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PubMed1999

Effectiveness of CSE to counterstain particles and dead bacterial cells with permeabilised membranes: application to viability assessment in waters.

The CSE dye (Chemunex, Maisons-Alfort, France) was combined with an activity marker to improve bacterial activity assessment in natural waters. Its effectiveness to counterstain dead cells with permeabilised membranes was investigated on live and dead cells of a variety of strains from collections or isolated from the natural environment. Cells were killed by heat treatment. For all strains tested, the fluorescent dye showed an intense staining of killed cells having permeabilised membranes while no significant signal was detected when applied to live cells. Furthermore, the CSE dye had no toxicity on viable cells. Then, CSE was combined with the ChemChrome V6 dye (Chemunex) to assess the activity of bacterial cells in different waters. Both fluorescences were analysed simultaneously by solid-phase cytometry. The active cell counts were sometimes lower when both dyes were combined suggesting that CSE was able to counterstain cells having a residual esterase activity and compromised membranes. These cells were subtracted from the active cell counts determined with ChemChrome V6. In most samples, active cell counts were congruent with those determined by the direct viable count method.

Bacteria↗

PubMed1993

Inhibition of HIV-1 replication in H9 cells by nystatin-A compared with other antiviral agents.

Nystatin A was compared in vitro with amphotericin B, AZT, or foscarnet for their respective abilities to inhibit the replication of human immunodeficiency virus type 1 (HIV-1) in H9 cells. HIV-1-infected H9 cells were cultured for 7 days in the presence of each of these drugs, at various concentrations. Reverse transcriptase activity and p24 antigen production were quantitated. Untreated, HIV-1-infected H9 cells served as the control. Nystatin A inhibited viral replication most effectively at 10 micrograms/ml, a concentration that did not affect cell viability. Nystatin-A treatment inhibited RT activity by 85% and p24 production by 90%. These levels of inhibition were comparable to that mediated by amphotericin B, AZT, or foscarnet at 10, 25, and 50 micrograms/ml, respectively. Western blot analysis of the HIV-1-infected H9 cells treated with these drugs did not detect any expression of viral proteins. These findings were further corroborated by indirect immunofluorescence studies using monoclonal anti-gp120 FITC-conjugated antibodies and by polymerase chain reaction for proviral DNA analysis, using a 32P-labeled probe. These results suggest that Nystatin A merits attention as an antiviral drug for the treatment of HIV-1 infection. In vivo drug delivery by liposome encapsulation to overcome problems of bioavailability is currently under study.

Amphotericin B↗

PubMed2001

Antibiotic susceptibility of attached and free-floating Helicobacter pylori.

Helicobacter pylori are found attached to mucous cells of the human stomach or under the mucous layer. Models mimicking the in vivo situation may be more suitable for H. pylori MIC determinations than traditional agar dilution methods. Megraud et al. (Antimicrobial Agents and Chemotherapy 1991, 35, 869-72) developed a model for measuring the susceptibility of attached and free-floating H. pylori. We have modified this model so that free-floating and attached H. pylori are treated in a more similar manner, before and after incubation with antibiotic, and performed additional controls to ensure H. pylori and tissue culture cells are not detrimentally affected and maintain their viability during the course of the experiment. We found only 10% of plate-grown H. pylori were competent for attachment to HEp-2 cells; however, all progeny of attached bacteria remained adherent. Killing curves were performed using 0, 0.001, 0.01, 0.1 and 1 mg/L amoxycillin, and 0, 0.0025, 0.0075 and 0.01 mg/L clarithromycin. H. pylori divided at concentrations <or= 0.01 mg/L amoxycillin and <or= 0.0025 mg/L clarithromycin. Contrary to the previous study, using our modified method we found that HEp-2 adherent and freefloating H. pylori are equally susceptible to amoxycillin (strains 26695, CCUG18943, CCUG19104 and CCUG19110) and clarithromycin (strain 26695). Therefore, we find no evidence that attachment of H. pylori to eukaryotic cells increases their resistance to antibiotics compared with non-attached bacteria. Nonetheless, these results confirm confidence in traditional MIC studies when a comparison is made between susceptible and resistant strains.

Amoxicillin↗

PubMed2002

In vitro activity of therapeutic drugs against Histomonas meleagridis (Smith, 1895).

Histomoniasis or blackhead is a life-threatening disease of turkeys that is caused by a flagellated protozoan, Histomonas meleagridis. The development of an assay to measure the sensitivity of drugs traditionally used against this parasite, as reputed to be effective against other protozoan parasites, is described. The in vitro minimum lethal concentrations (MLC), time for drug efficacy, and parasite viability after removal of residual drugs were determined. Three of the 10 tested drugs, fenbendazole, albendazole, and sulfadiazine, were found to be ineffective against H. meleagridis. Nifursol, the only compound still authorized as a feed additive in Europe, is an inhibiting agent but is not lethal in vitro. Roxarsone, an arsenical derivate similar to nitarsone (the only authorized drug in United States), is effective at high concentration (200 microg/mL) after a long exposure (48 h). The lethal activity of dimetridazole, metronidazole, ronidazole, tinidazole, and furazolidone in vitro was demonstrated. Dimetridazole (MLC = 25 microg/mL after 6 h of exposure), metronidazole (MLC = 50 microg/mL after 24 h), and furazolidone (MLC = 50 microg/mL after 24 h) are rapidly effective at low concentrations. These results confirm the effectiveness of dimetridazole, a drug that has been used in the treatment and prevention of blackhead. In May 2002 this compound was removed as feed additive in Europe.

Animals↗

PubMed2000

In vitro activities of a new lipopeptide antifungal agent, FK463, against a variety of clinically important fungi.

The in vitro antifungal activity and spectrum of FK463 were compared with those of amphotericin B, fluconazole, and itraconazole by using a broth microdilution method specified by National Committee for Clinical Laboratory Standards document M27-A (National Committee for Clinical Laboratory Standards, Wayne, Pa., 1997). FK463 exhibited broad-spectrum activity against clinically important pathogens including Candida species (MIC range, <==0.0039 to 2 microg/ml) and Aspergillus species (MIC range, <==0.0039 to 0.0313 microg/ml), and its MICs for such fungi were lower than those of the other antifungal agents tested. FK463 was also potently active against azole-resistant Candida albicans as well as azole-susceptible strains, and there was no cross-resistance with azoles. FK463 showed fungicidal activity against C. albicans, i.e., a 99% reduction in viability after a 24-h exposure at concentrations above 0.0156 microg/ml. The minimum fungicidal concentration (MFC) assays indicated that FK463 was fungicidal against most isolates of Candida species. In contrast, the MFCs of FK463 for A. fumigatus isolates were much higher than the MICs, indicating that its action is fungistatic against this species. FK463 had no activity against Cryptococcus neoformans, Trichosporon species, or Fusarium solani. Neither the test medium (kind and pH) nor the inoculum size greatly affected the MICs of FK463, while the addition of 4% human serum albumin increased the MICs for Candida species and A. fumigatus more than 32 times. Results from preclinical in vitro evaluations performed thus far indicate that FK463 should be a potent parenteral antifungal agent.

Antifungal Agents↗

PubMed2000

Antifungal activity of local anesthetics against Candida species.

OBJECTIVE: To evaluate the activity of benzydamine, lidocaine, and bupivacaine, three drugs with local anesthetic activity, against Candida albicans and non-albicans strains and to clarify their mechanism of activity. METHODS: The minimal inhibitory concentration (MIC) was determined for 20 Candida strains (18 clinical isolates and two American Type Culture Collection strains). The fungistatic activity was studied with the fluorescent probe FUN-1 and observation under epifluorescence microscopy and flow cytometry. The fungicidal activity of the three drugs was assayed by viability counts. Membrane alterations induced in the yeast cells were evaluated by staining with propidium iodide, by quantitation of intracellular K+ leakage and by transmission electron microscopy of intact yeast cells and prepared spheroplasts. RESULTS: The MIC ranged from 12.5-50.0 microg/mL, 5.0-40.0 mg/mL, and 2.5-10.0 mg/mL for benzydamine, lidocaine, and bupivacaine, respectively. The inhibitory activity of these concentrations could be detected with the fluorescent probe FUN-1 after incubation for 60 minutes. A very fast fungicidal activity was shown by 0.2, 50, and 30 mg/mL of benzydamine, lidocaine, and bupivacaine, respectively. CONCLUSIONS: At lower concentrations, the tested drugs have a fungistatic activity, due to yeast metabolic impairment, while at higher concentrations they are fungicidal, due to direct damage to the cytoplasmic membrane.

Agar↗

PubMed2004

Genetic interactions of DST1 in Saccharomyces cerevisiae suggest a role of TFIIS in the initiation-elongation transition.

TFIIS promotes the intrinsic ability of RNA polymerase II to cleave the 3'-end of the newly synthesized RNA. This stimulatory activity of TFIIS, which is dependent upon Rpb9, facilitates the resumption of transcription elongation when the polymerase stalls or arrests. While TFIIS has a pronounced effect on transcription elongation in vitro, the deletion of DST1 has no major effect on cell viability. In this work we used a genetic approach to increase our knowledge of the role of TFIIS in vivo. We showed that: (1) dst1 and rpb9 mutants have a synthetic growth defective phenotype when combined with fyv4, gim5, htz1, yal011w, ybr231c, soh1, vps71, and vps72 mutants that is exacerbated during germination or at high salt concentrations; (2) TFIIS and Rpb9 are essential when the cells are challenged with microtubule-destabilizing drugs; (3) among the SDO (synthetic with Dst one), SOH1 shows the strongest genetic interaction with DST1; (4) the presence of multiple copies of TAF14, SUA7, GAL11, RTS1, and TYS1 alleviate the growth phenotype of dst1 soh1 mutants; and (5) SRB5 and SIN4 genetically interact with DST1. We propose that TFIIS is required under stress conditions and that TFIIS is important for the transition between initiation and elongation in vivo.

Benomyl↗

PubMed1995

Firefly luciferase assay of adenosine triphosphate as a tool of quantitation of the viability of BCG vaccines.

The purpose of the present study was to compare conventional colony forming unit (CFU) enumeration with a bioluminescent measurement in order to quantitate living bacteria in BCG vaccine. Forty batches of BCG vaccine quantitated with respect to weight (10 each of concentration 120 mg/vial, 30 mg/vial, 1.5 mg/vial and 0.75 mg/vial, respectively) were tested. Adenosine triphosphate (ATP) was extracted using the boiling Tris-EDTA extraction method preceded by treatment with the ATP-hydrolysing enzyme apyrase, and the liberated ATP was assayed using a LKB 1251 luminometer. The sensitivity of the assay was 10(-11) M ATP which corresponded to 5 x 10(5) CFU/ml. A proportional relation between CFU and ATP content per vial was found up to 30 mg/vial, but for 120 mg/vial the CFU method seemed unreliable in contrast to the bioluminescence method. The measurements error for the logarithm (base 10) of ATP was 0.051 and this method was therefore more exact than the CFU method (0.097). Because the bioluminescence method is sensitive, reliable, time-saving and less expensive it seems preferable to CFU enumeration in the quality control of BCG vaccines.

Adenosine Triphosphate↗

PubMed2006

Effects of extracts of miswak and derum on proliferation of Balb/C 3T3 fibroblasts and viability of cariogenic bacteria.

OBJECTIVES: This study examined the effects of extracts of two chewing sticks on proliferation of fibroblasts and viability of cariogenic bacteria. METHODS: Aqueous extracts of miswak (Salvadora persica; Arak tree) and derum (Juglans regia; walnut tree) were prepared and their effects investigated on growth of Balb/C 3T3 mouse fibroblasts by measuring the mitochondrial succinic dehydrogenase activity. Furthermore, the effects on the viability of various cariogenic bacteria (Streptococcus mutans, Streptococcus salivarius, Lactobacillus casei and Actinomyces viscosus) was also determined. RESULTS: The data revealed that Balb/C 3T3 fibroblasts exposed to aqueous extracts of miswak or derum showed an increase in cell proliferation by 156% and 255%, respectively, in comparison with controls (p < 0.0001). Furthermore, extracts from both miswak and derum had adverse effects on the growth of the cariogenic microorganisms, with derum having significantly greater antimicrobial effects than miswak and at much lower concentrations against all the bacteria tested. The most sensitive organisms were A. viscosus, followed by S. mutans, S. salivarius, with L. casei being the most resistant. CONCLUSION: The results show that aqueous extracts of miswak and derum enhance the growth of fibroblasts and inhibit the growth of cariogenic bacteria, with the derum extract showing greater activity than miswak.

3T3 Cells↗

PubMed1982

Bactericidal activity of trimethoprim alone and in combination with sulfamethoxazole on susceptible and resistant Escherichia coli K-12.

The combined effects of trimethoprim and sulfamethoxazole on the viability of Escherichia coli K-12 and resistant strains possessing resistance plasmids were examined in minimal medium. When methionine, glycine, and adenine were present, sulfamethoxazole could enhance trimethoprim activity against E. coli K-12 so that the combination was bactericidal. However, this enhancement occurred over a narrow range of trimethoprim concentrations (0.04 to 0.2 mg liter-1) and only when the sulfamethoxazole concentration was more than 10 times that of trimethoprim. Under certain conditions, sulfamethoxazole enhanced trimethoprim bactericidal activity against E. coli K-12 carrying plasmid R1 at concentrations of sulfamethoxazole far below those required to inhibit the organism, but there was no such enhancement with the same host containing the SSu plasmid. Similar differences were found with strains possessing trimethoprim resistance plasmids R483 and R751. Sulfamethoxazole can promote a bactericidal response with trimethoprim in E. coli K-12 and some of its resistant derivatives, but only under a narrow range of concentrations.

Culture Media↗

PubMed2001

Dental unit waterline contamination and its possible implications during periodontal surgery.

BACKGROUND: Dental unit waterline contamination has become a concern to clinical dentistry. This concern arises from the fact that bacteria sloughed from established biofilms in dental unit waterlines increase heterotrophic bacteria counts in water exiting these units. METHODS: Scanning microscopy and bacterial viability staining were used to examine the sessile and planktonic biofilm present in dental unit waterlines and water samples, respectively. In addition, the limulus amebocyte assay was used to measure the lipopolysaccharide (LPS) levels in water samples. RESULTS: All dental unit waterlines were coated with a well-established biofilm made up of filamentous and bacillus-like microorganisms. Water samples collected from these dental units contained high numbers of individual bacteria and bacterial aggregates. A viability staining technique identified significantly more bacteria in water than could be cultured, and 64% of the total bacterial population stained as nonvital. Since the bacterial load (viable and nonviable) was high, we examined the LPS in dental unit water samples. The mean LPS levels in water collected from high-speed and air/water lines in use were 480 and 1,008 endotoxin units (EU)/ml. This was significantly higher than the mean level of 66 EU/ml found in water samples collected from adjacent clinic sinks. The LPS level at the start of the day (2,560 EU/ml) was reduced by 70% with 1 minute of flushing (800 EU/ml). Flushing times of 5 and 10 minutes were not able to reduce LPS levels to zero. CONCLUSION: The presence of high heterotrophic bacterial counts, sloughing biofilm, and high LPS levels are discussed in relation to patient risk and periodontal wound healing biology.

Analysis of Variance↗

PubMed1990

Effect of phosphate buffer agar on the number of UV-induced mutations to streptomycin resistance in Escherichia coli strains.

The number of UV-induced mutations to streptomycin resistance in Escherichia coli B/r cells depends on the post-irradiation incubation medium. When cells are incubated on phosphate buffer agar (PBA) as opposed to brain heart infusion agar (BHI), there is an irreversible loss in the number of potential streptomycin resistant mutants which develop. This decrease in mutant numbers is known to occur without a corresponding loss in overall cell viability and cannot be explained in terms of the kinetics of dimer excision. Recent studies have indicated that pyrimidine dimers are the substrate for this repair on PBA. The genetics of PBA repair was investigated by observing the influence of the post-irradiation incubation medium (PBA vs BHI) on the number of streptomycin resistant mutants which develop in E. coli strains deficient in various repair pathways. A recB and recC strain (WP3 recB trp-; WP7 recC trp-) showed repair of potential streptomycin resistant mutants similar to that of B/r when incubated on PBA following irradiation. The medium had no effect on the number of mutants expressed in an excision deficient uvrA- strain (WP2 uvrA trp-) or a polymerase I mutant (P3478 polA thy-). This was interpreted to mean that the loss of streptomycin resistant mutants on PBA involves the excision repair pathway and is dependent on the polA gene product, polymerase I.

Culture Media↗

PubMed1999

Stimulation of the multiplication of Micrococcus luteus by an autocrine growth factor.

Viable cells of Micrococcus luteus secrete a proteineous growth factor (Rpf) which promotes the resuscitation of dormant, nongrowing cells to yield normal, colony-forming bacteria. When washed M. luteus cells were used as an inoculum, there was a pronounced influence of Rpf on the true lag phase and cell growth on lactate minimal medium. In the absence of Rpf, there was no increase in colony-forming units for up to 10 days. When the inoculum contained less than 10(5) cells ml-1, macroscopically observable M. luteus growth was not obtained in succinate minimal medium unless Rpf was added. Incubation of M. luteus in the stationary phase for 100 h resulted in a failure of the cells to grow in lactate minimal medium from inocula of small size although the viability of these cells was close to 100% as estimated using agar plates made from lactate minimal medium or rich medium. The underestimation of viable cells by the most-probable-number (MPN) method in comparison with colony-forming units was equivalent to the requirement that at least 10(5) cells grown on succinate medium, 10(3) cells from old stationary phase, or approximately 10-500 washed cells are required per millilitre of inoculum for growth to lead to visible turbidity. The addition of Rpf in the MPN dilutions led to an increase of the viable cell numbers estimated to approximately the same levels as those determined by colony-forming units. Thus, a basic principle of microbiology - "one cell-one culture" - may not be applicable in some circumstances in which the metabolic activity of "starter" cells is not sufficient to produce enough autocrine growth factor to support cell multiplication.

Bacterial Proteins↗

PubMed1991

Influence of the isolation method on the stability of differentiated phenotype in cultured rat hepatocytes.

Primary cultures of adult rat hepatocytes were established using two different isolation procedures: a two-step collagenase perfusion and a method using ethylenediaminetetraacetate (EDTA) as the dissociating agent. Both techniques provided good yields of hepatocytes with comparable viability. The evolution of hepato-specific protein levels and several drug-metabolizing enzyme activities were followed for 8 days in cultured hepatocytes obtained by both methods. EDTA-isolated hepatocytes maintained a low gamma glutamyltransferase (GGT) activity, whereas collagenase-treated cells acquired a high GGT level. Transferrin secretion and tyrosine aminotransferase (TAT), alanine aminotransferase (ALT), and microsomal epoxide hydrolase (mEH) activities were stable in both EDTA- and collagenase-isolated hepatocytes, whereas albumin secretion, aspartate amino transferase (AST) activity, total cytochromes P-450 content, IA1 and IIB1 P-450 isoenzymes, NADPH-cytochrome P-450 reductase (EC 1.6.2.4) levels, and bilirubin glucuronidation decreased faster in collagenase-treated cells. The most important difference observed was the maintainance of the mixed-function oxidase system in EDTA-isolated hepatocytes. These results emphasize the critical role of isolation technique in stabilization of differentiated hepatocytes in primary culture.

Animals↗

PubMed2002

Experimental reproduction of the Jorge Lobo's disease in BAlb/c mice inoculated with Lacazia loboi obtained from a previously infected mouse.

Long-term maintenance of Lacazia loboi in the laboratory has not been reported. We report here the use BALB/c mice to maintain the Lacazia loboi for extended period of time. Eight to ten week-old mice were inoculated intradermally in both hind footpads with a fungal suspension from a macerated footpad obtained from an original mouse previously infected with the fungi and sacrificed 8 months after inoculation. The inoculated animals were sacrificed at different time intervals, footpads were excised, the right one was submitted to histopathological examination and the left one was macerated in sterile saline for fungal count and viability index determination. The inoculated animals presented the histopathological picture identical to the mice previously inoculated with material from human lesion. Granulomatous infiltrates with predominance of macrophages and giant cells were observed. The granulomas evolved progressively as observed in the different times of sacrifice. After 7 months of inoculation, macroscopic lesions were observed, and the number of fungi obtained from macerated footpads was higher than the number of inoculated fungi. The pattern of lesion development was similar to what was observed in animals infected with a fungal suspension obtained from a human lesion. Considering the histopathological findings, the clinical manifestations, and the finding of a higher number of fungi obtained than the inoculated into footpads of each mice, we believe the BALB/c mice strain is as an excellent way to maintain L. loboi in laboratory. Moreover, even after serial passages of the fungi, the granulomatous lesions are reproduced consistently in laboratory conditions.

Animals↗

PubMed1986

Evaluation of the bactericidal activity of beta-lactam antibiotics on slowly growing bacteria cultured in the chemostat.

The bactericidal activity of 23 beta-lactam antibiotics was compared in slowly growing bacteria cultured in a chemostat. In an attempt to mimic possible in vivo conditions, slowly growing cultures were produced by limitation of iron, glucose, phosphate, or magnesium. Only select antibiotics remained effectively bactericidal against slowly growing cells. For these compounds, the rate of antibiotic-induced loss of viability was a constant when killing was expressed per generation (in contrast to absolute time) in that slowly growing bacteria were killed proportionately more slowly. Individual antibiotics differed greatly, however, in their specific bactericidal activities against slowly growing cells, i.e., in the absolute degree of killing elicited during exposure of the bacteria to MIC equivalents of the drugs. Specific bactericidal activities varied not only with drug structure but also with the bacterial strains and, to a lesser extent, with the nature of the growth-limiting nutrient. In slowly growing cultures exposure to the low drug concentrations studied here (near MIC) caused killing without detectable lysis. Antibiotics with high specific bactericidal activities were capable of rapidly killing cultures of slowly growing pathogens despite extremely long generation times approaching those reported for in vivo growth rates.

Anti-Bacterial Agents↗

PubMed2003

Pentamidine is active in vitro against Fusarium species.

Fusariosis is an emerging opportunistic mycosis against which currently used antifungals have limited activity. Here, we investigated the in vitro activities of pentamidine (PNT) against 10 clinical isolates of Fusarium species (five Fusarium solani isolates and five non-F. solani isolates) by using the National Committee for Clinical Laboratory Standards microdilution method in three different media (RPMI, RPMI-2, and a yeast nitrogen base medium), disk diffusion testing, and viability dye staining. PNT had significant activities against all 10 Fusarium isolates. Non-F. solani isolates were more susceptible than F. solani isolates (P < 0.05). Additionally, PNT was fungicidal against all non-F. solani isolates, whereas it had fungistatic effects against four of the five F. solani isolates. PNT also exhibited greater activity against conidial than against hyphal development of the fungus. This fungicidal activity against non-F. solani Fusarium isolates was confirmed microscopically after staining of PNT-treated Fusarium oxysporum hyphae with the fluorescent viability dyes 5,(6)-carboxyfluorescein diacetate (CFDA) and bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC). The MICs at which 50% of the isolates were inhibited (2 micro g/ml for non-F. solani isolates and 4 micro g/ml for F. solani isolates) and the minimum fungicidal concentration at which 50% of the isolates were killed (8 micro g/ml for non-F. solani isolates) were much lower than the PNT tissue concentrations previously reported in humans using conventional daily intravenous PNT dosing. Finally, PNT was more active against Fusarium isolates in a hypoxic environment of in vitro growth (P < 0.05). This finding may be clinically significant, because Fusarium, an angiotropic mold, causes tissue infarcts with resultant low tissue perfusion. Our findings suggest that PNT may have a role in the management of Fusarium infections. Future in vivo studies are needed to verify these in vitro findings.

Aerobiosis↗

PubMed1997

Effect of aerosolization on culturability and viability of gram-negative bacteria.

Estimations of the bacterial content of air can be more easily made now than a decade ago, with colony formation the method of choice for enumeration of airborne bacteria. However, plate counts are subject to error because bacteria exposed to the air may remain viable yet lose the ability to form colonies, i.e., they become viable but nonculturable. If airborne bacteria exhibit this phenomenon, colony formation data will significantly underestimate the bacterial populations in air samples. The objective of the study reported here was to determine the effect of aerosolization on viability and colony-forming ability of Serratia marcescens, Klebsiella planticola, and Cytophaga allerginae. A collision nebulizer was used to spray bacterial suspensions into an aerosol chamber, after which duplicate samples were collected in all-glass impingers over a 4-h period. Humidity was maintained at ca. 20 to 25%, and temperature was maintained at 20 to 22 degrees C for each of two replicate trials per microorganism. Viability was determined by using a modified direct viable count method, employing nalidixic acid or aztreonam and p-iodonitrotetrazolium violet (INT). Cells were stained with acridine orange and observed by epifluorescence microscopy to enumerate total and viable cells. Viable cells were defined as those elongating in the presence of antibiotic and/or reducing INT. CFU were determined by plating on tryptic soy agar and R2A agar. It was found that culture techniques did not provide an adequate description of the bacterial burdens of indoor air (i.e., less than 10% of the aerosolized bacteria were capable of forming visible colonies). It is concluded that total cell count procedures provide a better approximation of the number of bacterial cells in air and that procedures other than plate counting are needed to enumerate bacteria in aerosol samples, especially if the public health quality of indoor air is to be estimated.

Aerosols↗