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Biomedical subjects

R C Cutlip

Publications and source records attributed to R C Cutlip.

At least 55 records · Page 3Linked to original sources

Effect of muramyl dipeptide on immunogenicity of Corynebacterium pseudotuberculosis whole-cell vaccines in mice and lambs.

Colostrum-deprived lambs and CF1 mice were vaccinated with water-in-oil emulsion vaccines containing nonviable whole cells (WC) of Corynebacterium pseudotuberculosis with and without muramyl dipeptide (MDP). Efficacy of vaccines was determined from the survival of mice and lesions in lambs after IV injection of 10(4) colony-forming units of C pseudotuberculosis. In mice, protection was related to the concentration of WC in the vaccine. At 50, 100, or 150 micrograms of WC, protection was good (78.8%). At 10 or 25 micrograms of WC, protection was considerably less (54.7%). At high WC concentrations, protection could only be moderately increased to 82.3% with high (50 and 100 micrograms) concentrations of MDP or increased to 90% protection with low (5 and 10 micrograms) concentrations of MDP. At low WC concentrations, protection significantly decreased to 32% (P less than 0.025) with high concentrations of MDP, but significantly increased to 72.5% (P less than 0.025) with low concentrations of MDP. Therefore, the amount of protection with lower concentrations of WC and MDP was comparable with the amount of protection with higher concentrations of WC without MDP. In lambs, high prechallenge antibody titers (geometric mean titers from 5.1 to 5.4 by day 35) were observed after vaccination with WC. Protection and vaccination site abscesses in lambs were related to the concentration of WC and MDP. Pulmonary or vaccination site abscesses were not observed in 4 of 4 lambs vaccinated with 1 mg of WC + 50 micrograms of MDP.

Acetylmuramyl-Alanyl-Isoglutamine↗

Evaluation of ethylenediaminetetraacetic acid, tetrasodium salt dihydrate (EDTA)-Tween 20 treatment versus protease digestion of formalin-fixed tissue sections for detection of bovine respiratory syncytial virus antigen in infected ovine lung.

The efficacy of protease and ethylenediaminetetraacetic acid, tetrasodium salt dihydrate (EDTA)-Tween 20 in unmasking bovine respiratory syncytial virus (BRSV) antigens in formalin-fixed lung tissue was compared using avidin-biotin immunoperoxidase procedure. Tissues were taken from experimentally infected lambs. BRSV antigen stained in both techniques. Treatment with EDTA-Tween 20 resulted in more intense staining of BRSV infected cells, more uniform cytoplasmic staining, less non-specific background, and superior cellular detail in comparison to protease digestion.

Animals↗

Encephalitozoonosis in arctic lemmings.

Encephalitozoonosis was identified for the first time in arctic lemmings (Dicrostonyx stevensoni and crosses of D. stevensoni and Dicrostonyx rubricatus). The most common clinical findings were circling, torticollis, posterior paralysis, depression, blindness and death related to parasitic granulomas in many tissues. Granulomas were most frequent in the central nervous system, but were seen in many other tissues throughout the body. Granulomas were characterized by collections of macrophages and varying numbers of lymphocytes and neutrophils with or without necrosis of parenchymal tissue. Most granulomas contained protozoon cysts with ultrastructural characteristics of Encephalitozoon cuniculi. Protozoon cysts were seen in vascular endothelium in many tissues accompanied by minimal or no inflammatory reaction.

Animals↗

Clinical and microbiologic findings in lambs inoculated with Pasteurella haemolytica after infection with ovine adenovirus type 6.

Colostrum-deprived lambs (10 to 20 days old) were inoculated with either ovine adenovirus type 6 (OAV-6; n = 6), Pasteurella haemolytica type A1 (n = 6), or OAV-6 followed by P haemolytica 5 days later (n = 10). Another group (n = 3) served as sham-inoculated controls. Lambs inoculated with OAV-6 or P haemolytica developed mild and moderate respiratory tract disease of 6 and 3 days' duration, respectively. Lambs inoculated with virus and bacteria developed clinical signs of respiratory tract disease of greater intensity and duration (9 days) than with either agent alone. Within 3 hours of bacterial inoculation, all lambs that received P haemolytica were anorectic, listless, and febrile, and had hyperpnea and dyspnea. Ovine adenovirus type 6 was isolated from all virus-inoculated lambs. Although P haemolytica was not recovered from all bacteria-inoculated lambs, it was recovered for a longer period in the group that received both agents. Antibody to OAV-6 was detected in virus-inoculated lambs as early as day 6 after inoculation. The control lambs remained clinically normal and neither virus nor bacteria were recovered at necropsy.

Adenoviridae↗

Effect of Pasteurella haemolytica (A1) capsular polysaccharide on sheep lung in vivo and on pulmonary surfactant in vitro.

Capsular polysaccharide (CP) of Pasteurella haemolytica (type A1) was first deposited by fiberoptic bronchoscopy into the lungs of sheep to examine lesions and changes in bronchoalveolar lavage cell populations and, later, was mixed with pulmonary surfactant to investigate alterations in physical properties or surface tension. At 22 hours after deposition, minimal lesions were seen in the lungs only at and contiguous to the site of CP deposition in 2 of 4 sheep. Microscopically, alveoli and interlobular septa were filled with edema fluid. Terminal airways and alveoli contained a moderate amount of neutrophils that varied between sheep. Significant differences in number or type of bronchoalveolar lavage cells were not observed in the weekly lavages between each group or among sheep within each group, either before or after deposition of CP or physiologic saline solution. After 6 hours of incubation at 37 C, CP-surfactant mixtures were examined with a surface tensiometer and centrifuged in sucrose gradients. The CP bound to surfactant, resulting in formation of a precipitate with a surface tension of 31.6 +/- 0.1 dynes/cm and a density of 1.07 to 1.08 g/ml. Lipopolysaccharide of P haemolytica, used as a control, also bound to surfactant, resulting in a complex with a surface tension of 57.7 +/- 0.4 dynes/cm and a density of 1.06 to 1.10 g/ml. Surfactant alone had a surface tension of 32.6 +/- 0.2 dynes/cm and density of 1.05 to 1.06 g/ml. The CP appears by itself not to be a direct major factor in the lung damage that develops in cases of pneumonic pasteurellosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Ovine progressive pneumonia (maedi-visna) in sheep.

Ovine progressive pneumonia (OPP) is a multi-systemic disease of sheep caused by a nononcogenic exogenous retrovirus belonging to the Lentiviridae subfamily. Characteristics of the disease are chronic lymphocytic pneumonitis, encephalitis, arthritis, mastitis and vasculitis associated with progressive wasting, dyspnea, lameness, indurated udder and, rarely, paralysis. Any one or all of the characteristics may be manifest. Transmission of the virus is predominantly through the colostrum to newborn lambs, however, transmission can occur by contact and in utero. Treatment of the disease is only symptomatic and prevention of infection is only by avoiding the virus.

Animals↗

Isolation and identification of mycoplasmas from the nasal cavity of sheep.

Mycoplasmas isolated from the nasal cavity of sheep in a ram test station were examined to determine their identity and prevalence. Specimens were obtained for mycoplasmal culture in 1980, 1982, and 1983 from 558 sheep, and mycoplasmas were isolated from 630 specimens from 320 sheep (57.3%). The isolates were characterized and differentiated into groups on the basis of sensitivity to digitonin, fermentation of glucose, and hydrolysis of arginine. Isolates in some groups were further characterized by use of additional diagnostic media, and their identity was confirmed by agglutination or growth inhibition with antiserum prepared from reference mycoplasmas. Of the 320 sheep with mycoplasmas, 293 had Mycoplasma ovipneumoniae, 12 had M arginini, and 1 had M capricolum. Two sheep had Acholeplasma spp, and 3 sheep had unidentified Mycoplasma spp. The remaining 9 sheep had M ovipneumoniae in combination with Acholeplasma spp (n = 3), M arginini (n = 3), M capricolum (n = 2), and an unidentified Mycoplasma spp (n = 1). The biochemical reactions of the M ovipneumoniae from the 293 sheep were similar, but varied in the degree of growth and fermentation in the basal medium containing glucose. The high prevalence of M ovipneumoniae indicated that it may be commensal in the upper respiratory tract of healthy sheep.

Animals↗

Failure of experimental vaccines to protect against infection with ovine progressive pneumonia (maedi-visna) virus.

Cell culture medium was harvested from cells infected with ovine progressive pneumonia (OPP) virus and used to prepare killed virus vaccines. Virus was inactivated by either heat, formalin, or ethyleneimine and used either without adjuvant, with Freund incomplete adjuvant, or with aluminum hydroxide adjuvant to vaccinate sheep. The sheep produced precipitating antibody against the virus but were not protected against infection when challenged with live OPP virus.

Adjuvants, Immunologic↗

Eradication of ovine progressive pneumonia from sheep flocks.

Two management methods for controlling ovine progressive pneumonia in sheep were evaluated. By a test and cull method, wherein seropositive sheep were culled from the flock, the causative virus was eradicated from a closed flock and controlled in an open flock. By an isolate-and-test method, wherein lambs were removed from infected ewes before nursing and reared in isolation, the virus was eradicated in 1 of 2 attempts to establish virus-free flocks. It was concluded that either method should be effective in controlling ovine progressive pneumonia. Results indicated that annual serologic monitoring of a virus-free flock would be necessary to ensure that the virus-free status is maintained.

Animals↗

Breed susceptibility to ovine progressive pneumonia (maedi/visna) virus.

In this retrospective study of breed differences in susceptibility to disease caused by ovine progressive pneumonia (OPP) virus, 29 Border Leicester sheep were compared with 46 Columbia sheep. As judged by frequency and severity of clinical signs and lesions attributable to the infection, Border Leicester sheep were markedly more susceptible than Columbia sheep and experimentally infected sheep were slightly more susceptible than naturally infected sheep. Differences in susceptibility to infection by the virus were not determined.

Animals↗

Complexing of bacterial lipopolysaccharide with lung surfactant.

Lipopolysaccharides (LPS) from Escherichia coli, Salmonella typhi, Klebsiella pneumoniae, Serratia marcescens, or Pseudomonas aeruginosa were mixed with pulmonary surfactant to investigate their in vitro interaction. After 6 h of incubation at 37 degrees C, LPS-surfactant mixtures were examined by sucrose density gradient centrifugation. The E. coli LPS-surfactant mixture was examined by immunoelectron microscopy with protein A-colloidal gold. The binding that occurred between LPS and the surfactant vesicles resulted in a complex with a density higher than the density of the surfactant alone. The protein A-colloidal gold identified LPS in the LPS-surfactant complexes. The toxicity of E. coli LPS was enhanced by complexing with the surfactant when compared with the intraperitoneal injection into CF1 mice, even at a 64:1 ratio of surfactant to LPS. The complexing of LPS and surfactant in the lung may alter the physiologic properties of surfactant that contribute to the physiopathological changes observed with some types of pneumonia.

Animals↗

Ultrastructure and frequency of mastitis caused by ovine progressive pneumonia virus infection in sheep.

Twenty-five sheep, experimentally (n = 15) or naturally (n = 6) infected with ovine progressive pneumonia virus and noninfected controls (n = 4), were evaluated for histological and ultrastructural lesions of mastitis. Histologically, nine of 15 experimentally infected sheep and all six naturally infected sheep had lympho-plasmacytic mastitis. Severity of the lesion increased with length of time after infection. Periductal lymphatic nodules were seen in five sheep experimentally infected for 2.8 years or longer and in five naturally infected sheep that were 3.7 years old or older. Ultrastructurally, responses to ovine progressive pneumonia virus were diffuse lympho-plasmacytic infiltrates in glandular interstitium, lymphocytic and occasional plasmacytic infiltrates in ductal walls and lumens, lymphoblasts surrounded by small lymphocytes in glandular interstitium, and degeneration of epithelium releasing cells and cellular debris into the lumen. Based on the prevalence of lesions, the mammary tissue was more susceptible to ovine progressive pneumonia virus than other target organs: lung, brain, and synovium. Lesions did not differ between breeds of sheep. Ovine progressive pneumonia virus was not seen in the mammary tissue but was isolated from 15 of 17 mammary glands.

Animals↗

Pulmonary lesions in lambs experimentally infected with ovine adenovirus 5 strain RTS-42.

Twelve lambs were inoculated transtracheally and intranasally with Mastadenovirus ovi 5 strain RTS-42 and killed sequentially. Pulmonary lesions were studied by light and electron microscopy. Four lambs served as sham inoculated controls. Pulmonary lesions consisted of multifocal areas of bronchiolitis and alveolitis associated with necrosis and sloughing of isolated type I and type II alveolar epithelial cells and nonciliated bronchiolar epithelial cells. This was followed rapidly by hyperplasia of the remaining epithelium and repair of the damage. A cellular infiltrate of neutrophils and macrophages began at 2 days after inoculation, peaked at 4 days after inoculation, gradually diminished until minimal at 12 days after inoculation, and was resolved at 21 days after inoculation. Surfactant was abundant and, along with debris, was removed from the alveoli by macrophages. Clinical disease was not seen, but lesions were believed to be sufficient to allow bacteria to colonize the lungs and cause severe disease.

Adenoviridae↗

Inoculation of lambs with ovine adenovirus 5 (Mastadenovirus ovi 5) strain RTS-42.

Twelve 1-week-old colostrum-deprived lambs inoculated with the RTS-42 strain of Mastadenovirus ovi 5 were killed and necropsied (2 lambs/day) on postinoculation days (PID) 2, 4, 6, 8, 12, and 21. Four noninoculated lambs were killed and necropsied (2/day) on PID 6 and 12. Virus was isolated from nasal secretions and feces on PID 1 to PID 6, from tracheal fluids and lung tissue of lambs necropsied on PID 2, 4, and 6, and from lung tissue from 1 lamb necropsied on PID 8. Virus was not recovered from liver, kidney, or small intestine of inoculated lambs or samples from noninoculated lambs. Serum antibody was first detected on PID 6 in the inoculated lambs. Noninoculated lambs remained seronegative. None of the lambs in the study developed clinical signs of infection although lesions were produced in the respiratory tract.

Adenoviridae↗

Susceptibility of colostrum-deprived swine of various ages to streptococcic lymphadenitis.

The effect of age on susceptibility of young pigs to streptococcic lymphadenitis was investigated. Twenty-nine cesarean-derived, colostrum-deprived pigs were allotted to 7 groups exposed to type IV group E Streptococcus (GES) at 5, 14, 28, 35, 56, 70, and 84 days of age. Four cesarean-derived, colostrum-deprived pigs were maintained as nonexposed controls. Six naturally farrowed, susceptible controls were exposed to GES at 63 to 84 days of age. All exposed pigs were killed and necropsied 28 days after exposure. Lesions of streptococcic lymphadenitis were not observed in pigs exposed at 5 or 14 days of age, except for 1 microabscess in a mandibular lymph node in a pig exposed at 14 days, but GES was recovered from 11% of lymph nodes examined from pigs of those age groups. Lesions and GES-positive lymph nodes were frequent in cesarean-derived, colostrum-deprived pigs exposed at 28 days and older and in susceptible controls. Serologic response to exposure, as determined by microtitration agglutination test and bactericidal test, was observed only in pigs exposed at 14 days and older. The absence of abscess development in pigs exposed at 5 or 14 days of age was not caused by antibody or failure of infecting organisms to reach the target organs.

Animals↗

Incubation of Pasteurella haemolytica and Pasteurella multocida lipopolysaccharide with sheep lung surfactant.

Lipopolysaccharides (LPS) were extracted from a serotype of each of 2 species of Pasteurella isolated from sheep with respiratory tract infections. Lipopolysaccharides from P haemolytica 82-25 (serotype 1A) or P multocida P-1573 (serotype 12) were mixed with sheep lung surfactant and were incubated for 6 hours at 37 C. After incubation, LPS-surfactant mixtures were centrifuged overnight in sucrose density gradients, and fractions were analyzed. Binding occurred between LPS and surfactant vesicles resulting in a stable complex with densities greater than those with the surfactant alone. The surfactant alone had a density of 1.052 to 1.060 g/ml. Diffuse bands of surfactant had a density of 1.075 to 1.092 when incubated with P haemolytica LPS and a density of 1.069 to 1.105 when incubated with P multocida LPS.

Animals↗

Preparation of sheep and cattle immunoglobulins with antibody activity by high-performance liquid chromatography.

High-performance anion-exchange chromatography (HPIEC) and high-performance size-exclusion chromatography (HPSEC) were used to purify serum IgM and IgG from sheep and cattle. Pooled serum from normal cattle and sheep and serum from sheep, infected with two different viruses, were prepared for HPIEC by chromatography on CM-Affi-Gel Blue. After HPIEC, fractions containing IgG and IgM were pooled and concentrated and further purified by HPSEC. The purity of fractions from HPIEC and HPSEC were evaluated by immunoelectrophoresis, protein-A-Sepharose affinity chromatography, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Monoclonal antibody specific for bovine IgG2 was used to assay for IgG2 and IgG2 contamination of other fractions. The antibody activity to ovine adenovirus 5 and to ovine progressive pneumonia virus was assayed by neutralization and immunodiffusion. Antibody activity was retained against both viruses in the fractions containing IgG1 and IgM. This high-performance liquid chromatography procedure was a rapid preparative method to produce specific immunoglobulins and could be used to evaluate the purity of immuno-reagents.

Animals↗

Immunogenicity of Corynebacterium pseudotuberculosis and the effect of adjuvants in mice.

Intraperitoneal inoculation of CF1 mice with 100 micrograms, 200 micrograms, or 300 micrograms whole cell (WC) or 250 micrograms, 500 micrograms, or 1000 micrograms cell wall (CW) of Corynebacterium pseudotuberculosis induced varying degrees of protection after intravenous challenge of immunity with 7.2 X 10(4) CFU of C. pseudotuberculosis. Generally, the degree of protection increased with the dose of WC or CW. However, intraperitoneal inoculation of mice with 100 micrograms, 200 micrograms, or 300 micrograms heat-killed Mycobacterium bovis BCG; 50 micrograms, 150 micrograms, or 300 micrograms of either muramyl dipeptide (MDP) or trehalose dimycolate (TDM); or 350 micrograms, 700 micrograms or 1400 micrograms Corynebacterium parvum did not induce resistance to intravenous inoculation of 7.4 X 10(4) CFU of C. pseudotuberculosis. The protection induced by 500 micrograms CW was enhanced by adding 100 micrograms BCG, 150 micrograms MDP, or 350 micrograms C. parvum but protection induced in mice by 300 micrograms of WC was not enhanced by adding any adjuvants.

Acetylmuramyl-Alanyl-Isoglutamine↗