Discrimination between sheep-associated and wildebeest-associated malignant catarrhal fever virus by means of a single-tube duplex nested PCR

A single-tube duplex nested polymerase chain reaction (sdn-PCR) was developed for the detection of and discrimination between ovine herpesvirus-2 (OvHV-2) and alcelaphine herpesvirus-1 (AIHV-1). These viruses respectively cause sheep- and wildebeest-associated malignant catarrhal fever (SA-MCF and WA-MCF). In the first step of the sdn-PCR, two primers with high annealing temperatures based on conserved regions of the tegument genes were used for DNA amplification. In the second step, two primer sets based on variable regions of the respective OvHV-2 and AIHV-1 genes and with annealing temperatures > 11 degrees C below the primers used in the first step, were used. Internal regions of different sizes from amplicons produced in the first step were amplified. This single-tube test obviates the need for two separate assays to detect both viral types, thereby reducing time, labour and cost.     

Influence of host diversity on development of epidemics: an evaluation and elaboration of mixture theory

ABSTRACT A spatiotemporal/integro-difference equation model was developed and utilized to study the progress of epidemics in spatially heterogeneous mixtures of susceptible and resistant host plants. The effects of different scales and patterns of host genotypes on the development of focal and general epidemics were investigated using potato late blight as a case study. Two different radial Laplace kernels and a two-dimensional Gaussian kernel were used for modeling the dispersal of spores. An analytical expression for the apparent infection rate, r, in general epidemics was tested by comparison with dynamic simulations. A genotype connectivity parameter, q, was introduced into the formula for r. This parameter quantifies the probability of pathogen inoculum produced on a certain host genotype unit reaching the same or another unit of the same genotype. The analytical expression for the apparent infection rate provided accurate predictions of realized r in the simulations of general epidemics. The relationship between r and the radial velocity of focus expansion, c, in focal epidemics, was linear in accordance with theory for homogeneous genotype mixtures. The findings suggest that genotype mixtures that are effective in reducing general epidemics of Phytophthora infestans will likewise curtail focal epidemics and vice versa.     

Public health importance of Brachiola algerae (Microsporidia)--an emerging pathogen of humans

Brachiola algerae, a parasite of Anopheles mosquitoes, has also been isolated from a human cornea, a cutaneous nodule and deep muscle tissue. All three human isolates of B. algerae are morphologically, serologically, and genetically similar to the mosquito-derived isolates including the original isolate of Vavra and Undeen. All of these isolates grew well in mammalian cell cultures at 37 degrees C and produced spores. Transmission electron microscopy revealed that all developmental stages including meronts, sporoblasts and spores were diplokaryotic and developed in direct contact with the host cell cytoplasm, a feature characteristic of the genus Brachiola. Spores of all isolates reacted well, in the immunofluorescence assay, with the rabbit anti-B. algerae serum. In the immunoblot assay, although the overall banding patterns of the human and mosquito isolates were similar, minor differences could be discerned. Sequencing of the PCR products of the amplified SSU rRNA gene revealed the existence of two distinct genotypes; the original mosquito (Undeen) isolate belonged to genotype 1 and the isolate from cornea and that from the deep muscle biopsy to genotype 2, whereas the isolates from a mosquito and one of the other two human isolates (one from skin abscess) had both genotypes, 1 and 2. It is known that spores of mosquito-derived B. algerae can not only proliferate in mammalian cell cultures at 37 degrees C but also can infect mice when injected into footpads or deposited on the corneal surface. These observations indicate that the spores have potential to be a risk factor for humans, especially those with immunodeficiency.     

Modeling Spatial Characteristics in the Biological Control of Fungi at Leaf Scale: Competitive Substrate Colonization by Botrytis cinerea and the Saprophytic Antagonist Ulocladium atrum

ABSTRACT A spatially explicit model describing saprophytic colonization of dead cyclamen leaf tissue by the plant-pathogenic fungus Botrytis cinerea and the saprophytic fungal antagonist Ulocladium atrum was constructed. Both fungi explore the leaf and utilize the resources it provides. Leaf tissue is represented by a two-dimensional grid of square grid cells. Fungal competition within grid cells is modeled using Lotka-Volterra equations. Spatial expansion into neighboring grid cells is assumed proportional to the mycelial density gradient between donor and receptor cell. Established fungal biomass is immobile. Radial growth rates of B. cinerea and U. atrum in dead cyclamen leaf tissue were measured to determine parameters describing the spatial dynamics of the fungi. At temperatures from 5 to 25 degrees C, B. cinerea colonies expanded twice as rapidly as U. atrum colonies. In practical biological control, the slower colonization of space by U. atrum thus needs to be compensated by a sufficiently dense and even distribution of conidia on the leaf. Simulation results confirm the importance of spatial expansion to the outcome of the competitive interaction between B. cinerea and U. atrum at leaf scale. A sensitivity analysis further emphasized the importance of a uniform high density cover of vital U. atrum conidia on target leaves.     

Predicting mortality of phosphine-resistant adults of Sitophilus oryzae (L) (Coleoptera: Curculionidae) in relation to changing phosphine concentration

Adults of a phosphine-resistant strain of Sitophilus oryzae (L) were exposed to constant phosphine concentrations of 0.0035-0.9mg litre(-1) for periods of between 20 and 168h at 25 degrees C, and the effects of time and concentration on mortality were quantified. Adults were also exposed to a series of treatments lasting 48, 72 or 168 h at 25 degrees C, during which the concentration of phosphine was varied. The aim of this study was to determine whether equations from experiments using constant concentrations could be used to predict the efficacy of changing phosphine concentrations against adults of S oryzae. A probit plane without interaction, in which the logarithms of time (t) and concentration (C) were variables, described the effects of concentration and time on mortality in experiments with constant concentrations. A derived equation of the form C(n)t = k gave excellent predictions of toxicity when applied to data from changing concentration experiments. The results suggest that for resistant S oryzae adults there is nothing inherently different between constant and changing concentration regimes, and that data collected from fixed concentrations can be used to develop equations for predicting mortality in fumigations in which phosphine concentration changes. This approach could simplify the prediction of efficacy of typical fumigations in which concentrations tend to rise and then fall over a period of days.     

In vitro study of heat production during power reduction of equine mandibular teeth

OBJECTIVE: To measure the amount of heat generated during 3 methods of equine dental reduction with power instruments. DESIGN: In vitro study. SAMPLE POPULATION: 30 premolar and molar teeth removed from mandibles of 8 equine heads collected at an abbatoir. PROCEDURE: 38-gauge copper-constantan thermocouples were inserted into the lingual side of each tooth 15 mm (proximal) and 25 mm (distal) from the occlusal surface, at a depth of 5 mm, which placed the tip close to the pulp chamber. Group-NC1 (n = 10) teeth were ground for 1 minute without coolant, group-NC2 (10) teeth were ground for 2 minutes without coolant, and group-C2 (10) teeth were ground for 2 minutes with water for coolant. RESULTS: Mean temperature increase was 1.2 degrees C at the distal thermocouple and 6.6 degrees C at the proximal thermocouple for group-NC1 teeth, 4.1 degrees C at the distal thermocouple and 24.3 degrees C at the proximal thermocouple for group-NC2 teeth, and 0.8 degrees C at the distal thermocouple and -0.1 degrees C at the proximal thermocouple for group-C2 teeth. CONCLUSIONS AND CLINICAL RELEVANCE: In general, an increase of 5 degrees C in human teeth is considered the maximum increase before there is permanent damage to tooth pulp. In group-NC2 teeth, temperature increased above this limit by several degrees, whereas in group-C2 teeth, there was little or no temperature increase. Our results suggest that major reduction of equine teeth by use of power instruments causes thermal changes that may cause irreversible pulp damage unless water cooling is used.     

Influence of concomitant quinidine administration on dextromethorphan disposition in rats

High doses of dextromethorphan (DM) have been clinically investigated for the treatment of multiple neuronal disorders including neuropathic pain. Several authors have suggested the concomitant administration of DM and a CYP2D6 reversible inhibitor in order to enhance the exposure of DM and limit the exposure to total dextrorphan (DX). The present study proposes to determine whether or not a single dose of quinidine is sufficient to enhance the plasma concentrations of DM in rats and keep those of DX at a minimal level. Oral doses of DM (50 mg/kg) were administered with increasing dose levels of quinidine (0, 2, 20, and 50 mg/kg) to male Sprague-Dawley rats and blood samples were collected over 24 h. Plasma concentrations of DM and total DX were determined using ESI-LC/MS/MS. Quinidine coadministration resulted in a more than twofold increase in the area under the curve of DM with an ED(50) of approximately 2 mg/kg whereas those of total DX were only increased by 21%. These results support the working hypothesis that a single dose of quinidine may enhance the plasma concentrations of DM relative to those of total DX and may therefore improve the treatment of neuropathic pain.     

Aplastic articular facets in a dog with intervertebral disk rupture of the 12th to 13th thoracic vertebral space

A 6-year-old, female spayed Pomeranian was presented with acute hind-limb paraplegia with the presence of deep pain perception and urinary incontinence. Myelography showed a Hansen type I herniation of the12th to 13th thoracic intervertebral space (T(12-13)). Articular facets of the T(12-13) and T(13) to first lumbar vertebra (L(1)) were absent. The spinal cord was decompressed using a bilateral T(12-13) modified lateral hemilaminectomy (pediculectomy). The aplastic sites were associated with minimal instability of the vertebral column, and stabilization of the vertebral column was not required. Familiarity with this condition is important, because articular facet aplasia may cause vertebral instability and may require an adjusted surgical approach or vertebral reduction and fusion following decompression.     

Clinical experience with butyl-2-cyanoacrylate adhesive in the management of canine and feline corneal disease

PURPOSE: To examine and evaluate clinical indications and postoperative outcome in a series of small animal patients in which corneal disease was managed by the application of butyl 2-cyanoacrylate adhesive. MATERIALS AND METHODS: In this retrospective study all small animal patients were identified that presented to the Royal Veterinary College, University of London over a 2-year period, in which corneal disease was managed by the application of butyl 2-cyanoacrylate. Indications for application, complicating factors prior to gluing, glue retention time, postoperative comfort, and extent of subsequent corneal reaction and scarring were noted for each case. Long-term follow-up data concerning visual and cosmetic outcome were obtained from owners and referring veterinarians. RESULTS: Thirty-seven patients (28 dogs and 9 cats), in which 39 eyes were treated, were identified. Indications for corneal gluing in this series included stromal ulceration (26/39 eyes); descemetocele (4/39 eyes); corneal laceration/foreign body (5/39 eyes); lamellar keratectomy (3/39 eyes) and superficial ulceration (1/39 eyes). At least one factor responsible for initiation, persistence or progression of the ulcer was identified in 66.7% of eyes prior to corneal gluing. These included keratomalacia; confirmed bacterial keratitis; corneal edema related to endothelial disease and keratoconjunctivitis sicca. Cyanoacrylate was generally well tolerated by patients with only 8/34 eyes demonstrating transient blepharospasm and increased lacrimation postoperatively. Retention time of cyanoacrylate varied widely from < 1 week to approximately 6 months, but was < 2 months in the majority (89%) of eyes. Exaggerated corneal vascularization was an infrequent postoperative complication, noted in only six canine eyes, and did not appear to be related to initial corneal disease, glue retention time or breed. CONCLUSIONS: Butyl 2-cyanoacrylate offers a convenient, economical and effective alternative to other treatment modalities, such as conjunctival grafts, in the management of corneal defects in canine and feline patients.