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Biomedical subjects

R T Greene

Publications and source records attributed to R T Greene.

At least 19 recordsLinked to original sources
PubMed1996

Clinical and serologic evaluation of cats with cryptococcosis.

OBJECTIVE: To evaluate the long-term clinical outcomes and serologic changes in cryptococcal antigen and antibody titers in cats with confirmed Cryptococcus neoformans infection. DESIGN: Prospective case series. ANIMALS: 47 cats with cryptococcosis. PROCEDURE: Cats included in this study were determined to have cryptococcosis on the basis of identification of C neoformans on histologic or cytologic examination, isolation of C neoformans in culture, or positive serologic test results for cryptococcal antigens. Information concerning the signalment, history, physical examination findings, FeLV and feline immunodeficiency virus status, serologic testing, treatment, and outcome for each cat was requested on a survey form. Follow-up measurements of serum cryptococcal antigen and antibody titers were requested for all surviving cats. RESULTS: Signalment and clinical signs of cats with cryptococcosis reported here were consistent with previous reports. Treatment consisted primarily of azole antifungal drugs. All cats were seronegative for cryptococcal antibody titers, whether tested initially or at follow-up examination. All but 1 cat tested were seropositive for cryptococcal antigens when initially tested. Cats with and without clinical signs of C neoformans infection were seropositive for cryptococcal antigens months to years after initial diagnosis of cryptococcosis. CLINICAL IMPLICATIONS: The results of this study indicate that serum titers to cryptococcal antigens in cats can persist with or without clinical signs for months to years after an initial diagnosis of cryptococcosis is made. Repeated evaluation of serum cryptococcal antigen titers is advised during the treatment of cats to monitor progress, evaluate prognosis, and guide cessation of treatment.

Animals↗
PubMed1996

Proficiency testing of selected antigen and antibody tests for use in dogs and cats.

OBJECTIVE: To determine the correlation of seroimmunologic test results between reference and nonreference laboratories. DESIGN: Retrospective data analysis. PROCEDURE: Serum samples obtained from naturally infected dogs and cats were distributed to reference and nonreference laboratories for seroimmunologic testing. Correlation of test results was evaluated by use of nonparametric analysis. RESULTS: Correlation coefficients were high between laboratory groups for samples tested for feline immunodeficiency virus antibodies, FeLV antigen, and toxoplasmosis antibodies in cats. Results for feline immunodeficiency virus antibody tests from reference laboratories were more likely to be positive than results from nonreference laboratories. Test results for feline infectious peritonitis antibodies, antinuclear antibodies, and Borrelia antibodies in cats were not significant. Coefficient correlations were significant for results of heart-worm antigen, Brucella antibodies, Toxoplasma antibodies, antinuclear antibodies, and rheumatoid factor in dogs. Results for Borrelia antibodies were not correlated between laboratory groups. CLINICAL IMPLICATIONS: Results were highly correlated between reference and nonreference laboratories for 8 of 14 seroimmunologic tests. Seroimmunologic tests for use in cats were less correlated as a group than those for use in dogs. Poor correlation of results between laboratories was attributed to variations in control agents, antigens, reagents, technical expertise, and cutoff values and end-point titers used for diagnosis.

Animals↗
PubMed1993

Staphylococcus intermedius: current knowledge on a pathogen of veterinary importance.

It has been 16 years since the identification of S. intermedius as a new species. Numerous investigations using cell wall and DNA analytic methods have now clearly demonstrated significant differences to warrant the creation of this new species. However, studies investigating virulence factors associated with S. intermedius have not uncovered evidence that differentiates virulent from non-virulent isolates. Therefore, at the present time, it is difficult for veterinary clinicians and microbiologists to determine the clinical significance of many S. intermedius isolates. Host-bacterial interactions and the hosts' immune status appear to be the factors most crucial in determining the outcome of infections, not the virulence of the organism. Continued research in the virulence factor field will hopefully lead to a better understanding on how we can differentiate virulent from nonvirulent isolates of S. intermedius.

Animals↗
PubMed1992

Surface hydrophobicity of Staphylococcus intermedius and Staphylococcus hyicus.

Surface hydrophobicity of 90 Staphylococcus intermedius and 55 S hyicus isolates was evaluated using the hexadecane adherence assay and the ammonium sulphate salt aggregation test. A strongly positive hydrocarbon adherence in the hexadecane adherence assay was demonstrated in 11 per cent of the S intermedius isolates and 7 per cent of the S hyicus isolates. Bacterial aggregation in 1.6 M, or less, ammonium sulphate was observed in 28 per cent of the S intermedius isolates and 37 per cent of the S hyicus isolates. There was no statistical correlation between the two assays. The adherence of both bacterial species to hexadecane was eliminated when the cells were first treated with pronase and trypsin, while it was mildly enhanced by prior heat treatment (60 degrees C and 95 degrees C for up to three hours). In contrast, aggregation of S intermedius in ammonium sulphate was not influenced by trypsin pretreatment, and aggregation of both bacterial species was diminished, or eliminated, with pronase or prior 95 degrees C heat treatment. Surface hydrophobicity, as measured in both assays, appeared to have no relationship with growth patterns in serum soft agar or production of slime. Similarly, the presence or absence of substantial surface receptor activity to fibrinogen, fibronectin or IgG did not appear to be related to surface hydrophobicity.

Agglutination Tests↗
PubMed1992

Small antibiotic resistance plasmids in Staphylococcus intermedius.

Antibiograms and plasmid profiles were evaluated for 116 Staphylococcus intermedius isolates collected from dogs in Germany and in the USA. Of the 26 S. intermedius isolates from Germany, 9 (34.6%) carried plasmids, while 20 (22.2%) of the 90 S. intermedius isolates from the USA were found to be plasmid-positive. Eight small resistance plasmids were identified and characterized using protoplast transformations and restriction endonuclease analyses. Five plasmids (3.8 and 3.9 kb) encoded for chloramphenicol resistance, 2 plasmids (each 2.5 kb) carried determinants for macrolide-lincosamide resistance, and one plasmid (4.5 kb) conferred resistance to tetracycline. Detailed restriction maps of these plasmids were constructed and served for structural comparisons with other small resistance plasmids found in staphylococci. These comparisons implied marked structural homologies with those prototype plasmids initially characterized in S. aureus of human origin.

Ampicillin Resistance↗
PubMed1992

Isolation and characterization of immunoglobulin binding proteins from Staphylococcus intermedius and Staphylococcus hyicus.

125-I-IgG binding activities were observed with 15 (17%) of 90 S. intermedius isolates from dogs and 39 (95%) of 41 S. hyicus isolates from pigs. Binding activities were not detected with S. hyicus isolates from cows. The IgG binding proteins of 2 S. intermedius, 2 S. hyicus, and protein A from S. aureus Cowan I were isolated from their cell surfaces. The proteins precipitated with IgG preparations from human, rabbit, pig, dog and horse, but not with IgG from cow, mouse and chicken. This indicated that these IgG binding proteins could be classified as type I receptors. In addition, the isolated proteins from all 3 staphylococcal species precipitated with polyclonal chicken anti-protein A antiserum. SDS-PAGE, Western blotting and gel isoelectric focussing of the proteins revealed numerous bands in the 42,000 D range and acid isoelectric points. The isoelectric point of the isolated proteins from both S. intermedius cultures was slightly more acidic than those from S. hyicus and S. aureus. The present results indicate a close functional and antigenic similarity, if not identity, between IgG binding proteins of S. intermedius and S. hyicus, and protein A of S. aureus.

Animals↗
PubMed1991

Comparison of an enzyme-linked immunosorbent assay to an indirect immunofluorescence assay for the detection of antibodies to Borrelia burgdorferi in the dog.

An enzyme-linked immunosorbent assay (ELISA) was compared to an indirect immunofluorescence assay (IFA) for detection of IgG antibodies to Borrelia burgdorferi in dog sera. The concordance of the two tests was 93.5% for sera from dogs from Maryland (n = 93), 98.0% for sera from dogs from North Carolina (n = 446), and 97.2% for the combined sample groups (n = 539). Twenty-five of the 27 samples with discordant or low positive results were tested, and showed immunoblot reactions to 1 to 10 different bands. Reaction patterns and intensity of the bands were quite variable, and did not explain a reason for the discordance.

Animals↗
PubMed1991

Canine Lyme borreliosis.

Lyme borreliosis is now the most commonly reported tick-transmitted disease in humans and is an important disease in dogs. Case reports of canine Lyme borreliosis have appeared in the literature during the last 6 years, but a complete description of the disease still is not available. Until an accurate diagnostic scheme is developed, the disease will remain incompletely understood. A nonlocalizing polyarthropathy is the most commonly described clinical manifestation of canine Lyme borreliosis, but other syndromes probably also exist. The difficulty in making a diagnosis is a result of the fact that dogs do not develop a characteristic skin lesion to mark the beginning of their disease, and many dogs become seropositive but never develop clinical manifestations. Also, Borrelia burgdorferi has been isolated from the blood of healthy dogs, which suggests that detecting a spirochetemia may not have diagnostic significance. Newer diagnostic tests are being evaluated, but at present the diagnosis of canine Lyme borreliosis should be one of exclusion. After other common illnesses are ruled out, serology and response to antibiotic therapy help suggest a diagnosis. Once the disease is accurately diagnosed, efficient therapeutic schemes will be developed based on randomized therapeutic trials. In addition, vaccines are being developed. Currently, without the ability to diagnose the disease accurately, their efficiency can not be demonstrated adequately. Future findings surely will change our understanding of this disease.

Animals↗
PubMed1991

Interlaboratory comparison of titers of antibody to Borrelia burgdorferi and evaluation of a commercial assay using canine sera.

Sixty canine serum samples were sent to 10 different diagnostic laboratories for anti-Borrelia burgdorferi antibody analysis. All laboratories knew of the study prior to receiving the samples. Agreement among all laboratories for all interpretations was 91% (546 of 600 samples). There was complete agreement among all the laboratories for only 32 (53%) of the samples. Most of the disagreements were due to differences reported by either one (15 samples) or two (7 samples) laboratories per sample. When discrepancies in interpretations existed, the interpretation reported by the majority of the laboratories was considered the standard for comparison. One laboratory had no discrepant interpretations from this standard, while the laboratory with the most discrepancies had 16. The median number of discrepancies per laboratory was five. By using pairwise comparisons between each laboratory and the majority standard, eight of the laboratories showed strong agreement and the remaining two showed fair to good agreement. The type of test used (enzyme-linked immunosorbent assay versus indirect immunofluorescence assay) did not appear to influence the number of discrepant interpretations reported. Sera considered to be positive by the majority of the laboratories usually reacted to more than five antigens in immunoblots, with at least three or more of those being intense reactions. For positive samples, reactivity was consistently present in the 60-, 41-, 31-, and 22- or 24-kDa regions. Samples considered negative usually reacted to fewer than three bands, with reactivity usually being faint. A commercially available, rapid dot blot assay showed strong agreement with the majority standard.

Animals↗
PubMed1991

Phage typing of Staphylococcus intermedius.

Staphylococcus intermedius, a coagulase-positive staphylococcal species, is a common canine pathogen and a rare human wound pathogen. A total of 145 strains of S. intermedius (ATCC 29663, 4 reference strains, 4 human isolates, 44 canine infection isolates, and 92 isolates from canine gingiva) were screened for lysogenic phage by a modified Fisk method. Nineteen phage preparations were prepared for preliminary typing experiments. Lytic activity was observed on 93 of 145 (64.1%) isolates, yielding 44 lytic patterns with individual strains susceptible to one or more phages. Five phages lysed only a single strain, but lytic patterns varied from 1 to 11 lytic phages per isolate. A distinct lytic pattern did not separate canine or human wound isolates from canine gingival isolates. All human wound isolates fell into the two most common canine gingival or wound patterns; the single human nasopharyngeal isolate was not lysed by any phage. Twenty-two of 44 (55%) canine wound isolates and 65 of 92 (71%) gingival isolates yielded lytic patterns. Lysogenic phages are common in S. intermedius. This preliminary study suggests that phage typing may be a useful tool in distinguishing epidemiologically related strains.

Animals↗
PubMed1990

Evaluation of four commercial anaerobic systems for identification of Eubacterium suis.

Four commercial anaerobic systems (CASs) were evaluated for usefulness in identification of Eubacterium suis. Twelve strains were evaluated in each system in triplicate, and results were interpreted independently by 5 individuals. Statistically significant differences (P less than 0.01) due to strain variation and reader interpretation accounted for discrepancies encountered. The reactivity, repeatability, and unique profiles generated made both CAS-1 and CAS-2 suitable adjuncts for identification of E. suis when colony morphology and Gram reaction were considered. Limited reactivity in CAS-3 limited its use as an aid in identification. Variability in test observations and the large number of numerical profiles generated precluded use of CAS-4.

Anaerobiosis↗
PubMed1989

Shared flagellar epitopes of Borrelia burgdorferi and Borrelia anserina.

Antigenic cross-reactivity between Borrelia burgdorferi and Borrelia anserina was studied using mouse immune sera and monoclonal antibodies. With immune sera, significant cross-reactivity between B. burgdorferi and B. anserina was demonstrated by indirect immunofluorescent assay. In immunoblots, most of the cross-reactivity was shown to be associated with the periplasmic flagella. Using monoclonal antibodies in immunoblots, it was shown that B. burgdorferi and B. anserina shared at least two flagellar epitopes, one of which was not shared with Borrelia hermsii or Borrelia coriaceae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of whole cell lysates and the use of a species-specific monoclonal antibody (H5332) which reacts with a major outer surface protein (Osp A) of B. burgdorferi readily differentiated the two species at the molecular level.

Animals↗
PubMed1988

Heterogeneity in immunoblot patterns obtained by using four strains of Borrelia burgdorferi and sera from naturally exposed dogs.

This study evaluated the differences in immunoblot patterns when serum specimens from seropositive dogs were reacted against four strains of Borrelia burgdorferi. Intense bands were consistently detected for the 83-, 66-, 61- to 60-, 41-, and 31-kilodalton regions for all four strains. Most variations were observed in the regions of 45 to 34 and 26 to 15 kilodaltons. Adsorption studies suggested that one reason for the observed variability was a difference in proteins among the organisms, rather than a difference in migration of proteins. Therefore, knowledge and consistency of the test antigen are essential when evaluating and comparing canine immunoblot patterns to B. burgdorferi, but for diagnostic purposes all of the serum samples would have been considered positive regardless of the strain used.

Animals↗
PubMed1988

Immunoblot analysis of immunoglobulin G response to the Lyme disease agent (Borrelia burgdorferi) in experimentally and naturally exposed dogs.

Immunoblots were used to study the immunoglobulin G response to Borrelia burgdorferi in experimentally and naturally exposed dogs. Adsorption studies confirmed that the antibodies were specific for B. burgdorferi. Experimentally exposed dogs were asymptomatic. Naturally exposed dogs included both asymptomatic animals and animals showing signs compatible with Lyme disease. Naturally exposed dogs were from four geographic regions of the country. No differences were detected between immunoblot patterns of naturally exposed symptomatic or asymptomatic dogs from different areas of the country. The immunoblot patterns obtained with sera from experimentally exposed dogs were different from those obtained with sera from naturally exposed dogs and were characterized by reactivity to fewer and different protein bands. Immunoblot analysis using an OspA-protein-producing Escherichia coli recombinant showed that experimentally exposed dogs produced antibodies to OspA, whereas naturally exposed dogs did not. Modifications of the immune response over time, different routes of antigen presentation, and strain variation are factors postulated to account for the observed differences.

Animals↗