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D A Mosier

Publications and source records attributed to D A Mosier.

At least 55 records · Page 3Linked to original sources
PubMed1989

Effects of Pasteurella haemolytica leukotoxin on isolated bovine neutrophils.

P. haemolytica leukotoxin caused rapid leakage of intracellular K+ (greater than 90% in 30 sec) from and cell swelling (approximately 100% increase in 15 min) of isolated bovine neutrophils. Incubation media made hypertonic by the addition of raffinose, dextran or inulin (carbohydrates with mol. wts of greater than or equal to 505) prevented leukotoxin-induced cell swelling, but not K+ leakage. Assuming that leukotoxin acts as a transmembrane molecular sieve, then the leukotoxin-induced functional transmembrane pore size in bovine neutrophil plasma membranes is slightly smaller than the molecular size of raffinose, i.e. 1.2 nm. Morphologically, leukotoxin caused bovine neutrophils to swell, lose their membrane ruffling, develop a finely porous surface, and form large plasma membrane defects. Exposure of neutrophils to leukotoxin caused slower (5-50 min) leakage of 80% of the cellular L-lactate: NAD oxidoreductase (E.C. 1.1.27, lactate dehydrogenase, LDH). Leukotoxin-induced K+ leakage and cell swelling developed in Ca2+-free medium, whereas leakage of lactate dehydrogenase develop only in medium containing Ca2+ and was inhibited by the addition of ethylene glycol-bis(B-aminoethyl ether)N,N,N1,N1-tetraacetic acid (EGTA). This sequence of leukotoxin-induced changes in neutrophils is compatible with the mechanism of action of pore-forming cytolysins.

Animals↗
PubMed1989

Serum IgG and IgM antibody response in cattle to antigens of Pasteurella haemolytica.

The serum IgG and IgM antibody responses of 48 cattle vaccinated with live Pasteurella haemolytica (LIVE), formalin-killed P. haemolytica in Freund's incomplete adjuvant (FIA), or formalin-killed P. haemolytica in aluminum hydroxide adjuvant (ALH) to a variety of P. haemolytica antigens were evaluated. Enzyme-linked immunosorbent assays (ELISAs) were used to determine the sequential and day 21 IgG and IgM antibody responses to whole P. haemolytica (WB), a capsular carbohydrate-protein subunit (CPS) extracted from the organism, P. haemolytica capsular carbohydrate (CC), and P. haemolytica leukotoxin (LT). LIVE and FIA vaccinates developed generally higher IgG and IgM responses to all antigens compared to ALH vaccinates. LIVE vaccinates developed IgG responses to LT which were significantly higher (P less than 0.05) than all other vaccinates. In contrast, FIA vaccinates developed significantly higher IgG responses to CPS than all other vaccinates. On the basis of the ELISA results, similar or cross reacting antigenic sites were present in preparations containing surface antigens (WB, CPS and CC), but not LT. Disease resistance, as determined by experimental lesions induced in the 48 calves by transthoracic challenge with P. haemolytica, was significantly greater in the LIVE and FIA vaccinates compared with ALH vaccinates. No significant difference in resistance was detected between LIVE and FIA vaccinates. Lesions in ALH vaccinates were not significantly different than those in phosphate-buffered saline (PBS) controls. Increased IgG responses to all antigens were significantly associated with resistance to experimental disease; however, IgG responses to CPS were most highly correlated with resistance. The only IgM response which was significantly correlated with resistance was the response to CPS. These studies indicate that serum IgG antibody responses to various surface antigens of P. haemolytica, as well as LT, can enhance resistance to experimental pneumonic pasteurellosis. Serum IgM responses, however, do not appear to play a major role in resistance to experimental disease.

Animals↗
PubMed1989

Transmembrane pore size and role of cell swelling in cytotoxicity caused by Pasteurella haemolytica leukotoxin.

Pasteurella haemolytica A1 leukotoxin causes rapid (5 to 15 min) leakage of intracellular K+ and cell swelling and slower (15 to 60 min), Ca2+-dependent formation of large plasma membrane defects (congruent to 100 nm) and leakage of lactate dehydrogenase from bovine lymphoma cells (BL3 cells) (K. D. Clinkenbeard, D. A. Mosier, A. L. Timko, and A. W. Confer, Am. J. Vet. Res., in press). Incubation of BL3 cells in medium made hypertonic by inclusion of 75 mM sucrose blocked leukotoxin-induced cell swelling, formation of large plasma membrane defects, and leakage of lactate dehydrogenase but did not block leukotoxin-induced leakage of intracellular K+. Carbohydrates with molecular weights less than that of sucrose, e.g., mannitol, did not block leukotoxin-induced cell swelling of BL3 cells. Increasing the concentration of mannitol to twice that of sucrose still resulted in no protective effect. Assuming that leukotoxin acts as a transmembrane molecular sieve, then the functional transmembrane pore size formed by leukotoxin in BL3 cells is slightly less than the size of sucrose, i.e., 0.9 nm. Exposure of BL3 cells to leukotoxin for 15 or 45 min followed by the addition of hypertonic sucrose to the incubation medium reversed leukotoxin-induced cell swelling and prevented further leakage of lactate dehydrogenase. Leukotoxin-induced leakage of lactate dehydrogenase required both cell swelling and Ca2+-dependent processes. The Ca2+-dependent steps can occur before or concurrent with cell swelling.

Animals↗
PubMed1989

Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis.

Antigens associated with whole Pasteurella haemolytica biotype A serotype 1, a capsular carbohydrate-protein extract of the organism, and P. haemolytica leukotoxin were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antigens of the electrophoresed preparations were detected by Western blotting (immunoblotting) with sera from cattle which were either nonvaccinated or vaccinated with live or killed P. haemolytica vaccines and had variable degrees of resistance to experimental pneumonic pasteurellosis. Distinct, easily recognizable antigens of these preparations were identified, and the antibody responses to these antigens were quantified by densitometry. To determine their importance to disease resistance, we then compared antibody responses with experimental lesion scores. Antibody reactivity to surface antigens which were significantly correlated with resistance and present in two or more of the preparations were detected at 86, 66, 51, 49, 34, 31, and 16 kilodaltons (kDa). Of these, antibody responses to antigens at 86, 49, and 31 kDa appeared most important based on their concentration and significance levels. Antibody reactivity to leukotoxin antigens which were significantly correlated with resistance and common with important surface antigens were detected at 86, 66, and 49 kDa. Antibody responses to unique leukotoxin antigens which were significantly correlated with resistance were present at 92 and 58 kDa.

Animals↗
PubMed1989

Comparison of the Pasteurella haemolytica A1 envelope proteins obtained by two cell disruption methods.

The French pressure cell and sonication methods of bacterial cell disruption were compared for the evaluation of surface proteins from Pasteurella haemolytica A1. Several protein bands were quantitatively different when compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and densitometry. With densitometry, sonicated preparations had higher concentrations of the 102-, 83-, 50-, 38-, and 30-kilodalton (kDa) proteins; French pressure cell preparations had higher concentrations of the 96-, 65-, and 42-kDa proteins. Qualitative differences between these two disruption methods were evident at the 102-, 96-, 91-, 50-, 38-, and 30-kDa protein bands. However, significant differences (P less than 0.05) were detected between the two methods at only the 102-, 96-, 91-, and 50-kDa bands.

Bacterial Proteins↗
PubMed1989

Serum antibody response to carbohydrate antigens of Pasteurella haemolytica serotype 1: relation to experimentally induced bovine pneumonic pasteurellosis.

The antibody responses to the capsular carbohydrate (CC) purified from Pasteurella haemolytica serotype 1 were determined by an ELISA, using 135 sera from 6 calves vaccinated with phosphate-buffered saline solution, formalin-killed P haemolytica bacterins, live P haemolytica, or an extract of P haemolytica referred to as carbohydrate-protein subunit (CPS). Calves vaccinated with live P haemolytica, bacterins, or CPS developed serum antibodies to CC. Bacterins containing Freund incomplete adjuvant or Freund complete adjuvant induced higher antibody responses than did bacterins containing aluminum hydroxide. In 4 of 6 experiments, high antibody responses to CC were significantly (P less than 0.05) correlated with resistance to transthoracic challenge exposure with P haemolytica. When calves were challenge exposed with a dose of P haemolytica that was 4.5 times greater than the standard challenge exposure dose or when calves that had been vaccinated with CPS were challenge exposed, antibody responses did not significantly (P greater than 0.05) correlate with resistance to challenge exposure. The amount of serum antibodies to CPS increased significantly (P less than 0.05) when calves were vaccinated with live or killed P haemolytica or with CPS, compared with that in calves given saline solution. In 5 of 6 experiments, correlation between high antibody responses and resistance to challenge exposure was significant (P less than 0.05). The correlation between those variables was not significant (P less than 0.07) for CPS-vaccinated calves. In the ELISA, treatment of CPS with sodium m-periodate, to oxidize periodate-sensitive carbohydrate epitopes, failed to markedly alter the antibody response to CPS.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

The evolution of vaccines for bovine pneumonic pasteurellosis.

Since the early 1900s bovine pneumonic pasteurellosis has been recognised as a major economic problem to European and North American cattle industries. Initial attempts to prevent the disease were complicated by incomplete knowledge of the causative organisms. Despite some early reports of vaccine-induced protection against disease, initial vaccines were of questionable protective value. From the late 1950s to the 1970s Pasteurella haemolytica and P multocida bacterins were the primary type of vaccine used commercially and experimentally. When viruses, most notably bovine herpesvirus 1 (infectious bovine rhinotracheitis virus) and parainfluenza-3 virus, were found to be associated with bovine respiratory disease, viral vaccines were used in attempts to prevent pneumonic pasteurellosis. Combinations of bacterins and viral vaccines were also developed and evaluated. Collectively, bacterins, viral vaccines and bacterin-virus combinations did not consistently reduce disease in experimental trials or field use. By the 1980s some studies using live vaccines were reportedly successful in reducing the incidence of pneumonic pasteurellosis. Current experimental studies revolve around the identification and incorporation of specific Pasteurella species antigen extracts into vaccines. The efficacy of these new extract vaccines is yet to be determined.

Animals↗
PubMed1989

Effects of Pasteurella haemolytica leukotoxin on cultured bovine lymphoma cells.

Leukotoxin activity from culture supernatants of Pasteurella haemolytica serotype 1 in logarithmic growth phase caused rapid (less than 5 min) release of intracellular K+, uptake of extracellular Ca2+, and swelling of cultured bovine lymphoma cells (BL3 cells). Release of 51CrO4(2-) and lactate dehydrogenase (LDH) from BL3 cells began after 15 minutes of incubation with leukotoxin at 37 C and was completed between 60 and 120 minutes of incubation. In addition, leukotoxin exposure of BL3 cells resulted in cell aggregation and adherence to glass surfaces. Scanning electron microscopy indicated that after 10 minutes of leukotoxin exposure, BL3 cells increased in size, and large membrane defects developed between 20 and 60 minutes of exposure. The rate of release of LDH from leukotoxin-exposed BL3 cells was proportional to the amount of leukotoxin added. At high cell concentrations, the activity of LDH released at completion was directly proportional to the amount of leukotoxin added. Leukotoxin-induced release of LDH required a divalent cation, whereas K+ release and cell swelling did not. The addition of Ca2+, Mn2+, and Ba2+ resulted in increased leukotoxin-induced release of LDH. Divalent cation concentrations of 0.5 to 2.5 mM resulted in 50% of maximal stimulation. Ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid blocked increased release of LDH caused by Ca2+ addition, but had no effect on K+ release or cell swelling. Leukotoxin action on BL3 cells (K+ release, cell swelling, Ca2+ uptake, and release of LDH) was prevented by incubation at 4 C.

Animals↗
PubMed1989

Direct effects of Pasteurella haemolytica lipopolysaccharide on bovine pulmonary endothelial cells in vitro.

Bovine pulmonary artery cells in cell culture were exposed to lipopolysaccharide (LPS) purified from Pasteurella haemolytica serotype Al. This resulted in severe membrane damage, which caused a time- and dose-dependent release of lactate dehydrogenase that was first detected 4 hours after exposure and reached a maximal mean release of 67% after 24 hours of exposure to 1 micrograms of LPS/ml. Mean release of 51chromium followed by a similar pattern and reached a maximum of 61% following 24 hours of exposure to 10 micrograms of LPS/ml. Morphologically, endothelial cells responded to LPS by marked cell membrane retraction, the formation of numerous cytoplasmic blebs, and ruffling of the cell membrane. Subsequently, the cells became round and detached. Cell detachment reached a mean of 95% following 8 hours of exposure to 1 micrograms of LPS/ml. These studies demonstrated that P haemolytica LPS is capable of causing direct damage to bovine pulmonary arterial endothelial cells, which may be important in the pathogenesis of bovine pneumonic pasteurellosis.

Animals↗
PubMed1986

Changing manifestations of a chronic feline haematopoietic proliferative disease during immunotherapy with staphylococcal protein A.

A cat with feline leukaemia virus-associated malignant disease was treated by ex vivo immunoadsorption using staphylococcal protein A coated filters. During the 12-week course of treatment, the morphological manifestations of the haematopoietic disease showed a progressive transition from erythroid to myeloerythroid to myeloid predominance, and the disease was preceded by and associated initially and terminally with a blast transformation of lymphoblastic morphology. Necropsy revealed massive meningo-cerebral, as well as hepatic, renal, myeloid, lymphoid, peritoneal and pelvic infiltrations largely consisting of lymphoblastic cells. Evidence of myeloid and erythroid differentiation was present in all the infiltrates. The several possible bases for this shift of morphological expression are considered.

Animals↗
PubMed1986

Enzyme-linked immunosorbent assay for detection of serum antibodies to Pasteurella haemolytica cytotoxin (leukotoxin) in cattle.

An enzyme-linked immunosorbent assay (ELISA) was developed for detection of bovine serum antibodies to the cytotoxin (leukotoxin) of Pasteurella haemolytica. A partially purified, cytotoxic, and immunogenic protein obtained from supernatants of logarithmic-phase P. haemolytica was used as the ELISA antigen. Preadsorption of sera with various cytotoxic, somatic, and capsular antigen preparations demonstrated that the assay was specific for anticytotoxin antibodies. ELISA anticytotoxin titers had a strong, significant correlation to cytotoxin-neutralizing-antibody titers. The ELISA, however, was more rapid and allowed for greater numbers of samples to be run than did the neutralization technique. ELISA anticytotoxin titers were high in cattle vaccinated with a live P. haemolytica vaccine, whereas unvaccinated cattle and cattle receiving a P. haemolytica bacterin had low ELISA anticytotoxin titers. A significant positive correlation between ELISA titers and resistance to experimental bovine pneumonic pasteurellosis was present.

Animals↗
PubMed1986

Chromatographic separation and characterization of Pasteurella haemolytica cytotoxin.

Biochemical and immunologic properties of the cytotoxin (leukotoxin) produced by Pasteurella haemolytica were examined. Crude, bacteria-free supernatants from logarithmic phase P haemolytica were fractionated, using a series of column chromatographic techniques. Sequential anion exchange chromatography, gel-filtration chromatography, and chromatofocusing resulted in a cytotoxic substance (cytotoxin-C) of approximately 160 kilodaltons (kD), as determined by use of gel-filtration chromatography. Polyacrylamide-gel electrophoresis of cytotoxin-C yielded 3 protein bands with relative mobilities of 0.37, 0.42, and 0.63. On the basis of immunoblotting with a cytotoxin-neutralizing bovine immunoglobulin for antigen detection, the 2 low-mobility bands shared a strong region of immunogenicity. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, principal protein constituents of cytotoxin-C were found at 160, 66, 57, and 23 kD. Using immunoblotting with cytotoxin-neutralizing immunoglobulin, strong, distinct reactions with the 66- and 57-kD bands were detected. Immunization of rabbits and mice with cytotoxin-C resulted in sera that reacted strongly with cytotoxin-C in enzyme-linked immunosorbent assays and immunodiffusion assays. The major immunogenic proteins also were detected by use of immunoblotting with anticytotoxin-C sera from rabbits and mice. Postinoculation rabbit sera neutralized crude cytotoxin.

Animals↗
PubMed1986

Serum antibodies to Pasteurella haemolytica lipopolysaccharide: relationship to experimental bovine pneumonic pasteurellosis.

An enzyme-linked immunosorbent assay was used to determine the serum antibody response to Pasteurella haemolytica lipopolysaccharide (LPS) for calves vaccinated with saline solution, a formalin-killed P haemolytica bacterin, or live P haemolytica. Bacterin-vaccinated calves had a lower antibody response to LPS than did calves vaccinated with live P haemolytica. Calves vaccinated with either saline solution or the bacterin were more susceptible to intrapulmonic challenge exposure with P haemolytica than were calves vaccinated with liver organisms. Serum antibody responses to P haemolytica LPS did not seem important for resistance to challenge exposure, because there was no significant correlation (P greater than 0.05) between the lung lesion score and antibody response to P haemolytica LPS. There was a highly significant correlation (P less than 0.001) between antibody detected against P haemolytica LPS and that against formalin-killed P haemolytica. Competitive binding studies indicated that P haemolytica LPS is a major antigenic determinant on the surface of P haemolytica. There did not seem to be substantial cross-reaction between LPS from P haemolytica and that from Escherichia coli (serotype O26:B6).

Animals↗
PubMed1986

Comparison of the antigens associated with saline solution, potassium thiocyanate, and sodium salicylate extracts of Pasteurella haemolytica serotype 1.

Pasteurella haemolytica antigenic extracts were made, using saline solution, potassium thiocyanate (KSCN), and sodium salicylate (SS) extraction procedures. Of the 3 techniques, saline solution extraction resulted in the lowest protein concentration and lowest ribonucleic acid-to-protein ratio. The extracts varied in protein:carbohydrate ratios, with the KSCN extract being highest and the saline solution extract the lowest. Each extract contained lipopolysaccharide, as determined by detectable quantities of 2-keto, 3-deoxyoctonate. The saline solution extract contained the fewest protein bands by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis, but contained the highest molecular weight proteins. All 3 extracts were reasonably similar antigenically, as detected by immunoblotting. Many of the protein bands present in the KSCN or SS extracts did not seem to be antigenic. Each extract was subjected to chromatofocusing, and the greatest antigenic peak, for each extract, failed to bind to the exchanger. These highly antigenic peaks, designated as saline solution, KSCN, or SS antigens, were similarly high in carbohydrate content, had similar antigenic-profiles, and contained high molecular weight (greater than 200,000) antigenic material, most likely carbohydrate in nature, as detected by immunoblotting. Inoculation of mice with 1 of the 3 extracts or the saline solution antigen resulted in marked antibody responses; however, protection against intraperitoneal challenge exposure to P haemolytica was minimal.

Animals↗
PubMed1985

Saline-extracted antigens of Pasteurella haemolytica: separation by chromatofocusing, preliminary characterization, and evaluation of immunogenicity.

Saline extracts of logarithmic-phase Pasteurella haemolytica, serotype 1, were separated by chromatofocusing. The resulting fractions were analyzed by immunodiffusion and an enzyme-linked immunosorbant assay, and six antigen groups (AG's) were identified. AG 1 did not bind to the column, AG's 2, 3 and 4 were eluted with a decreasing pH gradient, and AG's 5 and 6 were eluted with an increasing NaC1 gradient. Fractions containing each AG were pooled and further purified by gel filtration. The AG's were subsequently characterized as to protein, carbohydrate and 2-keto-3-deoxyoctanate (KDO) content. AG's 1, 5, and 6 had higher carbohydrate contents than AG's 2, 3 and 4. Only AG 5 contained detectable levels of KDO. The AG's were also analyzed by SDS-PAGE and immunoblotting. Each AG produced a characteristic pattern of proteins and antigens, although two antigenic proteins were common to all AG's. AG 1 contained the greatest number of antigenic proteins. Immunization of mice with each AG in Freund's incomplete adjuvant resulted in a strong antibody response to the homologous AG for four of the six AG's. Limited protection against a P. haemolytica challenge was observed in mice that were immunized with AG 2 or 4.

Animals↗
PubMed1984

Rhinosporidiosis in a dog.

Rhinosporidiosis was diagnosed in a black and tan coonhound. Clinical signs were characterized by 6 to 8 months of intermittent sneezing. Numerous polypoid nodules were removed from the nasal cavity. Histologic examination of the nodules revealed chronic pyogranulomatous inflammation caused by infection with Rhinosporidium seeberi. Rhinosporidiosis has been reported in human beings, dogs, and several other domestic species. It is an uncommon disease in the United States.

Animals↗
PubMedDate not supplied

Healing of canine autogenous cancellous bone graft donor sites.

Autogenous cancellous bone was procured from the proximal ends of one humerus and the contralateral tibia of eight adult mixed-breed dogs. Greater weights of bone were consistently harvested from the humerus than from the tibia. Restoration of cancellous bone was more rapid and complete in the humerus than in the tibia. The tibia harvest site filled primarily with fibrous tissue rather than with cancellous bone.

Animals↗