroscopy at the time of fiberoptic bronchoscopy should be available to the clinician, or alternative diagnostic methods should be considered.

Inhibition of ergosterol biosynthesis is not accompanied by a change in fatty acid composition in Saccharomyces cerevisiae treated with the antifungal agent 6-amino-2-n-pentylthiobenzothiazole.

The antifungal agent 6-amino-2-n-pentylthiobenzothiazole at a concentration of 40 microM lowered the specific growth rate of exponentially growing cultures of Saccharomyces cerevisiae by 36%. Treatment with 6-amino-2-n-pentylthiobenzothiazole inhibited the biosynthesis of ergosterol and caused an accumulation of the methylated sterol precursors ergosta-5,7-dienol and squalene, but had no significant effect on the composition and the rate of biosynthesis of fatty acids. The results indicate that neither the inhibition of ergosterol biosynthesis, nor the slowing-down of culture growth by this antifungal agent, led to a compensatory alteration in the pattern of fatty-acyl chains in membrane lipids. This finding contradicts the accepted wisdom for the action of a number of antifungal agents.

Isolation of mucoid strains of Pseudomonas aeruginosa from non-cystic-fibrosis patients and characterisation of the structure of their secreted alginate.

When the incubation period of primary isolation plates was extended to 48 h, mucoid strains of Pseudomonas aeruginosa were found in specimens from various infected sites in patients who did not have cystic fibrosis. The 17 mucoid isolates were characterised in terms of mucoid type, pyocin type, and their sensitivity or resistance to seven beta-lactam and two aminoglycoside antibiotics. The carbohydrate, uronic acid (alginate) and protein content of the water-soluble extracellular material of 15 strains was determined. This material was fractionated by ion-exchange chromatography, and the presence of alginate confirmed by the chemical assay of uronic acids and their quantitation by gas-liquid chromatography. Uronic acids were absent from a non-mucoid revertant of one strain. The strains produced alginate with a high content of mannuronic acid and substituted with O-acetyl groups. By proton nuclear magnetic resonance (1H-nmr) analysis the alginate from three strains was shown to lack polyguluronate blocks in its structure. These properties are also found in the alginate of mucoid P. aeruginosa strains from patients with cystic fibrosis.

Dextromethorphan attenuates post-ischemic hypoperfusion following incomplete global ischemia in the anesthetized rat.

The effects of dextromethorphan (DM) were tested in an in vivo model of incomplete global cerebral ischemia. Anesthetized rats were divided into 4 groups: Group 1 (saline); Group 2 (DM pre-treatment, 20 mg/kg i.v. bolus followed by 10 mg/kg/h DM infusion); Group 3 (DM post-treatment, 2 mg/kg i.v. bolus followed by 10 mg/kg/h DM infusion at the onset of post-ischemic hypoperfusion); and Group 4 (sham-operated, drug-treated). Groups 1-3 underwent 15 min of 4-vessel occlusion followed by 3 h of reperfusion. Administration of DM in sham-operated animals (Group 4) had no effect on cerebral blood flow or electroencephalographic (EEG) activity. In contrast, when compared to the Group 1 saline controls, significant attenuation of post-ischemic hypoperfusion and EEG dysfunction was demonstrated in ischemic rats treated with DM (both pre- and post-treatment), suggesting an ability of DM to improve cerebral blood flow (CBF) and brain function in cerebral ischemia.

Opiates inhibit the discharges of fine afferent units from inflamed knee joint of the cat.

The spontaneous discharges in 15 out of 19 small-diameter afferent units from inflamed knee joints of anaesthetized cats were significantly inhibited by one or several opiates (morphine in the dose range 1.0-5.0 mg/kg; gly-ol 0.5-5.0 mg/kg; U50488 1.0-10.0 mg/kg; ethylketocyclazacine 0.5-4.0 mg/kg administered by close arterial injection into the joint). In the majority of cases a subsequent injection of naloxone (1 mg/kg i.a.) significantly reversed this effect. These data provide an electrophysiological demonstration that opiates may act on opiate receptors located at peripheral sites of primary afferent fibres and hence exert a peripheral 'analgesic' effect.

Antibodies to calcitonin-gene related peptide reduce inflammation induced by topical mustard oil but not that due to carrageenin in the rat.

The influence of prior immunization to calcitonin gene-related peptide (CGRP) was examined on responses in two models of inflammation plasma: protein extravasation following topical application of mustard oil to the hind paw, and changes in paw volume following subplantar injection of carrageenin. In rats with antibodies to CGRP, there was an approximately 50% decrease compared to the control animals in extravasation, measured by the Evans blue method after topical mustard oil. In contrast, there was no difference in the inflammatory response to carrageenin in control and immunized rats. The results indicate that CGRP, which is probably released from the peripheral endings of cutaneous primary afferents, plays a role in the inflammatory response to mustard oil, but not carrageenin.

Pulsed high electric field causes 'all or nothing' membrane damage in Listeria monocytogenes and Salmonella typhimurium, but membrane H+-ATPase is not a primary target.

Salmonella typhimurium (CRA 1005) was more sensitive than Listeria monocytogenes (NCTC 11994) to pulsed high electric field (PHEF) treatment in distilled water (10, 15 and 20 kV/cm), 10 mM Tris-maleate buffer, pH 7.4 (15 kV/cm) and model beef broth (0.75%, w/v; 15 kV/cm). Sublethal injury could not be detected using a selective medium plating technique, indicating that bacterial inactivation by PHEF may be an 'all or nothing' event. PHEF-induced membrane permeabilisation resulted in an increase in the leakage of UV-absorbing material from the bacteria (UV-leakage) and a decreased ability of L. monocytogenes to maintain a pH gradient. A lack of correlation between the inhibition of H+-ATPase activity and PHEF treatment, cell viability or UV-leakage indicates that this enzyme is probably not a primary site of bacterial inactivation despite its role in the maintenance of internal pH.