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Autoproteolysis rate of rainbow trout muscle proteins.

Effect of some physicochemical agents (pH, temperature and incubation time) and iced storage of farmed rainbow trout (Oncorhynchus mykiss) on autoproteolysis of muscle proteins and quality (sensory assessment, instrumental texture, total bacterial counts) were determined. The results showed that the autoproteolysis rate proceeded with its highest intensity at pH 4.2 and temperature about 55 degrees C. The rate of protein hydrolysis and protease activity, as well as microbial contamination, gradually increased with prolonged periods of iced storage of fish. Properly iced trout immediately after its evisceration was acceptable in quality for up to 10 days.

Animals↗

Antibacterial effect of crude water-soluble arrowroot (Puerariae radix) tea extracts on food-borne pathogens in liquid medium.

AIMS: To evaluate the effect of crude water-soluble arrowroot tea extracts on microbial growth of food-borne pathogens in liquid medium and to confirm the damage to bacterial cells using Transmission Electronic Microscopy (TEM). METHODS AND RESULTS: Inhibition of growth of Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Listeria monocytogenes and Staphylococcus aureus was investigated using Brain Heart Infusion (BHI) broth containing 0 (control), 0.63, 1.25, 2.5 and 5.0% (w/v) arrowroot tea. Bacterial cell counts were performed on specific selective agar on days 0, 1, 3 and 5. BHI containing 5.0% arrowroot tea extract showed a 6-7 log suppression of growth for all test strains on days 3 and 5, compared with the control. Even 0.63% arrowroot tea effectively inhibited microbial growth of all test strains on day 5. TEM images of the samples treated with 5.0% arrowroot tea revealed the rupture of cell walls and nonhomogeneous disposition of cytoplasmic materials within treated bacteria. CONCLUSIONS: Crude water-soluble arrowroot tea extract strongly inhibited microbial growth of all test pathogens in liquid medium. SIGNIFICANCE AND IMPACT OF THE STUDY: Water-soluble arrowroot tea extract has the potential to be used directly on foods or as a spray on the surfaces of food handling and processing facilities in order to prevent microbial growth of both Gram-negative and Gram-positive bacteria.

Anti-Bacterial Agents↗

Effects of defaunation and various nitrogen supplementation regimens on microbial numbers and activity in the rumen of sheep.

Five sheep (average BW 62 kg) were fed 65% roughage: 35% concentrate diets (CP = 15%) in a 5 x 5 Latin square design to study the effects of combinations of defaunation and N supplements (soybean meal [SBM], corn gluten meal [CGM], blood meal [BM], urea, and casein) differing in ruminal degradation on ruminal microbial numbers and activity. Diets were fed twice daily (DM intake 1,759 g/d). Defaunation was accomplished with doses of 30 ml of alkanate 3SL3.sheep-1.d-1 for 3 d with 2 d of fasting. Treatment 1 (control) involved feeding faunated sheep a diet in which the supplemental N was 67% SBM N and 33% urea N. Treatment 2 involved feeding defaunated sheep the same diet as the control. Treatments 3, 4, and 5 involved feeding defaunated sheep diets in which the supplemental N source was either 67% CGM-BM N (CGM and BM combined on a 1:1 N ratio): 33% urea N, or 33% CGM-BM N:67% urea N or 33% CGM-BM N:33% urea N:33% casein N, respectively. Compared with the faunated control, defaunation (Treatments 2, 3, 4, and 5) increased (P less than .05) total direct counts of ruminal bacteria (2.7 vs 1.3 x 10(11)/ml), fungal zoospores (2.8 vs 1.4 x 10(5)/ml), and ruminal microbial protease activity (1.4 vs 1.0 mg azocasein/[ml ruminal fluid.h]). Defaunation did not have a consistent effect on ruminal microbial deaminase activity. Compared with the control, defaunation resulted in lower (P less than .05) total perchloric acid-soluble amino N in ruminal fluid at 4 and 10 h after the morning feeding. Defaunation did not decrease (P greater than .05) total free amino acid concentrations in ruminal fluid, but it altered the profile of free amino acids. Although defaunation increased (P less than .05) ruminal bacterial numbers, no increases in total microbial CP or OM concentrations in ruminal contents were observed.

Amino Acids↗

Non-lactic acid, contaminating microbial flora in ready-to-eat foods: a potential food-quality index.

The bacteriological profile of 87 samples of commercially available ready-to-eat (RTE) dairy and meat-products, packaged sandwiches and salads was obtained by testing for aerobic colony count, for lactic acid bacterial (LAB) count, for the presence and the extent of non-LAB microflora (contaminating microflora), and by testing for certain food-borne pathogens. The pathogens Listeria monocytogenes, Salmonella spp. and sulfite-reducing clostridia were not detected in any of the analysed samples. Whereas only three samples (3.4%) were deemed unacceptable for consumption for exceeding the established pathogen tolerance levels (for Staphylococcus aureus and Escherichia coli), several samples were found to contain non-lactic acid contaminating microflora of considerable magnitude. The log10 cfu g(-1) counts for contaminating microflora in the food categories examined were as follows: hard cheeses 4.85 (SD 1.17); semi-hard cheeses 5.39 (SD 1.37); soft cheeses 5.13 (SD 1.03); whey cheeses 6.55 (1.24); fermented meat-products 4.18 (SD 1.48); heat-treated meat-products 3.47 (SD 1.99); salads 3.37 (SD 1.56) and sandwiches 5.04 (SD 0.96). Approximately 1 in every 30 to 80 bacterial cells found on different types of cheeses and salads was a non-LAB microorganism; the respective ratios for fermented meat-products, heat-treated meat-products and sandwiches were 1 in 6, 2.5 and 15. The assessment of the contaminating microflora magnitude at various steps during the manufacture and distribution of RTE foods can serve as an index for monitoring the microbiological quality of the starting materials, the sanitation efficacy during processing and possible temperature abuse during processing, transportation or storage.

Colony Count, Microbial↗

The survival and recovery of bacteria in vacuum cleaner dust.

The possibility exists that environmental dust could be a source of gastro-intestinal infection in the domestic environment and that the causative microbes are collected during vacuum cleaning. This study examines the survival of total bacterial populations, Enterobacteriaceae and salmonella species in vacuum cleaner dust in vitro and in use. Total counts remain constant at around 10(6)-10(8) colony forming units (cfu) g-1 for at least 60 days. Enterobacteriaceae showed only a slight decline over the same period. Recovery of salmonellae artificially inoculated into vacuum dust was dose and time dependent: even relatively small inocula (< 100 cfu g-1) were recoverable more than one month after inoculation. Questionnaires and vacuum cleaner dust samples were received from 76 households. A significant association was found between total bacterial counts and whether the household was in a rural or urban situation and the number of people in the household. No association was found between total bacterial counts and the number of children in the household, the presence of pets, wearing shoes indoors, the makes, model and age of the vacuum cleaner, the type of dust collector or dust bag, how often the vacuum cleaner was used and the length of time between sample collection and microbiological testing. Salmonella species were isolated from vacuum cleaner dust from three of the 76 households although no factors could be identified linking the homes. This study shows that vacuum cleaners are effective collectors and reservoirs of microbial contamination and that these contaminants are able to survive for up to two months. Vacuum cleaner dust could thus be a useful indicator of environmental contamination in the home.

Colony Count, Microbial↗

Enhancement of humoral and protective immune response induced by live-attenuated Salmonella typhi by ampicillin.

Efficacy of live-attenuated Salmonella vaccines delivered by the mucosal route is limited by the dose and interference from mucosal flora of the alimentary tract. In a mouse model, the total antibody response towards lipopolysaccharide of S. typhi was significantly enhanced at day 21 post-immunization with live-attenuated S. typhi (Ty21a) when ampicillin was concomitantly administered (p<0.005), and the lethal dose 50 of mice in the ampicillin and control groups immunized with Ty21a after wild-type S. typhi challenge on day 24 was 4x10(7) and 1x10(7), respectively. The faecal bacterial counts of the ampicillin group at days 1 and 3 were significantly lower than those of the control group (p<0.01 and <0.05). On day 1, the number of mice with > or =10 Ty21a colonies isolated from the spleen was significantly higher in the ampicillin group than the control group (p<0.05). Furthermore, on the same day, Ty21a was isolated from the faeces of three mice from the ampicillin group, but only one from the control group. We conclude that ampicillin may have enhanced the humoral and protective immune responses by giving the Ty21a a selective advantage over the normal bacterial flora. This concept of antibiotic enhancement of immunization could have important implications for other live-attenuated vaccines, as well as the delivery of microbial antigens and DNA vaccines by live-attenuated Salmonella carriers.

Ampicillin↗

Influence of human wet cerumen on the growth of common and pathogenic bacteria of the ear.

The available data on the effect of human wet cerumen on bacterial growth are not conclusive. Nevertheless it is widely accepted that cerumen has a bactericidal effect. In this study the activity of human wet cerumen on bacterial growth was assessed by applying cerumen suspensions to bacterial cultures. Bacterial counts were performed before and after application of cerumen suspensions. A total of 383 assays was carried out with 73 pools of cerumen that were tested against cultures of Staphylococcus aureus, Staphylococcus epidermidis, Corynebacterium spp., Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa and Serratia marcescens. An increase in growth occurred much more frequently than a decrease in growth in almost every microrganism tested, with the mean increase percentage being much higher than the mean decrease percentage, except in the case of S. aureus. The largest average growth increase was obtained with E. coli. The largest average decrease in bacterial growth was recorded with S. marcescens. Our study does not support the conception of a decrease in bacterial growth produced by humen wet cerumen. In vitro, the most observable effect was in fact an increase in microbial growth.

Adolescent↗

Use of microelectrodes to investigate the effects of 2-chlorophenol on microbial activities in biofilms.

In order to assess the applicability of using microelectrodes as a tool for inhibition tests, temporal and spatial inhibitory effects of 2-chlorophenol (2-CP) on O(2) respiration and nitrification activities in municipal wastewater biofilms were investigated using microelectrodes for O(2) and NH(4)(+). The time-course microelectrode measurements demonstrated that 2-CP inhibited O(2) respiration and nitrification activities within 6-18 min. The microbial activities were inhibited only in the upper 400 microm of the biofilms by 2-CP, and the bacteria present in the deeper parts of the biofilms were still active, probably due to limited penetration of 2-CP. These results could reasonably explain the difference in inhibitory ratios of the O(2) respiration and nitrification activities in the biofilms. O(2) respiration activity was incompletely inhibited, which was attributed to the presence of O(2) respiration activities in the deeper parts of the biofilm. In contrast, nitrification activity was significantly inhibited because ammonia-oxidizing bacteria were present in the upper parts of the biofilm. These results indicate that the microelectrodes with a very quick response time and a high spatial resolution are useful tools to study temporal and spatial inhibitory effects of inhibitors on in situ microbial activities in biofilms.

Bacterial Physiological Phenomena↗

The effect of clindamycin gel insert in periodontal pockets, as observed on smears and cultures.

This study is aimed at the evaluation of a 1% clindamycin hydrochloride containing gel on the microbial flora of periodontal pockets deeper than 5 mm. In order to achieve that purpose, 20 patients with pocketing in the premolar-molar regions were selected. Active and placebo gel were inserted once during the first 2 weeks of this experimental study. Microbial samplings were performed 1, 2, 4 and 12 weeks after the experiment started. The samples were submitted to microscopic examination and also to culture. Changes in the microbial content of the periodontal pockets treated by subgingival scaling and clindamycin 1% gel were significant, compared with those obtained with subgingival scaling and placebo gel, particularly with respect to anaerobic black-pigmented bacteria and the motile gram-negative flora. However, after 3 months, most of the treated cases were recolonized by the same initial species, though never at pre-clindamycin levels. In the light of this study, it will be concluded that the use of a small amount of clindamycin hydrochloride inserted into a periodontal pocket, once a week for 2 weeks as a complement to periodontal subgingival scaling, is beneficial in the treatment of adult periodontitis, by eliminating more effectively the microbial pocket colonization.

Administration, Topical↗

Microbial assemblages in soil microbial succession after glacial retreat in Svalbard (high arctic).

Microbial community composition (cyanobacteria and eukaryotic microalgae abundance and diversity, bacterial abundance, and soil respiration) was studied in subglacial and periglacial habitats on five glaciers near Ny-Alesund, Svalbard (79 degrees N). Soil microbial communities from nonvegetated sites (subglacial, recently deglaciated, and cryoconite sediments) and sites with plant cover (deglaciated some hundreds of years ago) were analyzed. Physicochemical analyses (pH, texture, water content, organic matter, total C and N content) were also performed on the samples. In total, 57 taxa of 23 genera of cyanobacteriaand algae were identified. Algae from the class Chlorophyceae (25 species) and cyanobacteria (23 species) were richest in biodiversity. The numbers of identified species in single habitat types were 23 in subglacial, 39 inbarren, 22 in cryoconite, and 24 in vegetated soils. The highest cyanobacterial and algal biovolume and cell numbers, respectively, were present in cryoconite (13x10(4) microm3 mg-1 soil and 508 cells per mg of soil), followed by barren (5.7x10(4) and 188), vegetated (2.6x10(4) and 120), and subglacial (0.1x10(4) and 5) soils. Cyanobacteria prevailed in all soil samples. Algae (mainly green algae) were present only as accessory organisms. The density of bacteria showed a slightly different trend to that of the cyanobacterial and algal assemblages. The highest number of bacteria was present in vegetated (mean: 13,722x10(8) cells per mg of soil dry wt.), followed by cryoconite (3802x10(8)), barren (654x10(8)), and subglacial (78x10(8)) soils. Response of cyanobacteria and algae to physical parameters showed that soil texture and water content are important for biomass development. In addition, it is shown that nitrogen and water content are the main factors affecting bacterial abundance and overall soil respiration. Redundancy analysis (RDA) with forward selection was used to create a model explaining variability in cyanobacterial, algal, and bacterial abundance. Cryoconites accounted for most of the variation in cyanobacteria and algae biovolume, followed by barren soils. Oscillatoriales, desmids, and green coccoid algae preferred cryoconites, whereas Nostocales and Chroococcales occurred mostly in barren soils. From the data obtained, it is evident that of the studied habitats cryoconite sediments are the most suitable ones for the development of microbial assemblages. Although subglacial sediments do not provide as good conditions as cryoconites, they support the survival of microbial communities. Both mentioned habitats are potential sources for the microbial recolonization of freshly deglaciated soil after the glacier retreat.

Arctic Regions↗

Diversity of Saccharomyces strains on grapes and winery surfaces: analysis of their contribution to fermentative flora of Malbec wine from Mendoza (Argentina) during two consecutive years.

Spontaneous fermentations are still conducted by several wineries in different regions of Argentina as a common practice. Native Saccharomyces strains associated with winery equipment, grape and spontaneous fermentations of Malbec musts from "Zona Alta del Río Mendoza" region (Argentina) were investigated during 2001 and 2002 in the same cellar. Low occurrence of Saccharomyces on grapes and their limited participation during fermentation were confirmed. Strain sequential substitution during fermentation was observed. Between 30% and 60% of yeast population at the end of fermentation was coming from yeasts already present in the winery. A stable and resident Saccharomyces micro-flora in the winery was confirmed. It exhibited a dynamic behaviour during season and between years. Commercial strains were found during fermentation in different percentages, but their presence on winery equipment was low. The present work represents a first approach to winery yeast and spontaneous fermentation Saccharomyces population dynamics in an important viticultural region from Argentina that has never been characterized before. The results obtained have an important significance for the local industry, showing for the first time the real situation of the microbial ecology of alcoholic fermentation in an industrial winery from Mendoza, Argentina.

Argentina↗

Role of discontinuous chlorination on microbial production by drinking water biofilms.

Microbial quality in water distribution systems is strongly affected by the development of microbial biofilms. Production and release of microbial cells by the biofilm affect microbial levels in the water column and in some cases this fact constitutes a public health concern. In this study, we attempt to analyze in which way the existence of different episodes of chlorine depletion affects both biofilm formation and microbial load of an artificial laboratory system. The work was carried out using two parallel packed bed reactors both supplied with running tap water. One of the reactors was used as a control and was permanently exposed to the action of chlorine. In the other reactor, chlorine was neutralized at selected times during the experiment and for periods of variable length. During the experiment the concentration of total and viable cells from the effluent was monitored at the exit of each of the reactors. The data obtained were used to estimate microbial production from the biofilms. As an average, release of microbial cells to the water phase increased tenfold in the absence of chlorine. The results also indicate that disinfectant efficiency against the biofilm was not recovered when chlorine returned to normal levels after each event of chlorine neutralization. Cell viability in the water phase in the presence of chlorine was low at the beginning of the experiment but increased 4 orders of magnitude after five neutralization periods. Therefore, subsequent episodes of chlorine depletion may accelerate the development of microbial communities with reduced susceptibility to disinfection in real drinking water systems.

Biofilms↗

Bacterial colonization of the internal part of two-stage implants. An in vivo study.

This study aimed to investigate the presence of microorganisms in the inner thread of the fixture (endosteal implant part) of the Brånemark system. In 9 volunteers the apical part of 2 abutment screws that had been in place for 3 months was examined by means of differential phase-contrast microscopy. All screws harboured a significant quantity of microorganisms, mainly coccoid cells (86.2%) and nonmotile rods (12.3%). Motile organisms (1.3%) or spirochetes (0.1%) were only sporadically registered. A microbial leakage at the abutment/fixture interface probably is the most probable origin for this contamination.

Bacteria↗

Chairside sensor for rapid monitoring of Enterococcus faecalis activity.

In this study, optical spectroscopy was used to monitor a chromogenic, enzyme-substrate reaction for the rapid identification of Enterococcus faecalis. The detection system, comprising a miniature spectrophotometer and an accompanying data acquisition system, was placed in an incubator. During testing, a 3-ml test sample was placed in a cuvette within the spectrophotometer. This permitted online, real-time, and remote analysis of spectral signature needed to monitor the bacteria. It was observed that the absorption peak intensity increased conspicuously 3.5 h after inoculation and through the entire period of testing. A linear-regression analysis demonstrated a significant correlation between the increase in absorption peak intensity at 610 nm (r = 0.9389) and 653 nm (r = 0.9387) with the formation of colony-forming units. Optical spectroscopy-based sensing systems can pave the way for rapid, nonlaboratory-based approaches to monitor microbial status quantitatively and qualitatively from clinical samples.

Biosensing Techniques↗

Symbiotic bacteria direct expression of an intestinal bactericidal lectin.

The mammalian intestine harbors complex societies of beneficial bacteria that are maintained in the lumen with minimal penetration of mucosal surfaces. Microbial colonization of germ-free mice triggers epithelial expression of RegIIIgamma, a secreted C-type lectin. RegIIIgamma binds intestinal bacteria but lacks the complement recruitment domains present in other microbe-binding mammalian C-type lectins. We show that RegIIIgamma and its human counterpart, HIP/PAP, are directly antimicrobial proteins that bind their bacterial targets via interactions with peptidoglycan carbohydrate. We propose that these proteins represent an evolutionarily primitive form of lectin-mediated innate immunity, and that they reveal intestinal strategies for maintaining symbiotic host-microbial relationships.

Animals↗

Indigenous bacteria with antagonistic and plant-growth-promoting activities improve slow-filtration efficiency in soilless cultivation.

In tomato soilless culture, slow filtration allows one to control the development of diseases caused by pathogenic microorganisms. During the disinfecting process, microbial elimination is ensured by mechanical and biological factors. In this study, system efficacy was enhanced further to a biological activation of filter by inoculating the pozzolana grains contained in the filtering unit with 5 selected bacteria. Three strains identified as Pseudomonas putida and 2 as Bacillus cereus came from a filter whose high efficiency to eliminate pathogens has been proven over years. These 5 bacteria displayed either a plant growth promoting activity (P. putida strains) or antagonistic properties (B. cereus strains). Over the first months following their introduction in the filter, the bacterial colonisation of pozzolana grains was particularly high as compared to the one observed in the control filter. Conversely to Bacillus spp. populations, Pseudomonas spp. ones remained abundant throughout the whole cultural season. The biological activation of filter unit very significantly enhanced fungal elimination with respect to the one displayed by the control filter. Indeed, the 6-month period needed by the control filter to reach its best efficacy against Fusarium oxysporum was shortened for the bacteria-amended filter; in addition, a high efficacy filtration was got as soon as the first month. Fast colonization of pozzolana grains by selected bacteria and their subsequent interaction with F. oxysporum are likely responsible for filter efficiency. Our results suggest that Pseudomonas spp. act by competition for nutrients, and Bacillus spp. by antibiosis and (or) direct parasitism. Elimination of other fungal pathogens, i.e., Pythium spp., seems to differ from that of Fusarium since both filters demonstrated a high efficacy at the experiment start. Pythium spp. elimination appears to mainly rely on physical factors. It is worth noting that a certain percentage of the 5 pozzolana-inoculated bacteria failed to colonise the filter unit and were, thus, driven to the plants by the nutrient solution. Their contribution to the establishment of a beneficial microbial community in the rhizosphere is discussed.

Antibiosis↗

Total RNA concentration as an index of microbial activity and oxygen supply in an oxidation ditch.

Total RNA and chromosomal DNA concentrations at a municipal wastewater treatment plant with an oxidation ditch (OD) were monitored for 1.5 years using commercial extraction kits for DNA and RNA. No parameters correlated with the chromosomal DNA concentration. The total RNA concentration exhibited better correlation than the solids retention time and the mixed liquor suspended solids with the removal rate of total organic carbon, and can be regarded as an index of microbial activity. The total RNA concentration varied with a cycle of one year and increased at lower water temperatures in this OD. When diffusion theory was taken into account, it was found that the oxygen dissolution rate increased at lower temperature, and a small change in the oxygen dissolution rate caused a large variation in microbial activity and also affected nitrification and denitrification. The information was insufficient to clarify the various reaction relationships, but total RNA concentration will likely be useful as an index of microbial activity in actual wastewater treatment reactors.

Bacteria, Aerobic↗

Effect of hot water and hydrogen peroxide treatments on survival of salmonella and microbial quality of whole and fresh-cut cantaloupe.

Cantaloupe melon has been associated with outbreaks of salmonellosis. Contamination might be introduced into the flesh from the rind by cutting or by contact of cut pieces with contaminated rinds. Our objectives were to investigate the efficacy of hot water or hot 5% hydrogen peroxide treatments in reducing the population of native microflora and inoculated Salmonella on cantaloupe rind and transfer to fresh-cut tissue during cutting. Whole cantaloupes, inoculated with a cocktail of Salmonella serovars to give 4.6 log CFU/cm2 and stored at 5 or 20 degrees C for up to 5 days, were treated with hot water (70 or 97 degrees C) or 5% hydrogen peroxide (70 degrees C) for 1 min at 0, 1, 3, or 5 days postinoculation. Aerobic mesophilic bacteria and yeast and mold on treated whole melon and fresh-cut pieces were significantly (P < 0.05) reduced by all three treatments. Treatments with hot water (70 and 97 degrees C) caused a 2.0- and 3.4-log CFU/cm2 reduction of Salmonella on whole cantaloupe surfaces irrespective of days of postinoculation storage prior to treatment up to 5 days at 5 or 20 degrees C, respectively. Treatment with 5% hydrogen peroxide (70 degrees C) caused a 3.8-log CFU/cm2 reduction of Salmonella. Fresh-cut pieces prepared from untreated inoculated melons and those treated with 70 degrees C hot water were positive for Salmonella. However, fresh-cut pieces prepared from inoculated whole melon dipped in water (97 degrees C) or hydrogen peroxide (70 degrees C) for 60 s were negative for Salmonella, as determined by dilution plating onto agar medium, but were positive after enrichment at days 3 and 5 of storage at 5 degrees C. The ability to detect Salmonella in fresh-cut pieces was dependent on the initial level of inoculation. The results of this study indicate that the use of hot water (97 degrees C) or heated hydrogen peroxide to reduce the population of Salmonella on contaminated whole cantaloupes will enhance the microbial safety of the fresh-cut product.

Anti-Infective Agents, Local↗