Detecting anti-Treponema hyodysenteriae antibodies in swine serum using immunofluorometry.

An indirect fluorescent antibody test was adapted for measuring serum anti-Treponema hyodysenteriae antibodies with a fluorometer. The immunofluorescence was recorded as fluorescent signal units. Cultures of T. hyodysenteriae and Treponema innocens were used as antigen. There was a significant (P less than 0.01) correlation between the immunofluorescence recorded with the fluorometer and that evaluated visually with a microscope. The swine exposed orally to swine dysentery infective inoculum and subsequently hyperimmunized by the intravenous inoculation of live cultures of T. hyodysenteriae had the highest average fluorescent signal unit, which was 104.5. There was a significant (P less than 0.01) correlation between the level of anti-T. hyodysenteriae antibody and the interval length between the last day of diarrhea and the day of bleeding. However, in measuring fluorescent signal units in serum from swine infected with nonpathogenic large spirochetes, (T. innocens), there was also a significant (P less than 0.01) correlation between T. hyodysenteriae and T. innocens as antigen. The coefficient of variation of the average fluorescent signal unit for a highly positive serum and a highly negative serum between 16 runs of assays were 5.7% and 19% respectively; the coefficient of variation of the average fluorescent signal unit for duplicate samples on 358 serum samples tested was 5.8%.     

Studies on a fluorescent insect growth regulator metabolite complex with house-fly microsomal cytochrome P-450

The fluorescent insect growth regulator 5[[[5-(dimethylamino)-1-naphthalenyl]amino]-1,3-benzodioxole (DNSAB) forms a metabolite complex with house-fly microsomal cytochrome P-450. Formation of the metabolite complex is dependent on the presence of NADPH and O₂; NADH supports the reaction at a reduced rate. The presence of antibodies to house-fly cytochrome c (P-450) reductase in reaction mixtures inhibits the complex formation, indicating that the reductase is necessary for transfer of electrons from NADPH to cytochrome P-450 to complete the reaction. In the oxidized form, the metabolite complex has a single absorbance maximum at 431 nm, whereas the reduced form has two absorbance maxima at 426 (major) and 455 nm (minor). The pH of the media affects the extinction of the 426- and 455-nm Soret bands; increased pH decreases the extinction of the 426-nm band and increases the extinction of 455-nm band. Formation of the DNSAB metabolite-cytochrome P-450 complex decreases the amount of CO-reactive cytochrome P-450 by 24%. The metabolite complex is not dissociable by treatment with ferricyanide or by using centrifugation techniques. Dissociation is accomplished by addition of DNSAB to the oxidized metabolite complex. Kinetic analysis of the complex formation gives apparent K_m and V_max values at 2.55 ± 1.0 μM and 1.1 ± 0.4 × 10^?2 ΔA min^?1 nmol^?1 cytochrome P-450, respectively. Addition of juvenile hormone [(E,E)-cis-methyl-10,11-epoxy-7-ethyl-3,11-dimethyl-2,6-tridecadienoate; JH] to the reaction medium competitively inhibits the formation of the metabolite complex giving an inhibition constant of 16 μM. DNSAB synergized the lethal effects of JH against Aedes aegypti larvae threefold; however, JH did not synergize DNSAB. These data suggest that DNSAB may acquire its hormonal qualities by complexing a species of cytochrome P-450 that metabolizes JH, thereby prolonging the in vivo lifetime of this hormone.     

Bovine leukemia virus-associated antigens in lymphocyte cultures.

Short-term lymphocyte cultures from bovine leukemia virus (BLV)-infected cattle were tested for BLV-associated antigens at various times after incubation. Several immunologic methods were used, including fluorescent antibody tests, immunodiffusion, and radial immunodiffusion. Antigens were not detected in uncultured lymphocytes. The BLV-associated antigens were detected as early as 3 hours, with maximum antigen production occurring at 18 to 24 hours after incubation. These results indicate that culturing of lymphocytes in vitro is necessary for the expression of the virus.     

Chronology of development of serum antispirochete antibody in swine experimentally exposed to swine dysentery: preliminary report.

By using an indirect fluorescent antibody test (with immunofluorescence of large spirochetes as positive), serum antispirochete antibodies were detected initially at 4 weeks after onset of diarrhea in swine exposed to infective swine dysentery inoculum. Antibodies continued to be present in serum of swine tested 5 months after onset of diarrhea.     

Effects of maternal protein-energy malnutrition and cold stress on neutrophil function of bovine neonates

The effects of maternal protein or calorie deprivation (or both) on the bactericidal activity of neutrophils and sera from newborn calves subjected to cold stress were studied. Nutritional deficiencies in the dam had little effect on in vitro bactericidal activity of neutrophils and base-line sera taken at birth. Neutrophils obtained at birth destroyed Staphylococcus aureus but not Escherichia coli when incubated with either unheated or heated autologous base-line sera. Heat treatment of base-line sera to inactivate complement did not alter bacterial growth. When incubated in the presence of autologous base-line sera, neutrophils from 3-day-old calves were no more active in the destruction of either bacterium than were neutrophils from newborn calves. However, addition of day 3 (immunoglobulin-containing) sera enabled day 3 neutrophils to destroy E coli (P < 0.0001). The increased destruction of E coli by day 3 neutrophils and day 3 sera was not affected by heat treatment of the sera. Maternal protein deficiency significantly increased (P < 0.05) destruction of E coli by day 3 neutrophils and sera. This effect was independent of energy levels. There were no differences observed in the bactericidal activity of neutrophils and sera taken from calves exposed to 1 C or 21 C environmental chambers for 3 days. Also, cold stress-nutritional stress interactions were not detected.