Adaptation of an automated assay for determination of beta-hydroxybutyrate in dogs using a random access analyzer

An automated method for measuring beta-hydroxybutyrate was adapted to the Ciba-Corning 550 Express trade mark random access analyzer. The assay was based on a kinetic reaction utilizing hydroxybutyrate-dehydrogenase. Beta-hydroxybutyrate concentration (mmol/L) was calculated ratiometrically using a 1.0 mmol/l standard. Canine serum, plasma, and urine were used without prior deproteinization and only a 30-microliter sample was required. The method demonstrated good linearity between 0 to 2 mmol/l of beta-hydroxybutyrate. Analytical recovery (accuracy) within these concentrations ranged from 85.8 to 113.3%. Both within-run and day-to-day precision were determined, as was specificity of the assay in the presence of a variety of interfering substances. The automated assay was rapid and economical, with reagent stability maintained for at least 2 weeks at 4 degrees C. This assay can readily be applied toward the assessment of ketoacidosis in dogs, and with further validation, other species.     

Health and urban living

The majority of people now live in urban areas and will do so for the foreseeable future. As a force in the demographic and health transition, urbanization is associated with falling birth and death rates and with the shift in burden of illness from acute childhood infections to chronic, noncommunicable diseases of adults. Urban inhabitants enjoy better health on average than their rural counterparts, but the benefits are usually greater for the rich than for the poor, thus magnifying the differences between them. Subject to better evidence, I suggest that the main obstacles to improving urban health are not technical or even financial, but rather are related to governance and the organization of civil society.     

An olfactometer for measuring the repellent effect of chemicals on the stable fly Stomoxys calcitrans (L.)

An olfactometer is described that permits the evaluation of chemicals acting in the vapour phase against the stable fly Stomoxys calcitrans (L.). Hungry flies, held in a metal frame cage covered with soft gauze, are attracted to a target held adjacent to one side of the cage, and emitting an attractive airflow. Flies landing on the target are counted at intervals of 15s. Flies starved for 24h were tested in the olfactometer when 2, 3 and 4 days old. With the exception of the first exposure of the 4-day-old flies, all the flies of the three ages showed greater than 80% attraction to the olfactometer target after exposure for 2 min, for each of four successive exposures at 10-min intervals. For all ages there was a significant increase in attraction at the second exposure but no further increase with subsequent exposures. It is likely that the sensillae, stimulated by the first exposure, subsequently responded faster. Eight chemicals were incorporated into the airflow behind the target to test their effect on flies. Permethrin (a low vapour pressure pyrethroid) and crotoxyphos (a low vapour pressure organophosphate) did not act in the vapour phase. Empenthrin (a high vapour pressure pyrethroid) and dichlorvos (a high vapour pressure organophosphate) both exhibited the toxic effect of knock-down on the flies without repellency. Oil of citronella and citronellol, two known fly repellents, gave a general reduction of attraction that was dose dependent; fewer flies found the target and those that did stayed a shorter length of time. Natural pyrethrum gave a similar effect to these two at lower concentrations, but at higher concentrations, it also showed a repellent effect. N,N-Diethyl-m-toluamide gave a low reduction in attraction. The olfactometer shows potential for use with other flying insects, both to observe the effects of an insecticide and also to study the behavioural responses in the absence of insecticides.     

African horse sickness

African horse sickness virus (AHSV) causes a non-contagious, infectious insect-borne disease of equids and is endemic in many areas of sub-Saharan Africa and possibly Yemen in the Arabian Peninsula. However, periodically the virus makes excursions beyond its endemic areas and has at times extended as far as India and Pakistan in the east and Spain and Portugal in the west. The vectors are certain species of Culicoides biting midge the most important of which is the Afro-Asiatic species C. imicola. This paper describes the effects that AHSV has on its equid hosts, aspects of its epidemiology, and present and future prospects for control. The distribution of AHSV seems to be governed by a number of factors including the efficiency of control measures, the presence or absence of a long term vertebrate reservoir and, most importantly, the prevalence and seasonal incidence of the major vector which is controlled by climate. However, with the advent of climate-change the major vector, C. imicola, has now significantly extended its range northwards to include much of Portugal, Spain, Italy and Greece and has even been recorded from southern Switzerland. Furthermore, in many of these new locations the insect is present and active throughout the entire year. With the related bluetongue virus, which utilises the same vector species of Culicoides this has, since 1998, precipitated the worst outbreaks of bluetongue disease ever recorded with the virus extending further north in Europe than ever before and apparently becoming endemic in that continent. The prospects for similar changes in the epidemiology and distribution of AHSV are discussed.     

Recent developments in research into the Cyathostominae and Anoplocephala perfoliata

Intestinal helminths are an important cause of equine disease. Of these parasites, the Cyathostominae are the commonest group that infect horses. These nematodes consist of a complex tribe of 51 species, although individual horses tend to harbour 10 or so common species, in addition to a few rarer species. The Cyathostominae can be extremely pathogenic, and high levels of infection result in clinical symptoms ranging from chronic weight loss to colic, diarrhoea and death. As part of their life cycle, immature cyathostomins penetrate the large intestinal wall, where they can enter a state of inhibited larval development. These larvae can exist in this state for months to years, after which they subsequently re-emerge. If larvae re-emerge in large numbers (i.e. several million), severe pathological consequences ensue. The inhibited larvae are also relatively refractory to several of the currently available anthelmintics, so that horses treated previously with anthelmintics can still carry life-threatening burdens of these parasitic stages. Little is known about the cyathostomin larvae during their mucosal phase, and current research efforts are focused on investigating the biology of these stages. Much of the research described here highlights this area of research and details studies aimed at investigating the host immune responses that the mucosal larvae invoke. As part of this research effort, molecular tools have been developed to facilitate the identification of larval and egg stages of cyathostomins. These molecular tools are now proving very useful in the investigation of the relative contributions that individual, common cyathostomin species make to the pathology and epidemiology of mixed helminth infections. At the more applied level, research is also in progress to develop an immunodiagnostic test that will allow numbers of mucosal larvae to be estimated. This test utilises antigen-specific IgG(T) serum antibody responses as markers of infection. As anthelmintic resistance will be the major constraint on the future control of the Cyathostominae, researchers are now actively investigating this area and studies aimed at elucidating the molecular mechanisms of drug resistance are described. Another parasite which has assumed a clinically important role in horses is the tapeworm, Anoplocephala perfoliata. This parasite is prevalent world-wide and has been shown to be a significant cause of equine colic. Because previous methods of estimating the infection intensity of tapeworm were inaccurate, recent research has been directed at developing an immunodiagnostic ELISA for these cestodes. Specific IgG(T) responses to antigens secreted by adult tapeworms have been shown to provide a reasonable indication of infection intensity. An ELISA based on these responses is now commercially available. The steps involved in the development of this ELISA are described here. In addition to these recent advances in research, this review also outlines the principle areas for future research into these important equine parasites.