Studies of Microbial Populations Artificially Localized in Vivo. I. Multiplication of Bacteria and Distribution of Drugs in Agar Loci 1

Werner, Charles A.; Knight, Vernon; McDermott, Walsh; Adams, Carol; DuBois, Rebeckah

doi:10.1172/jci102943

Cited by 16 publications

(15 citation statements)

References 6 publications

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“…Although a fibrin-leucocyte membrane formed about disks in approximately 24 hours, disk integrity was maintained, and leucocytes did not gain access to the agar. The latter findings confirm those of Werner, Knight, and McDermott (13).…”

Section: Resultssupporting

confidence: 89%

“…The techniques employed were patterned after those described by Werner, Knight, and McDermott (13). A teria-containing suspension from reaching the wall of the mold; the desired result was easily achieved because the 0.2 ml.…”

Section: Methodsmentioning

confidence: 99%

“…Although there is also evidence that when it is administered in the initial phase of experimental infections due to susceptible organisms penicillin is bactericidal (10,11), elucidation of its effect in vivo has been complicated by the participation of host cellular defenses which, under certain circumstances, may in themselves be extremely efficient (12). The recent description by Werner, Knight, and McDermott of a method by which bacteria can be localized in tivo in a cellfree milieu (13) suggested the means by which the effect of penicillin on group A streptococci, in the absence of leucocytes, could be assayed at various stages of growth in the peritoneal cavity of the rabbit.…”

mentioning

confidence: 99%

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Effect of Penicillin on Group a Streptococci in Vivo in the Absence of Leucocytes 1

Darnell¹,

Pesch²,

Glaser³

1955

J. Clin. Invest.

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Studies of penicillin action in vitro have indicated that, under given experimental conditions, the antibiotic is bactericidal rather than bacteriostatic (1-5), and it has been further demonstrated that it exerts its bactericidal effect on susceptible microorganisms only when they are actively metabolizing, a state indicated usually by rapid growth (6-9). Although there is also evidence that when it is administered in the initial phase of experimental infections due to susceptible organisms penicillin is bactericidal (10, 11), elucidation of its effect in vivo has been complicated by the participation of host cellular defenses which, under certain circumstances, may in themselves be extremely efficient (12 tact with the hardened basal portion. Two ml. of plain agar were added as a cover to complete the disk, and after the agar had hardened, the aluminum ring was removed.Preparation of inoculum. In the experiments herein described, a single strain of group A streptococcus, S23 Burbank, was employed. The organism was obtained originally from Dr. Maclyn McCarty of the Rockefeller Institute for Medical Research; identical stock samples were prepared by concentrating cultures of the organism, grown in beef infusion broth with 10 per cent sheep serum and 0.5 per cent dextrose (BIB-SD), and storing 0.2 ml.aliquots at -720 C. For the various studies described, 5 ml. of BIB-SD at pH 7.8 were inoculated with 2 loopfuls of rapidly thawed stock culture and incubated at 370 C for 16 hours, after which 0.5 ml. was transferred to 5 ml. of fresh BIB-SD and incubated for 4 hours. Tenfold dilutions were made in tryptose phosphate broth, and 1.0 ml. of the 105 dilution was added to 9.0 ml. of melted agar at 400 C and thoroughly mixed. The 0.2 ml. portion of this mixture, employed in the disk, contained 350 to 400 chains.Implantation of disks. New Zealand rabbits, obtained from a single breeder, and free of demonstrable infection were used. The animals weighed 2.5 to 3.0 Kg., and were maintained under standard laboratory conditions. Preoperative preparation included careful depilatation of the abdomen over a rectangular area approximately 12 X 8 cm.; this area was then scrubbed vigorously with hexachloroprene soap (pHisoderm@). Laparotomy was performed under local anesthesia with 1 per cent procaine hydrochloride; aseptic technique was employed. A 6 to 8 cn. midline incision was made, and the disks placed so that contact between their surfaces was prevented. It was possible to insert as many as six disks on either side of the peritoneal cavity without difficulty. After implantation was completed, the peritoneum and muscle layers were closed with a continuous silk suture and the skin with Michele clips.Disks were removed through the same incision, and ground separately, without sand, in sterile mortars. In order to bring the final volume to 8.0 ml., 4.8 ml. of tryptose phosphate broth were added, and poured blood agar plates were made with 1 ml. aliquots of suitable tenfold dilutions of the ground disk suspensions for determination ...

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Section: Resultssupporting

confidence: 89%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Effect of Penicillin on Group a Streptococci in Vivo in the Absence of Leucocytes 1

Darnell¹,

Pesch²,

Glaser³

1955

J. Clin. Invest.

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“…Werner, Knight, and McDermott (18) showed that antibiotics were able to penetrate into "artificial abscesses" implanted into mice, thus reinforcing the idea that antibiotics did penetrate to the site where they were needed, but once there, were for some reason ineffective.…”

Section: Tmentioning

confidence: 92%

Interaction of Intraleukocytic Bacteria and Antibiotics

Mandell¹

1973

J. Clin. Invest.

198

110

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A B S T R A C T Bacteria that survive inside polymorphonuclear neutrophils (PMN) following phagocytosis are protected from the bactericidal action of most antibiotics. Two possible explanations are altered metabolism by intraleukocytic bacteria or failure of antibiotics to enter the phagosome. The oxygen consumption of intraleukocytic and extraleukocytic bacteria was measured as an index of bacterial metabolism. PMN respiration and bactericidal activity were suppressed with large doses of hydrocortisone and extraleukocytic bacterial oxygen consumption was abolished by the addition of lysostaphin. Intraleukocytic bacterial continued to consume oxygen suggesting that surviving ingested microorganisms are metabolically active. Neither penicillin (which cannot kill intraleukocytic bacteria) nor rifampin (which can kill intraleukocytic bacteria) was bactericidal for staphylococci at 5°C. Thus, rifampin is not uniquely able to kill "resting" bacteria. The total water space in the pellets was quantitated using tritium water and the extracellular water space was measured using Na2'5SO4. All penicillin associated with the cell pellet could be accounted for in extracellular water. Thus penicillin was completely excluded from the leukocytes. Rifampin was concentrated in the cell pellet 2.2 times when compared with the supernatant concentration.These studies suggest that a likely explanation for the survival of phagocytized bacteria in the presence of high concentrations of most antibiotics is the inability of the Dr. Mandell is the holder of a Research Career Development Award GM-49520 from the National Institute of General Medical Sciences.

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“…A metho(d was devised for localization of bacterial populatiosii in tilueniriche(d agar (liscs which were stuccessfullvy maintained in the peritoneal cavities of animial hosts for varying periods of time (12). As Six to eight 5-ml agar discs were introduced into the peritoneal cavity with care to ensure the contact of the flat surfaces of the agar discs with the peritoneum and to prevent-apposition of the discs with each other.…”

mentioning

confidence: 99%

Delivery of Antimicrobial Drugs Across Inflammatory Membrane in Rabbits

McCune¹

1960

J. Clin. Invest.

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Studies of Microbial Populations Artificially Localized in Vivo. I. Multiplication of Bacteria and Distribution of Drugs in Agar Loci 1

Cited by 16 publications

References 6 publications

Effect of Penicillin on Group a Streptococci in Vivo in the Absence of Leucocytes 1

Effect of Penicillin on Group a Streptococci in Vivo in the Absence of Leucocytes 1

Interaction of Intraleukocytic Bacteria and Antibiotics

Delivery of Antimicrobial Drugs Across Inflammatory Membrane in Rabbits

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