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Biomedical subjects

M T Collins

Publications and source records attributed to M T Collins.

At least 109 records · Page 6Linked to original sources

Crossed immunoelectrophoretic analysis of Legionella pneumophila serogroup 1 antigens.

By crossed immunoelectrophoresis, 85 different antigens were demonstrated in sonicated preparations of Legionella pneumophila serogroup 1 (Lp1). The precipitin patterns of 82 anodic-migrating antigens were numbered and were designated the Lp1 reference system. Eleven antigens were stable to boiling, and seven of these were shown to be surface antigens. One heat-stable surface antigen (antigen no. 61) was highly reactive with limulus amoebocyte lysates and formed a precipitin resembling lipopolysaccharide. Serum from an isolation confirmed case of Lp1 infection and serogroup-specific rabbit antiserum reacted specifically with antigen no. 61, which was designated the serogroup-specific antigen. Normal human and rabbit sera commonly had antibodies to antigen no. 66 of the Lp1 reference system. This antigen is antigenically related to the "common antigen" of Pseudomonas aeruginosa.

Animals↗

Cross-reactions between Legionella pneumophila (serogroup 1) and twenty-eight other bacterial species, including other members of the family Legionellaceae.

Cross-reactions between Legionella pneumophila serogroup 1 and 28 other bacterial species were studied by various quantitative immunoelectrophoretic techniques. A sonicated L. pneumophila antigen and purified homologous rabbit antibody were used as a reference system. Few antigens (0 to 6) cross-reacted with non-Legionellaceae, but two were found in nearly all gram-negative bacteria tested (antigens no. 1 and 66). Antigen no. 66 of the L. pneumophila reference system was shown to be antigenically similar to the "common antigen" of Pseudomonas aeruginosa reported in many gram-negative bacteria. Greater than 85% of the antigens from L. pneumophila serogroup 1 cross-reacted with the other six serogroups of L. pneumophila. By contrast, Fluoribacter (Legionella) bozemanae, F. (L.) dumoffii, F. (L.) gormanii, and Tatlockia (Legionella) micdadei cross-reacted with only 45, 53, 39, and 43% of the reference system antigens, respectively. The antigenic relatedness of members of the Legionellaceae, expressed as a matching coefficient, is discussed in terms of its taxonomic significance. Serogroup-, genus-, and family-specific antigens are identified in the L. pneumophila reference system.

Antigens, Bacterial↗

Detection of mycobacteria by radiometric and standard plate procedures.

A group of 89 smear-positive sputum specimens were evaluated by radiometric and standard plate procedures to determine the methodology which would provide the earliest detection of mycobacteria and maximum test sensitivity. Digested non-decontaminated specimens were concentrated and inoculated into modified selective BACTEC radiometric 7H12 broth and Mitchison selective 7H10 agar. Sodium hydroxide (1.5% final concentration) was then used to decontaminate these specimens. They were then concentrated and inoculated into both selective and nonselective 7H12 radiometric broths and into selective 7H10 and nonselective Middlebrook 7H11 agar media. The specimen processing and media combinations providing the earliest detection were non-decontaminated specimens with modified selective 7H12 BACTEC broth and decontaminated specimens with 7H12 BACTEC broths. Maximum sensitivity (percent positive) was obtained by using non-decontaminated specimens on Mitchison selective 7H10 Agar (98%) or decontaminated specimens in 7H12 BACTEC broth (95%). The decontamination process was found to reduce significantly the number of mycobacteria in clinical specimens, particularly the mycobacteria other than Mycobacterium tuberculosis. The specimen processing-media combinations providing the earliest detection and maximum recovery of mycobacteria (100%) were non-decontaminated specimens with modified selective 7H12 BACTEC broth or Mitchison selective agar and decontaminated specimens with 7H12 BACTEC broth or 7H11 agar.

Culture Media↗

Legionella microagglutination test. A serological study of Danish patients.

A microagglutination (MA) test for antibodies to 10 different Legionella antigens was evaluated by comparison with an indirect fluorescent antibody (IFA) test. Sera from 227 patients were investigated. There was good agreement between the two tests, especially when testing sera from patients with a rise in titre. The MA is promising as an easy, quick, cheap and reproducible test for antibodies to Legionella.

Agglutination Tests↗

Experimental infections of horses with Legionella pneumophila.

Attempts to infect horses with Legionella pneumophila were undertaken to determine pathogenicity and to evaluate the possibility that horses serve as a reservoir for the organism. A previous study showed that the prevalence of antibodies to L pneumophila in the equine population exceeded 30% of over 600 sera examined. Horses were infected experimentally with the Philadelphia 1 or Bloomington 2 strain of L pneumophila IV or by aerosolization. Signs of clinical illness were restricted to a transient febrile response. A transient decrease in circulating lymphocytes occurred 2 days after inoculation. At necropsy, only moderate generalized lymphadenopathy was noted. Histologically, the lungs contained evidence of a low-grade inflammatory response characterized by focal proliferation of alveolar lining cells, with few neutrophils and eosinophils. Lymph nodes had evidence of reactive hyperplasia. The tissue response to Bloomington 2 strain was slightly more pronounced than that to Philadelphia 1. Attempts to reisolate L pneumophila from blood and nasal or pharyngeal swabs were unsuccessful. The organism was not isolated by culturing tissues obtained at necropsy, nor was it demonstrated by tissue-staining techniques. However, all horses exhibited a marked increase in agglutinating antibodies to L pneumophila serogroups (SG) 1 and 3 as early as 4 days after inoculation. The serologic response was confirmed by indirect immunofluorescence and was shown to consist predominantly of immunoglobulin M by 2-mercaptoethanol treatment. Agglutinating antibodies persisted at least 4 months after infection. On the basis of these studies, the pathogenicity of L pneumophila SG 1 and 3 for the horse appears to be low. There is no evidence to support a role for the horse in the maintenance of these organisms in nature. Horses may be exposed in the environment and maintain a relatively long-lived serologic response to L pneumophila. However, it is also possible that they become infected with other strains of L pneumophila or Legionella-like organisms more pathogenic for horses, or other non-Legionella bacteria, which elicit a cross-reacting serologic response to L pneumophila SG 1 to 4.

Animals↗

Membrane filter technique for the isolation of Yersinia enterocolitica.

A membrane filter procedure was developed for the isolation of Yersinia enterocolitica from aquatic environments. Primary differentiation was based on the fermentation of sorbitol, the absence of lysine decarboxylase and arginine decarboxylase-dihydrolase activities, and the production of urease. Sodium deoxycholate was incorporated as an inhibitor of background organisms. The presumptive identification of Y. enterocolitica was accomplished in 50 h, and the rate of identity confirmation of typical colonies was 88%. The mean recovery rate of 15 strains from phosphate buffer suspensions was 91%, and quantitative recovery was demonstrated for low populations of the organism in both laboratory-prepared and naturally occurring mixed cultures. The technique was used to isolate 33 strains of Y. enterocolitica from 15 of 27 river water samples and from prechlorinated sewage effluent. Nine (27%) of the isolates were rhamnose positive, and only five (15%) were serotypable. Two isolates were identified as serotype O:4 (or O:4,32), two were O:17, and the fifth was O:40.

Animals↗

Prevalence of antibodies to Legionella pneumophila in animal populations.

We examined more than 2,800 human and animal sera for antibodies to four serogroups of Legionella pneumophila by using the microagglutination test. Antibody titers of greater than or equal to 1:64 were considered positive. The occurrence of positive equine sera (31.4%) was significantly higher than the occurrence of positive sera in cattle (5.1%), swine (2.9%), sheep (1.9%), dogs (1.9%), goats (0.5%), wildlife (0%), and humans (0.4%). The highest titer measured in horses was 1:512. The occurrence of positive sera in horses was related directly to age. In horses less than or equal to 1, 2 to 3, 4 to 7, 8 to 12, and greater than or equal to 13 years old, the percentages of positive sera were 0, 10.1, 30.3, 44.9 and 58.1%, respectively. When we compared age-specific serogroup-specific rates in horses from Colorado and Pennsylvania, we found differences. With horses 8 to 12 and greater than or equal to 13 years old, there was a significantly higher (P less than 0.05) occurrence of sera that reacted to serogroups II and III in horses from Pennsylvania. Of 242 positive sera, 43.8% reacted to a single serogroup (serogroup III or I most commonly), and 56.2% reacted to multiple serogroups (serogroups II and III or serogroups I, II, and III most commonly). A high percentage of seropositive horses suggested that horses are commonly infected with L. pneumophila or related organisms, and the age-specific rates of occurrence indicated that infection was related directly to duration of exposure. A definitive demonstration of equine infection will depend on isolation of the agent and repetition of this serological study with antigens obtained from organisms isolated from horses.

Age Factors↗

Urease testing of mycobacteria with BACTEC radiometric instrumentation.

A total of 140 mycobacterial isolates from patients treated at Fitzsimons Army Medical Center or the National Jewish Hospital and Research Center and from animal specimens submitted to the National Veterinary Services Laboratory were tested by using a urease procedure modified for use with a BACTEC model 301. Mycobacterial suspensions were prepared by using Middlebrook 7H10 Tween broth. Of the 98 mycobacteria isolates which were urease positive utilizing standard methodology, all were positive using the radiometric procedures. Similarly, all 42 urease-negative isolates were also negative employing the new methodology. Although maximum radiometric readings were observed at 48 h, all positive strains were readily identified 24 h after inoculation without sacrificing either test sensitivity or specificity. Thus, urease testing of mycobacteria, using the modified BACTEC radiometric methodology, was as sensitive, as specific, and more rapid than conventional methods.

Bacteriological Techniques↗

Evaluation of seven commercial oxidase test products with Pasteurella.

Seven commercial oxidase reagents were tested with 50 isolates each of Pasteurella multocida and P haemolytica. Each group of organisms consisted of a variety of serotypes from many locations and animal sources. Pasteurella multocida and P haemolytica were expected to be 90% oxidase-positive; however, only 2 commercial reagents were positive for greater than 90% of P multocida isolates. These were the Taxo N Dics and the Bacto-Differentiation Discs Oxidase. Only the Taxo N Discs were positive for greater than 90% of P haemolytica isolates. The 5 other commercial reagents demonstrated a variety of results. Pathotec Cytochrome Oxidase Strips were not positive with any of the 100 isolates tested. Correlation was not observed between dimethyl-p-phenylenediamine vs tetramethyl-p-phenylenediamine and the percentage of positive reactions.

Bacteriological Techniques↗

Seroepidemiology of Leptospira interrogans serovar hardjo in Colorado antelope and cattle.

A seroepidemiologic study was conducted to determine the prevalence of Leptospira interrogans serovar hardjo infection in Colorado cattle and antelope. Sera were tested by the microscopic agglutination test, using a standard battery of 12 antigens as well as 4 additional serovars of the Hebdomadis serogroup: hardjo, szwajizak, sejroe, and balcanica. Sera from 1,856 cattle and 544 antelope were tested. In both populations, the predominant serovar detected was hardjo; hardjo titers greater than or equal to 1:400 were found in 3.2% of the cattle and 3.6% of antelope. A positive correlation was found to exist between serologic evidence of infection in cattle and the presence of antelope. In counties with presumably infected antelope, 5.6% of cattle sera reacted to hardjo; in counties with uninfected antelope, 2.6% of cattle tested were seropositive for hardjo; where no antelope existed, titers of greater than or equal to 1:400 were not detected.

Animals↗

Identification of Pasteurella multocida and Pasteurella haemolytica by API 20E, Minitek, and Oxi/Ferm systems.

Fifty serotyped isolates each of Pasteurella multocida and Pasteurella haemolytica were tested on the API 20E strip (Analytab Products, Plainview, N.Y.), the Oxi/Ferm tube (Roche Diagnostics, Nutley, N.J.), and the Minitek system (BBL Microbiology Systems, Cockeysville, Md.). None of the rapid test systems reliable identified these organisms. With the API system, discrepancies between expected and actual results for the oxidase test and nitrate test frequently resulted in misidentification or no identification. The Minitek system misidentified 68% of the P. haemolytica isolates. The Minitek identification of Pasteurella depends on 100% positive xylose reactions, whereas only 56% of the P. haemolytica strains were positive for xylose fermentation. The Oxy/Ferm system, instead of giving a definitive identification, in most instances merely placed Pasteurella in a category of similar organisms.

Bacteriological Techniques↗

Use of the API 20E system to identify non-Enterobacteriaceae from veterinary medical sources.

The capability of the API 20E system to identify gram-negative nonfermenters and nonenteric fermenters was evaluated for 272 isolates from veterinary sources. Two different methods were used for interpreting the carbohydrate fermentation reactions on the strip. In method I, weakly fermented (yellow-green) carbohydrates were considered positive for all oxidase-positive organisms, and in method II, yellow-green carbohydrates were considered positive for all organisms requiring incubation for 48 hours. By both methods, the API system correctly identified 62% of the isolates. With method I, 31% of the isolates were misidentified and 6% were not identified. With method II, 21% of the isolates were misidentified and 17% received no identification. Organisms most affected by these 2 methods of interpretation were Pasteurella and Actinobacillus. Identifications reached by the API system were also compared with identifications made by veterinary diagnostic laboratories. The frequency of identifications agreements was not significantly affected by the method of API carbohydrate fermentation reaction interpretation. Generally, 30% of the identifications agreed (diagnostic laboratories vs API) when using only the API Index, whereas 51% agreed when the entire API computer data base identifications were included. The type of identification disagreements between diagnostic laboratories and the API system, however, was significantly affected by the method of API strip interpretation. With method I, 42% of the identifications were different and 6% were not in the API data base. With method II, 33% of the identifications were different and 17% were not in the API data base. Biotype differences between human and veterinary isolates were also compared. Significant differences between the predicted and actual reactions were noted for Pseudomonas aeruginosa and Bordetella bronchiseptica; however, these differences did not affect their correct identification to the API Index. For Pasteurella multocida, most profile numbers were not listed in the API Index because of differences in the actual vs predicted oxidase and nitrate reduction reactions; however, they were correctly identified with the total computer data base.

Animals↗

Use of the API 20E system to identify veterinary Enterobacteriaceae.

A total of 503 veterinary enteric bacterial pathogens obtained from state veterinary diagnostic laboratories were tested on API 20E strips to determine whether this rapid microidentification system could be utilized for veterinary clinical microbiology. The API 20E strip accurately identified 96% of the veterinary isolates and misidentified 3%. Identifications by the API system and the diagnostic laboratories were in agreement in 85% of the isolates, disagreement on 16% of the isolates, and 1% were not identified by the API strip. Differences in identification occurred primarily in distinguishing between Klebsiella and Enterobacter and between Enterobacter and Escherichia coli. These disagreements were most often due to incorrect identifications by the diagnostic laboratory rather than by the API system. Biotype differences between human and veterinary isolates were compared. Significant differences were noted in several biochemical reactions. The main differences observed for E. coli isolates were in ornithine decarboxylase production and melibiose fermentation. The largest differences for Salmonella occurred in arginine dihydrolase production, citrate utilization, and inositol fermentation, whereas for Klebsiella pneumoniae the main differences were noted in urease production and nitrate reduction. These biotype differences, however, did not affect the accurate identification of organisms on the API strip.

Animal Population Groups↗

Immune response of channel catfish under different environmental conditions.

Channel catfish were maintained under conditions of low (15.4 C) and fluctuating (15.4 to 26.9 C/24 hours) temperatures, low (126 mg/L NO3-N) and high (289 mg/L NO3-N) nitrate in recirculating systems, crowding (171 g body mass/L), and fasting. They were vaccinated with formalin-killed enteric red-mouth bacterium, and antibody titers were monitored weekly for 10 weeks. Only those fish maintained in low or fluctuating temperature environments had significant (P less than 0.01) immunosuppression. The other environmental conditions studies, which are commonly encountered in intensive fish culture operations, did not compromise the humoral immune response of channel catfish.

Animals↗

Mycobacterium paratuberculosis. Factors that influence mycobactin dependence.

Mycobacterium paratuberculosis does not produce any detectable mycobactin, an iron-binding compound that is synthesized by most Mycobacterium spp. and necessary for the growth of all mycobacteria. This study examined the influence of various culture conditions on mycobactin dependence in M. paratuberculosis. Using a radiometric growth assay, we found the minimal concentration of mycobactin J necessary for growth of M. paratuberculosis to be 0.006 microM, whereas 1.2 microM (1 microgram/ml) was required for optimal growth. In media without mycobactin at iron concentrations less than or equal to 100 microM, growth of M. paratuberculosis occurred at pH 5.0, but not pH 6.8. Iron concentrations greater than 100 microM did not significantly increase growth at pH 5.0, but at pH 6.8 the growth rate increased with increasing amounts of iron reaching a rate equal to control cultures containing mycobactin. Mycobacterium paratuberculosis appeared to lose mycobactin dependence when subcultured; however, this was subsequently shown to be a result of mycobactin carried over from primary medium. Removal of this contaminating cell-wall-associated mycobactin reestablished mycobactin dependence. We conclude that mycobactin dependence must be carefully determined because it is a key test used in identification of M. paratuberculosis and may be easily influenced by media pH, iron concentration, and mycobactin carryover from primary media.

Animals↗

Apolipoprotein E type 4 allele and cerebral glucose metabolism in relatives at risk for familial Alzheimer disease.

OBJECTIVE: Cerebral parietal hypometabolism and left-right asymmetry occur early in the course of Alzheimer disease (AD), and the apolipoprotein E type 4 allele (APOE epsilon 4) is a risk factor for familial AD. To determine if APOE epsilon 4 is associated with lowered brain function in nondemented relatives at risk for familial AD, we studied 12 relatives with APOE epsilon 4 and 19 relatives without APOE epsilon 4. We also compared them with seven patients with probable AD. DESIGN: After grouping subjects according to diagnosis and genotype, brain function measures were compared among groups. SETTING: University medical center. PATIENTS: At risk subjects had mild memory complaints, normal cognitive performance, and at least two relatives with AD. Subjects with APOE epsilon 4 did not differ from those without APOE epsilon 4 in mean age at examination (56.4 vs 55.5 years) or in neuropsychological performance (mean Mini-Mental State Examination score, 28.8 vs 29.3). MAIN OUTCOME MEASURES: Cerebral glucose metabolism was measured using positron emission tomography and fludeoxyglucose F 18. RESULTS: Parietal metabolism was significantly lower and left-right parietal asymmetry was significantly higher in at-risk subjects with APOE epsilon 4 compared with those without APOE epsilon 4. Patients with dementia had significantly lower parietal metabolism than did at-risk subjects with APOE epsilon 4. CONCLUSIONS: These results suggest that the inheritance of APOE epsilon 4 is associated with reduced cerebral parietal metabolism and increased asymmetry in non-demented relatives at risk for probable AD. Longitudinal study will determine if glucose metabolic measures provide a means to monitor experimental treatment responses during the early phases of the disorder.

Adult↗