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PubMed2005

Combined effects of coating, modified atmosphere packaging, and gamma irradiation on quality maintenance of ready-to-use carrots (Daucus carota).

The objective of this study was to evaluate the effect of an edible coating combined with modified atmosphere (MA; 60% O2, 30% CO2, and 10% N2) packaging and gamma irradiation on the microbiological stability and physicochemical quality of minimally processed carrots. A coating based on calcium caseinate and whey protein isolates was used. Coated and uncoated peeled minicarrots were packed under the MA or air (78.1% N2, 20.9% O2, and 0.036% CO2), irradiated at 0.5 or 1 kGy, and stored at 4 +/- 1 degrees C for 21 days. Samples were evaluated periodically for aerobic plates counts (APCs) and physicochemical properties (firmness, white discoloration, and whiteness index). Gamma irradiation did not significantly affect the physicochemical properties of the carrots (P > 0.05). Microbiological analysis revealed that for uncoated carrots irradiation at 0.5 and 1 kGy under air and MA reduced the APCs by 3.5 and 4 log CFU/g and by 4 and 4.5 log CFU/g, respectively. For coated carrots, irradiation at 0.5 and 1 kGy under air and MA reduced the APCs by 4 and 4.5 log CFU/g and by 3 and 4.25 log CFU/g, respectively. The coating was able to protect carrots against dehydration during storage under air. Coating and irradiation at 1 kGy were also able to protect carrot firmness during storage under air. MA packaging retarded whitening of uncoated carrots but had a detrimental effect on firmness. The edible coating used in this study did not significantly inhibit (P > 0.05) microbial growth on carrots.

Carbon Dioxide↗
PubMed2005

Effects of community versus single strain inoculants on the biocontrol of Salmonella and microbial community dynamics in alfalfa sprouts.

Potential biological control inoculants, Pseudomonas fluorescens 2-79 and microbial communities derived from market sprouts or laboratory-grown alfalfa sprouts, were introduced into alfalfa seeds with and without a Salmonella inoculum. We examined their ability to inhibit the growth of this foodborne pathogen and assess the relative effects of the inoculants on the alfalfa microbial community structure and function. Alfalfa seeds contaminated with a Salmonella cocktail were soaked for 2 h in bacterial suspensions from each inoculant tested. Inoculated alfalfa seeds were grown for 7 days and sampled during days 1, 3, and 7. At each sampling, alfalfa sprouts were sonicated for 7 min to recover microflora from the surface, and the resulting suspensions were diluted and plated on selective and nonselective media. Total bacterial counts were obtained using acridine orange staining, and the percentage culturability was calculated. Phenotypic potential of sprout-associated microbial communities inoculated with biocontrol treatments was assessed using community-level physiological profiles based on patterns of use of 95 separate carbon sources in Biolog plates. Community-level physiological profiles were also determined using oxygen-sensitive fluorophore in BD microtiter plates to examine functional patterns in these communities. No significant differences in total and mesophilic aerobe microbial cell density or microbial richness resulting from the introduction of inoculants on alfalfa seeds with and without Salmonella were observed. P. fluorescens 2-79 exhibited the greatest reduction in the growth of Salmonella early during alfalfa growth (4.22 log at day 1), while the market sprout inoculum had the reverse effect, resulting in a maximum log reduction (5.48) of Salmonella on day 7. Community-level physiological profiles analyses revealed that market sprout communities peaked higher and faster compared with the other inoculants tested. These results suggest that different modes of actions of single versus microbial consortia biocontrol treatments may be involved.

Colony Count, Microbial↗
PubMed2006

On the lag phase and initial decline of microbial growth curves.

The lag phase is generally thought to be a period during which the cells adjust to a new environment before the onset of exponential growth. Characterizing the lag phase in microbial growth curves has importance in food sciences, environmental sciences, bioremediation and in understanding basic cellular processes. The goal of this work is to extend the analysis of cell growth curves and to better estimate the duration of the lag phase. A non-autonomous model is presented that includes actively duplicating cells and two subclasses of non-duplicating cells. The growth curves depend on the growth and death rate of these three subpopulations and on the initial proportion of each. A deterministic and a stochastic model are both developed and give the same results. A notable feature of the model is the decline of cells during the early stage of the growth curve, and the range of parameters when this decline occurs is identified. A limited growth model is also presented that accounts for the lag, exponential growth and stationary phase of microbial growth curves.

Bacteria↗
PubMed1999

Comparative efficiency of nitrocellulose membranes versus RODAC plates in microbial sampling on surfaces.

The efficiency of nitrocelluose membranes in collecting microbial samples from rigid and flat surfaces has been studied and compared with that of RODAC plates. The experimental design was based on multiple samplings in close succession in the same place. The median efficiency of membrane filters (78.55) was higher than that of RODAC plates (65.37). The data demonstrate the reliability of membranes and show that membranes are more efficient than RODAC plates as a sampling method for measuring surface microbial contamination.

Collodion↗
PubMed1998

Five-year incidence of caries, salivary and microbial conditions in 60-, 70- and 80-year-old Swedish individuals.

The 5-year incidence of dental caries in a random sample of 60-, 70- and 80-year-old inhabitants of Goteborg was related to salivary and microbial conditions. Of the 208 persons examined at baseline, 148 (71%) participated in the follow-up examination; 69, 51 and 28, respectively, in the different age groups. The study revealed that 27% of the participants had not developed any carious lesions during the 5-year period and that the incidence of coronal and root caries increased with age. In the 60-year-olds, 2.5% of the susceptible coronal and root surfaces, respectively, had decayed, while the corresponding figures for the 80-year-olds were 8.8% for coronal surfaces and 9.8% for root surfaces. In all, 18% had an unstimulated saliva secretion rate of below 0.1 ml/min and 14% had a stimulated secretion rate of <0.7 ml/min, with a mean rate which decreased with increasing age from 2.0 to 1.6 and 1.3 ml/min (p = 0.02). The overall salivary counts of lactobacilli and mutans streptococci, particularly the subspecies of Streptococcus sobrinus, had increased during the period. In the respective age groups of 60, 70 and 80 years, 15, 39 and 39% had a mutans streptococci count of > or = 10(6) CFU/ml in saliva and the corresponding figures for > or = 10(5) lactobacilli counts were 22, 31 and 43%. In the stepwise regression analysis, it was found that age, salivary levels of mutans streptococci and lactobacilli and number of teeth were the best predictors of the incidence of root caries. In conclusion, these observations indicate that there is an increased risk of dental caries with age owing to unfavourable microbial and salivary conditions.

Age Factors↗
PubMed2001

Natural attenuation potential of cyanide via microbial activity in mine tailings.

Biological removal by indigenous microflora of cyanide, contained in old (6-9 years) and fresh tailings (3 months), was studied in order to assess its natural attenuation potential via biodegradation. To investigate the presence of indigenous microflora in tailings, total heterotrophic and cyanide resistant bacteria were counted using the spread-plate method. The free cyanide mineralization potential was estimated using K14CN in the presence of various unlabeled cyanide concentrations (0, 5, and 10 mg CN/kg). The biodegradation of cyanide contained initially in the samples was also investigated by monitoring formate, formamide, ammonia and total cyanide (CNT) concentrations over 111 days. The enumeration of total heterotrophic and cyanide-resistant bacteria in old tailings showed an average population of 105 cfu/g. However, no growth was detected in fresh tailings. Nevertheless, cyanide mineralization tests indicated the presence, in both old and fresh tailings, of a cyanide-degrading microflora. In old tailings, maximum mineralization percentages of free cyanide ranging from 85% to 100% were obtained after 65 days at all concentrations tested. A mineralization percentage of 83% after 170 days was also observed in fresh tailings. No decrease of total cyanide concentration in old tailings was observed when the biodegradation of endogenous cyanide was tested whereas a significant decrease was recorded in fresh tailings after 96 days. The presence of strong metal-cyanide complexes resistant to biodegradation could explain the absence of biodegradation in old tailings. This study demonstrated the presence of an indigenous free cyanide-degrading microflora in both old and fresh tailings, and suggests that natural attenuation of cyanide in gold mine tailings is likely to occur via microbial activity.

Bacteria↗
PubMed2001

Chemostat modeling of Escherichia coli persistence in conventionalized mono-associated and streptomycin-treated mice.

We have previously shown that Escherichia coli BJ4 has similar doubling time in mice that are mono-associated (having only the inoculated E. coli BJ4) or streptomycin-treated (having mainly gram-positive bacteria plus the inoculated E. coli BJ4). We also showed that when the mice were conventionalized (fed cecum homogenate from conventional mice or ones with a complete microbial flora), the introduction of complete flora in both cases increased the in vivo doubling time, while decreasing the colony counts in fecal samples. To determine whether the increase in doubling time could explain the decrease in colony counts, we analyzed our previous results by a chemostat model. The analysis shows that the increasing doubling time alone is sufficient to explain the decrease in colony counts in mono-associated mice, but not in the streptomycin-treated mice. The observed decreasing rate in colony counts in streptomycin-treated mice is slower than predicted. Furthermore, whereas the model predicted a decrease to extinction in both mice, the E. coli persist at a frequency 10-80 times higher in streptomycin-treated mice than in mono-associated mice. Thus, while a chemostat model is able to explain some of the population dynamics of intestinal bacteria in mice, additional factors not included in the model are stabilizing the system. Because we find that E. coli declines more slowly and to a higher stabilization frequency in streptomycin-treated mice, which have a more diverse flora before conventionalization, we take these results to suggest that the persistence of E. coli populations is promoted by species diversity. We propose that a mechanism for the persistence may be the presence of new E. coli niches created by keystone species in the more diverse flora.

Animals↗
PubMed1996

Bacterial and fungal flora of dust deposits in a pig building.

OBJECTIVE: The purpose of the study was to investigate the bacterial and fungal flora of dust deposits in a newly built pig grower finisher building. Viable bacterial counts and microbial species found in a barn which had never housed pigs were compared with those in a barn housing 144 pigs. METHODS: The quantitative streak plate method was used to measure viable bacterial counts on nutrient agar or sheep blood agar. Viable bacterial counts of the dust deposits were expressed as the number of colony forming units (CFUs)/mg of dust. Gram positive cocci and Gram negative bacilli were identified by an automated system. Identifications with a confidence interval > 90% were accepted at the species level. Fungi were identified to the genus level with slide culture preparations on cereal agar. RESULTS: The lowest viable bacterial count (4.8 x 10(4)/mg of dust) was found in the barn with no pigs. In the barn with pigs the highest viable bacterial count (2.1 x 10(6)/mg of dust) was in dust from the top of a partition close to pig activity. Six species of bacteria or fungi were found in dust from the room with no pigs, whereas 22 different microorganisms were detected in dust from the room with pigs. With the exception of Enterobacter agglomerans no other species of the family Enterobacteriaceae was found in dust deposits in this new pig building. Twelve species of Gram positive bacteria were found in the room housing pigs. CONCLUSIONS: The pig is not only a source but also a disperser of airborne bacteria in pig buildings. Speciation of the microbial flora in dust from the pig building suggests that many of the microorganisms were either of human or environmental origin. Nevertheless as some of these microorganisms are known opportunistic pathogens or allergens and because of the documented increased incidence of chronic respiratory symptoms in pig workers, precautions to reduce inhalation of microbial or dust particles by pig workers seem prudent.

Animals↗
PubMed2001

A preliminary investigation of the microbiology and endotoxin content in the water reservoirs of benchtop non-vacuum autoclaves.

UNLABELLED: To determine the microbial content and endotoxin concentration in the water reservoirs of benchtop autoclaves used in general dental practice. DESIGN: The study was done in two stages. Firstly water samples were taken daily from the reservoirs of 20 autoclaves used in general dental practices for six days. The microbial content and endotoxin concentration was determined. Second the practitioners were instructed to wash and clean the reservoirs of the autoclaves each morning prior to refilling with fresh water. The reservoirs were sampled in the evenings after a day's use and the microbial content and endotoxin concentration determined. RESULTS: The total viable count of bacteria before cleaning varied from 2,300 - 8 x 10(4) cfu/ml and after from 0-40 cfu/ml. The endotoxin concentrations before cleaning varied from 360-2,200 EU/ml and after 0-80 EU/ml. CONCLUSIONS: The reservoirs of non-vacuum benchtop autoclaves can become severely contaminated with micro-organisms particularly Gram-negative bacteria. The endotoxin content of the reservoirs can get to high and unacceptable concentrations. Draining the water reservoir and cleaning on a daily basis reduces the microbial content and the endotoxin concentration to acceptable levels.

Colony Count, Microbial↗
PubMed1995

Effects of surgical treatment on the microbial flora in residual periodontal pockets.

The aim of this study was to investigate the effects of modified Widman flap surgery on the microbial flora in deep residual periodontal pockets remaining after non-surgical treatment. Micro-organisms from samples of subgingival plaque obtained from adult periodontitis sites in six patients were cultured aerobically and anaerobically, on a variety of media and identified. Non-surgical treatment produced little clinical improvement and had little effect on the total microbial count. Subsequent modified Widman flap surgery resulted in significant clinical improvements which were accompanied by an overall reduction in microbial counts. Regular reviews for scaling and oral hygiene maintained the clinical improvements and balance in favour of beneficial microbial species during the 1 year of the study.

Adult↗
PubMed2005

Monochloramine versus sodium hypochlorite as antimicrobial agents for reducing populations of bacteria on broiler chicken carcasses.

Studies were conducted to compare the effect of sodium hypochlorite (SH) versus monochloramine (MON) on bacterial populations associated with broiler chicken carcasses. In study 1, nominal populations (6.5 to 7.5 log CFU) of Escherichia coli, Listeria monocytogenes, Pseudomonas fluorescens, Salmonella serovars, Shewanella putrefaciens, and Staphylococcus aureus were exposed to sterilized chiller water (controls) or sterilized chiller water containing 50 ppm SH or MON. SH at 50 ppm eliminated all (6.5 to 7.5 log CFU) viable E. coli, L. monocytogenes, and Salmonella serovars; 1.2 log CFU of P. fluorescens; and 5.5 log CFU of S. putrefaciens. MON eliminated all (6.5 to 7.5 log CFU) viable E. coli, L. monocytogenes, S. putrefaciens, and Salmonella serovars and 4.2 log CFU of P. fluorescens. In study 2, chicken carcasses were inoculated with P. fluorescens or nalidixic acid-resistant Salmonella serovars or were temperature abused at 25 degrees C for 2 h to increase the populations of naturally occurring E. coli. The groups of Salmonella serovar-inoculated or temperature-abused E. coli carcasses were immersed separately in pilot-scale poultry chillers and exposed to tap water (controls) or tap water containing 20 ppm SH or 20 ppm MON for 1 h. The P. fluorescens-inoculated group was immersed in pilot-scale poultry chillers and exposed to tap water (controls) or tap water containing 50 ppm SH or 50 ppm MON for 1 h. Carcasses exposed to the SH treatment had nominal increases (0.22 log CFU) in E. coli counts compared with controls, whereas exposure to MON resulted in a 0.89-log reduction. Similarly, average nalidixic acid-resistant Salmonella serovar counts increased nominally by 34% (41 to 55 CFU/ml) compared with controls on carcasses exposed to SH, whereas exposure to MON resulted in an average nominal decrease of 80% (41 to 8 CFU/ml). P. fluorescens decreased by 0.64 log CFU on carcasses exposed to SH and decreased by 0.87 log CFU on carcasses exposed to MON. In study 3, SH or MON was applied to the chiller in a commercial poultry processing facility. E. coli counts (for carcass halves emerging from both saddle and front-half chillers) and Salmonella prevalence were evaluated. Data from carcasses exposed to SH during an 84-day historical (Hist) and a 9-day prepilot (Pre) period were evaluated. Other carcasses were exposed to MON and tested during a 27-day period (Test). E. coli counts for samples collected from the saddle chiller were 25.7, 25.2, and 8.6 CFU/ml for Hist, Pre, and Test, respectively. E. coli counts for samples collected from the front-half chiller were 6.7, 6.9, and 2.5 CFU/ml for Hist, Pre, and Test, respectively. Salmonella prevalence was reduced from 8.7% (Hist + Pre) to 4% (Test). These studies indicate that MON is superior to SH in reducing microbial populations in poultry chiller water.

Animals↗
PubMed2004

Granulocyte colony-stimulating factor and an in vitro whole blood model of melioidosis.

The study reported here was conducted in order to explore the mechanism of action of granulocyte colony-stimulating factor (G-CSF) in the treatment of Burkholderia pseudomallei infections (otherwise known as melioidosis). Use of G-CSF as an adjunct to antibiotics has been associated with decreasing mortality among patients with melioidosis in the tropical region of the Northern Territory of Australia. However, using an in vitro whole blood assay, no significant difference was detected in the bactericidal activity of samples obtained from dialysis patients, patients with type 2 diabetes mellitus and healthy controls, and there was no improvement following coincubation with G-CSF.

Australia↗
PubMed2006

Confirmation and prevention of intestinal barrier dysfunction and bacterial translocation caused by methotrexate.

When intestinal barrier function is damaged bacterial translocation (BT) can occur. The injury to intestinal barrier function caused by chemotherapy has been investigated in some studies, however, definitive evidence of BT caused by chemotherapy is lacking. The aim of this study was to investigate small intestinal barrier dysfunction and BT and to evaluate the preventive effect of granulocyte colony-stimulating factor (G-CSF) on intestinal barrier dysfunction and BT in a rat model of chemotherapy. Sprague-Dawley rats were treated with methotrexate (MTX; 3.5 mg/kg) for 3 days to induce intestinal barrier dysfunction and BT, gavaged Escherichia coli TG1 labeled with green fluorescent protein for 2 days to track BT, and G-CSF (10 microg/kg) for 4 days to prevent intestinal barrier dysfunction and BT. Intestinal permeability was measured by the urinary excretion rate of lactulose and mannitol following administration by gavage. Representative tissue specimens from the mesenteric lymph nodes, spleen, liver, and kidney were aseptically harvested for bacteria culture in ampicillin-supplemented medium. Light microscopy was performed on intestinal samples. MTX induced significant mucosal and villous atrophy in ileum and significantly increased intestinal permeability. MTX also induced noticeable BT. G-CSF significantly increased the mucosal thickness and villous height of the ileum and decreased intestinal permeability and BT. In conclusion, MTX caused intestinal barrier dysfunction and BT, and G-CSF prevented intestinal barrier dysfunction and BT.

Animals↗
PubMed2004

Anaerobic biodegradation of no. 2 diesel fuel in soil: a soil column study.

Soil and sediments are contaminated with petroleum hydrocarbons in many parts of the world. Anaerobic degradation of petroleum hydrocarbon is very relevant in removing oil spills in the anaerobic zones of soil and sediments. This research investigates the possibility of degrading no. diesel fuel under anaerobic conditions. Anaerobic packed soil columns were used to simulate and study in situ bioremediation of soil contaminated with diesel fuel. Several anaerobic conditions were evaluated in soil columns, including sulfate reducing, nitrate reducing, methanogenic, and mixed electron acceptor conditions. The objectives were to determine the extent of diesel fuel degradation in soil columns under various anaerobic conditions and identify the best conditions for efficient removal of diesel fuel. Diesel fuels were degraded significantly under all conditions compared to no electron supplemented soil column (natural attenuation). However, the rate of diesel degradation was the highest under mixed electron acceptor conditions followed in order by sulfate reducing, nitrate reducing, and methanogenic conditions. Under mixed electron acceptor condition 81% of diesel fuel was degraded within 310 days. While under sulfate reducing condition 54.5% degradation of diesel fuel was observed for the same period. This study showed evidence for diesel fuel metabolism in a mixed microbial population system similar to any contaminated field sites, where heterogeneous microbial population exists.

Anaerobiosis↗
PubMed2006

Intramuscular immunization with a monogenic plasmid DNA tuberculosis vaccine: Enhanced immunogenicity by electroporation and co-expression of GM-CSF transgene.

Plasmid DNA vaccine has been widely explored for tuberculosis immunization but there is a need to develop the ways to improve its immunogenicity. In this study, we have constructed a plasmid DNA vaccine coding for Ag85A alone or for both Ag85A and GM-CSF and investigated the immune adjuvant effects of electroporation and GM-CSF co-expression, alone or in combination, on CD4 and CD8 T cell IFN-gamma responses, CTL activities and immune protection from pulmonary Mycobacterium tuberculosis challenge in a Balb/c mouse model. We have found that use of electroporation allows a single intramuscular (i.m.) DNA injection to be as effective as repeated i.m. DNA injections in activation of both Ag85A-specific CD4 and CD8 T cells. Co-expression of immune-enhancing cytokine GM-CSF by the same plasmid DNA TB vaccine could further enhance T cell activation including CTL activities on top of electroporation. With regard to immune protection from pulmonary M. tb challenge, use of electroporation also allows a single i.m. DNA injection to be as effective as repeated i.m. DNA injections. Co-expression of GM-CSF transgene also moderately enhances immune protection and such effect is more evident for systemic protection. However, GM-CSF expression has little added effect on immune protection by electroporation-aided immunization protocols. Our findings thus will help with the development of future DNA TB immunization strategies.

Adjuvants, Immunologic↗
PubMed2003

The influence of process parameters for the inactivation of Listeria monocytogenes by pulsed electric fields.

The influence of the electric field strength, the treatment time, the total specific energy and the conductivity of the treatment medium on the Listeria monocytogenes inactivation by pulsed electric fields (PEF) has been investigated. L. monocytogenes inactivation increased with the field strength, treatment time and specific energy. A maximum inactivation of 4.77 log(10) cycles was observed after a treatment of 28 kV/cm, 2000 micros and 3490 kJ/kg. The lethal effect of PEF treatments on L. monocytogenes was not influenced by the conductivity of the treatment medium in a range of 2, 3 and 4 mS/cm when the total specific energy was used as a PEF control parameter. A mathematical model based on the Weibull distribution was fitted to the experimental data when the field strength (15-28 kV/cm), treatment time (0-2000 micros) and specific energy (0-3490 kJ/kg) were used as PEF control parameters. A linear relationship was obtained between the log(10) of the scale factor (b) and the electric field strength when the treatment time and the total specific energy were used to control the process. The total specific energy, in addition to the electric field strength and the treatment time, should be reported in order to evaluate the microbial inactivation by PEF.

Colony Count, Microbial↗
PubMed2004

Impairment of growth of Listeria monocytogenes in THP-1 macrophages by granulocyte macrophage colony-stimulating factor: release of tumor necrosis factor-alpha and nitric oxide.

BACKGROUND: Listeria monocytogenes tends to survive in phagocytes. Granulocyte macrophage colony-stimulating factor (GM-CSF) protects mice against L. monocytogenes infection, and mice knocked out for the GM-CSF gene are more susceptible to these infections. METHODS: THP-1 cells were used to characterize the GM-CSF receptor (binding isotherms; STAT5 phosphorylation), measure the intracellular growth of L. monocytogenes (5 h after phagocytosis), examine the influence of a 24-h incubation with GM-CSF before infection, measure the production of tumor necrosis factor (TNF)-alpha and the expression of nitric oxide synthase (iNOS), and evaluate the influence of anti-GM-CSF receptor (GM-CSFR alpha ) and anti-TNF-alpha antibodies and the addition of N(omega)-nitro-L-arginine methyl ester (L-NAME) and catalase. RESULTS: THP-1 cells display functional GM-CSFR alpha. GM-CSF impairs the intracellular growth of L. monocytogenes to approximately 65% of its value in unstimulated cells. This effect is abolished by anti-GM-CSFR alpha, anti-TNF-alpha antibodies, and catalase (and, to a lesser extent, by L-NAME). GM-CSF stimulates the release of TNF-alpha and the expression of iNOS. TNF-alpha added to unstimulated cells (even in large amounts) does not fully reproduce the impairment in the growth of L. monocytogenes caused by GM-CSF. CONCLUSIONS: GM-CSF impairs the intracellular growth of L. monocytogenes by a synergistic action of the GM-CSF-triggered release of autocrine TNF-alpha and hydrogen peroxide and the production of NO (associated with the stimulation of the expression of iNOS).

Blotting, Western↗