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PubMed1997

Effect of human newborn BCG immunization on monocyte viability and function at 3 months of age.

SETTING: Immune response induced by BCG vaccination seems to reflect the development of T-cell immunity and monocyte activation. Participants were recruited from a large prospective study in infants from a suburb in Santiago, Chile. OBJECTIVE: To determine whether newborn BCG immunization changes the innate ability of cultured monocyte-macrophages to ingest and kill virulent mycobacteria in the absence of lymphocytes. DESIGN: The study population consisted of 15 three-month-old, tuberculin-positive infants immunized with BCG (Japanese) at birth, 13 randomly-selected, age-matched tuberculin-nonreactive infants in whom BCG immunization was postponed until one year of age, and five BCG-immunized, tuberculin-reactive adults. Adherent cells were cultured for 48 h. Monocyte-macrophage viability and number and viability of intracellular Mycobacterium tuberculosis bacilli were assessed after an additional 2 h and 4 and 7 days of incubation. RESULTS: There was no difference in the mean number of adherent cells present after 48 h among the three study groups. Adherent cells from BCG-immunized infants and adults had a significantly higher viability after 7 days in culture than adherent cells from non-immunized infants. The percentage of cells ingesting M. tuberculosis and the number of bacilli per cell after 2 h and 4 days was significantly higher in immunized infants and adults than in non-immunized infants. However, there was no evidence for increased killing of mycobacteria by cells from immunized infants and adults. CONCLUSION: These results suggest that BCG vaccination increases monocyte viability and the uptake of M. tuberculosis without enhancing the ability to kill ingested M. tuberculosis in the absence of lymphocytes.

Adult↗

PubMed1995

Comparison of antigen detection, polymerase chain reaction and culture for detection of Chlamydia trachomatis in genital infection.

An in-house polymerase chain reaction (PCR) test using ribosomal RNA gene primers was compared with chlamydia antigen detection (DIF) and culture for the detection of Chlamydia trachomatis. Five hundred and forty-eight fresh (unstored) genital swabs and 174 urines (collected at the same time) from patients attending a sexually-transmitted diseases clinic were examined. PCR, DIF and culture detected chlamydia in 43, 35 and 42 swabs respectively from the 43 resolved positive cases. The specificity on the resolved negative specimens was 100% for each of the tests. From the urines, PCR and DIF detected the organism in 16 and 15 cases respectively of the 23 resolved positive males tested but in only 2 and 3 cases respectively of the 9 resolved positive females tested. Specificities were 100% in all cases. Both of the non-culture tests manifested problems with urine due to inhibitory activity (in PCR test) or excessive debris (in DIF test) in about 5% of the specimens. Culture of the urines yielded sensitivities of 40% in the males and 22% in the females. Overall PCR was more sensitive than either culture or DIF on both urethral and cervical swabs and urines. The urines yielded less than three-quarters the number of positives that was obtained from the swabs and were considered to be an unsatisfactory specimen for chlamydial diagnosis. It is concluded that PCR is a satisfactory alternative to culture on genital swabs and may be preferable in situations where the viability of the organisms is in question. DIF remains useful because of its speed and simplicity but is insufficiently sensitive to be relied upon by itself.

Adolescent↗

PubMed1997

The effect of quinolones on the intracellular killing of Staphylococcus aureus in neutrophil granulocytes.

The effect of ciprofloxacin, lomefloxacin, fleroxacin and ofloxacin on the intracellular killing of Staphylococcus aureus in human neutrophil granulocytes was studied. Each drug was tested in concentrations of 0.25, 1, 4, 16 and 64 times the MIC and the intracellular killing was measured for up to 5 h of incubation. All four quinolones increased the intracellular killing in a concentration- and time-dependent manner. When compared at concentrations of 4 x MIC, ofloxacin increased the killing significantly more than the other quinolones. All four quinolones increased the killing significantly more than the beta-lactam dicloxacillin. All five antibiotics increased the killing significantly relative to the control without antibiotics. None of the antibiotics affected the viability of the granulocytes or their ability to generate superoxide anion. In conclusion, the quinolones increased the intracellular killing of S. aureus in neutrophil granulocytes.

Adult↗

PubMed2006

Efficacy of iron chelators on Campylobacter concentrations in turkey semen.

Campylobacter is a leading bacterial cause of human foodborne infections in the United States. Recent studies suggest that the organism is highly prevalent in poultry semen and may contribute to vertical transmission between the breeder hen and offspring. Because Campylobacter requires iron for its growth and survival, the objective of this study was to determine if the addition of natural and synthetic chelators such as ovotransferrin, desferrioxaime, EDTA, or 2,2'-dipyridyl could reduce or eliminate Campylobacter in turkey semen. In a preliminary study without semen, a commercial poultry semen extender was supplemented with various concentrations of ovotransferrin, desferrioxaime, EDTA, or 2,2'-dipyridyl and inoculated with an average of 10(8) cfu/mL of a wild-type Campylobacter coli turkey semen isolate. At 6 and 24 h of storage at 4 degrees C, a sample was taken from each treatment group and enumerated for Campylobacter. In all 3 trials, Campylobacter was undetectable (< 10(2)) in the commercial poultry semen extender supplemented with 20 mg/mL of 2,2'-dipyridyl. There were no differences observed in Campylobacter concentrations in the commercial poultry semen extender supplemented with ovotransferrin, desferrioxaime, or EDTA compared with unsupplemented controls. In a follow-up study, pooled semen samples were randomly collected from toms, diluted with a commercial poultry semen extender supplemented with 5, 10, or 20 mg/mL of 2,2'-dipyridyl and inoculated with an average of 10(8) cfu/mL of a wild-type C. coli turkey semen isolate. At 6 and 24 h of storage at 4 degrees C, samples were taken from each treatment group, enumerated for Campylobacter, and evaluated for sperm viability. In all 3 trials, supplementing the commercial poultry semen extender with 20 mg/mL of 2,2'-dipyryidyl significantly reduced (3 to 4 logs) Campylobacter concentrations when compared with the positive controls. Sperm viability was also reduced with this treatment, and, therefore, the use of 2,2'-dipyridyl may not be a practical treatment for reducing Campylobacter in poultry semen.

2,2'-Dipyridyl↗

PubMed1991

Isolation and characterization of a lipolytic bacterium capable of growing in a low-water-content oil-water emulsion.

A unique lipolytic bacterium was isolated in a selective growth system consisting of 99% triglycerides and a 1% water phase. The bacterium, termed Pseudomonas aeruginosa YS-7, was able to grow in an environment of low water content and could also survive amphipathic, osmotic, and matrical water stress in a triglyceride-rich culture. The isolated strain was identified as P. aeruginosa on the basis of standard physiological, biochemical, and serological assays. The strain is a gram-negative motile rod, aerobic, pigment forming, and capable of growing at 42 degrees C. It is highly tolerant of high concentrations of the cationic detergent cetyltrimethylammonium bromide and of the fatty acid salts derived from bacterial hydrolysis of the oil. Growth of the bacterium in a pure culture in a 99% triglyceride medium lasted until most of the water was evaporated or consumed. Growth was accompanied by triglyceride hydrolysis, which continued to occur even after growth saturation until the water was totally depleted. No loss of viability was observed when the culture was maintained under water-depleted conditions for an additional 40 h. A second cycle of bacterial growth and triglyceride hydrolysis was immediately initiated upon the addition of 1% (vol/vol) water to the culture. Lipase activity was stable regardless of changes in culture conditions. The isolated strain is uniquely resistant to severe water stress in a triglyceride-rich medium or under cold acetone precipitation compared with 12 other microbial strains, including bacteria and yeasts. Among these 12, only the lipolytic strains grew in the 99% triglyceride medium, but they reached a cell mass fourfold smaller than that of P. aeruginosa YS-7.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetone↗

PubMed2005

Fluorescent multivalent opsonophagocytic assay for measurement of functional antibodies to Streptococcus pneumoniae.

We developed fluorescent mono- and multivalent opsonophagocytic assays (fOPA and fmOPA, respectively) specific for seven Streptococcus pneumoniae serotypes (4, 6B, 9V, 14, 18C, 19F, and 23F). Bacterial survival was quantitated with alamar blue, a fluorescent metabolic indicator. Both fOPA and fmOPA allow for determination of viability endpoints for up to seven serotypes with high levels of agreement to the reference method. The fmOPA eliminates colony counting, reduces serum volume, and produces results in 1 day.

Antibodies, Bacterial↗

PubMed2001

Validation of an in vitro biofilm model of supragingival plaque.

The study of biofilm structure and function mandates the use of model systems for which a host of environmental variables can be rigorously controlled. We describe a model of supragingival plaque containing Actinomyces naeslundii, Veillonella dispar, Fusobacterium nucleatum, Streptococcus sobrinus, and Streptococcus oralis wherein cells are cultivated anaerobically in a saliva-based medium on hydroxyapatite discs coated with a salivary pellicle, with material and pieces of apparatus common to all microbiology laboratories. After 0.5 hr, 16.5 hrs, 40.5 hrs, and 64.5 hrs, the composition of adherent biofilms was analyzed by culture techniques, live/dead fluorescence staining, and confocal laser scanning microscopy. Repeated independent trials demonstrated the repeatability of biofilm formation after 40.5 hrs and 64.5 hrs. Brief exposures of biofilms to chlorhexidine or Triclosan produced losses in viability similar to those observed in vivo. This biofilm model should prove very useful for pre-clinical testing of prospective anti-plaque agents at clinically relevant concentrations.

Actinomyces↗

PubMed2003

Bactericidal effects of Lactobacillus reuteri and allyl isothiocyanate on Escherichia coli O157:H7 in refrigerated ground beef.

Two naturally occurring antimicrobial agents were tested in packages of refrigerated ground beef for their ability to reduce the viability of Escherichia coli O157:H7 during storage. Allyl isothiocyanate (AITC) and Lactobacillus reuteri were tested separately and together for their action against a cocktail of five strains of E. coli O157:H7 in ground beef held at 4 degrees C for 25 days. Ground beef prepared from whole, raw inside round beef roasts was inoculated with low (3 log CFU/g) or high (6 log CFU/g) levels of the E. coli O157:H7 mixture. The beef was treated with AITC (about 1,300 ppm), L. reuteri, or both, along with 250 mM of glycerol per kg of meat at two levels (3 and 6 log CFU/g) and according to a design that yielded 8 controls plus 10 different treatments. Samples were analyzed for E. coli O157:H7 survivors, numbers of total bacteria, and lactic acid bacteria on days 0 to 25 at 5-day intervals. L. reuteri at both input levels with glycerol killed E. coli O157:H7 at both inoculated levels before day 20. AITC completely eliminated E. coli O157:H7 at the low-inoculum level (3 log CFU/g) and reduced viability >4.5 log CFU/g at the high-inoculum level (6 log CFU/g) by the end of the storage period. The combination of L. reuteri and AITC did not yield an additive effect against E. coli O157:H7 viability. L. reuteri in the presence of glycerol was highly effective against E. coli O157:H7 in ground beef during refrigerated storage (4 degrees C) in modified atmosphere packages. Sensory testing is planned to evaluate effects of treatments.

Cold Temperature↗

PubMed2002

Growth studies of potentially probiotic lactic acid bacteria in cereal-based substrates.

AIMS: The overall growth kinetics of four potentially probiotic strains (Lactobacillus fermentum, Lact. reuteri, Lact. acidophilus and Lact. plantarum) cultured in malt, barley and wheat media were investigated. The objectives were to identify the main factors influencing the growth and metabolic activity of each strain in association with the cereal substrate. METHODS AND RESULTS: All fermentations were performed without pH control. A logistic-type equation, which included a growth inhibition term, was used to describe the experimental data. In the malt medium, all strains attained high maximum cell populations (8.10-10.11 log10 cfu ml(-1), depending on the strain), probably due to the availability of maltose, sucrose, glucose, fructose (approx. 15 g l(-1) total fermentable sugars) and free amino nitrogen (approx. 80 mg l(-1)). The consumption of sugars during the exponential phase (10-12 h) resulted in the accumulation of lactic acid (1.06-1.99 g l(-1)) and acetic acid (0.29-0.59 g l(-1)), which progressively decreased the pH of the medium. Each strain demonstrated a specific preference for one or more sugars. Since small amounts of sugars were consumed by the end of the exponential phase (17-43%), the decisive growth-limiting factor was probably the pH, which at that time ranged between 3.40 and 3.77 for all of the strains. Analysis of the metabolic products confirmed the heterofermentative or homofermentative nature of the strains used, except in the case of Lact. acidophilus which demonstrated a shift towards the heterofermentative pathway. All strains produced acetic acid during the exponential phase, which could be attributed to the presence of oxygen. Lactobacillus plantarum, Lact. reuteri and Lact. fermentum continued to consume the remaining sugars and accumulate metabolic products in the medium, probably due to energy requirements for cell viability, while Lact. acidophilus entered directly into the decline phase. In the barley and wheat media all strains, especially Lact. acidophilus and Lact. reuteri, attained lower maximum cell populations (7.20-9.43 log10 cfu ml(-1)) than in the malt medium. This could be attributed to the low sugar content (3-4 g l(-1) total fermentable sugar for each medium) and the low free amino nitrogen concentration (15.3-26.6 mg l(-1)). In all fermentations, the microbial growth ceased at pH values (3.73-4.88, depending on the strain) lower than those observed for malt fermentations, which suggests that substrate deficiency in sugars and free amino nitrogen contributed to growth limitation. CONCLUSIONS: The malt medium supported the growth of all strains more than barley and wheat media due to its chemical composition, while Lact. plantarum and Lact. fermentum appeared to be less fastidious and more resistant to acidic conditions than Lact. acidophilus and Lact. reuteri. SIGNIFICANCE AND IMPACT OF THE STUDY: Cereals are suitable substrates for the growth of potentially probiotic lactic acid bacteria.

Culture Media↗

PubMedDate not supplied

[The effect of subinhibitory concentrations of antibiotics on the adhesion of enterobacteria].

It is impossible to determine rigidly a net result of the influence of antibiotics on the interaction between parasite and host cells, as many factors participating in this process are not studied. Adhesion of microorganisms is one of the essential mechanisms of the above interaction. Antibiotics with a different mechanism of action in the subinhibitory concentrations affecting viability of microbes either slightly or nowise have been studied for their effect on adhesion on a model of the intestine section of human embryos and experimental animals. Most of antibiotics influenced differently adhesion of the microorganisms, that also depended on the species attribution of the latter. The accelerated selection of resistance during a successive passage via the suggested adhesion system was observed. The data obtained elucidated certain mechanisms of the effect of antibiotics on the microbial populations at the initial phase of the infectious process and under the primary contamination of mucosa.

Animals↗

PubMed1998

Microbial relatives of seed storage proteins: conservation of motifs in a functionally diverse superfamily of enzymes

Plant storage proteins comprise a major part of the human diet. Sequence analysis has revealed that these proteins probably share a common ancestor with a fungal oxalate decarboxylase and/or related bacterial genes. Additionally, all these proteins share a central core sequence with several other functionally diverse enzymes and binding proteins, many of which are associated with synthesis of the extracellular matrix during sporulation/encystment. A possible prokaryotic relative of this sequence is a bacterial protein (SASP) known to bind to DNA and thereby protect spores from extreme environmental conditions. This ability to maintain cell viability during periods of dehydration in spores and seeds may relate to absolute conservation of residues involved in structure determination.

Journal Article↗

PubMed2002

Early formation of Streptococcus sobrinus biofilm on various dental restorative materials.

OBJECTIVES: To examine the formation of dental biofilm by Streptococcus sobrinus on different types of restorative materials, using a model consisting of host and bacterial constituents. METHODS: The adsorption pattern of saliva to the restorative material was determined by means of gel electrophoresis coupled with computerized densitometry techniques. The amount of salivary proteins adsorbed onto the surfaces was measured using the Bradford method. Sucrose-dependent bacterial adhesion to the saliva-coated restorative material was tested by radioactive-labelled Streptococcus sobrinus, and viable counts of these bacteria in the biofilm was determined using bacterial culture techniques. RESULTS: Different adsorption patterns by salivary proteins to restorative materials were recorded. Durafil and acrylic dental materials demonstrated the most affinity to salivary proteins. A surface dependent adhesion profile was recorded, showing a high affinity of albumin and amylase to Acrylic and Durafil materials. Bacterial accumulation was the highest with Fuji LC and Fuji GC, which also demonstrated the highest bacterial viability. CONCLUSIONS: Our study demonstrates the specificity of biofilm formation on different brands of dental restorative materials. Formation of a variety of dental biofilms has a significant impact on the progression of dental diseases in the oral cavity.

Acrylic Resins↗

PubMed1990

Ethanol fermentation by nystatin-resistant strains of Saccharomyces cerevisiae.

Nystatin-resistant mutants of haploid and polyploid strains of Saccharomyces cerevisiae were isolated by plating on gradient plates with increasing nystatin concentrations (60-3000 U/ml). Some of the mutants were defective in ergosterol biosynthesis, and produced zymosterol and cholestatetraenol-like sterols. Those mutants which do not form ergosterol produce less ethanol than the parent strains. They also had lower viability during fermentation of glucose solutions (8-13% vs. 33-47%). This became more pronounced in fermentations of higher concentrations of glucose. A nystatin-resistant but ergosterol-forming mutant had a similar fermentation capacity to the parent strain.

Chromatography, Thin Layer↗

PubMed1995

High-throughput screen for detecting antimycobacterial agents.

A simple, robust assay system which can be used to screen for inhibitors of mycobacterial growth has been developed. A strain of the rapidly growing saprophyte Mycobacterium aurum is used as the test organism. Inhibition of its growth is highly predictive of activity against Mycobacterium tuberculosis, which cannot itself be used in screening because of its growth characteristics and highly infectious nature. The viability of M. aurum in the presence of a test sample is monitored by measuring the uptake of radiolabelled uracil into the cells. In a microtiter plate format, the screen has the potential for testing several thousand samples per day.

Antitubercular Agents↗

PubMed1993

Fate of Enterobacter cloacae JP120 and Alcaligenes eutrophus AEO106(pRO101) in soil during water stress: effects on culturability and viability.

A sandy loam soil near field capacity moisture content (psi = -0.050 MPa) or air dried (psi = -300 MPa) was inoculated with about 3 x 10(7) CFU of Enterobacter cloacae JP120 and Alcaligenes eutrophus AEO106(pRO101) per g and incubated in 40-g portions at 17 degrees C in closed or open Erlenmeyer flasks. In the field-moist soil, selective plating, direct viable counts, and DNA hybridization showed only minor changes in the numbers of E. cloacae and A. eutrophus cells with time (14 days), and the results obtained with the three detection methods generally agreed. In the air-dried soil, the majority of both bacteria were found as intact DNA-carrying cells that were neither culturable nor viable by the methods employed in this study. The numbers of culturable E. cloacae and A. eutrophus cells dropped to 10(5) and 10(2) CFU/g, respectively, 2 h after inoculation. Direct viable counts showed that only about 1% of the cells detected by immunofluorescence microscopy were viable, but a fraction of viable nonculturable cells of both bacteria was present. A. eutrophus did not tolerate desiccation as well as E. cloacae. Only a minor fraction of the two test organisms regained their culturability or viability after rewetting of the air-dried soil; the number of total heterotrophic culturable bacteria, however, increased more than 10-fold and reached 73% of the level found in the field-moist soil at day 14.

Alcaligenes↗

PubMed1990

Vanadate-resistant mutants of Saccharomyces cerevisiae show alterations in protein phosphorylation and growth control.

This work describes two spontaneous vanadate-resistant mutants of Saccharomyces cerevisiae with constitutive alterations in protein phosphorylation, growth control, and sporulation. Vanadate has been shown by a number of studies to be an efficient competitor of phosphate in biochemical reactions, especially those that involve phosphoproteins as intermediates or substrates. Resistance to toxic concentrations of vanadate can arise in S. cerevisiae by both recessive and dominant spontaneous mutations in a large number of loci. Mutations in two of the recessive loci, van1-18 and van2-93, resulted in alterations in the phosphorylation of a number of proteins. The mutant van1-18 gene also showed an increase in plasma membrane ATPase activity in vitro and a lowered basal phosphatase activity under alkaline conditions. Cells containing the van2-93 mutant allele had normal levels of plasma membrane ATPase activity, but this activity was not inhibited by vanadate. Both of these mutants failed to enter stationary phase, were heat shock sensitive, showed lowered long-term viability, and sporulated on rich medium in the presence of 2% glucose. The wild-type VAN1 gene was isolated and sequenced. The open reading frame predicts a protein of 522 amino acids, with no significant homology to any genes that have been identified. Diploid cells that contained two mutant alleles of this gene demonstrated defects in spore viability. These data suggest that the VAN1 gene product is involved in regulation of the phosphorylation of a number of proteins, some of which appear to be important in cell growth control.

Adenosine Triphosphatases↗

PubMed1983

Pivmecillinam in treatment of Staphylococcus saprophyticus urinary tract infections.

In a non-randomised trial 15 women with Staphylococcus saprophyticus bacteriuria were treated with pivmecillinam 200 mg t.i.d. for 1 week. At follow-up 2-4 weeks post treatment only 11 patients were cured. As a complement to the study the occurrence of Staph. saprophyticus bacteriuria was analysed according to age, sex and season. Mecillinam MIC's and viability curves of Staph. saprophyticus were investigated. It is concluded that mecillinam should only be used to treat urinary tract infections caused by bacteria sensitive in vitro.

Age Factors↗

PubMed1983

Studies on a new antibiotic M-92 produced by Micromonospora. IV. Bactericidal action of the component VA-2.

The action of VA-2, the most active component of antibiotic M-92, against S. aureus is bactericidal but not bacteriolytic. The bactericidal action is markedly affected by incubation temperature, whether bacterial cells are prolific or resting. The bactericidal kinetics of VA-2 is biphasic, since addition of VA-2 caused rapid and straight decrease in viability curve and reached a plateau after several minutes. The bactericidal activity of VA-2 is blocked by 2,4-dinitrophenol. Alike to many membrane-active bacteriocins, VA-2 seems to exert its action through two stages.

2,4-Dinitrophenol↗