Vitamin B6-dependent Streptococcus mimicking fungi in a patient with endocarditis.

A patient was referred to our hospital with a tentative diagnosis of fungal endocarditis based upon clinical symptoms, suggestive travel history, and microscopic visualization in blood cultures of gram-negative bulbous filaments that appeared to be fungal elements. Subcultures of the blood culture bottles were unsuccessful on all media with the exception of blood agar plates, which had been cross-streaked with Staphylococcus aureus. These plates grew vitamin B6-dependent streptococci. This nutritionally variant organism was determined by biochemical tests to be Streptococcus mitis (mitior). It had a penicillin MIC and MBC of 0.015 micrograms/mL and 0.03 micrograms/mL, respectively and streptomycin MIC and MBC of 0.78 micrograms/mL and 1.56 micrograms/mL, respectively. The patient was treated with these two agents and recovered. We stress the importance of suspecting vitamin B6-dependent streptococci, even when gram stains may suggest presence of other microorganisms.

Pathogenesis, lethality, and immunizing effect of experimental cutaneous cryptococcosis.

Mice were subcutaneously inoculated with small numbers of virulent Cryptococcus neoformans and divided into groups. Numbers of viable yeasts at the site were estimated at weekly intervals for 5 weeks on the basis of cultures of minced tissue excised from sacrificed animals. Organisms multiplied at the site for at least 4 weeks and were still detectable after the 5th week, although in reduced numbers. Agglutinins appeared within a week, but these antibodies were not detectable during the 2nd through the 5th week. Cryptococcal polysaccharide began to appear in the sera at 3 weeks, persisting through the duration of 5 weeks. All animals appeared healthy, but a few sickened after many months and died of systemic cryptococcosis. All of these events were observed in many separate experiments. The immunizing capacity of a cutaneous lesion was tested by challenging some of the above animals with viable C. neoformans after various intervals of time, either subcutaneously at a site distant from that of the vaccination or intravenously. Although we were unable to demonstrate reduced multiplication of yeasts in the brains, lungs, and spleens of intravenously challenged animals, it was possible to show that multiplication was inhibited at the site of subcutaneous challenge. It was noted also that vaccinated animals lived longer after lethal intravenous challenge than did nonvaccinated animals. The latter protection was observed, however, only when challenge followed vaccination by 3 weeks or longer, and it was effective only against a relatively low challenge dose. Mice were protected against a higher dose if they had previously received killed cryptococci, alternating subcutaneous and intraperitoneal inoculations, one of which contained a microbial adjuvant. No protection was observed in animals that were subcutaneously vaccinated with inert materials such as chitin, latex spheres, or even cryptococcal cell walls themselves.

Resistance to challenge and macrophage activity in mice previously vaccinated with formalin-killed Cryptococcus neoformans.

Mice were vaccinated with formalin-killed Cryptococcus neoformans either intranasally, intravenously, or intraperitoneally. Resistance to lethal challenge and in vitro phagocytic activity of alveolar and peritoneal macrophages was studied. Increased resistance to lethal challenge was seen following vaccination by each of the three routes but increased phagocytic activity was seen only in alveolar macrophages from mice vaccinated intranasally.

Pulmonary cell response in mice following intranasal instillation with Cryptococcus neoformans.

Cryptococcus neoformans was instilled intranasally into mice which were sacrificed to determine the fate of instilled cryptococci and the pulmonary cellular response. During the first 12 h, instilled cryptococci decreased significantly in numbers and did not disseminate to extrapulmonary sites. There was a transient increase in neutrophils which may have been due to an influx of bacteria which accompanied the process of intranasal instillation.

Bacterial contamination in irreversible hydrocolloid impression material and gingival retraction cord.

This study tested two dental materials in factory-sealed containers for the presence of bacteria that may be a source of infection. Twenty samples of two dental materials found to have contamination in a pilot study were taken from unopened containers using a sterile technique. The samples were inoculated onto chocolate agar plates and into thioglycolate broths with appropriate controls. Plates were examined, colonies were enumerated, Gram stained, and identified. The resulting contamination frequencies were compared for statistical significance using Fisher's exact test. Organisms were isolated from 10% of the negative inoculation control agar plates, while none of the control broths showed contamination. The alginate (irreversible hydrocolloid) showed contamination in 50% of the plates and in 65% of the broths (p less than 0.05). The retraction cord had a small sample size and yielded organisms on 5% of the sample plates and in 20% of the thioglycolate broths (p greater than 0.05).

Comparison of media and techniques for detection of group A streptococci in throat swab specimens.

Detection of group A streptococci in primary throat cultures was compared by using aerobic and anerobic incubation with selective nonselective media. Sheep blood agar plates incubated anaerobically detected 98% of the group A streptococci, whereas aerobically incubated blood agar plates which had been stabbed at the time of inoculation detected only 63%. Blood agar plates containing sulfamethoxazole and trimethoprim (23.75 and 1.25 mirograms per ml, respectively) detected only 70% of group A streptocci when incubated aerobically and 84% when incubated anaerobically.

Capsule size of Cryptococcus neoformans: control and relationship to virulence.

Capsule size of five isolates of Cryptococcus neoformans was controlled by cultivation in media containing varying amounts of sugar. High concentrations of sugar (e.g., 16%) suppressed encapsulation whereas low concentrations (e.g., 1%) allowed maximal encapsulation. Suppression of capsule size was attributed at least in part to the increased osmolarity of the medium because a medium with low sugar concentration but having high osmolarity (by virtue of added sodium chloride) also produced cells having small capsules. The extent of control was more marked with certain of the isolates than with others. Mice were intravenously inoculated with cells of a single isolate cultivated so as to have either small or large capsules, and virulence was measured by comparing death rates. Results indicate that virulence after such an inoculation is a constant characteristic of an isolate and is not affected by size of the capsule of the cells in the inoculum.

Microbial contamination in two antimicrobial and four control brands of alginate impression material.

Previous investigations have revealed commercial alginate impression material to be contaminated with viable microorganisms. Some manufacturers are now producing alginate materials that contain antimicrobial agents. The purpose of this study was to test and compare two antimicrobial and four control brands without antimicrobial agents of commercial dental alginate impression material for the presence of viable microorganisms. Forty-eight or 96 measured samples of each brand were taken from previously unopened containers using a sterile technique. The samples were placed on chocolate agar plates and in thioglycolate broth tubes and were incubated along with appropriate parallel controls. After incubation, colonies were enumerated, gram-stained, and identified using standard microbiologic methods. The two antimicrobial brands contained viable organisms in 12.5% of the samples incubated on agar media and also contained such organisms from 0% to 16.7% of the samples incubated in thioglycolate media. The four control brands contained viable organisms in from 29.2% to 100% of the samples incubated on agar media and also contained these organisms in from 25% to 79.2% of the samples incubated in thioglycolate media. There was a statistically significant difference (p less than 0.05) in contamination frequencies among some brands. Contamination frequencies of the top and middle portions of the containers did not differ significantly. The concentration of organisms in contaminated samples was 2.8 colony-formed units (CFUs) per gram for the antimicrobial alginates, and from 9 to 161.1 CFUs per gram for the control brands.(ABSTRACT TRUNCATED AT 250 WORDS)

Microbial contamination in four brands of irreversible hydrocolloid impression materials.

A previous investigation showed unopened irreversible hydrocolloid impression material to be contaminated with viable microorganisms. This study tested and compared four brands of commercial irreversible hydrocolloid impression material in factory-sealed containers for the presence of viable microorganisms. Twenty-four measured samples of each brand were taken from previously unopened containers using a sterile technique. The samples were placed on chocolate agar plates or in thioglycolate broth tubes and were incubated along with appropriate parallel controls. After incubation, colonies were enumerated, stained with Gram's stain and identified using standard microbiologic methods. The four brands contained viable organisms in 50% to 100% of the samples incubated of agar media, and in 12% to 67% of the samples incubated in thioglycolate media. Samples from the top and middle portions of the containers had approximately equal contamination frequencies. The concentration of organisms varied from 12 to 82 colony-formed units per gram of contaminated sample. Most organisms isolated were common environmental contaminants. These samples contained viable microorganisms which, during routine use, may present a hazard to immunocompromised patients.

Intraoral vesicles occurring after alginate impressions.

In 47 of 227 dental students intraoral vesicles developed after multiple alginate impressions. The lesions were generally solitary and clear, and appeared within 24 to 48 hours after the impression. They were most frequently located inside the vermilion border of the lips and resolved spontaneously in 2 to 5 days. The purpose of this study was to determine the cause of these reactions. Histopathologically one lesion was suggestive of a contact allergy. Cutaneous patch tests, which proved negative, were performed on 14 students to determine whether an allergy to the alginate flavoring existed. The surface of three lesions were cultured and the organisms identified. Contamination studies were carried out on seven unopened containers of the alginate powder and resulted in the isolation of some organisms similar to the mucosal cultures; however, no relationship can be proved. These findings indicate that the cause of the vesicles remains unknown, and further studies are necessary to establish the cause.