The Demonstration of Capsules About Hemolytic Streptococci with India Ink or Azo Blue

Butt, Emma; Bonynge, C. W.; Joyce, R. L.

doi:10.1093/infdis/58.1.5

Cited by 52 publications

(11 citation statements)

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“…Strain $23G, as already stated, produces no detectable M protein even in the absence of trypsin. Capsular studies performed with India ink (23) showed that the trypsin did not visibly alter the capsules of any of the four strains (see Tables I and II, and Fig. 1).…”

Section: Methodsmentioning

confidence: 90%

Studies on the Pathogenicity of Group a Streptococci

Foley

Wood

1959

The Journal of Experimental Medicine

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There is abundant experimental evidence that both the M protein and the capsular gel of Group A streptococci play significant roles in pathogenicity (1-7). That they enhance virulence by interfering with surface phagocytosis is indicated by the observations described in the preceding paper (8).Despite the general agreement that both the capsules and the M protein of Group A streptococci are antiphagocytic (9, 10), their precise relationship to virulence has remained obscure. Numerous strains have been described, for example, which appear to produce abundant amounts of M protein and yet are relatively avirulent for mice (1,(11)(12)(13)(14). Others possessing large capsules have, likewise, been found to be lacking in virulence (3, 15), as illustrated by the glossy variant of the $23 strain referred to in the accompanying paper (8). Indeed, it has been shown that streptococci which produce both capsules and M substance also may lack pathogenicity for mice. The T14 strain, used in this and the previous study, is just such an organism (8).In the case of pneumococcal and Friediander's bacillus infections, the situation is less complex (16), inasmuch as these organisms are protected against leucocytes by a single antiphagocytic factor (i.e., a carbohydrate capsule) (9, 17, 18). As has been emphasized elsewhere, however, the quantitative aspects of capsule formation have a direct bearing upon the virulence of organisms such as Type III Pneumococcus (19,20). This fact, taken in conjunction with the knowledge that a double antiphagocytic mechanism is involved in Group A streptococcal infections, has prompted us to restudy the quantitative and combined effects of the M protein and the hyaluronate capsule upon phagocytosis. The results appear to clarify the relationship of both factors to virulence.

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Section: Methodsmentioning

confidence: 90%

Studies on the Pathogenicity of Group a Streptococci

Foley

Wood

1959

The Journal of Experimental Medicine

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“…A sample of each strain grown on blood agar plates was examined for presence of capsule by the india ink method described by Butt et al (1938). Methylene blue was used as a counter stain and slides were examined by phase contrast microscopy.…”

Section: India Inkmentioning

confidence: 99%

Encapsulation of coagulase-negative staphylococci of bovine origin

Matthews¹,

Oliver²,

Guidry³

1991

J Appl Microbiol

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1991. Capsule expression was assessed in six coagulase-negative staphylococcal strains in serum-soft agar and by india ink and electron microscopy. Classification of strains as encapsulated by serum-soft agar and india ink methods differed. Staphylococcus chromogenes, Staph. hyicus, and Staph. simulans grew as diffuse colonies in serum-soft agar and unstained halos were detected in india ink preparations. Staphylococcus hominis and Staph. simulans grew as diffuse colonies in serum-soft agar but no unstained halo was seen in india ink preparations. Staphylococcus hyicus was the only strain that gave negative results with serum-soft agar and india ink assays. Conventional electron microscopy revealed the presence of capsular polysaccharides on the cell surface of Staph. chromogenes, Staph. hominis and Staph. hyicus. Conventional electron microscopic technique used to examine the surface of cells was detrimental to capsule structure. During dehydration the capsule collapsed and appeared as electron dense aggregates at the surface of cells. T o confirm results of conventional electron microscopy and to visualize clearly the cell surface, encapsulated Staph. hyicus and unencapsulated Staph. simulans were observed after freeze-fracture and etching by scanning electron microscopy. The fibrous nature of capsular polysaccharides surrounding cells of Staph. hyicus were distinct and confirmed observation by conventional electron microscopy. A rapid transmission electron microscopic technique is described also for observation of capsule. Results of the rapid T E M method agreed with conventional T E M and SEM. The finding that coagulase-negative staphylococci isolated from bovine milk are capable of capsule production may be important when investigating pathogenicity of these micro-organisms.

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“…These isolates, along with the strains MS11 and F62 which have been serially passaged in this laboratory for over 4 yr, were incubated at 35°C in 5% CO2. Cultures were also frozen at -70°C in Trypticase Soy Broth (BBL) containing 40% (vol/vol) glycerol to maintain stocks.Capsule stains that were used as originally described include the following: India ink wet mount (12); India ink smear and stain (13,14); Hiss copper sulfate (15); Hiss potassium carbonate (15); sodium caseinate-methyl violet (16); Wright's stain (17); eosin-serum (18); flagella-capsule stain (19); Congo red-methylene blue (20); Alcian blue negative stain (21); Moller's negative stain (22); and Gram capsule stain (23). The Alcian blue positive capsule stain (24) was modified by conversion of bound Alcian blue to Monastral fast blue (25,26).…”

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The capsule of the gonococcus.

James¹,

Swanson²

1977

The Journal of Experimental Medicine

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The presence or absence of capsules on Neisseria gonorrhoeae has been argued for many years. In 1921, Israeli (1) summarized opinions on encapsulation of gonococci as follows: "Some hold that it does possess a capsule, some question its existence, and some flatly deny that this organism has a capsule." In the intervening years, the existence of a gonococcal capsule has had both proponents (1-7) and opponents (8-11) but no definite resolution of the question. The present study was undertaken to try to clarify the controversy by utilizing numerous staining techniques for light microscopic evaluation of the presence or absence of capsules in several recently isolated as well as long-term serially passaged strains of gonococci. Our findings confirm the presence of capsules on Neisseria gonorrhoeae. Materials and MethodsClinical isolates ofN. gonorrhoeae were obtained as cultures on Thayer-Martin agar from the Salt Lake City County Health Department. The organisms were subsequently passaged on typing agar (GC agar base formulation of Baltimore Biological Laboratory (BBL) modified to contain half the recommended amount of Trypticase Peptone) plus 1% IsoVitaleX (BBL) and were identified as gonococci by Gram stain, oxidase reaction, and sugar oxidation. These isolates, along with the strains MS11 and F62 which have been serially passaged in this laboratory for over 4 yr, were incubated at 35°C in 5% CO2. Cultures were also frozen at -70°C in Trypticase Soy Broth (BBL) containing 40% (vol/vol) glycerol to maintain stocks.Capsule stains that were used as originally described include the following: India ink wet mount (12); India ink smear and stain (13,14); Hiss copper sulfate (15) (25,26). Both Pelikan (Gunther Wagner) and Higgins (Faber-Castell) India inks were used in India ink wet mounts and both were satisfactory.Several types of shearing forces were used to remove capsules. These include ejecting suspensions of gonococci through a 25-gauge needle, shaking suspensions of organisms with glass beads, and sonication. The affect of these treatments was assessed in India ink wet mounts (12) or by Congo red negative staining (20). ResultsIn all 16 strains of gonococci isolated from clinical specimens for this study and in the two multiply passaged laboratory strains, capsules are demonstrable. Of

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The Demonstration of Capsules About Hemolytic Streptococci with India Ink or Azo Blue

Cited by 52 publications

References 0 publications

Studies on the Pathogenicity of Group a Streptococci

Studies on the Pathogenicity of Group a Streptococci

Encapsulation of coagulase-negative staphylococci of bovine origin

The capsule of the gonococcus.

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