Preparation of a Glycoprotein Fraction From Pooled Human Plasma and Its Evaluation as a Substrate for the Assay of Clostridium Welchii (C. Perfringens) Neuraminidase

Fraser, Aileen; Smith, J. K.

doi:10.1099/00222615-8-2-235

Cited by 9 publications

(23 citation statements)

References 31 publications

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“…ThepH optimum for neuraminidases may vary considerably with different buffer systems and different substrates (Fraser and Smith, 1975;Rosenberg and Schengrund, 1976) but our standard assay with human glycoprotein substrate FVII with acetate buffer at pH 5.1 also gave good results with the B. fragilis enzyme (see the figure). The neuraminidase produced by certain species, e.g., V. cholerae, is calcium dependent but added Ca2+ is not required in our assays for the enzyme produced by clostridia (Fraser, 1978) or Bacteroides species (table 11).…”

Section: Discussionmentioning

confidence: 80%

“…Neuraminidases (EC 3.2.1.18) are produced by many species of aerobic and Anaerobic culture methods were those described by Fraser and Smith (1975). Cultures were prepared from lyophilised stock and maintained in CMB.…”

Section: Introductionmentioning

confidence: 99%

“…Human glycoprotein substrate FVII (Fraser and Smith, 1975) was used at a. final concentration of 140-1 70 pg N-acetyl neuraminic acid (NANA)/ml reaction mixture (Fraser, 1978).…”

Section: Introductionmentioning

confidence: 99%

“…Neuraminidase assays were performed in a range of sodium-acetate buffers as described by Fraser and Smith (1975). The reaction mixture of 1.0-ml volume consisted of: 0.1 ml of the bulk cell extract of B. fragilis NCTC9344 (see above) diluted 1 in 40 in distilled water; 0.65 ml of the appropriate acetate buffer; and 0.25 ml of substrate FVII diluted in distilled water to give a final concentration in the reaction mixture of 160 pg NANA/ml.…”

Section: Introductionmentioning

confidence: 99%

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Neuraminidase production by Bacteroidaceae

Fraser¹,

Brown²

1981

Journal of Medical Microbiology

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SUMMARY. The production of neuraminidase (EC 3.2.1.18) by 77 strains of Bacteroidaceae was investigated by techniques previously used to study neuraminidase production by clostridia. Conditions for culture and assay of Bacteroides fragilis neuraminidase were characterised. The enzyme is predominantly cell associated; it is not calcium dependent and the pH optimum for its production is c. 4-5.Most neuraminidase-positive Bacteroides strains produced the enzyme well in the test media but a few strains failed to'produce it consistently in one or other of the media. Because of these occasional variations, strains were grown and tested in at least two media before being defined as neuraminidase negative.Within the B. fragilis group of species, B. fragilis, B. vulgatus, B. distasonis, B. ovatus, B. thetaiotaomicron and B. variabilis were neuraminidase positive while B. eggerthii, B. uniformis and B. splanchnicus were negative. Two subspecies of B. melaninogenicus (ss. melaninogenicus and ss. levii) were positive but the other (ss. intermedius) was negative. Strains of B. oralis and B. bivius produced the enzyme while B. ruminicola, B. disiens, B. asaccharolyticus and B. corrodens did not. The microaerophilic B. ochraceus were also positive. None of the Fusobacterium or Leptotrichia species tested produced neuraminidase.Our results for neuraminidase production are consistent for all strains of each species examined and we suggest that tests for neuraminidase production would be a valuable addition to biochemical tests currently used in taxonomic studies of the Bacteroidaceae.

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Section: Discussionmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 99%

“…Human glycoprotein substrate FVII (Fraser and Smith, 1975) was used at a. final concentration of 140-1 70 pg N-acetyl neuraminic acid (NANA)/ml reaction mixture (Fraser, 1978).…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Neuraminidase production by Bacteroidaceae

Fraser¹,

Brown²

1981

Journal of Medical Microbiology

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“…These workers confirmed that the reference British heat-resistant food-poisoning strains examined by them did not produce neuraminidase, but they showed that one of five American food-poisoning strains of C. welchii that could not otherwise be differentiated from these strains was neuraminidase positive. American foodpoisoning strains generally include many that are haemolytic or heat-sensitive or both; the majority of American food-poisoning strains examined by Moss et al (1967) did produce neuraminidase.In the present study, the more sensitive neuraminidase-assay procedure described by Fraser and Smith (1975) was used for a further investigation of neuraminidase production in a range of heat-resistant and heat-sensitive British food-poisoning strains of C. welchii. …”

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confidence: 99%

The Production of Neuraminidase by Food-Poisoning Strains of Clostridium Welchii (C. Perfringens)

Fraser

Collee

1975

Journal of Medical Microbiology

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THERE is confusion in the literature regarding the production of neuraminidase (EC 3.2.1.18) by food-poisoning strains of Clostridium welchii (C. perfringens) type A ; this partly reflects the changing definition of these food-poisoning strains. Many episodes of food poisoning caused by C. welchii result from ingestion of strains that produce very heat-resistant spores, i.e., spores that survive boiling in faecal suspension for 1 h or more (Hobbs et al., 1953). These strains, which are also non-haemolytic, have been considered as " typical food-poisoning" strains. Although such strains are most likely to survive cooking procedures, it is now well recognised that food poisoning can also be caused by strains that produce relatively heat-sensitive spores, and these strains may be haemolytic or non-haemolytic (Hauschild and Thatcher, 1967 ;Sutton and Hobbs, 1968;Hobbs, 1969). Food-poisoning strains of C. welchii are serotyped with two sets of antisera, one for the typical heat-resistant strains-Hobbs' types 1-24-and the other for the so-called heat-sensitive strains-types i-xviii (Hobbs et al., 1973).Collee was unable to show neuraminidase production in cultures of four British typical food-poisoning strains but, because culture supernates of two strains produced a myxovirusreceptor-inactivating effect on red cells (Collee, 1965a), he concluded that " more extensive investigation is required before it can be claimed that neuraminidase is never produced by typical food-poisoning strains " (Collee, 19653).Moss, Schekter and Cherry (1963, working in the United States, surveyed many strains of C. welchii for neuraminidase activity. These workers confirmed that the reference British heat-resistant food-poisoning strains examined by them did not produce neuraminidase, but they showed that one of five American food-poisoning strains of C. welchii that could not otherwise be differentiated from these strains was neuraminidase positive. American foodpoisoning strains generally include many that are haemolytic or heat-sensitive or both; the majority of American food-poisoning strains examined by Moss et al. (1967) did produce neuraminidase.In the present study, the more sensitive neuraminidase-assay procedure described by Fraser and Smith (1975) was used for a further investigation of neuraminidase production in a range of heat-resistant and heat-sensitive British food-poisoning strains of C. welchii. Strains of C. welchii.Four classical haemolytic strains of C. welchii type A were used. Strain L2Ab was described in the previous paper (Fraser and Smith, 1975); strain L3A was also obtained from Professor C. L. Oakley, School of Medicine, University of Leeds. Strains C1 and 032 were laboratory strains originally isolated from wound infections in Edinburgh.Typical non-haemolytic, heat-resistant, food-poisoning strains of C. welchii type A of Hobbs' types 1-4 (Nos. 8359, 8238, 8239 and 8247 respectively) were obtained from the National Collection of Type Cultures, Colindale, London; strains of Hobbs' types 5-24 were kindly provi...

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