Immunocytochemical labelling of Bacteroides nodosus pili using an immunogold technique.

An immunogold labelling technique was used to label the pili of the bacterium Bacteroides nodosus. The labelling was distinct and highly specific, and individual pili could be recognised beneath the gold probe. The labelling of somatic antigens could be distinguished from that of pilus antigens. Furthermore, labelling of fragments of cytoplasm released by cell lysis and trapped in the pili could be distinguished from pilus labelling. An antiserum that had been raised against strain 80200 (serotype N) labelled pili of strain 215 (serotype B). Double labelling experiments with this antiserum and the antiserum against strain 215 (serotype B) showed that both antisera label the same pili bundles. The ease of detection of the immunocytochemical reaction should enable this technique to be used as a routine screen for pilus antigens. It also possesses the potential for much wider applications for immunolabelling other antigens, such as viruses, that can be obtained in suspension.

Onset and remission of ovine footrot.

Sheep exposed to uncontrolled outbreaks of footrot were observed on one farm for 3 successive years and on another for one year. Observations were made from mid-winter to early summer. In each outbreak about 70% or more of sheep exposed to footrot acquired the disease. Natural remission occurred in each outbreak and some cases healed while new cases continued to develop. A feature of all outbreaks observed was that a much higher proportion of late onset cases healed than cases which developed early in the outbreaks. By contrast, 90% of sheep already affected before spring had lesions which persisted into next summer. It is suggested that there is a spectrum of resistance to footrot among sheep. This spectrum includes animals which remain chronically infected and those that do not become infected at all. Intermediate levels of resistance are reflected firstly in the time taken by different groups of sheep, to become affected after exposure and secondly, to heal after becoming affected.

Comparison of alum-absorbed or non-alum-absorbed oil emulsion vaccines containing either pilate or non-pilate Bacteroides nodosus cells in inducing and maintaining resistance of sheep to experimental foot rot.

Highly pilate (P) or non-pilate (NP) cells of Bacteroides nodosus were compounded into oil emulsion (O) either with or without prior absorption onto alum (A). The abilities of these four preparations (referred to as PAO, NPAO, PO and NPO vaccines) to stimulate antibody production and to protect sheep from foot rot were compared. Two injections of PAO vaccine protected sheep against homologous challenge 12 weeks after the second dose by PO, NPO and NPAO vaccines were less effective. Sheep were protected against homologous challenge for 14 weeks after a single dose of PAO vaccine and for 22 weeks after three doses; an ameliorative effect was still evident 40 weeks after the third dose. Protection against challenge with two heterologous strains was demonstrated at six weeks after three doses of vaccine. A numerical system of scoring the lesions also confirmed that foot rot in vaccinated sheep challenged outside the 'protective' period of the vaccine was somewhat less severe than in controls. PAO vaccine induced much higher and more persistent titres of agglutinins than the other vaccines tested. There was a relationship between agglutinin titres and resistance to homologous challenge.

Effect of alum-precipitated or oil-adjuvant Bacteroides nodosus vaccines on the resistance of sheep to experimental foot rot.

The feet of sheep which had been given two doses of oil adjuvant foot rot vaccine were completely resistant to direct challenge with a virulent homologous culture of Bacteroides nodosus four weeks after vaccination, partially resistant after eight weeks but completely susceptible after 12 weeks. Sheep given two doses of alum precipitated vaccine, or a dose of oil emulsion vaccine followed by a dose of alum precipitated vaccine, were resistant four weeks but not eight weeks later. Foot rot in affected, vaccinated sheep was more severe in those which had received the alum precipitated vaccine than in those given the oil emulsion vaccine, but vaccinated sheep were always less severely affected than controls. Where oil emulsion vaccine provided the primary stimulation, agglutinin titres were 18,000 four weeks after revaccination with either vaccine, falling to 4000 after a further four weeks. Significantly poorer titres were observed when two doses of alum precipitated vaccine only were given.

Effect immunization with highly purified alpha- and beta-toxins on staphylococcal mastitis in rabbits.

Experiments were carried out to determine whether immunization of female rabbits with highly purified staphylococcal alpha- or beta-toxins would protect them against intramammary challenge with staphylococci. High circulating anti-alpha-toxin titers reduced the lethal hemorrhagic edematous form of the disease ("blue-breast") produced by strains BB and Compton 201 to a localized chronic abscess form. No such protection was afforded by high anti-beta-toxin titers. Immunization with alpha- or beta-toxins produced no change in the clinical picture of the disease produced by CN.6708, a strain of Staphylococcus responsible for a natural outbreak of abscess-type rabbit mastitis. From these experiments it would appear that alpha-toxin is a key antigen in the blue-breast form of rabbit mastitis. Since the abscess form of the disease was not prevented by immunization with either alpha- or beta-toxin, other virulence factors must be acting to produce this more localized disease.