Preparation and testing of a polyvalent conjugate for direct fluorescent-antibody detection ofLegionella pneumophila

Thomason, Berenice M.; Harris, P P; Lewallen, Karen R.; McKinney, Roger M.

doi:10.1007/bf02602876

Cited by 22 publications

(14 citation statements)

References 7 publications

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“…the smears were fued with Kukpatrick fixative (60% ethanol, 30% chloroform, 10% Formalin), stained with the globulin, and counter-stained with a fluorescent anti-rabbit globulin (GIBCO Diagnostics, Grand Island, New York), as described by Goldman (8). After rinsing in phosphate-buffered saline (pH 8.0) and distilled water, the smears were air-dried and mounted to 0.5 M carbonate-buffered (pH 9.0) glycerin (21). They were observed with a Zeiss fluorescent microscope (X40 objective) equipped with BG 12 excitation and 50-barrier filters.…”

Section: Bacterialmentioning

confidence: 99%

Pseudomonas pseudoalcaligenes subsp. citrulli subsp. nov.

Schaad

Sowell

Goth

et al. 1978

International Journal of Systematic Bacteriology

119

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Ten nonfluorescent Pseudomonas strains isolated from water-soaked lesions on cotyledons of plants of five Citrullus lanatus (watermelon) plant introductions were characterized and compared phenotypically with 22 other pseudomonads. The strains were distinguished phenotypically from other known plant pathogenic pseudomonads. The watermelon bacterium was aerobic. Cells were rod-shaped, gram negative, and motile by means of a single polar flagellum. They were nonfluorescent and grew at 41°C but not at 4°C. Oxidase production and the 2-ketogluconate reaction were positive. The 10 strains utilized p-alanine, rA-leucine, D-serine, n-propanal, ethanol, ethanolamine, citrate, and fructose for growth. No growth occurred with sucrose or glucose. Their deoxyribonucleic acid base composition was 66 1 mol% guanine plus cytosine. The bacterium is phenotypically similar to P. pseudoalcaligenes but differs from it in being pathogenic to watermelon, Cucumis melo (cantaloupe), Cucumis sativus (cucumber), and Cucurbita pep0 (squash). The name P. pseudoalcaligenes subsp. citrulli is proposed for the new subspecies, of which strain C-42 (= ATCC 29625) is the type strain.For several years a bacterial disease of watermelons, characteAed by water-soaked lesions of cotyledons, has been observed on certain plant introductions at the Regional Plant Introduction Station, Experiment, Ga. A disease with similar symptoms on watermelon seedlings was reported to be caused by Pseudomonas lachrymans (9), but we could not isolate a pathogenic fluorescent pseudomonad. Our preliminary investigations showed that the causal bacterium was similar to the unidentified nonfluorescent pseudomonad isolated by Webb and Goth (22) from plants of watermelon plant introductions 174103 and 174104. The purpose of this investigation was to identify the causal bacterium isolated from diseased watermelon seedlings and to compare it with several other fluorescent and nonfluorescent pseudomonads, including the strain of P. lachrymans isolated from watermelon (9). Isolation and cultural properties. Seeds from different watermelon plant introductions were sown in trays (30 by 50 cm) of soil in a greenhouse with a day/night .ternpertwe of approximately 30/21 "C. Cotyledons and the first true leaves showing greasy, dark green lesions were collected, and the surfaces were disinfested in a freshly prepared 0.5% solution of sodium hypochlorite for 3 min. After rinsing for 2 min in sterile distilled water, the tissue from the lesions was removed and cut into smaller pieces in 0.1 ml of sterile distilled water, using a sterile scalpel. The suspension was streaked onto King medium B (KB; 11) and incubated for 5 days at 27°C (colonies of nonpathogenic pseudomonads were usually present after 2 days). Colonies of the watermelon bacterium were removed from the agar with a transfer loop and purified by streaking onto plates of KB. Stock cultures were maintained on KB slants. The media used for determining cultural characteristics were KB, yeast extract-dextrose-CaC03 (22), and Difco...

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Section: Bacterialmentioning

confidence: 99%

Pseudomonas pseudoalcaligenes subsp. citrulli subsp. nov.

Schaad

Sowell

Goth

et al. 1978

International Journal of Systematic Bacteriology

119

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“…Application of immunofluorescence for presumptive mass screening and monitoring of water supplies offers a practical, and perhaps economic, alternative to conventional techniques. It has been field-tested for use in rapid detection of enteropathogenic E. coli (Cherry et al, 1960;Pugsley and Evison,1974;Abshire, 1976), Shigella flexneri and S. sonnei , and Salmonella typhi (Thomason and Wells, 1971) in fecal smears and for identification of typhoid carriers (Thomason and McWhorter, 1965). Using polyvalent FAs for salmonellae, Cherry et al (1972) screened enrichments of several surface waters and found 60% more positive samples than they could detect by conventional culture methods.…”

Section: Enterobacteriaceaementioning

confidence: 99%

The Immunofluorescence Approach in Microbial Ecology

Bohlool

Schmidt

1980

Advances in Microbial Ecology

165

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“…Antiserum and anti-rabbit globulin were diluted 1:4 and 1:16, as determined by results of a block test (12). After rinsing in phosphate-buffered saline (pH 8.0) and distilled water, the slides were air-dried and mounted in 0.5 M carbonate-buffered (pH 9.0) glycerin (41). Slides were observed with a Zeiss fluorescence microscope (40X objective) equipped with a BG 12 excitation and 50 barrier fdter.…”

Section: Serological Testsmentioning

confidence: 99%

Synonymy of Pseudomonas coronafaciens, Pseudomonas coronafaciens pathovar zeae, Pseudomonas coronafaciens subsp. atropurpurea, and Pseudomonas striafaciens

Schaad

Cunfer

1979

International Journal of Systematic Bacteriology

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Eleven strains of Pseudomonas coronafaciens, four strains of P. coronafaciens pathovar zeae, six strains of P. coronafaciens subsp. atropurpurea, two strains of P. striafaciens, and one Pseudomonas sp. were compared by physiological, biochemical, serological, and pathological tests. All strains tested utilized sucrose, glucose, fructose, glycerol, and caprate as sole carbon sources, hydrolyzed Tween 80, reduced litmus milk, and grew at 0°C. All strains were arginine dihydrolase and oxidase negative and all strains failed to hydrolyze starch, reduce nitrate, or grow at 41OC. Cells of all strains were immunofluorescent positive when stained by the indirect immunofluorescence test with antiserum to cells of P. coronafaciens or P. coronafaciens subsp. atropurpurea. No differences among strains were detected in test tube agglutination tests. No significant differences were detected in the susceptibility of Avena sativa (oats), Secale cereale (rye), Triticum aestivum (wheat), Hordeum vulgare (barley), Bromus inermis (smooth bromegrass), B. japonicus (Japanese brome), B. secalinus (chess brome), B. tectorum (cheatgrass), Agropyron repens (quackgrass), Zea mays (maize), or Phleum pratense (timothy) to the different strains. All strains produced toxin in Wooley medium except P. striafaciens and strains of P. coronafaciens subsp. atropurpurea from Italian ryegrass. The present division of strains of P. coronafaciens based on minor differences in pathogenicity and symptomology is untenable .

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Preparation and testing of a polyvalent conjugate for direct fluorescent-antibody detection ofLegionella pneumophila

Cited by 22 publications

References 7 publications

Pseudomonas pseudoalcaligenes subsp. citrulli subsp. nov.

Pseudomonas pseudoalcaligenes subsp. citrulli subsp. nov.

The Immunofluorescence Approach in Microbial Ecology

Synonymy of Pseudomonas coronafaciens, Pseudomonas coronafaciens pathovar zeae, Pseudomonas coronafaciens subsp. atropurpurea, and Pseudomonas striafaciens

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