The suspensions were decimally diluted in the same medium. those with juvenile periodontitis and severe forms of adult periodontitis the carriage rate is often much higher (14, 15, 23), The majority of individuals (over 90%) with juvenile periodontitis have raised antibody titres Ebrotidine to a potent leukotoxin produced by leukotoxic strains (20). Ebrotidine Production of leukotoxin may represent a virulence mechanism by destroying the hosts Rabbit Polyclonal to LIPB1 polymorphonuclear leukocytes (PMNs) and monocytes that migrate locally in response to the microbial assault. Non-human primates harbor leukotoxic strains of (16) and have detectable serum antibody titers against the leukotoxin which, with macaque monkeys. We have studies the acquisition of leukotoxic strains of and the dynamics of the immune response to the leukotoxin. The results may eventually become useful in explaining why seems to be commensal in most primates but a potential pathogen in humans. Material and methods Animals One group of 75 monkeys was used in a cross-sectional study to determine the relationship between the age at which they acquire and the level of humoral anti-leukotoxin antibodies. The animals ranged from 2 day-old neonates to adults more than 10 years. Those less than 12 weeks aged were breast-fed, the remainder were fed Ebrotidine a variety of diet programs; Mazuri Primate Diet (Special Ebrotidine Diet Solutions Limited, Witham, Essex, UK), a high-sucrose cariogenic sandwich diet (3) or a starch-based maintenance diet (2). A second group of 10 neonatal monkeys was used to characterize changes in serum anti-leukotoxin antibody levels during the 1st 6 months after birth. These animals were breast-fed for the first 3C4 weeks after which they were fed the Mazuri Primate Diet. All animals experienced free access to water. Sera Blood was from the femoral artery and sera were separated and freezing at ?20C. For shipment from your U.K. to Philadelphia small quantities of sera were freeze-dried and upon introduction reconstituted with distilled water. Enumeration and recognition of A. actinomycetemcomitans Sterile scalpel blades were used to remove plaque from teeth and the mucosal surfaces were sampled with sterile cottonwool swabs. The bacteria were dispersed in 2 ml of Ebrotidine Thioglycollate medium without dextrose or indication, (TG medium, Difco Laboratories, Western Molesey, Surrey, UK), by strenuous vortexing in the presence of sterile glass beads for 15 s. The suspensions were decimally diluted in the same medium. Duplicate, 0.1 ml aliquots of right dilutions on to horse blood agar base (Oxoid, Ltd., Basingstoke, Hampshire, UK) supplemented with 5% (v/v) horse blood and menadione at 0.5 g per ml and incubated anaerobically at 37C for 5 days. was isolated within the medium described by Slot machines (12), which we (data not shown) as well as others (21) have found to be superior to that reported by Mandell and Socransky (9). Colonies having the characteristic appearance of (12, 13) were enumerated by microscopic examination of the selective medium and usually 5 presumptive colonies were subcultured from each sample. These isolates were evaluated for acid production from glucose, sucrose, lactose, fructose, xylose, mannitol, mannose and dextrin, for nitrate reduction, for catalase production and for growth activation by X and V factors (4, 13). The number of was indicated as a percentage of the total colony count on the nonselective medium. Cross-sectional study of in each sample was identified. Intra-oral localization of at each site identified. Estimation of anti-leukotoxin antibody activity in sera The anti-leukotoxin titers of sera were identified from 46 monkeys investigated.
