agar and Pleural-Effusion

This study was aimed to evaluate the performance of blood agar for the growth of mycobacteria from clinical specimens sent to Mycobacteriology Laboratory of Samsun Chest Diseases Hospital. One hundred fifty six clinical specimens including 123 sputum, 28 bronchoalveolar lavage (BAL) and 5 pleural fluid specimens were inoculated in Löwenstein-Jensen (LJ), BACTEC MGIT 960 system (Becton Dickinson, USA) and blood agar following decontamination process. The specimens were also simultaneously examined for the presence of acid-fast bacilli (AFB). Thirty five mycobacteria strains (33 Mycobacterium tuberculosis and 2 atypical mycobacteria) grew in blood agar, 38 (36 M.tuberculosis and 2 atypical mycobacteria) in LJ media and 46 (44 M.tuberculosis and 2 atypical mycobacteria) in BACTEC MGIT 960 system. Among 29 AFB negative specimens, 20 revealed growth in both blood agar and LJ medium and 27 in MGIT system. AFB positive 20 samples yielded growth in 15 samples in blood agar, 18 in LJ medium and 19 in MGIT system. Among the total of 156 samples, contamination was observed in 15 (9.6%) samples in blood agar, 16 (10.2%) in LJ medium and 18 (11.5%) in MGIT system. Growth time was 5-35 days (mean 18 ± 7.4), 11-35 days (mean 19 ± 5.9) and 5-15 days (mean 10 ± 2.4) for blood agar, LJ medium and BACTEC MGIT 960 system, respectively. The three samples which revealed contamination in BACTEC MGIT 960 system, grew successfully in both blood agar and LJ medium without contamination. In one sample, growth was observed only in LJ medium but neither in blood agar nor BACTEC MGIT 960 system. However, in another sample, growth was observed only in blood agar while no growth was detected in LJ or BACTEC MGIT 960 system. Six samples yielded mycobacteria only in BACTEC MGIT 960 system. These results indicated that simultaneous use of one liquid and one solid medium to grow mycobacteria from the clinical samples seemed to be complementary. Blood agar was a promising choice since it was found to be as effective as LJ medium for the growth of mycobacteria, however, this issue needs to be further evaluated in a multicentre study with a larger specimen collection.

Topics: Agar; Blood; Bronchoalveolar Lavage Fluid; Culture Media; Humans; Mycobacterium Infections, Nontuberculous; Mycobacterium tuberculosis; Nontuberculous Mycobacteria; Pleural Effusion; Sputum; Tuberculosis

Prior attempts to create an animal model of empyema by direct inoculation of bacteria alone into the pleural space have been unsuccessful. The animals either died of overwhelming sepsis or cleared the infection from the pleural space without development of an empyema. We hypothesized that injection of bacteria with a nutrient agar into the pleural space would allow the bacteria to remain in the pleural space for an extended time period, permitting an empyema to develop. The bacterium Pasteurella multocida in brain heart infusion (BHI) agar was injected into the right hemithorax of 12 New Zealand white male rabbits. Our preliminary studies showed that the animals died in less than 7 days if they were not given parenteral antibiotics. For this reason, the rabbits were given penicillin, 200,000 U, IM, every 24 h starting 24 h after bacterial injection. Pleural fluid was sampled by thoracentesis at 12, 24, 48, 72, and 96 h after bacterial injection. Pleural fluid pH, glucose, lactate dehydrogenase (LDH), leukocyte count, and Gram's stain and culture (in one half of the animals) were obtained at each time point. Pleural biopsy specimens were obtained at autopsy after 96 h. The mean pleural fluid pH reached a nadir of 7.01 at 24 h and remained less than 7.1 throughout the experiment. The mean pleural fluid glucose level reached a nadir of 10 mg/dL at 24 h. The mean pleural fluid LDH peaked at 21,000 IU/L at 24 h and the mean pleural fluid leukocyte count peaked at 12 h with a value of 67,000 cells per cubic millimeter. Gram's stains revealed organisms and cultures were positive for growth in all animals at 12 and 24 h. Some animals had positive Gram's stains and growth on cultures up to 72 h after bacterial injection. At autopsy, all rabbits injected with bacteria had gross pus in the right pleural space and had developed a thick pleural peel. Microscopic specimens of the pleura revealed large numbers of leukocytes (primarily polymorphonuclear lymphocytes) with invasion of the adjacent lung and chest wall. In conclusion, this model more closely mimics the empyema that occurs in humans, relative to previous animal models. This model appears appropriate for additional randomized studies in which different methods for the treatment of empyema can be evaluated.

Topics: Agar; Animals; Biopsy; Culture Media; Disease Models, Animal; Empyema, Pleural; Glucose; Hydrogen-Ion Concentration; Injections, Intramuscular; L-Lactate Dehydrogenase; Leukocyte Count; Lung; Male; Neutrophils; Pasteurella Infections; Pasteurella multocida; Penicillins; Pleura; Pleural Effusion; Rabbits; Thorax

Topics: Agar; Antineoplastic Agents; Ascites; Cell Survival; Cells, Cultured; Clone Cells; Colony-Forming Units Assay; Doxorubicin; Drug Resistance; Female; Humans; Ovarian Neoplasms; Pleural Effusion; Tumor Stem Cell Assay

Topics: Agar; Biopsy; Breast Neoplasms; Cell Survival; Clone Cells; Culture Techniques; Female; Humans; Lung Neoplasms; Male; Melanoma; Methylcellulose; Microscopy, Electron; Neoplasms; Pleural Effusion

Two cases of Legionnaires' disease were diagnosed by direct isolation of the organism, from pleural fluid obtained before death in one case and lung tissue obtained after death in both cases. The organisms were recovered on a commercially prepared, enriched chocolate agar (Gibco, Madison, Wisconsin). Subcultures grew on commercially prepared, enriched chocolate agar (Baltimore Biological Laboratories, Cockeysville, Maryland) and on in-house enriched chocolate agar prepared with GC Medium Base (Difco, Detroit, Michigan). No growth was obtained on enriched chocolate agar prepared with trypticase soy agar. The organisms were poorly visualized in Gram-stained sections of formalin-fixed lung tissue. In Giemsa-stained sections poorly stained intracellular and extracellular slender rods were seen. However, with a silver impregnation stain, either a modified Dieterle or a modified Warthin-Starry procedure, many large, blunt-ended rods were seen. Smears prepared from minced formalin-fixed lung tissue and stained with a fluorescent antibody conjugate contained large numbers of well-stained organisms.

Topics: Adult; Agar; Aged; Bacteria; Female; Humans; Legionnaires' Disease; Lung; Male; Pleural Effusion; Staining and Labeling

Fourteen breast cancer lines (8 human, 5 rat, and 1 mouse) have been studied in terms of their ability to form multicellular tumor spheroids (MTS) with the agar-base method. Only 8 of the lines formed MTS in contrast to a 100% efficiency in a series of 11 varied tumors reported in the initial studies with this method. We have compared the lines that do and do not form MTS in terms of a variety of characteristics (e.g., estrogen receptors, time in serial passage, growth in nude mice, etc.), and only one characteristic, the source of the original tumor cells, was predictive of MTS-forming ability. All 8 of the breast cancer lines (and the original 11 lines) that formed MTS had been obtained from solid growths (primaries or metastases), while the 6 breast cancer lines that did not form MTS were all derived from pleural effusions. Similarly, artificial selection for an ascites variant of the MTS-forming rat 13762 adenocarcinoma line produced the 13762-A line, which could no longer form MTS. These results suggest that breast cancer cells derived from pleural effusions are genetically different from the bulk of the tumor cells in solid breast cancer samples, that they are unable to grow in true solid form, and that these differences persist in spite of prolonged propagation in tissue culture.

Topics: Adaptation, Physiological; Agar; Animals; Breast Neoplasms; Cell Aggregation; Cell Line; Culture Techniques; Female; Humans; Mammary Neoplasms, Experimental; Mice; Neoplasm Metastasis; Neoplasms, Experimental; Pleural Effusion; Rats

Topics: Agar; Electrophoresis; Gels; Humans; Isoenzymes; L-Lactate Dehydrogenase; Pleural Effusion

Topics: Adsorption; Agar; Animals; Antigens; Ascitic Fluid; Barium Sulfate; C-Reactive Protein; Calcium; Chemical Precipitation; Edetic Acid; Filtration; Gels; Humans; Immune Sera; Immunoelectrophoresis; Methods; Pleural Effusion; Polysaccharides; Polysaccharides, Bacterial; Quaternary Ammonium Compounds; Rabbits; Streptococcus pneumoniae

Topics: Agar; Anemia, Hemolytic; Blood; Clinical Enzyme Tests; Electrophoresis; Histocytochemistry; Humans; In Vitro Techniques; Kidney Diseases; L-Lactate Dehydrogenase; Liver Diseases; Myocardial Infarction; Neoplasms; Pleural Effusion