Fate of Escherichia coli and Escherichia coli O157 in soils and drainage water following cattle slurry application at 3 sites in southern Scotland

Abstract. Slurry from farm animals may contaminate water supplies, rivers and bathing waters with faecal coliforms, such as Escherichia coli . Where animals harbour the O157 strain the hazard to human health is particularly high, but both the hazard level, and the low incidence and sporadic nature of the excretion of E. coli O157 make it difficult to study this strain under field conditions. The survival of total E. coli and of E. coli O157 were compared in the laboratory for two soils under controlled temperature and moisture. E. coli O157 die-off rate was the same as or quicker than for total E. coli . This result meant that field experiments studying the fate of total E. coli should give a satisfactory evaluation of the risk of water contamination by the O157 strain. In four field experiments at three sites, slurry containing total E. coli numbers of 2.2 × 10⁴ to 5.7 × 10⁵ colony forming units per mL (c.f.u. mL^–1) was applied to drained field plots. Field die-off was faster than expected from laboratory experiments, especially in one experiment where two weeks dry weather followed application. In all but this experiment, the first drain flow events after slurry application led to very high E. coli concentrations in the drains (10³ to 10⁴ c.f.u. mL^–1). E. coli O157 was present in the slurry used for two of the experiments (33 c.f.u. per 100 mL in each case). However the proportion of E.coli O157 was very low (about 1 in 10⁵) and it was not detected in the drainage water. After the first week E. coli drainage water numbers decreased rapidly but they were 1–10 c.f.u. mL^–1 for much of the sampling period after slurry application (1–3 months).     

Phalangeal and navicular bone hypoplasia and hoof malformation in the hind limbs of a foal

Anatomical anomalies in the hind feet of a seven month old Appaloosa foal were identified and investigated through the use of gross anatomical dissection, radiography and angiography. Abnormalities were restricted to the distal aspect of both hind legs, the right hind leg being more severely affected. Anatomically the right foot resembled that of an equine fetus of approximately 120 days gestational age. Disruption of vascular perfusion to hoof structures was evident in both hind legs and was related to areas of abnormal bone conformation as well as to areas of abnormal ossification and calcification. Phalangeal and navicular bone hypoplasia were apparent as were soft tissue and joint anomalies. Although the etiology of the defects identified remains obscure, several theories are suggested, namely heritability, acquired defects and the possible teratogenic effects of clenbuterol.     

The F38-Like Group,a New Group of Caprine Mycoplasmas?

This paper concerns the taxonomic status of the F38-like group (MacOwan), a prime determinant of contagious caprine pleuropneumonia (CCPP). Extensive biochemical and serological investigations on strain F38 are reported. Some complex serological relationships with other mycoplasma species are revealed. The results, taken in conjunction with earlier published work on geno-typic characters, lead to the conclusion that final classification of these organisms should await further comparative studies of a number of field strains with a related group of strains classified as M. capri-colum.The characterization of F38 confirms its partial relationship to the “M. mycoides group” of ovine/caprine/bovine mycoplasmas, and has also revealed a very close phenotypic relationship to the bovine mycoplasma serogroup 7, a finding of potential diagnostic and epidemiological importance.Key words: mycoplasmas, classification, F38-like group     

Kinematics of the equine thoracolumbar spine

At least three types of movement take place in the joint complexes of the equine thoracolumbar spine: dorsoventral flexion and extension, axial rotation and lateral bending. Using the standard right-handed Cartesian coordinate system, these movements may be defined as rotation about the x, y and z axes respectively. Except in cases of intervertebral fusion, all three types of movement occur in each joint complex of the equine back. The greatest amount of dorsoventral movement takes place at the lumbosacral and the first thoracic intervertebral joints. The greatest amount of axial rotation and lateral bending was measured in the mid-thoracolumbar spine at the level of the 11th or 12th thoracic intervertebral joints. The caudal thoracic and the lumbar spine is the least mobile region of the equine back. In the mid-thoracic spine, lateral bending was always accompanied by a "coupled" axial rotation. The presence of the rib cage stabilised the cranial thoracic vertebrae against axial rotation.     

A scanning electron microscopic study of the vascular system of the bovine hind limb claw

The vascular organisation of the hind limb claws of clinically normal cattle was studied using scanning electron micrographs of plastic corrosion casts. The microvasculature of the dermal papillae and lamellae was demonstrated. A consistent vascular pattern was observed in the dermal papillae of the periople, coronary margin, sole and bulb. Ultrastructurally, the papillar vasculature consisted of a central artery and vein surrounded by a fine network of interconnected capillaries and venules. Arteriovenous anastomoses, connecting the central artery and vein, and peripherally situated arteriovenous loops were recognised in the terminal papillae of the white zone only. Arteriovenous anastomoses, focal enlargements and distension of the capillary bed were located within the interior half of the dermal lamellae. Numerous arteriovenous anastomoses were present throughout the entire dermis of the claw and were situated predominantly at the base of the dermal papillae and lamellae. The anastomoses and focal capillary enlargements may influence the blood flow in normal conditions and may play a significant role in the pathophysiology of bovine laminitis and sole ulceration.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001