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Improvement of garri quality by the inoculation of microorganisms into cassava mash.

Lactobacillus delbruckii, Lactobacillus coryneformis, and a Saccharomyces sp., previously found among 214 isolates to be the highest producers of linamarase, amylase, and lysine were inoculated separately or mixed into cassava mash and fermented with, or without dewatering, for 24, 48, 72, 96 and 120 h. At the end of the fermentation period, the mash was converted to garri by heating over a gas burner. The mash and the garri resulting from it were assessed for lysine and residual cyanide, while the garri was also studied for its organoleptic properties. The inoculation of the microorganisms into cassava mash produced a sharp drop in the cyanide content of the mash, particularly when the mixture of organisms was inoculated into undewatered cassava mash rather the single organisms. After 24 h, while the cyanide content of the control (uninoculated mash) mash was 3.06 microg/g in the dewatered mash, and 4.24 microg/g in the undewatered mash, the inoculation of a mixture of the three organisms caused the cyanide content in the inoculated mashes to drop by 150% to 1.96 microg/g (dewatered mash) and by 300% to 1.43 microg/g (undewatered mash). It also appears that the process of producing the garri from the mash by heating, caused a further reduction of the cyanide content: thus the garri always contained less cyanide than the mash from which it was made. The lysine content of the mash was also highest when all three organisms were mixed; it also tended to increase with increasing length of fermentation of the mash. Whereas the single organism most effective in reducing the cyanide content of mash was Lactobacillus delbruckii, in the case of lysine production, it was the yeast. The organoleptic properties of the garri which were assessed were flavour, texture, colour and general acceptability. In general these properties were superior at the P < 0.01 level in garri made from the undewatered mash in comparison with that from dewatered mash, especially when the mixture of organisms was used. The inoculation of the mixture of the three organisms produced a dramatic reduction in the time (24 h) taken for the highest 'general acceptability' score to be given by the tasters when compared with the singly inoculated organisms and the control, which attained this characteristic after about 96 h of incubation in mash.

Cyanides↗

Concentration of carbon dioxide in the water-phase as a parameter to model the effect of a modified atmosphere on microorganisms.

The effect of modified atmosphere packaging can mainly be attributed to the bacteriostatic action of CO2. The dissolved CO2 in the water-phase of a food product is strongly dependent on several intrinsic and extrinsic parameters and will determine the effectiveness of a modified atmosphere packaging configuration. The effect of pH, gas/product ratio, initial %CO2 in the gas-phase, lard content and storage temperature on the amount of dissolved CO2 was screened in a preliminary experiment. The initial CO2-concentration in the gas-phase and the gas/product ratio turned out to be the two major factors determining the amount of dissolved CO2. The initial pH also determined significantly the final CO2-concentration in the broth. Temperature and lard content were shown to have only a minor effect on the amount of dissolved CO2 compared to the above mentioned parameters. This demonstrates the importance of the packaging configuration in the effectiveness of a modified atmosphere. In a second step, a model was constructed to predict the amount of dissolved carbon dioxide in modified BHI-broth as a function of the gas/product ratio, the initial CO2-concentration and the temperature by means of Response Surface Methodology (RSM). A second equation was also derived based on Henry's law and was shown to be a powerful tool in the quantification of the effect of intrinsic and extrinsic parameters on the CO2-solubility in food products. The possibility of the use of the concentration of dissolved CO2 in the water-phase as a determinative factor for the inhibitory effect of modified atmospheres was examined on Pseudomonas fluorescens. Growth curves at 7 degrees C of P. fluorescens in different packaging configurations (initial %CO2 and gas/product ratio) resulting in equal amounts of dissolved CO2 were compared. P. fluorescens was shown to be similarly inhibited by equal amounts of dissolved CO2-concentrations, independent of the packaging configuration. This demonstrates the potential of the application of the concentration of dissolved CO2 in the water-phase as a parameter to characterise a modified atmosphere and its inhibition of certain microorganisms.

Carbon Dioxide↗

Methodology for detection and typing of foodborne microorganisms.

Over the past decade many improvements have been seen in both conventional and modern methods for the detection of pathogenic bacteria in foods. Modifications and automation of conventional methods in food microbiology include sample preparation, plating techniques, counting and identification test kits. ATP bioluminescence techniques are increasingly used for measuring the efficiency of cleaning surfaces and utensils. Cell counting methods, including flow cytometry and the direct epifluorescent filter technique are suitable techniques for rapid detection of microorganisms, especially in fluids. Automated systems based on impedimetry are able to screen high numbers of samples based on total bacterial counts within 1 day. Immunoassays in a wide range of formats make rapid detection of many pathogens possible. Recently, there have been important developments in the use of nucleic acid-based assays for the detection and subtyping of foodborne pathogens. The sensitivity of these methods has been significantly increased by the use of the polymerase chain reaction and other amplification techniques. Alternative and rapid methods must meet several requirements concerning accuracy, validation, speed, automation, sample matrix, etc. Both conventional and rapid methods are used within hazard analysis critical control point programs. Further improvements especially in immunoassays and genetic methods can be expected, including the use of biosensors and DNA chip technology.

Bacterial Typing Techniques↗

Nickel uptake and utilization by microorganisms.

Nickel is an essential nutrient for selected microorganisms where it participates in a variety of cellular processes. Many microbes are capable of sensing cellular nickel ion concentrations and taking up this nutrient via nickel-specific permeases or ATP-binding cassette-type transport systems. The metal ion is specifically incorporated into nickel-dependent enzymes, often via complex assembly processes requiring accessory proteins and additional non-protein components, in some cases accompanied by nucleotide triphosphate hydrolysis. To date, nine nickel-containing enzymes are known: urease, NiFe-hydrogenase, carbon monoxide dehydrogenase, acetyl-CoA decarbonylase/synthase, methyl coenzyme M reductase, certain superoxide dismutases, some glyoxylases, aci-reductone dioxygenase, and methylenediurease. Seven of these enzymes have been structurally characterized, revealing distinct metallocenter environments in each case.

Acetyl Coenzyme A↗

Detection of hydrogen peroxide produced by microorganisms on an ABTS peroxidase medium.

Previously published methods to detect the production of hydrogen peroxide by microorganisms have been of limited usefulness for routine diagnosis because the sensitivity of some of these procedures does not allow the isolation of single H2O2-producing colonies. The new medium containing 2,2'-azino-di (3-ethyl benzthiazoline-6-sulphonic acid) and horseradish peroxidase is more sensitive and faster than established methods.

Benzothiazoles↗

Resistance to the migration of microorganisms of a needle-free disinfectable connector.

The aim of this study was to assess the efficacy of a disinfectable needle-free connector in reducing the pass of micro-organisms to the lumen of the catheter. A prospective, controlled, experimental trial was performed in which a laboratory model simulated the insertion of a peripheral venous catheter. Catheters inserted in sterile conditions in a hemoculture bottle were closed with the disinfectable needle-free connector (study group) or with a cap (control group). After 9 days of contamination and manipulation of the connector and cap external surfaces, 100% of bottles in the control group were contaminated whereas 60% remained sterile in the study group. The disinfectable needle-free connector showed more resistance to the pass of microorganisms than the conventional cap according to our experimental model.

Catheterization, Central Venous↗

Levels of naturally occurring microorganisms on surgical instruments after clinical use and after washing.

Surgical instruments exposed to sterile body sites should be contaminated with relatively low levels of microbial contamination or bioburden; however, few studies in the literature have determined the quantitative level and types of contamination. A study was conducted at 2 clinical sites to determine the level of microbial contamination of surgical instruments after clinical use and after washing. Quantitative assays showed that bioburden levels were in the range of 0 to 4415 colony forming units per instrument after clinical use, and 88% of the instruments had bioburden levels lower than 1000. As expected, a reduction in counts occurred after washing; however, in some cases, higher counts were found on the instruments after the washing process. Although the washing procedure is effective in reducing the microbial levels deposited on the surgical instruments during use, a recontamination process occurs that results in increased counts after washing. The low bioburden level after washing consists of predominantly vegetative microorganisms that present a relatively low challenge to sterilization and disinfection systems.

Colony Count, Microbial↗

Antimicrobial susceptibility of microorganisms recovered from intraabdominal infections at Belo Horizonte, Brazil.

We evaluated antimicrobial susceptibility patterns of microorganisms isolated from intraabdominal infection of Brazilian patients, by agar dilution, agar diffusion, and E test. Among the strictly anaerobes, 57.7% were resistant to penicillin, 28.2% to clindamycin, and 9.9% to metronidazole. The majority of Escherichia coli and Staphylococcus were sensitive and resistant to almost all drugs, respectively. Half of Candida samples were resistant to itraconazole. Our data reinforce the importance of this kind of study to support rational antimicrobial therapy.

Anti-Infective Agents↗

Elemental characterization of microorganism granules by EFTEM in the tube wall of a deep-sea vent invertebrate.

Microorganisms colonizing the exoskeletons of the tube worm Riftia pachyptila are described at the ultrastructural level. The prokaryotic cells from the worm tube wall differ from those colonizing the exoskeleton outer surface in the presence of an electron dense granule. The morphology and distribution of these bacteria-like cells are described. Prokaryotic organisms are assembled in nodules which increased in size in the oldest part of the exoskeleton. The aspect, location and elemental composition of the intracellular granules are determined. Most of them (100 nm in diameter) are located close to the cell membrane and exhibit a homogeneous and amorphous content. EDX and EFTEM microanalyses show that these structures contain phosphorus, oxygen and iron. All together these data suggest that these granules are iron polyphosphates. These structures may act as energy sources for making ATP during anoxic conditions as existing in hydrothermal environments.

Adenosine Triphosphate↗

[Cutaneous signs of Rickettsia and related microorganisms].

INTRODUCTION: Rickettsiosis are emerging, life-threatening infectious diseases that are world-wide distributed and have various manifestations. Most of the time, skin involvement is the prominent feature of clinical manifestations. Clinicians must therefore be well aware of these various clinical pictures. CURRENT KNOWLEDGE AND KEY POINTS: Knowledge about rickettsial infections has recently improved due to either the identification of new vectors and reservoirs, the description of new systemic and cutaneous signs and symptoms, or the amelioration of diagnosis methods. Cutaneous manifestations mainly depend on the type of microorganism and on the host characteristics. In some cases, they are so typical that their only identification would lead to the diagnosis. FUTURE PROSPECTS AND PROJECTS: As growing population movements may lead to an increase in the disease incidence, detailed knowledge of clinical, particularly cutaneous, manifestations of rickettsiosis is required in order to improve both diagnosis and treatment. When identified, cutaneous manifestations of rickettsiosis allow the clinician to confirm rapidly the diagnosis, using polymerase chain reaction or immunofluorescence and to institute prompt and specific treatment, thus reducing unfavorable disease outcome.

Diagnosis, Differential↗

Experimental evaluation of the processes resulting from the introduction of the transgenic microorganism Escherichia coli Z905/pPHL7 (lux+) into aquatic microcosms.

The processes resulting from the introduction of the tranagenic microorganism (TM) E. coli Z905/pPHL7 into aquatic microcosms have been modeled experimentally. It has been shown that the TM E. coli is able to adapt to a long co-existence with indigenous heterotrophic microflora in variously structured microcosms. In more complex microcosms the numerical dynamics of the introduced E. coli Z905/pPHL7 population is more stable. In the TM populations staying in the microcosms for a prolonged time, changes are recorded in the phenotypic expression of plasmid genes (ampicillin resistance and the luminescence level) and chromosome genes (morphological and physiological traits). However, in our study microcosms, the recombinant plasmid persisted in the TM cells for 6 years after the introduction, and as the population adapts to the conditions of the microcosms, the efficiency of the cloned gene expression in the cells is restored. In the microcosms with high microalgal counts (10(7) cells/ml), cells with a high threshold of sensitivity to ampicillin dominate in the population of the TM E. coli Z905/pPHL7.

Adaptation, Physiological↗

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass.

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 1/2-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estunate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.

Air Microbiology↗

Modelling of genetically engineered microorganisms introduction in closed artificial microcosms.

The possibility of introducing genetically engineered microorganisms (GEM) into simple biotic cycles of laboratory water microcosms was investigated. The survival of the recombinant strain Escherichia coli Z905 (Apr, Lux+) in microcosms depends on the type of model ecosystems. During the absence of algae blooming in the model ecosystem, the part of plasmid-containing cells E. coli decreased fast, and the structure of the plasmid was also modified. In conditions of algae blooming (Ankistrodesmus sp.) an almost total maintenance of plasmid-containing cells was observed in E. coli population. A mathematics model of GEM's behavior in water ecosystems with different level of complexity has been formulated. Mechanisms causing the difference in luminescent exhibition of different species are discussed, and attempts are made to forecast the GEM's behavior in water ecosystems.

Animals↗

Gravitaxis in unicellular microorganisms.

Orientation of organisms with respect to the gravitational field of the Earth has been studied for more than 100 years in a number of unicellular microorganisms including flagellates and ciliates. Several hypotheses have been developed how the weak stimulus is perceived. Intracellular statoliths have been found to be involved in gravitaxis of Loxodes, while no specialized organelles have been detected in other ciliates, e.g. Paramecium. Also in the slime mold Physarum no specialized gravireceptors have been identified yet. In the flagellate Euglena gracilis the whole cell body, which is denser than the surrounding medium, seems to act as a statolith pressing onto the lower membrane where it activates mechanosensitive ion channels. Similar results were obtained for the ciliate Paramecium. In contrast to the flagellate Euglena, several ciliates have been found to show gravikinesis, which is defined as a dependence of the swimming velocity on the direction of movement in the gravity field.

Animals↗

A physical and chemical characterization of Martian permafrost as a possible habitat for viable microorganisms.

Data from experiments with model samples show that ion transfer coefficients in the water-rich permafrost on Mars must be three orders of magnitude less than those of terrestrial permafrost. The effects of low temperatures and of carbon dioxide have been accounted for. Exchange between cells and the environment is impeded in Martian permafrost. The microscopic distributional heterogeneity of concentration, pH, Eh, and other physicochemical parameters may be more pronounced in the permafrost of Mars. We present a classification of unfrozen water types in the permafrost that is based on the structures of unfrozen water films and on their functions with respect to cells. Any viable microorganisms on Mars probably exist with minimum metabolism in compact zones with energy carriers and high transfer coefficients. These zones may be microvolumes of unfrozen water in which cells accumulate.

Atmospheric Pressure↗

A primitive cyanobacterium as pioneer microorganism for terraforming Mars.

The primitive characteristics of the cyanobacterium Chroococcidiopsis suggest that it represents a very ancient type of the group. Its morphology is simple but shows a wide range of variability, and it resembles certain Proterozoic microfossils. Chroococcidiopsis is probably the most desiccation-resistant cyanobacterium, the sole photosynthetic organism in extreme arid habitats. It is also present in a wide range of other extreme environments, from Antarctic rocks to thermal springs and hypersaline habitats, but it is unable to compete with more specialized organisms. Genetic evidence suggests that all forms belong to a single species. Its remarkable tolerance of environmental extremes makes Chroococcidiopsis a prime candidate for use as a pioneer photosynthetic microorganism for terraforming of Mars. The hypolithic microbial growth form (which lives under stones of a desert pavement) could be used as a model for development of technologies for large-scale Martian farming.

Agriculture↗

Hydrogenases of phototrophic microorganisms.

This review surveys recent work done in the laboratory of the author and related laboratories on the properties and possible practical applications of hydrogenases of phototrophic microorganisms. Homogeneous hydrogenase preparations were obtained from purple non-sulfur (Rhodospirillum rubrum S1, Rhodobacter capsulatus B10) and purple sulfur (Chromatium vinosum D, Thiocapsa roseopersicina BBS) bacteria, and from the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum L; highly purified hydrogenase samples were prepared from the cyanobacterium Anabaena cylindrica and from the green alga Chlamydomonas reinhardii. It was shown that hydrogenases of R. capsulatus and T. roseopersicina contain Ni and Fe-S cluster. The cytochromes of the c or b type serve as native electron acceptors for the hydrogenases of the purple bacteria and cyanobacteria; rubredoxin or cytochrome c for the hydrogenase of the green sulfur bacterium; and ferredoxin for Ch. reinhardii hydrogenase. The hydrogenase of T. roseopersicina BBS reversibly activates H2 at Eh less than -290 mV (pH 7), whereas those from R. capsulatus and from C. limicola f. thiosulfatophilum exhibit their maximum activity at Eh greater than -300 mV and are thus favourable only for the H2 uptake. Hydrogenase synthesis in different phototrophs depends on pO2, H2 concentrations and organic substrates. Organic compounds, which serve as electron donors and carbon sources, repress hydrogenase synthesis in R. rubrum, R. capsulatus and in Ectothiorhodospira shaposhnikovii when present at high concentrations. The synthesis of T. roseopersicina hydrogenase is constitutive. H2 notably stimulates hydrogenase activity in R. capsulatus. The synthesis of hydrogenase in R. sphaeroides 2R occurs only in the presence of H2 and does not depend on the presence of organic compounds in the medium.

Bacteria↗

A model for the influence of the greenhouse effect on insect and microorganism geographical distribution and population dynamics.

A model for the influence of the greenhouse effect on insect and microorganism geographical distribution and population dynamics using cellular automata is presented. Based on this model, an algorithm has been developed and used to determine the geographical distribution and population dynamics of a hypothetical species in an scenario of global warming. The species' initial population distribution is assumed to be Gaussian. After the initiation of global warming, the population moves and after a few decades the population distribution is no longer Gaussian. Larger populations are found in the direction of population movement.

Algorithms↗