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Biomedical subjects

R F Rosenbusch

Publications and source records attributed to R F Rosenbusch.

At least 19 recordsLinked to original sources
PubMed2004

Destruction of Mycobacterium paratuberculosis, Salmonella spp., and Mycoplasma spp. in raw milk by a commercial on-farm high-temperature, short-time pasteurizer.

The 2002 NAHM's Dairy Survey indicated that 87.2% of dairy farms in the United States feed waste milk to their neonatal calves. Although cost-effective, this practice can lead to increased calf morbidity and mortality due to ingestion of pathogenic agents. In an effort to reduce the risk of infection, dairy producers are implementing on-farm pasteurization of the waste milk as a control procedure before feeding the milk to calves. In the present study, the efficacy of a commercial high-temperature, short-time (HTST) on-farm pasteurizer unit to destroy Mycobacterium paratuberculosis, Salmonella enterica spp., and Mycoplasma spp. in raw milk was evaluated. Replicate experiments were run for 3 isolates of M. paratuberculosis, 3 serovars of Salmonella (derby, dublin, typhimurium); and 4 species of Mycoplasma (bovis, californicum, canadense, serogroup 7) at 2 different levels of experimental inoculation. In addition, HTST pasteurization experiments were performed on colostrum experimentally inoculated with M. paratuberculosis. After culture of the pasteurized milk samples, no viable M. paratuberculosis, Salmonella, or Mycoplasma were recovered, regardless of species, strain, or isolate. Pasteurization of colostrum was also effective in the destruction of M. paratuberculosis but resulted in an average 25% reduction in colostral immunoglobulin. These results suggest that HTST pasteurization is effective in generating a safer product to feed to young calves.

Animals↗
PubMed2001

Use of arbitrarily primed polymerase chain reaction to investigate Mycoplasma bovis outbreaks.

Colony lineages from three Mycoplasma bovis outbreaks representing different husbandry conditions in the United States were characterized with arbitrarily primed polymerase chain reaction (AP-PCR). Cases studied included a closed beef herd, a dairy calf ranch, and a feedlot. The DNA was obtained from colony lineages and used for AP-PCR with primers REP1R-I and REP2-I. Case A and C lineages were uniform by AP-PCR analysis. Lineages from case B showed heterogeneity with AP-PCR. Outbreaks A and C were therefore both infected by one source, while the ranch (case B) was infected by multiple calf shipments. The AP-PCR typing method provides genotypic epidemiological information to successfully characterize M. bovis from sequential sampling of outbreaks and different husbandry conditions.

Animal Husbandry↗
PubMed2001

Cross-reactive proteins among eight bovine mycoplasmas detected by monoclonal antibodies.

Whole cell proteins of eight bovine mycoplasmas (M. bovoculi, M. bovis, M. dispar, M. bovirhinis, M. arginini, M. verecundum, M. canadense, M. alkalescens) were separated by SDS-PAGE and transferred to nitrocellulose paper. Rabbit anti-M. bovoculi serum was found to react with immunoblots of all mycoplasma species tested. These cross-reactive proteins were in the range of 35,000-100,000 molecular weight. Monoclonal antibody MA25.5 developed against a M. bovoculi 94 kDa surface protein cross-reacted with a band of 62 kDa from M. dispar and three bands of 89, 85 and 74 kDa from M. arginini only while MA18.13 that recognized a band of 57 kDa from M. bovoculi did not react with the other species. The role of MA25.5 monoclonal antibody in inhibiting the growth of M. bovoculi, M. dispar and M. arginini was tested using the metabolic-inhibition (MI) test. Monoclonal antibody MA25.5 inhibited the growth of M. bovoculi and also inhibited M. dispar growth but at lower MI titers, while it showed no effect on the growth of M. arginini.

Animals↗
PubMed2001

Detection of Mycoplasma bovis in preservative-treated field milk samples.

Control of mycoplasmal mastitis requires individual cow milk sampling for culture and identification of Mycoplasma bovis. This sampling is time-consuming and expensive. Currently, some herds sample cows monthly with the dairy herd improvement (DHI) program, but a preservative is added to this milk that kills M. bovis. In this paper, a nested polymerase chain reaction (PCR) procedure that allows for rapid testing of preservative-treated milk is validated. The specificity of the nested PCR assay was confirmed by testing isolated nucleic acids of other organisms phylogenetically related to M. bovis or common to milk. A comparison against blind-passage culture on 53 field milk samples determined its sensitivity. Exposure of seeded milk samples to the procedure resulted in a sensitivity of 5.1 cfu equivalents per milliliter. Analysis of these results proved that the nested PCR assay was as sensitive as traditional culture and can be used on preservative-treated milk.

Animals↗
PubMed2000

Pasteurization of discard mycoplasma mastitic milk used to feed calves: thermal effects on various mycoplasma.

Discard milk from sick or antibiotic-treated cows is often used as an economical alternative to milk replacer at dairy farms. This practice poses a health risk to calves if the discard milk is from cows with mycoplasma mastitis. Mycoplasma bovis, Mycoplasma californicum, and Mycoplasma canadense are among the agents known to cause contagious mastitis in cattle and occasionally pneumonia, otitis media, or arthritis in calves. This report describes a recent outbreak of calf polyarthritis and respiratory disease on a midwest dairy farm. The farm fed discard mycoplasma mastitic milk to its calves. On-the-farm pasteurization of the discard milk to 65 degrees C for 1 h before feeding prevented additional illness in the calves. Discard milk samples were collected before and after heating and tested for mycoplasma by culture. Only samples collected before pasteurization yielded live cultures. Common mastitic mycoplasma agents were also tested for sensitivity to heat. It was determined that 65 degrees C killed M. bovis and M. californicum after 2 min of exposure, while M. canadense remained viable for up to 10 min. Exposure to 70 degrees C inactivated M. bovis and M. californicum after 1 min, but M. canadense samples were positive for up to 3 min. Thus, M. canadense appears to be more heat resistant than M. bovis and M. californicum. Heat treatment that results in the destruction of M. canadense should be used for the pasteurization of discard mycoplasma mastitic milk.

Animal Feed↗
PubMed1999

Interactions of Mycoplasma bovoculi with erythrocytes: role of p94 surface protein.

The attachment of two strains of Mycoplasma bovoculi to erythrocytes was measured using 35S-methionine-labelled organisms. Receptor sites of M. bovoculi involved in this attachment are trypsin-sensitive, since mild trypsin treatment of the intact organisms abolished this process completely. Pretreatment of erythrocytes with trypsin or increasing concentrations of neuraminidase resulted in no measurable effect. Monoclonal antibody MA25.5 directed against a M. bovoculi surface antigen of 94 kDa termed p94 blocked 40% of the attachment, while MA18.13 directed against a 57 kDa protein band of M. bovoculi had no effect on the attachment process. Other properties of M. bovoculi were tested using six strains of the mycoplasma and erythrocytes from several animal species. None of the strains showed haemagglutinating or haemadsorbing activities.

Animals↗
PubMed1999

Differential serologic response to Mycoplasma ovipneumoniae and Mycoplasma arginini in lambs affected with chronic respiratory disease.

An enzyme-linked immunoabsorbent assay (ELISA) was used to evaluate the levels of antibodies to Mycoplasma ovipneumoniae and M. arginini in lambs with chronic respiratory disease. Sera were obtained from lambs in several flocks at various stages of the clinical disease and tested with sodium dodecyl sulfate (SDS)-treated M. ovipneumoniae and M. arginini whole cells and a crude capsular extract of M. ovipneumoniae as the antigens. There were low levels of antibody to M. ovipneumoniae in flocks sampled at the early stages of infection, whereas increased levels of antibody were present in lambs from flocks that had apparently recovered from the clinical disease. Slowly rising titers of circulating antibodies to M. ovipneumoniae were confirmed by sequential bleeding of lambs during the course of the clinical disease. However, antibody levels of M. arginini were more likely to increase earlier in the disease process. There was significant cross-reactivity between the 2 SDS-treated antigens in both the ELISA test and western immunoblotting. In contrast, the crude capsular extract was specific for detecting antibodies to M. ovipneumoniae.

Animals↗
PubMed1998

Occurrence of autoantibodies to cilia in lambs with a 'coughing syndrome'.

A respiratory disease of lambs that has been termed the 'coughing syndrome' has been observed in the mid-western region of the United States of America. Mycoplasma ovipneumoniae (M. ovipneumoniae) and Mycoplasma arginini (M. arginini) were routinely isolated from the respiratory tract of lambs with this disease. A high level of antibodies reactive with ovine cilia of the upper respiratory tract was detected in the sera from many of the lambs in affected flocks but not in sera of lambs from unaffected flocks. The reactivity of these antibodies with cilia was demonstrated by ELISA and confirmed by indirect immunofluorescent staining and western immunoblotting. These antibodies were predominantly of the IgG isotype. They were distinct from cold or warm agglutinins and could be absorbed from the sera with cilia but not with antigens of common bacterial pathogens of the sheep respiratory tract including M. ovipneumoniae, M. arginini, Pasteurella haemolytica, Pasteurella multocida or Neisseria ovis. In addition, their occurrence appeared to be independent of the specific antibodies to M. ovipneumoniae and M. arginini. Western immunoblotting indicated that the antibodies were directed primarily against an antigen with apparent molecular weight of 50 kDa. In one flock from which serial serum samples were collected from the same lambs over a 10-month period, antibodies to ovine cilia developed before the onset of the clinical disease and persisted for a period of several months until most of the lambs had apparently recovered. However, colonization of the respiratory tract of the lambs by M. ovipneumoniae preceded the production of these antibodies. Sequential serum samples taken from another flock, with no known history of this coughing, showed no such antibodies throughout the sampling period. It is suggested that an immunopathologic mechanism involving production of autoantibodies directed against a ciliary antigen of the lambs could be a contributing factor to the pathogenesis of this clinical disease.

Animals↗
PubMed1998

Identification and localization of a 94 kDa membrane protein found in Mycoplasma bovoculi strains.

Six isolates of Mycoplasma bovoculi obtained from cattle herds with bovine keratoconjunctivitis were analyzed by gel electrophoresis and immunoblotting techniques. All six strains showed similarity in their protein profiles although no two patterns were identical. Antigenic differences between strains were detected in immunoblots reacted with post-exposure calf serum. A common 94 kDa protein band designated p94 was detected in all six strains reacted with monoclonal antibody MA25.5 developed to one of the strains. The p94 was also recognized in these strains by the calf serum. Trypsin treatment of intact mycoplasma cells resulted in the removal of p94 from immunoblots reacted with MA or hyperimmune rabbit serum. Other trypsin-resistant antigens shared between strains or being strain-specific in nature were identified when trypsin-treated mycoplasma cells were reacted with hyperimmune rabbit serum. The p94 antigen was shown to be of mycoplasmal origin by radio-immunoprecipitation using the MA or hyperimmune rabbit serum. These studies identify the presence of a surface antigen (p94) on M. bovoculi membrane in all strains examined that is trypsin sensitive by the use of monoclonal antibody, calf serum and hyperimmune rabbit serum.

Animals↗
PubMed1998

Field isolates of Mycoplasma ovipneumoniae exhibit distinct cytopathic effects in ovine tracheal organ cultures.

Ovine tracheal ring explants were infected with four different Mycoplasma ovipneumoniae and one M. arginini field isolate and their ability to induce cytopathic effects was tested by measuring ciliary activity and intracellular calmodulin release. Infected tracheal rings showed significantly decreased ciliary activity as compared to the non-infected control rings. There were, however, marked differences between isolates in the onset and severity of the effects which correlated with their ability to produce hydrogen peroxide. Infected tracheal rings released more calmodulin than the non-infected controls. The amount of calmodulin released also varied between isolates, and somewhat reflected the degree of loss of ciliary activity in the corresponding rings induced by the different isolates. Light and electron microscopic examinations of infected tracheal rings revealed disorganisation and sloughing of the epithelium, and association of mycoplasmas only with the cilia. Following repeated in vitro passages, the organisms had reduced ability to inhibit ciliary activity which correlated with decreased hydrogen peroxide production. Addition of catalase to the organ cultures delayed loss of ciliary activity. These results suggest that M. ovipneumoniae induced ciliostasis in ovine tracheal ring explants which correlated with hydrogen peroxide production. Furthermore, these M. ovipneumoniae-induced injuries to respiratory epithelial cells could contribute to the role that this organism may play in sheep respiratory disease.

Animals↗
PubMed1998

Demonstration of a capsule on Mycoplasma ovipneumoniae.

OBJECTIVE: To examine Mycoplasma ovipneumoniae for presence of a capsule and its potential role in adherence. SAMPLE POPULATION: 17 isolates of M ovipneumoniae and 2 isolates of M arginini, recovered from sheep with respiratory tract disease. PROCEDURE: Mycoplasmas were cultured in modified Fills broth medium, ovine fetal lung cells, or ovine tracheal ring explants. Pelleted mycoplasmas or ring cultures infected with mycoplasmas were treated with ruthenium red or polycationic ferritin and visualized by transmission electron microscopy. Reactivity of several lectins with the mycoplasmas was studied by use of a microtitration plate agglutination test. RESULTS: Electron microscopy revealed a large number of M ovipneumoniae cells covered with an electron dense-stained amorphous material suggesting that it was a capsule. Multiple passages of the microorganisms in modified Friis broth medium decreased thickness of the capsule, but not percentage of cells encapsulated. Marked differences were observed when M ovipeumoniae isolates grown in modified Friis broth medium or co-cultured with ovine fetal lung cells were compared for capsular thickness or percentage of encapsulation. In thin sections of ruthenium red-stained tracheal ring cultures, the mycoplasmas appeared to be in close contact with cilia through their capsule. All isolates of M ovipneumoniae reacted strongly with wheat germ agglutinin lectin. CONCLUSIONS: Mycoplasma ovipneumoniae produces a polysaccharide capsule with variable thickness that is dependent on culture conditions and strain. Morphologic observations suggest that this capsule facilitates adherence of the organism to ciliated epithelium.

Agglutination↗
PubMed1997

Expression of functions by normal sheep alveolar macrophages and their alteration by interaction with Mycoplasma ovipneumoniae.

Normal sheep alveolar macrophages collected by bronchial lavage were exposed to live or heat-killed Mycoplasma ovipneumoniae organisms, and their capability to ingest Staphylococcus aureus and to elicit antibody-dependent cellular cytotoxicity against sensitized chicken red blood cells was tested. Controls consisted of non-infected macrophages in M199 medium. In addition, the effect of M. ovipneumoniae on expression of surface molecules on these sheep alveolar macrophages was determined. The percentage of S. aureus ingested by nontreated sheep alveolar macrophages was significantly higher than that of infected macrophages. Live mycoplasmas were more effective in suppressing the ingestion of S. aureus by these macrophages than killed mycoplasmas. Both live and killed mycoplasmas suppressed the cytolytic effect of the sheep alveolar macrophages to a similar degree. About 78% and 45% of the normal sheep alveolar macrophages had IgG and complement receptors, respectively. Infection of these macrophages with M. ovipneumoniae decreased significantly the expression of IgG receptors but had no effects on complement receptors. There were substantial increases in the expression of both MHC class I and class II by the mycoplasma-induced macrophages as compared with unstimulated macrophages. Live mycoplasmas were more effective in inducing expression of both classes than killed mycoplasmas. The results, taken together, suggest that M. ovipneumoniae induced alterations in macrophage activities and this may be a contributing factor in the pathogenesis of respiratory disease induced by the organism.

Animals↗
PubMed1996

Mycoplasma bovis-associated pneumonia and arthritis complicated with pyogranulomatous tenosynovitis in calves.

Immunohistochemistry and bacteriologic culturing were used to detect Mycoplasma bovis in tissue specimens from feedlot calves affected with pneumonia and arthritis. Two herds with 110 Charolais calves and 25 Angus calves were examined. Clinical signs included severe respiratory distress, anorexia, pyrexia, and lameness, which affected nearly a third of the calves. Lung lesions were characterized by numerous abscesses. Synovial lesions of the limbs included pyogranulomatous tenosynovitis, bursitis, and synovitis, particularly in the areas of the carpal and elbow joints. Abscesses in lung and synovial tissues contained accumulations of M bovis antigens, as revealed by immunohistochemistry. The findings of this report indicate that infection with M bovis may result in a pneumonia-arthritis syndrome with pyogranulomatous lesions in calves.

Abscess↗
PubMed1995

Immune responses to the capsular polysaccharide of Mycoplasma dispar in calves and mice.

Humoral and cell-mediated immune responses to the capsular polysaccharide (CPS) of M. dispar and polygalacturonic acid (pGaIU--a structurally similar polysaccharide) were investigated in calves experimentally infected with Mycoplasma dispar and in mice immunized with CPS or pGaIU. Sera, tracheobronchial lavage and nasal fluids, collected before and after infection in calves, were checked for the presence of anti-CPS and anti-pGaIU antibodies. The sera from mice injected with CPS or pGaIU were checked for different classes of anti-CPS and anti-pGaIU antibodies. Peripheral blood lymphocytes from calves and splenic lymphocytes from mice were monitored for specific proliferative responses to CPS and pGaIU. At about 2 weeks post-infection, anti-CPS IgM response in serum, anti-CPS and anti-pGaIU IgM and IgA response in lavage fluid and lymphocyte proliferative response was seen in the calves. Mice immunized with CPS and pGaIU gave exclusively IgM responses. No secondary response was seen in mice immunized with CPS in contrast to mice immunized with pGaIU. Antibodies cross-reactive with pGaIU were present in the sera of CPS-immunized mice but antibodies cross-reactive with CPS were not found in pGaIU-immunized mice. No significant blastogenic response was shown by mouse splenocytes to CPS or pGaIU.

Animals↗