Admission inflammatory markers and isolation of a causative organism in patients with spontaneous spinal infection.

INTRODUCTION: The purpose of this study was to investigate the significance of the inflammatory markers on admission in the isolation of a causative pathogen in patients with spinal infection. Spinal infection is treated frequently at spinal units and can encompass a broad range of clinical entities. Its diagnosis is often delayed because of the difficulty of identifying the responsible pathogen. METHODS: Patients with spinal infection treated in our institution over a period of eight years were identified and their notes studied retrospectively. Admission C-reactive protein (CRP), white cell count (WCC) as well as co-morbidities and mode of pathogen identification were recorded. Overall, 96 patients were included in the study. RESULTS: The CRP levels on admission were correlated significantly with the overall potential for isolation of a pathogen (p<0.0001) and positive biopsy cultures (p=0.0016). Admission WCC levels were associated significantly with the overall potential for isolation of a pathogen (p=0.0003) and positive biopsy cultures (p=0.0023). Both CRP and WCC levels were significantly negatively correlated with the duration of the preceding symptoms (p=0.0003 and p<0.0001 respectively). Delay in presentation was significantly negatively correlated with organism isolation (p=0.0001). Multivariate analyses identified the delay in presentation as the strongest independent variable for organism isolation (p=0.014) in cases of spontaneous spinal infection when compared with the admission CRP level (p=0.031) and WCC (p=0.056). CONCLUSIONS: In spontaneous spinal infection, delay in presentation is the strongest independent variable for organism isolation. High inflammatory marker levels on admission are a useful prognostic marker for the overall potential of isolating a causative organism either by blood cultures or by biopsy in patients with negative blood cultures. Furthermore, the admission inflammatory marker levels allow for treating surgeons to counsel their patients of the likelihood of achieving a positive microbiological yield from biopsy.

The value of radiographs obtained during forced traction under general anaesthesia in predicting flexibility in idiopathic scoliosis with Cobb angles exceeding 60 degree.

Our aim in this prospective radiological study was to determine whether the flexibility rate calculated from radiographs obtained during forced traction under general anaesthesia, was better than that of fulcrum-bending radiographs before corrective surgery in predicting the extent of the available correction in patients with idiopathic scoliosis. We evaluated 33 patients with a Cobb angle > 60 degrees on a standing posteroanterior radiograph, who had been treated by posterior correction. Pre-operative standing fulcrum-bending radiographs and those with forced-traction under general anaesthesia were obtained. Post-operative standing radiographs were taken after surgical correction. The mean forced-traction flexibility rate was 55% (SD 11.3) which was significantly higher than the mean fulcrum-bending flexibility rate of 32% (SD 16.1) (p < 0.001). We found no correlation between either the forced-traction or fulcrum-bending flexibility rates and the correction rate post-operatively (p = 0.24 and p = 0.44, respectively). Radiographs obtained during forced traction under general anaesthesia were better at predicting the flexibility of the curve than fulcrum-bending radiographs in curves with a Cobb angle > 60 degrees in the standing position and may identify those patients for whom supplementary anterior surgery can be avoided.

Additional morphological and physiological heterogeneity within the midgut of larval Aedes aegypti (Diptera: Culicidae) revealed by histology, electrophysiology, and effects of Bacillus thuringiensis endotoxin.

Analysis of larval Aedes aegypti midgut using scanning electron microscopy, nuclear and mitochondrial dyes, response to Bacillus thuringiensis israelensis CryIVB toxin, and electrophysiology is described. The anterior ventriculus ("stomach") region is found to have much lower mitochondrial densities than other midgut regions. The transitional region is distinguished by apical surface architecture, and by region-specific effects of CryIVB endotoxin. In this region CryIVB causes holes ranging from 1.0 to 7.0 microm in diameter (mean 3.3+/-0.53 microm, N=12), blisters 16.9+/-1.54 microm in diameter (N=10), and separation of adjacent cells. The holes are not consistent with damage due to the colloid osmotic lysis model of delta-endotoxin activity. The posterior ventriculus possesses a distinctive cellular architecture consisting of hemispherical, domed apical membranes surrounded by deep clefts. Functional and morphological heterogeneity is revealed within the posterior ventriculus, with the anterior end dominating the electrical profile of isolated, perfused preparations and showing the greatest response to serotonin. Hyperpolarization of the transepithelial potential by serotonin occurred in conjunction with a decrease in the space constant lambda, ruling out closure of ion channels as the mechanism of action of serotonin.

Confirmatory method for the analysis of carbadox and olaquindox in porcine feedingstuffs using LC-electrospray MS-MS.

A method is described for the quantitative determination of the two feed additives carbadox and olaquindox in porcine feedingstuffs. The use of these agents in feedingstuffs was prohibited in the European Union as a result of concerns about their toxicity. Regulatory laboratories are required to have suitably validated analytical methods to ensure compliance with the ban. The analytes were extracted from finished feedingstuffs into acetonitrile:chloroform (1:1, v/v), and aliquots (1.0 ml) of the extract were dried down under a stream of nitrogen at 65 degrees C. All residues were re-dissolved in HPLC mobile phase containing acetonitrile/water/formic acid. Analysis was based on LC coupled to positive-ion electrospray MS-MS, with daughter ions for carbadox at m/z 231 and 90, and for olaquindox at m/z 212 and 143 being monitored. The method was validated by analysing feed samples fortified with carbadox and olaquindox at 0.5, 2.5 and 5 mg kg(-1) on three separate occasions. Sample preparation was simple, thus allowing the confirmation of these compounds in large numbers of samples.

Confirmation of carbadox and olaquindox metabolites in porcine liver using liquid chromatography-electrospray, tandem mass spectrometry.

A method is described for the quantitative determination of quinoxaline-2-carboxylic acid (QCA) and methyl-3-quinoxaline-2-carboxylic acid (MQCA), the metabolites that have been designated as the marker residues for the veterinary drugs, carbadox and olaquindox, respectively, in swine tissue. The method is suitable for use as a confirmatory method under EU National Surveillance Schemes. Porcine liver samples were subjected to protease digestion followed by liquid-liquid extraction. Further clean-up was performed by automated solid phase extraction (SPE) and was followed by a final liquid-liquid extraction step. Analysis was performed using a narrow bore column HPLC coupled to electrospray MS/MS, operated in positive ion mode. MS/MS product ions were monitored at m/z 102 and 75 amu for QCA, m/z 145 and 102 amu for MQCA and at m/z 106 and 152 amu for the d(4)-QCA and d(7)-MQCA internal standards, respectively. The method has been validated at 3.0, 10, 50 and 150 microg kg(-1) for both metabolites. The method performance characteristics-the decision limit (CCalpha) and the detection capability (CCbeta) have been determined for QCA at 0.4 and 1.2 microg kg(-1), respectively, and for MQCA at 0.7 and 3.6 microg kg(-1), respectively.

Quinoxaline-2-carboxylic acid in pigs: criteria to distinguish between the illegal use of carbadox and environmental contamination.

Carbadox cannot be used in food-producing animals within the European Union following the adoption of Commission Regulation EC 2788/98/EC. Monitoring of the longest remaining residue--quinoxaline-2-car-boxylic acid (QCA)--is the most effective way of enforcing the prohibition on its use. The study was under taken to determine if QCA could be passed from pig to pig following the exposure of unmedicated animals to housing that had previously contained medicated animals. Drug-withdrawal studies were also carried out on medicated animals. Distinction between treated animals and those exposed to QCA might be required by competent national authorities to determine whether a positive result for QCA in tissue is truly 'violative'. Comparison of the ratio concentrations of QCA in tissues and body fluids was made to determine if they, could be used as criteria for discrimination between illegally treated animals and environmental contamination.

Development and validation of an improved method for confirmation of the carbadox metabolite, quinoxaline-2-carboxylic acid, in porcine liver using LC-electrospray MS-MS according to revised EU criteria for veterinary drug residue analysis.

A method is described for the quantitative determination of quinoxaline-2-carboxylic acid (QCA), the marker residue for the veterinary drug carbadox, in swine liver. Tissue is subjected to alkaline hydrolysis followed by liquid-liquid extraction. QCA residues are cleaned up using automated solid phase extraction (SPE), before a final liquid-liquid extraction step. Analysis is based on LC coupled to positive ion electrospray MS-MS, monitoring product ions at m/z 129, 102 and 75 for QCA and at m/z 106 for the internal standard (d4-QCA). The method has been validated according to draft revised EU criteria for analysis of veterinary drug residues, and is suitable for monitoring tissues taken under national surveillance schemes. The method has been validated at 3, 10, 30, 100 and 300 microg kg(-1). The method performance characteristics, CCalpha (decision limit) and CCbeta (detection limit) were determined to be 0.16 and 0.27 microg kg(-1), respectively. The described method, which is relatively rapid and applicable to large sample numbers, correlates well (r2 = 0.9799) with a widely used GC-MS assay for QCA.

Accumulation and persistence of flea larvicidal activity in the immediate environment of cats treated with imidacloprid.

To investigate the persistence of flea larvicidal activity in the immediate environment of cats treated with imidacloprid, eggs of the cat flea Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), from untreated donor cats, were incubated on samples of fleece blanket taken from the floor of cages used by treated or untreated cats for a total of 10 or 20 6-h periods over 2-4 weeks, respectively. Sufficient imidacloprid accumulated during these periods to reduce the emergence of adult fleas by 94.7-97.6% when the blankets were tested after 18 weeks' storage at room temperature. A typical laundry procedure (washing with detergent at 50 degrees C and low temperature tumble drying) removed this biological activity. Unwashed control blankets did not support the flea life-cycle as effectively as washed blankets or a sand substrate.

Establishment of the cat flea (Ctenocephalides felis felis) on the ferret (Mustela putorius furo) and its control with imidacloprid.

As the ferret, Mustela putorius furo L. (Carnivora: Mustelidae), is becoming increasingly popular as a pet animal and as it is susceptible to the cat-flea, Ctenocephalides felis felis Bouché (Siphonaptera: Pulicidae), an experimental model was established for evaluating insecticidal treatments on this host. A high establishment rate (76.7-91.8%) was recorded when 60 unfed adult C. felis were placed on ferrets. This provided an adequate infestation for chemotherapeutic evaluation without causing undue discomfort to the host. Twelve ferrets were allocated to two groups matched for sex and individual ability to sustain a flea population. One group was treated topically with an imidacloprid spot-on formulation at a dose rate of 10 mg/kg body-weight on Day 0. All ferrets were infested with C. felis on Days -1, 7, 14, 21 and 28, and flea counts were performed 8 and 24 h post-treatment and one day after each subsequent infestation. Fleas were removed at all but the 8 h count (when they were returned to their host). Flea burdens were reduced by 95.3% (P < 0.001) within 8 h of treatment and 100% efficacy was recorded at 24 h. At 1, 2, 3 and 4 weeks post-treatment, protection against re-infestation was 92.9% (P < 0.001), 55.7% (P < 0.02), 18.3% (NS) and 7.4% (NS), respectively. Thus, at this dose rate, imidacloprid gave excellent efficacy against a resident C. felis population and provided a high level of residual activity for at least one week after treatment.

Control of flea populations in a simulated home environment model using lufenuron, imidacloprid or fipronil.

Control strategies were evaluated over a 6-month period in a home simulation model comprising a series of similar carpeted pens, housing matched groups of six cats, in which the life-cycle of the flea Ctenocephalides felis felis Bouche (Siphonaptera: Pulicidae) had been established. Additional adult fleas were placed on the cats at intervals to mimic acquisition of extraneous fleas from outside the home. Treatment strategies included a single subcutaneous deposition of injectable lufenuron supported by initial treatments with a short-acting insecticidal spray, or monthly topical applications of imidacloprid or fipronil. An untreated control group indicated that conditions were suitable for flea replication and development. Controls had to be combed on 18 occasions to remove excessive flea burdens and two developed allergic reactions. Lufenuron cats were combed once and required two insecticidal treatments in the first month to achieve control. Even so, small flea burdens were constantly present thereafter. Imidacloprid and fipronil treatments appeared to give virtually complete control throughout. Single fleas were found on imidacloprid cats on two occasions, whereas none were recovered from fipronil cats at any time after the first treatment. Tracer cats were used to monitor re-infestation rates at the end of the trial period. Small numbers of host-seeking fleas were demonstrated in all treatment pens, indicating that total eradication had not been accomplished. It is concluded that the home environment simulation model incorporating tracer animals could provide a powerful tool for studying flea population dynamics under controlled conditions but improved techniques are needed for quantifying other off-host life-cycle stages.

Inhibition of immature Ctenocephalides felis felis (Siphonaptera: Pulicidae) development in the immediate environment of cats treated with imidacloprid.

The hypothesis that dermally applied imidacloprid may transfer from treated cats, Felis catus L., to their immediate environment in quantities sufficient to have a significant effect on developing immature cat fleas, Ctenocephalides felis (Bouché), was tested in a controlled experiment. Flea eggs harvested from untreated donor cats were incubated on replicated samples from blankets used by treated or untreated cats under standardized conditions. As compared with controls, the percentage of adult flea emergence on blankets used by treated animals was reduced by 100% in the 1st wk after treatment and by 84, 60, and 74% in subsequent weeks (P < 0.001).

Kapandjii's procedure in a former far eastern prisoner of war with poliomyelitis of the affected limb.

A case of traumatic radio-ulnar dislocation in a limb previously affected by poliomyelitis is presented. Treatment with Kapandjii's procedure is described and discussed.

Evaluation of flea control strategies using fipronil on cats in a controlled simulated home environment.

Three groups of six cats were kept in similar carpeted pens in which a self-replicating population of Ctenocephalides felis had been established. One group was left untreated, but the other groups were treated every 28th day with 0.5 ml of a 10 per cent fipronil spot-on formulation, and the cats in one of the treated groups also wore a methoprene collar. No fleas were found on any of the treated cats, either during the first 13 weeks of the study, when heavy flea burdens were developing in the control pen, or over the next 11 weeks when a declining number of fleas was present on the control group.

Comparison of flea control strategies using imidacloprid or lufenuron on cats in a controlled simulated home environment.

OBJECTIVE: To compare the effect of monthly treatments with imidacloprid (an adulticide) or lufenuron (an insect development inhibitor) for protecting cats against Ctenocephalides felis felis in a simulated home environment. ANIMALS: 3 matched groups of 4 cats each. PROCEDURE: A self-propagating flea life cycle continuously exposing cats to 'natural' infestation was established in 3 pens. Small artificial infestations were later superimposed to mimic the effect of a cat roaming outdoors and acquiring extraneous fleas. One pen housed an untreated control group, and the other 2 pens housed cats treated every 28th day with an imidacloprid spot-on formulation or lufenuron suspension, respectively. Flea counts were performed at 14-day intervals for 112 days. RESULTS: Flea numbers increased on control cats around day 42 when mean counts on cats in the imidacloprid and lufenuron groups decreased by 100 and 86.8 percent, respectively. Fleas were not found on any imidacloprid-treated cat, but lufenuron-treated cats were consistently parasitized. CONCLUSIONS: Imidacloprid administered at monthly intervals maintained flea burdens below the limit of detection, whereas clinically important flea populations developed in the lufenuron treatment pen. CLINICAL RELEVANCE: Results from this experimental model suggest that flea populations within a home may be controlled by carefully timed on-host treatments with potent long-acting insecticides such as imidacloprid.

Inhibitory effect of GA3 on the development of thidiazuron-induced somatic embryogenesis in geranium (Pelargonium xhortorum Bailey) hypocotyl cultures.

Somatic embryogenesis in geranium (Pelargonium xhortorum Bailey cv 'Scarlet Orbit Improved') can be achieved by incubating hypocotyl explants on MS medium supplemented with thidiazuron (TDZ; 10 µM for 3 days followed by subculture on medium devoid of any plant growth regulators. The presence of gibberellins (GAs) during both the induction and expression phases of embryogenesis was significantly detrimental to somatic embryo formation on the hypocotyl explants. The addition of the GA-synthesis inhibitors paclobutrazol, uniconazole or ancymidol during the period of growth and differentiation of somatic embryos increased the number of somatic embryos formed on each explant. However, paclobutrazol added during the period of induction had no significant influence on somatic embryo formation. Results suggest that both exogenously supplied as well as endogenous GAs play a role, albeit a negative one, on somatic embryogenesis of geranium.

A novel approach to flea control on cats, using pyriproxyfen.

Pyriproxyfen is a new insect growth regulator with potent activity against developing stages of the cat flea, Ctenocephalides felis. To determine its duration of action after application as a spot-on topical treatment, 20 cats were allocated to two groups. One group acted as untreated controls, and each cat in the other was given a single dose of 1 mg/kg pyriproxyfen. Each cat was infected with fleas at weekly intervals and caged. Eggs were collected and their viability determined as a measure of the effect of the agent on adult fleas. The effect of the transfer of pyriproxyfen from each cat to its immediate surroundings was monitored by observing the development of fertile flea eggs placed on samples of blanket taken from each cage. Results from untreated control cats showed that the experimental conditions were favourable for the lifecycle of the flea. Flea reproduction on treated cats was inhibited completely for seven weeks and substantially for at least a further two weeks. Blankets used by cats treated up to two weeks previously prevented flea development entirely and substantial control was still achieved up to four weeks after treatment.

Evaluation of the effect of the fenbendazole sustained-release intraruminal device on the immunity of calves to lungworm.

Each of 10 set-stocked calves was given a fenbendazole sustained-release intraruminal bolus at turnout for the control of parasitic bronchitis while a group of 10 similar calves was left untreated. The respiratory rates of the control calves were not greatly increased during the grazing season, but persistent coughing was evident from early July when they all had patent lungworm infections. Only occasional coughing was reported from the bolus-treated calves except for a transient increase in its frequency in late September. In mid-August, one of the treated calves was passing lungworm larvae and when they were housed six of the 10 had patent infections. In August tracer calves picked up an average of 23.5 lungworms per day from the control paddock but only 2.1 from the paddock grazed by the treated calves. In September the corresponding figures were 7.6 and 19.2 lungworms per day, respectively, from the control and 'treated' paddocks. After housing, the post mortem worm counts after an experimental challenge with Dictyocaulus viviparus larvae were reduced by 99.2 and 98.7 per cent (P < 0.0001), respectively, for the control and bolus-treated calves in comparison with weight-matched parasite-naive calves. Thus, despite a relatively low level of challenge during the grazing season, the treated animals had developed a considerable degree of protective immunity.