Alterococcus agarolyticus, gen.nov., sp.nov., a halophilic thermophilic bacterium capable of agar degradation

Five strains of facultatively anaerobic moderately thermophilic bacteria were isolated from two hot springs in the intertidal zone of Lutao, Taiwan. They produced extracellular agarase on agar medium, yielding reducing sugars and organic acids as the end products under either aerobic or anaerobic conditions. The growth temperature range was approximately 38-58 degrees C with an optimal temperature of about 48 degrees C. The five strains tolerated a relatively narrow pH range from 7.0 to 8.5. They were Gram-negative halophiles growing optimally at 2.0-2.5% NaCl (ca. 0.34-0.43 M). They were capable of anaerobic growth by fermenting glucose and producing various organic acids such as butyrate, propionate, formate, lactate, and acetate. Cells grown in liquid medium were motile monotrichous cocci, normally 0.8-0.9 micron in diameter. They possessed saturated anteiso-15-carbon acid (anteiso-C15:0) as the most abundant cellular fatty acid (46.0-51.3 mo1%) and had G + C contents ranging from 65.5 to 67.0 mo1%. They are the first thermophiles found to degrade agar and also the first halophilic thermophilic bacteria known to be capable of both aerobic and anaerobic fermentative growth. These bacteria are considered to represent a new genus that we named Alterococcus, and Alterococcus agarolyticus is the type species.     

Karyotyping of Saccharomyces strains with different temperature profiles

This study examined the karyotype, the fermentation performance and the optimum growth temperature (Topt) of 28 yeast strains all identified as species belonging to Saccharomyces sensu stricto. The strains were isolated from fermented musts, which had not been inoculated, at two temperature ranges: 20-40 degrees C and approximately 0-6 degrees C. The results demonstrated a correlation between the Topt and the chromosome organization. In particular, strains with Topt of less than 30 degrees C showed only two bands in the region between 365 and 225 kb, while those with a Topt greater than 30 degrees C had three bands in this size range. From a taxonomic viewpoint, the Topt is a better indicator for the Saccharomyces sp. than the ceiling temperature of 37 degrees C currently used to differentiate cryotolerant Saccharomyces bayanus and S. pastorianus from non-cryotolerant S. cerevisiae and S. paradoxus strains.     

Magnetic resonance (MR) imaging and MR spectroscopy of nerve regeneration and target muscle energy metabolism in a model of prosthesis-guided reinnervation in rats

The effect of fluctuating temperatures on microbial growth is important in the passage of foods from production to consumption. Suspensions of Salmonella typhimurium have been subjected to sinusoidally time-varying temperatures of periods from 60 to 240 min between 4 degrees and 22 degrees C, that is within and below the growth temperature range. The suspensions were prepared with two concentrations of sodium chloride and adjusted to two different values of pH. The change in the numbers of viable bacteria was measured with time and the experimental growth curves and average generation times compared with predictions based on isothermal growth data. Generally, the experimental average generation times exceeded the predictions by not more than 10%. In enumerating viable bacteria in the suspensions containing 3.5% (w/v) sodium chloride it was necessary to use sodium chloride in the diluent and recovery medium in order to recover the bacteria quantitatively.     

Management of mania in the elderly: an update

Twenty-eight genetic loci have been physically mapped to specific large restriction fragments of the Streptococcus mutans GS-5 chromosome by hybridization with probes of cloned genes or, for transposon-generated amino acid auxotrophs, with probes for Tn916. In addition, restriction fragments generated by one low-frequency-cleavage enzyme were used as probes to identify overlapping fragments generated by other restriction enzymes. The approach allowed construction of a low resolution physical map of the S. mutans GS-5 genome using restriction enzymes ApaI (5'-GGGCC/C), SmaI (5'-CCC/GGG), and NotI (5'-GC/GGCCGC).     

Interleukin-1beta and interleukin-8 concentrations in the lower uterine segment during parturition at term

In order to study strain-specific differences in their growth behaviour at different, and particularly lower, temperatures, generation times for 45 strains of Salmonella enteritidis isolated from food were determined impedimetrically over a temperature range from 7 to 42 degrees C. In practical terms, 7 degrees C is the minimum requirement for Salm. enteritidis growth, and generation time variability increases markedly as this temperature is reached. Reports in the literature describing psychrotrophic behaviour and multiplication at lower temperatures cannot be confirmed. Generation time variability increased as temperature moved away from the optimal range with variation coefficients tending to rise as temperature fell. The great variability of multiplication parameters near the growth limit found in Salm. enteritidis may also be a characteristic of other bacterial species. It is therefore imperative to commence studies on larger numbers of strains to allow prediction of their behaviour at lower temperatures.     

Kinetics of heat inactivation of Venezuelan equine encephalomyelitis virus

Thermal inactivation of Venezuelan Equine Encephalomyelitis Virus (VEEV) was studied at temperatures from 26 degrees to 55 degrees C. Inactivation of infectivity took place by two thermodynamically different reactions, one of which predominated at temperatures below 44 degrees C and the other at higher temperatures. The presence of 1 or 2 M NaCl stabilized the VEE virus at low temperatures but enhanced the inactivation at high temperatures. This latter effect at temperatures higher than 50 degrees C, is associated with the occurrence of two-component survival curves. The different effects of hypertonic NaCl concentrations at the two ranges of temperature, are related to different mechanisms of inactivation operating at each range (protein denaturation and nucleic acid-RNA breakdown). Different kinetics of thermal inactivation at 55 degrees C were observed between virus strains with different virulence. However, no significant correlations was found between the virulence of the eleven VEE virus strains studied and their thermostability at 37 degres and 55 degrees C.     

Thermal sensitivity of growth and feeding in Manduca sexta caterpillars

We explore how the thermal sensitivity of organismic performance emerges from the thermal sensitivity of the underlying component processes involved, using growth and feeding of Manduca sexta caterpillars as a model system. We measured thermal performance curves for the short-term rates of growth, consumption, protein (casein) digestion, amino acid (methionine) uptake, and respiration in fifth-instar caterpillars over a biologically realistic temperature range from 14 degrees to 42 degrees C. Growth and consumption rates increased between 14 degrees and 26 degrees C, reached a maximum value near 34 degrees C, and declined rapidly above 38 degrees C. In contrast, protein digestion rate and respiration rate increased monotonically over the entire temperature range, and amino acid uptake rate increased with temperatures up to 38 degrees C and then leveled off between 38 degrees and 42 degrees C. These results suggest that the shape and position of the thermal performance curve for growth rate--in particular the maximum at 34 degrees C and rapid decline above 38 degrees C--was most closely correlated with the thermal sensitivity of consumption rate; the declining growth performance above 38 degrees C was not associated with declines in digestion or uptake rates or with accelerated respiration rates at these temperatures.     

Violent versus nonviolent stalkers

The sgp gene from Streptococcus mutans has been previously isolated, characterized, and demonstrated to encode a G-protein. In order to investigate the function of this gene, a novel antisense RNA strategy was developed. Expression of sgp antisense RNA in Escherichia coli led to transient inhibition of growth. In addition, sgp antisense RNA expression in S. mutans resulted in decreased growth under environmental stress conditions (44 degrees C, acidic pH, and high osmolarity). Therefore, these results suggest that the sgp gene plays a role in modulating the stress responses of S. mutans. This approach could be applicable for investigating the function of essential genes in other organisms for which mutants are not available.     

Physiopathology of bedsores

Listeria monocytogenes is a food-borne pathogenic bacterium that can be found in soft cheese. At the beginning of cheese ripening, the pH is about 4.85-4.90. The aim of this work was to study the influence of temperature, preincubation temperature (temperature at which the inoculum was cultivated) and initial bacterial concentration on the survival of L. monocytogenes (strain Scott A) at pH 4.8. It was demonstrated in an earlier study that these factors did influence growth kinetics. Survival studies of L. monocytogenes were done in a laboratory broth simulating cheese composition. Four test temperatures (2, 6, 10 and 14 degrees C) and two preincubation temperatures were studied (30 degrees C or the test temperature). Listeria monocytogenes (strain Scott A) was unable to grow at pH 4.8 under all conditions tested. The time for 10% survival was about 11 and 2 d, at 2 degrees C with preincubation at 2 degrees C and 30 degrees C, respectively; 9 d at 6 degrees C with preincubation at 6 degrees C; 4 d at 6 degrees C with preincubation at 30 degrees C; and 1 d at 14 degrees C with preincubation at 14 degrees C or at 30 degrees C. The results show that survival of L. monocytogenes (strain Scott A) at pH 4.8 is not dependent on initial bacterial concentration but on both the test and preincubation temperatures.     

Thermally induced molecular disorder in human stratum corneum lipids compared with a model phospholipid system; FT-Raman spectroscopy

The molecular basis of lipid packing in human stratum corneum and a model phospholipid system has been studied as a function of temperature using Fourier Transform (FT) Raman spectroscopy. Thermally induced molecular rearrangements of the model lipid system, dipalmitoylphosphatidyl choline (DPPC), and stratum corneum were investigated using FT Raman spectroscopy coupled to a heating chamber. Spectra were recorded for a range of sample temperatures and the results for the two systems were compared, producing previously unreported information of the thermal behaviour for the different systems. Discrete thermal events were recorded for both systems by plotting band separation of the lipid v(CH2) symmetric and asymmetric stretching modes against temperature. The main thermal events observed for DPPC included a 'pre-melting' between 37 and 39 degrees C, the main transition observed between 41 and 42 degrees C, a 'post-transition' between 42 and 43 degrees C and three minor transitions at 58-60, 65-70 and 75-80 degrees C. No evidence was found for the pre-transition of DPPC, previously observed at 34-35 degrees C. The main transitions for human stratum corneum were observed at 35-45, 55, 72 and 83 degrees C, measured from lipid CH2 stretching and bending vibrations. The keratin thermal transition at about 100 degrees C exerted little effect on the lipid bands and no characterisable structural changes were reflected in the keratotic bands.     

Evaluation of Candida acidothermophilum in ethanol production from lignocellulosic biomass

A Saccharomyces-cerevisiae-based simultaneous saccharification and fermentation (SSF) of lignocellulosic biomass is limited to an operating temperature of about 37 degrees C, and even a small increase in temperature can have a deleterious effect. This points to a need for a more thermotolerant yeast. To this end, S. cerevisiae D5A and a thermotolerant yeast, Candida acidothermophilum, were tested at 37 degrees C, 40 degrees C, and 42 degrees C using dilute-acid-pretreated poplar as substrate. At 40 degrees C, C. acidothermophilum produced 80% of the theoretical ethanol yield, which was higher than the yield from S. cerevisiae D5A at either 37 degrees C or 40 degrees C. At 42 degrees C, C. acidothermophilum showed a slight drop in performance. On the basis of preliminary estimates, SSF with C. acidothermophilum at 40 degrees C can reduce cellulase costs by about 16%. Proportionately greater savings can be realized at higher temperatures if such a high-temperature SSF is feasible. This demonstrates the advantage of using thermophilic or thermotolerant yeasts.     

Heat resistance of Listeria monocytogenes by two recovery media used with and without cold preincubation

Three strains of Listeria monocytogenes were heat-treated at three temperatures in physiological saline by a capillary tube method. Recovery of heat-treated bacteria was performed on blood agar and on tryptose phosphate agar with ferric citrate and aesculin (TPA-FE). Both media were used in two ways: (1) incubation at 37 degrees C for 7 d, and (2) preincubation at 4 degrees C for 5 d in order to obtain repair of heat-injured bacteria, followed by incubation at 37 degrees C for 7 d. D and z values were determined. In both incubation procedures, better average recovery was obtained on blood agar than on TPA-FE. Thus, higher D values were recorded when blood agar was used. In most cases the differences were statistically significant. Repair at 4 degrees C of heat-injured bacteria occurred on both media but the proportions of repaired bacteria were higher on blood agar. The repair on this medium was generally reflected in higher D values for preincubated samples. Some significant differences in heat resistance were noted between the strains.     

L-(+)-lactate dehydrogenase deficiency is lethal in Streptococcus mutans

The previously cloned gene for L-(+)-lactate dehydrogenase (LDH) from Streptococcus mutans was mutagenized in vitro. An Escherichia coli transformant which expressed a thermolabile LDH activity was identified. The ldh(Ts) gene was introduced into S. mutans on a suicide vector to create a heterodiploid expressing both wild-type and thermolabile LDH activities. Self-recombinants which had only one ldh gene were isolated. One of these clones expressed only the thermolabile LDH activity. This isolate grew well at 30 degrees C but did not grow at 42 degrees C under a variety of cultivation conditions, thereby proving that LDH deficiency is lethal in S. mutans in the absence of compensatory mutations.     

Beta-adrenoceptor stimulation-mediated preconditioning-like cardioprotection in perfused rat hearts

The metabolic rates of six female grey seal (Halichoerus grypus) pups were measured during their postweaning fast at air temperatures between -15 degrees and 30 degrees C. The composite of their individual thermal neutral zones extended from a mean lower critical temperature of -7.1 degrees +/- 0.7 degree C to a mean upper critical temperature of 23.0 degrees +/- 0.4 degree C. Within the thermal neutral zone, mean standard metabolic rate of the fasting animals was 1,265 +/- 82 kcal d-1, or about 1.1 times the value predicted for an adult animal of similar body mass (mean mass = 40.9 +/- 1.2 kg). For those grey seal populations that reproduce during winter months in the eastern and western Atlantic and in the Baltic Sea, the lower critical temperature of fasting pups corresponds closely with the coldest mean monthly air temperature at the northern end of their breeding ranges. This observation supports the hypothesis that cold ambient air temperatures limit the northern breeding distribution of grey seals, primarily through their thermoregulatory effects on small, fasting pups before they enter the water.     

Molecular characterization of the mutable flaked allele for flower variegation in the common morning glory

Force-velocity data at different temperatures (range, 10-35 degrees C) from intact fibre bundles are analysed to determine the temperature dependence of the maximal mechanical power output of fast and slow rat muscles. At 35 degrees C, the maximal mechanical power was approximately 250 kW m(-3) (=250 microW mm(-3)) in fast (probably an underestimate) and approximately 100 kW m(-3) in slow muscle. Within the more physiological temperature range (25-35 degrees C), the temperature coefficient (Q10) of maximum power was 2-2.5. In both muscles, the maximal power at 10 degrees C was only about 3-5% of that at 35 degrees C, the decrease being particularly pronounced at temperatures below 20 degrees C (Q10 of 5-7).     

Inhibitory effect of 0 degree C storage on the proliferation of Yersinia enterocolitica in donated blood

BACKGROUND: Yersinia enterocolitica is frequently identified in cases of bacterial sepsis due to red cell transfusion. One of the features that makes Y. enterocolitica particularly dangerous is that, unlike most other bacterial contaminants of blood components, this organism can actively multiply in currently recommended refrigerator temperatures (1-6 degrees C). The effect of a colder than normal storage temperature on Y. enterocolitica growth was investigated to determine whether bacteria growth could be reduced or inhibited at 0 degree C. STUDY DESIGN AND METHODS: Twenty-four units of freshly collected donated blood were obtained. Three sets of 7 units each were inoculated with Y. enterocolitica O:3, Y. enterocolitica O:20, and Y. enterocolitica O:5, 27, respectively. The remaining 3 units served as uninoculated controls. Each of the 24 bags was split into two equal aliquots, with one aliquot stored at 4 degrees C and the other at 0 degree C. Bacteria growth was measured twice weekly for 6 weeks. Endotoxin and hemoglobin levels were also measured at selected intervals. RESULTS: Bacteria growth was detected earlier and in higher concentrations in the aliquots stored at 4 degrees C. Twenty-two of the 42 inoculated aliquots had measureable bacteria growth. Thirteen aliquots had been maintained at 4 degrees C, and nine had been stored at 0 degree C. Sixteen of these 22 aliquots were matched pairs. Exponential growth was detected after 14 to 32 days in the 4 degrees C aliquots and after 28 to 39 days in the 0 degree C aliquots. Final bacteria counts were much higher in the 4 degrees C aliquots (10(5)-10(14) colony-forming units/mL) than in the 0 degree C aliquots (10(1)-10(4) colony-forming units/mL) on Day 42. Endotoxin was present in all 13 of the 4 degrees C aliquots with actively growing Y. enterocolitica. CONCLUSION: Storage of red cells at 0 degree C markedly prolongs the time required for Y. enterocolitica to achieve exponential grwoth and results in lower concentrations of bacteria.     

What physicians should currently observe in the area of dental health

Several factors govern the development of human dental caries. The actual caries incidence in children depends greatly on fluoride consumption, reduction of fermentable sugar intake (in favour of the use of sugar substitutes), dental health habits and regular dental check-ups. A significant reduction of dental caries in children and young adults is achieved in many countries following these principles. However, there remains a group, about 25%, with a particularly high incidence of caries. Today, efforts are directed at detecting these risk subjects. Mutans streptococci and lactobacilli are the main causative bacteria of human dental caries. The initiation of dental caries is preceded by colonisation of the dentition by S. mutans, usually in early childhood. The early establishment of these bacteria in the mouth of human infants is dependent on intrafamilial--mainly maternal--transmission of saliva. The higher the level of S. mutans in the maternal saliva, the more likely is the colonisation of primary dentition by cariogenic bacteria. The earlier the colonisation, the higher the incidence of caries. Today we have new diagnostic aids: Simple microbiological tests to detect the individuals with high salivary levels of these cariogenic bacteria, so called SM-millionaires. The tests are also a good educational aid for motivating patients to individual prophylaxis. What could a paediatrician or gynaecologist do to help the dentist? He could inform the parents early enough to avoid saliva contacts. For example, the mother should not take the child's spoon or pacifier in her mouth. The dentist meets the children much later, at an age where they already have caries. The older methods, mentioned at the begin, are also still valuable.     

Heart rate during development in the turtle embryo: effect of temperature

Growth and development can occur over a wide range of physical conditions in reptiles. Cardiovascular function must be critical to this ability. However, information on cardiovascular function in developing reptiles is lacking. Previous work indicated that in reptiles the effects of temperature on growth and metabolism are largely restricted to early development. This study examined whether the previously observed effects of temperature and different perinatal patterns of metabolism observed in amniotic vertebrates are correlated with cardiovascular function. Embryonic and hatchling carcass mass, heart mass and heart rate (HR) were compared for snapping turtle eggs (Chelydra serpentina) incubated at 24 degrees and 29 degrees C. Incubation time was shorter at 29 degrees C (56.2 days) than at 24 degrees C (71.1 days). Carcass and heart growth showed a sigmoidal pattern at both temperatures. However, cardiac growth showed a relative decrease as incubation proceeded. Incubation temperature significantly affected the HR pattern during development. The HR of embryos incubated at 24 degrees C was constant for most of incubation (51.8 +/- 4.8 min-1). A small decrease was observed just prior to and a large decrease immediately following hatching (posthatch, 22.3 +/- 4.1 min-1). At 29 degrees C embryonic HR was greater than at 24 degrees C early in development (72.3 +/- 3 min-1). The HR steadily decreased to values equivalent to those at 24 degrees C. The HRs of 24 degrees C and 29 degrees C hatchlings were not different. Cardiac output (estimated as the product of heart mass and HR) increased rapidly during early development and then slowed dramatically at both temperatures. These data are consistent with the suggestion that temperature exerts its effects primarily early in development. Furthermore, the changes in cardiovascular function are correlated with metabolic changes in hatching vertebrates.     

Congenital nystagmus image motion: influence on visual acuity at different luminances

This research study was undertaken to examine the relationship between pulmonary artery blood temperature (regarded as the 'gold standard' measurement for core body temperature), axilla temperature using the Tempa.DOT Ax chemical thermometer and tympanic membrane temperature using the Diatek 9000 InstaTemp thermometer. Sixty adult intensive care patients had their temperatures monitored. A single set of five simultaneous temperatures, i.e. left and right axilla, left and right tympanic membrane (TM), and pulmonary artery (PA) blood were recorded. The mean difference between left and right TM temperatures was 0.58 degree C, and although both were moderately well correlated with PA temperature (r = 0.63 and 0.78, respectively) the mean differences between the two sites were clinically significant (0.85 degree C and 0.94 degree C, respectively). The range of differences between the sites was significant. Plotting limits of agreement showed that both left and right TM temperatures may be up to 1.2 degrees C above or 1.3 degrees C below PA blood temperature: a clinically unacceptable range. In particular, large temperature differences were recorded when patients were lying with one side of their head to a pillow. Fan therapy directed to the head was not found to affect these differences significantly. The mean difference between left and right axilla temperatures was 0.36 degree C, and although both were modestly correlated with PA temperature (r = 0.48 and 0.53, respectively) the mean differences between the two sites were clinically significant (0.47 degree C and 0.50 degree C, respectively). The range of differences between the sites was particularly significant. Plotting limits of agreement showed that both left and right axilla temperatures may be up to 1.2 degrees C above or 1.6 degrees C below PA blood temperature: a clinically unacceptable range. Because the range of temperature differences found between PA blood and the other sites was so great, it is concluded that neither the chemical axilla thermometer nor the tympanic membrane thermometer used in this study are clinically reliable tools for adult intensive care patients.     

Effect of temperature on growth and activity of a methanogenic culture utilising acetate

Studies with a methanogenic culture enriched for use of acetic acid showed that this culture had an optimum growth temperature of 35 degrees C, with only small differences for other temperatures between 30 and 40 degrees C. The optimum temperature was the same when determined on the basis of biomass production rate during the exponential (log) phase of growth (0.08-0.09 day-1, at 35 degrees C), amount of biomass present at the end of the log phase (100 mg/l), activity of the biomass (rate of conversion in millimoles per day per milligram (dry wt.) biomass present, 0.08 at end of log phase), or biomass yield (mg (dry wt.) biomass produced per millimole acetic acid converted, 1.0-1.1). Temperatures outside the range 30 to 40 degrees C caused marked reductions in the above parameters. The maximum temperature for growth was 42-44 degrees C; the minimum, below 15 degrees C, the lowest temperature studied. Acetic acid conversion to methane was 0.8-1.0 mol/mol, and was independent of temperature.