Medicine and Health Sciences Commons^™

Non-Opsonic Attachment Of Bordetella Bronchiseptica Mediated By Cd11/Cd18 And Cell Surface Carbohydrates, Karen B. Register, Mark R. Ackermann, Marcus E. Kehrli Jr.

Mark R. Ackermann

Porcine atrophic rhinitis associated with Bordetella bronchiseptica is characterized by a severe inflammatory response in the mucosa of the nasal turbinates. Initial infiltrates of polymorphonuclear leukocytes (PMN) are followed by accumulations of mononuclear cells. In this report, we have investigated the interaction between porcine PMN and B. bronchiseptica. PMN incubated in PBS with a fluorescently labeled hemagglutinating porcine isolate, but not a non-hemagglutinating variant, had high levels of cell-associated fluorescence as determined by flow cytometry. Light microscopy indicated that most cell-associated bacteria were ingested. Transmission electron microscopy confirmed the presence of intracellular bacteria, which were contained within membrane-bound phagosomes. A …

Pulmonary Lesions In Fetuses Exposed In Utero To Porcine Reproductive And Respiratory Syndrome Virus, Kelly M. Lager, Mark R. Ackermann

Mark R. Ackermann

The salient features of a porcine reproductive and respiratory syndrome (PRRS) epizootic include respiratory disease followed by reproductive failure. The respiratory component is usually recognized first in finishing pigs or in the breeding herd as a mild flu-like disease with a high morbidity for all ages and high mortality in younger pigs. In breeding herds, acute maternal reproductive failure often follows the onset of respiratory disease. PRRS is characterized by a sudden increase in early farrowings, late-term abortions, stillborn and mummified fetuses, weak neonates with high mortality, late returns to estrus, and repeat breeders. No consistent histopathology has been obseived …

Redistribution Of Insoluble Interphotoreceptor Matrix Components During Photoreceptor Differentiation In The Mouse Retina, Kristina Mieziewska, Agoston Szel, Theo Van Veen, Gustavo D. Aguirre, Nancy Philp

Gustavo D. Aguirre, VMD, PhD

Distribution Of Anti-Cd68 (Ebm11) Immunoreactivity In Formalin-Fixed, Paraffin-Embedded Bovine Tissues, Mark R. Ackermann, B. M. Debey, T. J. Stabel, J. H. Gold, K. B. Register, J. T. Meehan

Mark R. Ackermann

A commercially acquired anti-human macrophage antibody (anti-CD68; EBM11) was used in an immunocytochemical technique to detect macrophages in formalin-fixed, paraffin-embedded tissues from cattle, pigs, humans, rats, turkeys, dogs, and cats. In healthy cattle, the antibody labeled alveolar macrophages, pulmonary intravascular cells (presumably intravascular macrophages), and macrophage-like cells in other tissues. In bovine lungs infected with Pasteurella haemolytica, EBM11 antibody labeled 95% of alveolar macrophages and macrophages within alveolar septa but only 0–2% of streaming or “oat” leukocytes. Alveolar macrophages were also stained by EBM11 in pigs but not in rats, turkeys, dogs, and cats. The antibody also stained macrophage aggregates …

Phenotypic Expression Of Recombinant Plasmids Pkt107 And Phk11 In An Avirulent Avian Escherichia Coli, Richard E. Wooley, Lisa K. Nolan, John Brown, Penelope S. Gibbs, Denise I. Bounous

Lisa K. Nolan

An avirulent wild-type avian Escherichia coli strain (Av) was electrotransformed with plasmids coding for complement resistance (pKT107) and Colicin V (ColV) production (pHK11) in order to study the effects of complement resistance and ColV production on virulence. Transformants were also compared with the wild type for embryo lethality, uptake by macrophages, motility, growth rate, plasmid content, and hemolysis. Growth rates and complement resistance patterns of strain Av and transformant Av+pHK1 1 were similar, but Av+pHK11 caused a significantly greater number of deaths in embryos and acquired motility. Transformant Av+pKT107 had a lower rate of phagocytosis, a slower growth rate, and …

Characterization Of An Avirulent Mutant Of A Virulent Avian Escherichia Coli Isolate, Lisa K. Nolan, Richard E. Wooley, Catherine W. Giddings, John Brown

Lisa K. Nolan

A virulent, complement-resistant avian Escherichia co/lisolate and its avirulent, complement-sensitive, transposon-insertion mutant were compared for the purpose of revealing structures associated with complement resistance. Both had a smooth lipopolysaccharide layer, contained traT, and lacked a capsule, but the mutant possessed a 16.2-kilodalton outer-membrane protein (OMP) not present in the wild-type. This protein may be the product of a coding region interrupted by transposon insertion. Such results suggest that an OMP greater than 16.2 kilodaltons in size may be responsible for the complement resistance and virulence of this wild-type E. coli.

Infected Cell Types In Ovine Lung Following Exposure To Bovine Respiratory Syncytial Virus, J. T. Meehan, R. C. Cutlip, H. D. Lehmkuhl, J. P. Kluge, Mark R. Ackermann

Mark R. Ackermann

Sixteen adult sheep (ten females, six males obtained from a closed flock at National Animal Disease Center, Ames, IA) were experimentally infected with bovine respiratory syncytial virus strain 375 (BRSV), and lung tissues were stained for viral antigen. Two infected sheep were euthanatized at each of the following post-inoculation times: 12, 24, 36, 48, 72, 96, 144, and 192 hours. Lung, nasal turbinates, trachea, right cranial bronchial and mediastinal lymph nodes, liver, and spleen were collected for histologic evaluation. An indirect immunoperoxidase technique was performed on routine paraffin-embedded sections of lung tissue, trachea, turbinates, and bronchial and mediastinal lymph nodes …

Effect Of Normal Intestinal Flora Of Chickens On Colonization By Virulent Colicin V-Producing, Avirulent, And Mutant Colicin V-Producing Avian Escherichia Coli, Richard E. Wooley, John Brown, Penelope S. Gibbs, Lisa K. Nolan, Kathy R. Turner

Lisa K. Nolan

Colonization of the intestinal tracts of newly hatched chicks with Escherichia coli was attempted by swabbing test organisms onto the air-shell of 19-day-old embryos. Test organisms consisted of two virulent E. coli isolates, one avirulent isolate, and one laboratory-derived mutant of the avirulent isolate carrying a recombinant plasmid coding for Colicin V production. Chicks were cultured weekly for 3 weeks for total E. coliand for the test organisms using selective media. Control chicks were sampled on weeks 1 and 5, and the normal E. coli intestinal microflora were examined for the production of colicins. The two virulent E. coli isolates …

Plasma Lipid Changes In Prcd-Affected And Normal Miniature Poodles Given Oral Supplements Of Linseed Oil. Indications For The Involvement Of N-3 Fatty Acids In Inherited Retinal Degenerations, Robert E. Anderson, Maureen B. Maude, Gregory M. Acland, Gustavo D. Aguirre

Gustavo D. Aguirre, VMD, PhD

Decreased Opsin Mrna And Immunoreactivity In Progressive Rod-Cone Degeneration (Prcd): Cytochemical Studies Of Early Disease And Degeneration, Jun Huang, Marie-Françoise Chesselet, Gustavo Aguirre

Gustavo D. Aguirre, VMD, PhD

Association Of K-1 Capsule, Smooth Lipopolysaccharides, Trat Gene, And Colicin V Production With Complement Resistance And Virulence Of Avian Escherichia Coli, Richard E. Wooley, Lisa K. Nolan, John Brown, Penelope S. Gibbs, Catherine W. Giddings, Kathy S. Turner

Lisa K. Nolan

A group of complement-resistant, virulent avian Eschberichia coli isolates were compared with a group of complement-sensitive, avirulent avian isolates for the presence of K-1 capsule, smooth lipopolysaccharides (LPS), the traT gene, and Colicin V (ColV) production. These parameters were selected because of their reported association with complement resistance and virulence in E. coli. Lethality in chicken embryos has also been shown to be correlated with virulence of avian E. colifor chickens. The complement-resistant, virulent E. coli isolates did not possess a K-1 capsule. Production of ColV and the presence of smooth LPS were significantly correlated with embryo lethality. There was …

Identification Of Β2 Integrins In Bovine Neutrophils By Scanning Electron Microscopy In The Backscatter Mode And Transmission Electron Microscopy, Mark R. Ackermann, M. E. Kehrli Jr., H. K. Hawkins, J. L. Amenson, J. E. Gallagher

Mark R. Ackermann

Mac-I, LFA-I, and p150,95 are β2 integrin glycoproteins present in the plasma membranes of neutrophils, macrophages, and lymphocytes.

Diethylene Glycol Distearate (Dgd): A Versatile Embedding Medium For Retinal Cytochemistry, Jun C. Huang, Kristina Mieziewska, Nancy Philp, Theo Van Veen, Gustavo D. Aguirre

Gustavo D. Aguirre, VMD, PhD

Protection Of Pasteurella Multocida Dermonecrotic Toxin-Challenged Rats By Toxoid-Induced Antibody, R. K. Pettit, R. B. Rimler, Mark R. Ackermann

Mark R. Ackermann

Two different doses of glutaraldehyde-treated Pasteurella multocida dermonecrotic toxin (PMDT) were used to immunize rats. Rats developed serum IgG antibodies specific for native PMDT, and IgG titers increased with dose and number of toxoid immunizations. Survival rates in both active immunization and passive serum neutralization experiments were dependent on dose of toxoid vaccination and serum levels of anti-PMDT IgG. Vaccination with toxoid prevented weight loss but not leukocytosis and increased complement titers in toxin-challenged rats. Toxoid, itself, induced minimal leukocytosis but no alterations in complement titers or weight gain.

Heterophil Chemotaxis In Chickens With Natural Staphylococcal Infections, James R. Andreasen Jr., Claire B. Andreasen, Mohammad Anwer, Anita E. Sonn

Claire B. Andreasen

Heterophil chemotaxis using heterophils isolated from the peripheral blood of five commercial broiler chickens naturally infected with staphylococcal bacteria was compared by the modified Boyden-chamber technique with chemotaxis of heterophils from two chickens from the same flock not infected with Staphylococcus (field controls) and from four healthy laboratory control broiler chickens. The infected chickens had gross and histologic lesions of staphylococcal tenosynovitis and osteomyelitis. Staphylococci were isolated from the lesions. Hematologic parameters and histologic lesions of infected chickens also were examined. Compared with field and laboratory controls, Staphylococcus-infected chickens had heterophilic leukocytosis. The heterophils of Staphylococcus-infected chickens had significantly lower …

Chicken Heterophil Chemotaxis Using Staphylococcus-Generated Chemoattractants, James R. Andreasen Jr., Claire B. Andreasen, Mohammad Anwer, Anita E. Sonn

Claire B. Andreasen

Heterophil chemotaxis, in response to chemotactic factors generated by three different strains of staphylococcal bacteria, was measured using the modified Boyden-chamber technique. Heterophils were obtained from healthy 6-to-8-week-old broiler chickens. Each bacterial strain generated factors that were chemotactic for chicken heterophils. Factors generated by two pathogenic isolates of Staphylococcus aureus, however, induced significantly greater chemotaxis in chicken heterophils than those generated by a nonpathogenic Staphylococcus isolate.

Failure Of The Congo Red Dye Uptake Test To Discriminate Between Virulent And Avirulent Avian Escherichia Coli, Kathy R. Spears, Richard E. Wooley, John Brown, Lisa K. Nolan

Lisa K. Nolan

Twenty avian Escherichia coli isolates from normal and diseased chickens were compared by use of three virulence tests. These tests included the uptake of Congo red dye, an embryo lethality test, and a quantitative microtiter complement resistance test. A direct correlation was seen between the results of the complement resistance test and the embryo lethality test. The results of the Congo red test did not correlate with the two other tests.

Identification And Prevalence Of A Genetic Defect That Causes Leukocyte Adhesion Deficiency In Holstein Cattle, Dale E. Shuster, Marcus E. Kehrli Jr., Mark R. Ackermann, R. O. Gilbert

Mark R. Ackermann

Two point mutations were identified within the gene encoding bovine CD18 in a Holstein calf afflicted with leukocyte adhesion deficiency (LAD). One mutation causes an aspartic acid to glycine substitution at amino acid 128 (D128G) in the highly conserved extracellular region of this adhesion glycoprotein, a region where several mutations have been found to cause human LAD. The other mutation is silent. Twenty calves with clinical symptoms of LAD were tested, and all were homozygous for the D128G allele. In addition, two calves homozygous for the D128G allele were identified during widespread DNA testing, and both were subsequently found to …

Relationship Of Complement Resistance And Selected Virulence Factors In Pathogenic Avian Escherichia Coli, Richard E. Wooley, Kathy R. Spears, John Brown, Lisa K. Nolan, Oscar J. Fletcher

Lisa K. Nolan

Complement resistance, antibiotic resistance profiles, and virulence profiles of 80 Escherichia coli isolates from the intestines of normal chickens (40 isolates) and chickens diagnosed as having colisepticemia (40 isolates) were compared. Differences were observed between the two groups for antibiotic resistance, siderophore production, presence of type 1 pili, complement resistance, motility, and size of plasmids. The systemic isolates were more likely to have siderophores and type 1 pili, and to be complement-resistant and motile than were the intestinal isolates. No differences between the two groups were observed for colicin production. Further comparison of the 10 most complement-resistant isolates from the …

The Role Of Docosahexaenoic Acid In Retinal Degenerations, Maureen Maude, Gustavo Aguirre, Gregory Acland, Robert Anderson

Gustavo D. Aguirre, VMD, PhD

Characteristics Of Conjugative R-Plasmids From Pathogenic Avian Escherichia Coli, Richard E. Wooley, Kathy R. Spears, Lisa K. Nolan, John Brown, Mark A. Dekich

Lisa K. Nolan

Three of four virulent avian Escherichia coli isolates transferred a single large molecular-weight R-plasmid to two recipient E. coli strains. Antibiotic resistances transferred included streptomycin (two isolates) and streptomycin-tetracycline-sulfa (one isolate). Production of colicin and siderophores, complement resistance, and embryo lethality present in the virulent isolates were not transferred to recipient organisms. From the results, it appears that the R-plasmids of these virulent avian E. coli are not associated with virulence.

Animal Model Of Human Disease: Bovine Leukocyte Adhesion Deficiency: Beta2 Integrin Deficiency In Young Holstein Cattle, Marcus E. Kehrli Jr., Mark R. Ackermann, Dale E. Shuster, Martin J. Van Der Maaten, Frank C. Schmalstieg, Donald C. Anderson, Bonnie J. Hughes

Mark R. Ackermann

Leukocyte adhesion deficiency (LAD), is a rare, autosomal recessive disorder caused by a lack or partial absence of a family of leukocyte integrins, Mac-1, LFA-1, and p150,95.1 The leukocyte (32) integrins are glycoproteins essential for normal leukocyte-endothelial cell adherence and emigration. Affected children develop recurrent bacterial infections, persistent leukocytosis, severe hypoplasia of lymphoid tissues and isolated neutrophils, eosinophils, macrophages, and lymphocytes have abnormal function when tested in vitro. Most patients without bone marrow transplants die at an early age.

Use Of Rats To Compare Atrophic Rhinitis Vaccines For Protection Against Effects Of Heat-Labile Protein Toxin Produced By Pasteurella Multocida Serogroup D, J. R. Thurston, R. B. Rimler, Mark R. Ackermann, N. F. Cheville

Mark R. Ackermann

Four bacterin-toxoid and three bacterin commercial vaccines against atrophic rhinitis were tested in rats for their capacity to immunize against the lethal and systemic effects of purified heat-labile protein toxin (D-toxin) produced by Pasteurella multocida serogroup D. Only one bacterin-toxoid vaccine stimulated sufficient immunity to prevent the death of all rats challenged with D-toxin. None of the vaccines prevented weight loss, leukocytosis or increases in serum complement titers in rats challenged with D-toxin. Rats provide an inexpensive animal model for testing the capacity of vaccines to generate antitoxic immunity against the lethal and systemic effects of D-toxin.

Comparison Of A Complement Resistance Test, A Chicken Embryo Lethality Test, And The Chicken Lethality Test For Determining Virulence Of Avian Escherichia Coli, Lisa K. Nolan, Richard E. Wooley, John Brown, Kathy R. Spears, H. W. Dickerson, Mark Dekich

Lisa K. Nolan

Results with four pathogenic avian Escherichia coli isolates and one avirulent isolate in a complement resistance test, a chicken lethality test, and a chicken embryo lethality test were compared. Results of the complement resistance test with these isolates were highly correlated to results of the chicken lethality test of virulence. The chicken embryo test yielded results that were of a medium positive correlation with the chicken lethality results. The results of the complement resistance and chicken embryo lethality tests were highly correlated.

Transposon Mutagenesis Used To Study The Role Of Complement Resistance In The Virulence Of An Avian Escherichia Coli Isolate, Lisa K. Nolan, Richard E. Wooley, Richard K. Cooper

Lisa K. Nolan

The role of complement resistance in the virulence of an avian Escherichia coli isolate was examined with transposon mutagenesis. A suicide plasmid containing a kanamycin-encoding mini-transposon was used to transform a virulent complement-resistant avian E. coli isolate. A less resistant mutant was identified that contained a transposon insertion in a plasmid and in the chromosome. This loss of complement resistance was associated with a drop in virulence in an embryo assay. No other phenotypic changes were detected in the mutant. These results suggest that complement resistance is associated with the virulence of this organism.

Intestinal Adenocarcinoma Of The Ileocecal Junction In A Chicken, James R. Andreasen Jr., Claire B. Andreasen

Claire B. Andreasen

An 89-week-old male chicken was presented with signs of depression, emaciation, and weakness. At necropsy, a stricture was found at the ileocecal junction that resulted in blockage and dilation of the ileum proximal to the stricture. Histologically, neoplastic epithelial cells that contained mucin had invaded the intestinal wall and produced a fibrous connective tissue reaction. The lesion was diagnosed as scirrhous intestinal adenocarcinoma.

Light Microscopic And Ultrastructural Pathology Of Seminiferous Tubules Of Rats Given Multiple Doses Of Pasteurella Multocida Group D Protein Toxin, Mark R. Ackermann, James P. Tappe Jr., John R. Thurston, Richard B. Rimler, Dale E. Shuster, Norman F. Cheville

Mark R. Ackermann

Male Holtzman rats were given subcutaneous doses of a purified Pasteurella multocida group D heat-labile toxin on alternate days for up to 22 days. Rats were necropsied at 18 days or 36 days (14 days after last dose of toxin) or when moribund, and testicles were taken for histologic and ultrastructural examination. Other selected tissues, including liver and spleen, were taken for histologic examination. Histologically, testicular and splenic lesions occurred more consistently and at much smaller doses when compared with lesions in other target organs such as liver. Testicular and splenic lesions were present in all rats (6/6) given 0.8 …

Comparison Of A Quantitative Microtiter Method, A Quantitative Automated Method, And The Plate-Count Method For Determining Microbial Complement Resistance, Margie D. Lee, Richard E. Wooley, John Brown, Kathy R. Spears, Lisa K. Nolan, Emmett B. Shotts Jr.

Lisa K. Nolan

A quantitative microtiter method for determining the degree of complement resistance or sensitivity of microorganisms is described. The microtiter method is compared with a quantitative automated system and the standard plate-count technique. Data were accumulated from 30 avian Escherichia coli isolates incubated at 35 C with either chicken plasma or heat-inactivated chicken plasma. Analysis of data generated by the automated system and plate-count techniques resulted in a classification of the microorganisms into three groups: those sensitive to the action of complement; those of intermediate sensitivity to the action of complement; and those resistant to the action of complement. Although the …

Comparison Of Chicken Plasma And Guinea Pig Serum In A Quantitative Microtiter Method Of Determining Microbial Complement Resistance, Richard E. Wooley, John Brown, Kathy R. Spears, Lisa K. Nolan

Lisa K. Nolan

Y. A quantitative microtiter method using chicken plasma is described for determining the degree of complement resistance or sensitivity of avian Escherichia coli isolates. Results obtained with the microtiter method using chicken plasma were compared with results obtained using commercially available standardized guinea pig serum as the source of complement. The test organisms consisted of five isolates of E. coli isolated from chickens. Three isolates were from flocks with colisepticemia; one was from a flock with omphalitis; and one isolate was a non-pathogenic control. Data were accumulated from the five avian E. coli isolates incubated at 35 C with either …

Bronchiolar Metaplasia And Ulex Europaeus Agglutinin I (Uea-I) Affinity In Mycoplasma Hyopneumoniae-Infected Lungs Of Six Pigs, Mark R. Ackermann, M. C. Debey, B. M. Debey

Mark R. Ackermann

Mycoplasma hyopneumoniae adheres to the cilia and apical plasma membrane of cells that line the trachea, bronchi, and bron ch ioles. Colonization result s in ciliostasis and peribronchial and peribronchi olar nod ular aggregates of lymphocytes and plasma cells.">Altho ugh adhesion-receptor interaction s between the organism and respiratory epithelial cells have been examined, little attention has been given to the morphologic and phenotypic changes that may occur in cells lining bron chioles during infection. In this study, we describe morphologic cha nges and differenc es in lectin staining patt ern s in the bron chi oles from lungs of …